About this Journal Submit a Manuscript Table of Contents
International Journal of Cell Biology
Volume 2012 (2012), Article ID 176287, 16 pages
http://dx.doi.org/10.1155/2012/176287
Review Article

Endothelial Cells and Astrocytes: A Concerto en Duo in Ischemic Pathophysiology

1Université Lille Nord de France, 59000 Lille, France
2UArtois, LBHE, EA 2465, 62300 Lens, France
3IMPRT-IFR114, 59000 Lille, France
4Departments of Physiology, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA
5Departments of Pediatrics, Loma Linda University School of Medicine, Loma Linda, CA 92354, USA

Received 2 March 2012; Accepted 30 April 2012

Academic Editor: Carola Förster

Copyright © 2012 Vincent Berezowski et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The neurovascular/gliovascular unit has recently gained increased attention in cerebral ischemic research, especially regarding the cellular and molecular changes that occur in astrocytes and endothelial cells. In this paper we summarize the recent knowledge of these changes in association with edema formation, interactions with the basal lamina, and blood-brain barrier dysfunctions. We also review the involvement of astrocytes and endothelial cells with recombinant tissue plasminogen activator, which is the only FDA-approved thrombolytic drug after stroke. However, it has a narrow therapeutic time window and serious clinical side effects. Lastly, we provide alternative therapeutic targets for future ischemia drug developments such as peroxisome proliferator- activated receptors and inhibitors of the c-Jun N-terminal kinase pathway. Targeting the neurovascular unit to protect the blood-brain barrier instead of a classical neuron-centric approach in the development of neuroprotective drugs may result in improved clinical outcomes after stroke.

1. Introduction: Current Clinical Overview of Stroke

In the United States, stroke is the number one cause of chronic disability and the fourth leading cause of death, with approximately 7 million adults affected [1]. Annually there are approximately 800,000 strokes in the US, of which 87% are ischemic strokes, 10% are primary hemorrhages, and 3% are subarachnoid hemorrhages [1]. Together they cause the country a financial burden of approximately 62.7 billion dollars [2]. Cerebral ischemic stroke is caused by an occlusion of a cerebral blood vessel, typically by a thrombus, which causes a decrease in cerebral blood flow and thus limits the supply of oxygen and nutrients globally (in global ischemia) or to certain regions of the brain (in focal brain ischemia). This absence of blood flow in a brain region causes neuronal death in addition to damaging the vascular tree; the vascular tree is usually made more fragile during the ischemic period and damaged during reperfusion. Time is an important parameter in the evolution of brain injury. In 2006, Saver et al. have estimated the impact of stroke on the brain tissue [3] to be immense; the brain may lose up to 120 million neurons, 830 billion synapses and 714 km of myelinated fibers for each hour after stroke onset [3]. Ischemic stroke seems to accelerate aging of the brain at a rate of 3.6 years each time when the symptoms are not treated [3]. Therefore, the clinical goal of acute stroke treatment is to reduce brain damage by limiting the time of ischemia through thrombectomy (mechanical endovascular approach) or thrombolytic therapy, which consists of in lysing the blood clot in order to restore cerebral blood flow.

Recombinant tissue plasminogen activator (rtPA) is currently the only thrombolytic molecule administered during acute cerebral infarction that provides a clinical benefit in terms of survival and neurological outcome [4]. The rtPA administration must be within the first 4 hours 30 minutes after stroke onset to maintain the beneficial effects without substantially raising the side effects/risk [5, 6], which limits its use. Based on the organization of emergency care, only 5% of stroke patients are eligible for this therapy in this narrow time window, which leaves the remaining 95% of patients without any beneficial treatment available. The major risk of rtPA is the extension of the damage due to potential bleeding [7]. The need for drug development to prevent the neuronal loss has driven research on neuroprotective agents that aim to save viable neurons located in the ischemic penumbra area. However, all of the proposed neuroprotective treatments specifically targeting neurons that showed promise on the bench have failed in clinical trials [8].

In 2000, the neurovascular unit (NVU) was proposed as a physiological unit composed by neurons, astrocytes, and endothelial cells [9]; there is a growing interest in studying the changes of the NVU after stroke. In addition to cell death, ischemic stroke is characterized by changes in the properties of the blood-brain barrier (BBB) with physical disruption of the tight junctions contributing to aggravation of cerebral edema and consequently neuronal death. The new strategy for drug development is to have molecules with a broader spectrum targeting not just the neurons but the NVU as a whole entity. In the present paper, we will focus on some molecular and cellular mechanisms of astrocytes and endothelial cells. We will look specifically at: (1) the ways astrocytes and endothelial cells work in concert in stroke pathophysiology such as BBB disruption and edema formation, (2) how they could be affected after rtPA treatment, and (3) new drug developments in the future.

2. Definition of the Neurovascular/Gliovascular Unit

Several groups have proposed the NVU as a physiological unit composed of not only endothelial cells, astrocytes, and neurons but also pericytes, smooth muscle cells, and the interacting circulating peripheral immune cells [1012]. The term “gliovascular” emphasizes the importance of the interactions between astrocytes and cerebral blood vessels within the NVU [13], which are critical in cerebral blood flow regulation [14], brain energy metabolism [15], and also the maintenance of the BBB properties [13].

The BBB is located in the endothelial cells of brain vessels, with the presence of tight junctions and adherens junctions between the cells (Figure 1) that prevent paracellular diffusion and act as a unit to regulate ions and other molecules between peripheral blood flow and brain parenchyma. Tight junctions are composed of several protein families: trans-membrane proteins (claudins and occludins), cytoplasmic proteins, and zona occludens proteins. They bind the afore mentioned proteins with structural cytoskeletal proteins such as actin. Adherens junctions are formed by proteins such as platelet-endothelial cell adhesion molecule (PECAM) and vascular endothelial-cadherin, which contribute to the close physical contact between endothelial cells and facilitate the formation of tight junctions.

fig1
Figure 1: (a) Schematic drawing of the neurovascular unit (NVU) in the capillary bed composed by the neuron, astrocyte endfoot, basal lamina, pericyte, and endothelial cell. The endothelial cell is the first barrier between the blood stream and the nervous tissue. The presence of the tight junction composes the physical barrier and the movement of substrates is controlled by several transporters. The astrocyte endfeet are linked with the gap junction, allowing movement of several solutes in the astrocyte network. The basal lamina is composed of several proteins such as agrin, dystroglycan, and perlecan. (b) A close-up schematic drawing of the endothelial cells and astrocyte endfeet with some of the proteins involved in edema formation and resolution.

The brain endothelial cells of the BBB also present specific transport proteins located on the luminal and abluminal membranes for nutrients, ions, and toxins to cross the endothelial layer between the blood stream and brain [13, 16]. For example, energy molecules are transported by specific solute carriers such as glucose transporter 1 (GLUT 1) and monocarboxylate transporters 1 and 2 (MCT1, MCT2). Large molecular weight solutes (e.g., large proteins and peptides) are able to cross the BBB and enter the intact CNS via endocytotic mechanisms called receptor-mediated transcytosis, such as with insulin, or adsorptive-mediated transcytosis, exemplified by albumin. On the other hand, transport can also be achieved by the ATP-binding protein (ABC) family, which consumes ATP to effectively transport a wide range of lipid-soluble compounds from the brain endothelium. In the BBB examples of ABC transporters for efflux transport are P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP), and breast cancer resistance protein (BCRP) [16]. These efflux transporters are understood as gatekeepers of the brain because they keep tight control over which substances are allowed to enter the CNS through the endothelial cell barrier (Figure 1). Endothelial cells also present a metabolic barrier of the BBB, which functions to inactivate molecules capable of penetrating cerebral endothelial cells.

Quite recently it has been proposed that the primary barrier of the BBB may extend to the basal lamina, thus preventing the entry of immune cells into the parenchyma under normal brain conditions [12]. Historically the brain was thought to be an immune cell deficient organ, and the BBB was thought to prevent passage of any immune cells into the brain. However, peripheral immune cells from the blood have been observed to enter and be present in the brain at multiple time points during embryonic development [17] and in normal physiological conditions in adults [12]. Therefore, the theory of the CNS as an immune-independent organ has recently started to be reexamined and revised. Engelhardt and collaborators elegantly compare the perivascular space as a castle moat with perivascular antigen presenting cells floating as guards, confined by the inner and outer wall, which is the basement membrane of the astrocytic endfeet and the endothelial cell, respectively [12]. Endothelial cells and other cells, such as the astrocytes, may also contribute to the tight regulation of the movement of immune cells between the peripheral blood stream and the brain. However, the exact mechanisms by which peripheral cells enter the brain are still a matter of discussion. Moreover, rather than the BBB being a rigid wall, it provides a dynamic interface between the brain and the rest of the body.

As mentioned previously, the presence and the maintenance of these barrier properties are important for brain homeostasis and for neuronal functioning [13]. In fact, disruption of tight junctions leads to BBB disruption and extravasation of blood components and water, which contribute to vasogenic edema formation. We will cover these in more detail in the following section.

3. Edema Process after Stroke: Endothelium and Astrocyte, Concerto en Duo

3.1. BBB Disruption and Edema Formation

Cerebral edema has been traditionally divided into 2 major classes: cytotoxic and vasogenic [18] for cerebrovascular diseases and other brain pathologies. Cytotoxic edema is defined by intracellular accumulation of water coming from the extracellular space without BBB disruption. Vasogenic edema appears after BBB disruption, leading to a diffusion of proteins from the blood to the tissue followed by water accumulation in the extracellular space [18]. However, this division alone does not explain fully the diversity and the complexity of the edema process in brain ischemia as well as in the other brain injuries and disorders. Based on several recent advances in the understanding of the molecular mechanisms of edema formation and BBB properties, a third subtype of edematous processes was named ionic edema and described as a continuum between the cytotoxic to vasogenic edema in the cerebrovascular diseases [19, 20]. In fact, cytotoxic, or anoxic, edema occurs within the first few minutes after cerebral blood flow stoppage and is characterized as swelling of the astrocytes and neuronal dendrites [20, 21]. The cellular swelling within the first 10 minutes is a result of oxygen and glucose deprivation followed by a slow rise in extracellular [K+] [22]. The absence of oxygen and energy nutrients induces a disruption of the cellular ionic gradients and leads to entry of ions into cells. Water follows this ionic gradient into the cells and induces cellular swelling. Cytotoxic/anoxic edema may evolve quickly to become ionic edema because the absence of oxygen and nutrients further alters the energy balance in endothelial cells and the ionic gradients, including transcapillary flux of Na+ in these cells [19, 23]. The endothelial cells also require a large amount of ATP production, characterized by the high density of mitochondria, which are important for the regular homeostatic BBB functions such as maintenance of ionic gradients and membrane transporters [24, 25]. The absence of energy supplies for these cells would severely impair these functions. Reperfusion induces overpressure accompanied by shear stress on the nonperfused vascular tree that results in early transient leakage of the BBB [26, 27]. This leakage results in further entry of water through the endothelial cells resulting in brain swelling within 30 minutes after reperfusion [26, 27] and additional BBB permeability [27, 28]. This early opening of the BBB has also been described clinically in humans and is frequently associated with hemorrhagic transformation [29]. Early reperfusion probably mitigates the BBB alterations, but if it is delayed, reperfusion will exacerbate the amount of endothelial injury [3032]. The final step is the development of vasogenic edema, in which there is disruption of cerebrovascular endothelial tight junctions leading to increased permeability to albumin and other plasma proteins [18]. Another contributing factor of brain edema formation in addition to tight junction disruption is brain endothelial transcytosis [33]. BBB disruption is usually coupled with the inflammatory response and activation of matrix metalloproteinases (MMP) [34, 35]. In fact, vasogenic edema development is aggravated by MMP-9, which degrades basal lamina, the connection between astrocytic endfeet and endothelial cells [36].

In the clinic, diffusion-weighted imaging (DWI) and T2-weighted imaging (T2WI) magnetic resonance imaging (MRI) modalities are used extensively to assess postischemic edema [20, 37, 38]. T2 values represent water content and apparent diffusion coefficient (ADC) values derived from DWI images represent water mobility in the tissue [20, 37]. ADC values decrease rapidly after stroke onset, indicating restricting water movement, and are interpreted as evidence of ionic edema with the characteristic swelling of the brain cells causing a decrease in extracellular space as proposed in our classification mentioned before. T2 values increase at later time points, which are associated with vasogenic edema [20, 39].

The molecular mechanisms and temporal development of edema after stroke have been well studied. However, the cellular and molecular mechanisms involved in edema resolution are not well understood in stroke and other brain diseases. The healing of the endothelial cells with stabilization of the tight junctions may be a critical step to limit the entry of blood components into the brain. Thus, stabilizing the NVU may be an essential component of controlling edema formation and BBB breakdown after stroke.

Postischemic BBB disruption has been commonly believed to be biphasic [40], but recent work suggests that the BBB disruption may be continuous for up to 5 weeks after ischemia in rats [28]. BBB leakage was demonstrated using gadolinium and magnetic resonance imaging (MRI) at 25 min; 2, 4, 6, 12, 18, 24, 36, 48, and 72 hours; and 1, 2, 3, 4, and 5 weeks after ischemia [28]. Similarly, albumin leakage through the BBB, especially in the hippocampus, has also been observed in spontaneously hypertensive stroke prone rats long term [41]. Although these data do not completely rule out the possibility of a biphasic pattern in the opening of the BBB, the long-term leakage of the BBB is important to note from the standpoint of postischemic edema because this disruption could account for a prolonged vasogenic edema.

3.2. “Concerto en Duo”: Astrocyte Network in Edema Formation and Resolution

As part of the NVU the astrocyte endfeet in contact to the blood vessels are well known for to swell after stroke [4244]. The recent knowledge on the transporters and channels in this astrocyte subdomain gives new perspectives on the understanding of astrocyte swelling. In fact, aquaporin 4 (AQP4), a member of the family of 13 water channel proteins, is proposed to have an important role in edema formation [20, 45]. AQP4 is the most abundant water channel in the brain, in part due to its high concentration on astrocytic endfeet which are in contact with all the cerebral blood vessels [46, 47]. More recently, AQP1 has also been described in a subpopulation of astrocytes within the nonhuman primate but not in rodents, suggesting interspecies differences and a possible role in brain water homeostasis [48]. AQP1 has also been reported to be present in peripheral endothelia and primary rat brain endothelial cell cultures [49]. Interestingly, Dolman and collaborators observed that mRNA AQP1 levels were lower in cultured brain endothelial cells when cocultured with astrocytes [49], suggesting an inhibition effect of the astrocytes on the AQP expression in endothelia. In fact, there are publications reporting a low level of AQP in endothelial cells in vivo [50], although AQP is more abundant in astrocytes [49, 5154].

Currently, AQP4 is considered as a key player in the edema process by its location on the astrocyte endfeet [20, 55]. AQP4 is assembled in homotetramers where each individual aquaporin represents a water channel [56]. Interestingly, AQP4 is also organized in the astrocyte endfeet membrane in a larger geometric structure known as an orthogonal array of particles (OAPs), which has been described with freeze-fracture techniques and electron microscopy studies (Figure 2) [57]. OAPs are present in all astrocyte endfeet in contact with the blood vessels as well as the glia limitans. OAPs are formed with two isoforms of AQP4: long (AQP4-M1) and short splice variants (AQP4-M23). The ratio of AQP4-M1 to AQP4-M23 determines the size of these OAPs [57] in contact with the basal lamina of brain vessels (Figure 1). Experiments in oocytes showed that the AQP4-M23 isoform stabilizes the OAP structure [57, 58]. However, the exact functional roles of the OAPs remain unknown in normal and pathological conditions. Recently, AQP4-m1 mRNA and protein were found to increase quickly after stroke onset, while AQP4-m23 remained the same. The increase of AQP4-m1 early after ischemia could favor a shift toward M1 in the M1/M23 balance, which is known to favor small size OAPs [27]. In accordance with this work, previous studies have shown that early disorganization of OAPs on the astrocyte endfeet after global cerebral ischemia preceded astrocyte swelling [59]. Although a direct effect of the modification in the ratio of AQP4-M1 to AQP4-M23 on water permeability has not yet been directly investigated in vivo, in a preconditioning model, a strong increase in AQP4 expression and increase of AQP4-M1 were correlated with reduced edema and less water in the tissue, suggesting increased water diffusibility which resulted in the removal of excess liquid from the brain tissue [27]. Interestingly, it was recently proposed that the assemblage of 4 aquaporin molecules forms a central pore, through which water, ions, and gases may flow depending on the AQP subtype. For example, the central pore is permeable for O2, CO2, and possibly nitric oxide for AQP1, 4, and 5 [56, 60]. Thus, the disruption of the OAPs may also affect the diffusion of ions and gas through the central pore.

fig2
Figure 2: (a) Schematic drawing of the aquaporin homotetramer assembly within the lipid membrane: the central pore is proposed to be permeable to cations and gases (green arrows). Each individual aquaporin facilitates bidirectional water movement depending on the osmotic gradient (blue arrows). (b) AQP4 homotetramer is assembled in a higher structure named orthogonal array of particles (OAPs). Two isoforms of AQP4, AQP4-M1 (purple circles) and AQP4-M23 (blue circles) isoforms, contribute to the formation of OAPs. In vitro experiment showed that higher expression of AQP4-M23 contributes to the formation of larger OAPs. (c) Increase of AQP4-M1 induced disruption of OAPs. Recent knowledge on AQP leads us to hypothesize that the large OAPs contribute to gas and cation diffusion in the astrocyte membranes through central pores (green arrows).

Due to its location in the astrocyte endfeet in contact with the blood vessels, AQP4 has been proposed to be linked with BBB integrity [52, 61, 62] and cell adhesion [63]. In the epithelial cells of the eye lens AQP0 is present in the OAPs and participates in epithelial cells linkage; however it does not facilitate water flux [64]. In this case, the presence of AQP4 in the astrocyte endfeet membrane was dependent on the presence of proteins in the basal lamina such as agrin, -dystroglycan, and laminin [65, 66] in addition to syntrophin and dystrophin protein complexes [67, 68]. The connection of AQP4 to proteins in the basal lamina may explain the ability of astrocytes to maintain the integrity of the blood-brain barrier, suggesting a possible role for AQP4 as a structural molecule within the perivascular space [61]. However, reports using AQP4 knock out (AQP4-KO) mice show contradicting results regarding the modifications in the BBB structure suggesting that AQP4 may not be integral to the BBB structure [61, 69]. Similarly in our siRNA silencing studies, BBB permeability was not significantly changed at distance from the site of injection after injection of siRNA against AQP4, even though AQP4 expression was decreased [55]. We also showed that the upregulation of AQP4 in a preconditioning model did not prevent the early opening of the BBB after stroke [27].

Heparan sulfate proteoglycan is a large family of proteins with agrin and perlecan, involved in the basal lamina composition located between the astrocyte endfeet and endothelial cells [54, 70]. Agrin and dystroglycan seem to play an integral role in the maintenance of astrocyte polarity by the interaction with AQP4 in the astrocyte endfeet [54]. Specifically, agrin KO mice showed a significantly decreased density of OAP in the astrocyte endfeet when compared to wildype but overall immunoreactivity of AQP4 did not differ significantly [71]. Dysfunctions in the basal lamina are related to increase of the BBB disruption, promoting edema formation. In fact, a family of endopeptidases, matrix metalloproteinases (MMPs), has been shown to degrade the proteins of the basal lamina and contribute to vasogenic cerebral edema [36]. In the human brain, MMPs are usually very low in concentration under nonpathological conditions [72]. However, after injuries such as ischemic stroke, certain MMPs such as MMP-2, -3, and -7 and especially MMP-9 have been shown to be upregulated in the brain (reviewed in [72]). This layer between astrocytes and endothelial cells is a potential future target for the NVU protection. Recently, Dr. Bix and collaborators have shown that administration of perlecan domain V, which is the c-terminal fragment, administered 24 hours after ischemic stroke has beneficial effects by interacting with integrins [73]. Perlecan domain V increased expression of vascular endothelial growth factor (VEGF), thus promoting angiogenesis, and interestingly did not lead to increased BBB permeability [73] even though VEGF is known to increase BBB permeability after ischemia [74]. Perlecan has also been shown to modulate postischemic astrogliosis through interaction with dystroglycans and integrins in the astrocytes [75].

Astrocytic AQP4 is not only linked with the matrix proteins but also with several other channels present in higher concentration in the astrocyte endfeet such as potassium inner rectifying channel 4.1 (KIR4.1), connexins (Cx), and also chloride channel 2 (CIC-2) [76, 77]. Colocalization of AQP4 and KIR4.1 suggests that AQP4 may have a role in potassium homeostasis by facilitating water diffusion along the potassium gradient and AQP4-KO mice display a delay in potassium reuptake during electrical activity [76]. The decrease of AQP4 expression using siRNA showed an associative decrease of connexin 43 (Cx43), a protein involved in gap junction formation, and a decrease of CIC-2, involved in the regulatory volume decrease function of the astrocytes. Interestingly, gap junctions and AQP4 are morphologically closely associated [78] with the astrocyte endfeet. The gap junctions in the astrocyte contribute to the formation of a complex network named the astroglial network [79]. Intercellular and intracellular communication that facilitate the movement of second messengers, amino acids, nucleotides, energy metabolites, and small peptides [7982] in astrocyte processes occur through gap junctions, which are made up of a family of channel proteins called connexins [83, 84]. In astrocytes, Cx30 and Cx43 are predominant [8385]. However, it is also important to note that Cx43, along with Cx37, Cx40 [86, 87], and Cx45 [87], is also expressed in brain endothelial cells. The protein level of Cx40 and Cx45 was shown to increase in cerebral arteries, but no change in protein or mRNA was observed for brain endothelial Cx43 and Cx37 after a model of brain injury causing cerebral vascular dysfunction [87]. The effect of astrocytic Cx43 upregulation or downregulation after ischemia still remains controversial and there is no consensus as to what provides beneficial effects [88]. However, in humans, there are reports that show that Cx43 protein levels were increased in the penumbra [89]. And because Cx43 and Cx30 knockouts have been observed to be more edema prone [90], it is possible that the increase in Cx43 after ischemia may be a physiological response to decrease edema. The induction of Cx43 may be facilitating water flow throughout the astrocyte network to diversify and dissipate the accumulation of fluid from just one region. From these data we hypothesize that gap junction proteins, specifically Cx43 on astrocytes, are working with AQP4. Evidence for this also comes from a significant decrease of Cx43 observed in mouse astrocyte cell cultures after administration of small interference RNA against AQP4 [91]. Although direct functional data are still lacking, one possibility is that AQP4 and Cx43 is working together to direct water flow between astrocytes and could be controlling astrocytic swelling.

The role of AQP4 in cerebral edema formation and resolution has been studied in several models. However the precise role of AQP4 remains unclear and depends on the pathological model used [92, 93]. Indeed, the absence of AQP4 was shown to prevent the formation of edema in a permanent ischemia model in AQP4-KO mice [94]. Similarly, edema formation is prevented in -syntrophin knockout mice at 24 h after stroke [68]. This decrease of brain swelling was correlated with the loss of the perivascular AQP4 domain in -syntrophin-KO mice [68]. These results suggest that perivascular AQP4 has an important role in edema formation. However, the absence of AQP4 in AQP4-KO mice also prevents water clearance in an experiment of intrastriatal infusion of a saline solution, showing that AQP4 is critical for water removal from tissue [95]. Conversely, in a preconditioning stroke model, a higher induction of AQP4 was correlated with edema reduction [27]. However, this reduction of edema may be referring to vasogenic edema, in which case, AQP4 is said to aid in edema resolution by actively pumping out water from the cerebral tissue to peripheral blood [95]. The redistribution of the water in the astrocyte compartment through the astrocyte network would also be possible for the CSF compartments. This hypothesis is supported by a publication showing an increase of AQP4 in ependymal cells in the border of the ventricles in a traumatic brain injury model [96].

To summarize, the exact mechanism causing decreased edema formation is not yet fully understood, but AQP4 and the astrocyte network with the gap-junction proteins may certainly be contributing. Osmotic gradients can also play an important role, and recently, high AQP4 expression was observed in hypersaline treatment after stroke correlating with decreased edema formation at 48 hours [97].

4. rtPA: A Unique Drug for Stroke Treatment with Aversive Effects on the NVU

4.1. Clinical Evidence (from Bed to the Bench, Neurotoxicity of rtPA)

As discussed in the introduction, recombinant tissue plasminogen activator (rtPA) is currently the only thrombolytic molecule FDA approved for treatment of acute ischemic stroke [4]. The intact BBB is usually an obstacle for most neuropharmacological agents in healthy patients. The dysfunction of the BBB after ischemia could cause problems for the therapeutic function of rtPA. This protease targets fibrin-bound plasminogens and converts them into plasmins, which then cut the fibrin clot and lyse it. Intravenously infused at a dose of 0.9 mg/kg over one hour, rtPA provides increased survival and better neurological outcomes [4]. To be beneficial for the patient, rtPA must be administered within the first 4 h 30 min after stroke onset [5, 6]. Despite the organization of emergency care, only 5% of stroke patients are eligible for this therapy. In fact, late administration of rtPA translated to a higher risk of bleeding and extension of the lesion [7]. Higher doses of rtPA do not bind only the fibrin clot but also activate the circulating plasminogen activator (tPA). This activation contributes to a generalized fibrinolysis and fibrinogenolysis, which is suspected to be a cause of bleeding. But the mechanisms of the hemorrhagic transformation after rtPA treatment seem to be more complex than can be accounted for by the affinity of rtPA for fibrin alone. In fact, the enhanced fibrin specificity of tenecteplase and reteplase, two rtPA derivatives, resulted in no significant difference in terms of cerebral hemorrhage [98, 99].

Interestingly, the comparison with myocardial infarction shows a low incidence of cerebral hemorrhage after rtPA administration [100] suggesting a direct link between bleeding and the ischemic pathophysiology. Clinical studies showed that 80% of bleeding after cerebral thrombolysis occur preferentially in the ischemic territory [7].

4.2. Aversive Effects of rtPA Treatment on the NVU after Stroke

To have a better understanding of the aversive effects of rtPA its neurotoxic effects were examined. It is well known that endogenous tPA is present in the blood stream, endothelial cells, neurons, and microglial cells [101]. In the brain parenchyma, tPA activity was found to be pleiotropic and associated with synaptic plasticity and cell death [102104]. In fact, tPA interacts with several neuronal proteins such as N-methyl-D-aspartate (NMDA) receptors, one subtype of glutamatergic receptors, low-density lipoprotein-receptor-related protein (LRP), and Annexin-II [101, 105, 106]. tPA is synthesized in neurons, stored in presynaptic vesicles, and released following depolarization in synergy with the neurotransmitters. In the synaptic cleft, tPA binds and cleaves the NR1 subunit of NMDA receptors that causes an amplification of calcium influx in postsynaptic neurons and an increase of the glutamatergic response in physiological conditions. However, this physiological response becomes excitotoxic after ischemia and is magnified after rtPA injection [101, 107, 108]. The injection of antibodies against the NR1-subunit prevented these proexcitotoxic effects of endogenous tPA and reduced brain infarction and BBB leakage after stroke [109]. These data suggest that the NMDA receptor may be a protective drug target for the NVU after stroke and may provide a potential extension of the rtPA therapeutic window [109].

The presence of rtPA in the brain parenchyma has been explained by its passage through the BBB in several in vitro models with different proposed mechanisms.(i)rtPA diffuses into the brain parenchyma through an already opened BBB as a consequence of the ischemic process. As we discussed previously, the kinetics of the BBB opening is complex in the early stages after stroke and it is difficult to observe this with clinical imaging [29]. Interestingly, in vitro endothelial monolayer cultured with astrocytes enables us to observe the ability of rtPA to cross the intact BBB [110], which is increased under oxygen-glucose deprivation (OGD) [111]. Therefore, as rtPA potentially diffuses through an open or closed BBB in early time points after stroke onset, it may aggravate neuronal cell death as described previously.(ii)rtPA could cross the BBB by degrading the endothelium via its own proteolytic activity, but it is not a requirement in the intact BBB [110]. The ability of rtPA to cross the intact BBB at a thrombolytic dose suggests that this protease may interact first with the endothelial cells before the BBB breakdown. In fact, rtPA promotes breakdown of the BBB [112] by stimulating the synthesis activity of MMP-9 [113116] and other MMP isoforms [117] exacerbating the degradation of the basal lamina and subsequent vasogenic edema formation and hemorrhage. The thrombolytic products could exacerbate the proposed mechanism [118].(iii)Finally, LRP potentially contributes in trans-endothelial transport of the exogenous rtPA [106, 119, 120] and then activates the astrocytic MMP-9 and nuclear factor NF- B, which promotes the expression of inducible nitric oxide synthase (iNOS). This increase of NO results in increased BBB permeability [121].

With all these data together, Yepes and collaborators have proposed the following potential cellular and molecular events to explain the toxicity of the rtPA and tPA on the NVU [104].(1)Circulating endogenous tPA and rtPA cross the BBB (intact or damaged endothelial layer) and increase MMP-9 activity in the basal lamina soon after stroke onset which compromises the NVU integrity and makes it fragile. (2)Then tPA and rtPA bind to the astrocytic LRP, inducing the loss of the extracellular domain of LRP [122, 123] in the basal lamina, and release the intracellular domain of LRP in the astrocytic cytoplasm to activate NF- B. This NF- B activation increases iNOS and MMP9 expression and overall function in the whole NVU, causing separation of astrocytic endfeet from the basal lamina. This is usually observed at the later stages of BBB breakdown. However, it is tempting to speculate that this cascade, which involves the perivascular cells of the NVU, would be an accelerated pathological process resulting from the use of rtPA. It is possible that rtPA and tPA may also affect the phenotype of the astrocyte endfeet by changes in the level of expression of key proteins such as AQP4 and also Cx43.

4.3. New Therapeutic Strategies for rtPA Treatment after Stroke

The BBB is definitely not a barrier to rtPA in stroke but the BBB does become a serious barrier to the effective usage of this drug in clinic due to the neurotoxic effects and the risk of hemorrhagic transformation. Interestingly, tPA may be endogenously synthesized by the central nervous system in neurons and endothelial cells [124]. However, tPA and rtPA have effects on the endothelial cells, astrocytes, and neurons and possibly other glial cell types such as oligodendrocytes and microglia. In order to prevent the aversive effects of rtPA while maintaining the benefits of early reperfusion, several new therapeutic strategies have been examined to prevent the interaction of rtPA with the NMDA receptor within the NVU [104]. In fact, NMDA receptors are expressed not only in neurons but also in oligodendrocytes and endothelial cells [125, 126]. One of these strategies uses an LRP antagonist (RAP) to minimize the binding of rtPA with LRP in the endothelial cells. A second strategy uses the ATD-NR1 antibody to block rtPA binding of the NR1 subunit on neuronal NMDA receptors. The last one uses a mutation of the rtPA to decrease its adverse effects on the nervous tissue [104]. An example of a natural drug, desmoteplase, the vampire bat Desmodus Rotundus Salivary Plasminogen Activator (DSPA), is a thrombolytic agent under development. It shows little neurotoxicity and has the ability to interact with the BBB endothelium through the same receptor (LRP) as that of tPA [127, 128]. Unfortunately, the clinical trial of DIAS-2 (Desmoteplase In Acute ischemic Stroke) showed no benefit of the desmoteplase versus placebo [129]. Although the outcome of this clinical trial was disappointing, promising alternatives pathways are being investigated. In fact, Gleevec, a FDA approved drug for treatment of chronic myelogenous leukemia, was recently proposed to prevent the complications associated with rtPA treatment [130]. Gleevec inhibits the activation of platelet-derived growth factor alpha receptor (PDGFR). It was shown that tPA increases BBB permeability through the indirect activation of perivascular astrocytic PDGFR [130].

MMP inhibition is a good strategy based on reports of easy monitoring of MMP blood levels, defining them as potential biomarkers of brain damage [131, 132]. But because endogenous MMPs are also key mediators in stroke recovery by contributing to inflammatory and remodeling responses, pharmacological targeting must be accurately applied for acute stroke phases so; their beneficial effects are not compromised [133, 134]. Despite efforts to understand the complex link between BBB integrity and the hemorrhage risk [112], a better definition and understanding of NVU kinetics and the mechanisms underlying their dysfunction is still needed to better define eligibility criteria for rtPA treatment. Thus, alternative approaches other than MMP inhibition as mentioned before in some recent developments will offer interesting treatment strategies after stroke.

5. NVU Protection May Be the Future instead of Neuroprotection in Stroke Treatment

5.1. Preconditioning for Future Development of New Drugs

Given the small number of patients eligible for thrombolysis, many pharmaceutical compounds have been developed to limit the progression of brain injury by targeting different mechanisms leading to neuronal death [135]. Despite promising protective effects observed in preclinical studies, no compound to date has demonstrated benefit against stroke-induced neuronal death after facing the rigorous wall of clinical trials [136].

As mentioned in Section 1, research on brain diseases has focused on neuronal damage, as it was thought to be the major cause of cognitive deficits. However, ischemic stroke is a complex brain disease characterized by sudden onset of disabilities related to brain damage with a vascular origin. Because the development of many neuroprotective molecules for treatment over the last twenty years has been unsuccessful, researchers have switched gears towards investigating the natural endogenous neuroprotection of ischemic tolerance [137]. The purpose of the ischemic tolerance preconditioning is to induce endogenous defense mechanisms prior to the ischemic event that will attenuate the eventual consequences of ischemia. This resistance to ischemic damage can be achieved experimentally by several stimuli including ischemic preconditioning [138]. The concept and protocols were adapted from previous studies done in myocardial infarction. In fact, a short duration of coronary occlusion is unable to cause myocyte necrosis. However, when carried out before a prolonged occlusion, a short occlusion significantly reduced the final infarct volume of the myocardium [139]. This initial nonharmful ischemic insult triggered endogenous mechanisms that made the organ more resistant to the next attack for up to two periods of ischemic tolerance [139]. The first period of ischemic tolerance resulted from posttranscriptional responses and began minutes after preconditioning. The second, longer period, began 24 hours after preconditioning and lasted up to 7 days with maximal protection found at 3 days.

As with the cardiac preconditioning, ischemic tolerance in the brain also has delayed mechanisms leading to neuroprotection [140]. However, the mechanisms are complex and not well understood. The induction of ischemic tolerance likely depends on the coordinated responses at the genomic, molecular, cellular, and tissue levels [141143], which suggests the importance of the interactions between the astrocyte and endothelial cells in the NVU. Regarding neurovascular events in stroke pathophysiology, there has been a growing interest in vascular approaches to the preconditioning mechanisms. Protective effects of preconditioning were observed in vivo, demonstrating that endothelium function is preserved by improving cerebral blood flow during reperfusion in areas surrounding the lesion [144], and that BBB integrity is maintained with a reduction in edema formation [145]. The induced protection was again correlated not only with a decreased expression of MMP-9 [146] but also with a reduced neutrophil adhesion to endothelial cells through a decreased expression of ICAM-1 [147, 148]. These results were confirmed by in vitro studies that report a protective effect via preservation of BBB integrity, by both a decreased expression of the inflammatory molecules ICAM-1 and VCAM-1 [149, 150] and maintenance of tight junction structure [149]. Moreover, preconditioning also facilitates the increase of AQP4 expression at early time-points after stroke onset, which is associated with a decrease of the edema formation [27]. A recent study also reported the protective role of glial tissue preconditioning in severe stroke [151]. These recent observations suggest that future drug development must focus on drugs affecting the entire NVU instead of one cell type as was proposed in the 1990s with the development of calcium channel and NMDA inhibitors. Recently, some compounds like edaravone, an antioxidant, showed benefits in preclinical and clinical studies by protection of the NVU [152, 153]. But further trials are needed to confirm these promising preliminary results [154].

5.2. Protection of the NVU: Focus on PPARs

Preventive neuroprotection also involves management of risk factors, which is supported by studies showing that physical exercise [155] or lipid-lowering treatment reduces the occurrence and severity of stroke [156158]. In this context, the involvement of pharmacological agents that are activators of nuclear receptors like peroxisome proliferator-activated receptors (PPARs) could be a promising study. Present in three isoforms, , , and , these receptors exhibit pleiotropic activity in the sense that they can activate or repress the transcription of many genes involved in lipid and carbohydrate metabolism in addition to inflammation [159, 160]. PPARs are expressed in neurons, endothelial cells, and glial cells [161]. Activation of the PPARs has long-term effects lasting from hours to days, which correspond to an activation of gene transcription (named transactivation) as has been seen in lipid and carbohydrate metabolism. However, activation of the PPARs induce a cellular response within minutes to hours and this corresponds to an inhibition of gene transcription named transrepression [162]. The latter mechanism does not require binding to DNA, but rather protein-protein interaction involving other transcription factors like NF- B of STAT-3 and AP-1, to inhibit their activity as reported for inflammatory genes [163].

Independent of its lipid-lowering activity, PPAR- activation was found to be neuroprotective in several in vivo studies carried out in mice subjected to transient ischemia with preventive or curative treatments by agonists such as fenofibrate, WY-14643, and resveratrol (a polyphenol present in grapes) [164166]. The observed protection is the result of an anti-inflammatory mechanism, which decreases the expression of adhesion molecules, ICAM-1 and VCAM-1, in brain endothelial cells. Effects of antioxidants were also observed. However, a study using a BBB in vitro model combining endothelial cells with glial cells from wild-type or PPAR- knockout mice has demonstrated not only that the observed protection against OGD-induced hyperpermeability was dependent on this nuclear receptor activation but also that the ligand targeted specifically the endothelial cells without modulation of the classical PPAR- target genes associated with inflammation or metabolism [167]. Moreover, protective effects of PPAR- were not only reported through similar mechanisms [168] but also via an inhibition of NF B and TNF- pathways [169, 170] and macrophages/microglial cells activation, thus preventing cytokine production [171]. One study also suggests that PPAR- agonists could inhibit excitotoxicity-induced neuronal death [172].

Statins are HMG-CoA reductase inhibitors. This enzyme catalyzes the conversion of HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) to mevalonate, a precursor of cholesterol. As lipid lowering agents statins also exert pleiotropic effects at the vascular level [173]. In addition to protection against excitotoxicity in cultured neurons [174], statins have demonstrated preservation of BBB endothelial cells’ integrity against glutamate excitotoxic challenge in vitro [175]. These compounds also enabled the reduction of MMP-9 synthesis in rtPA-activated astrocytes [176]. The effects of statins may involve nuclear receptors, through an increase in both expression and activity of PPAR- [177179]. More recently, brain endothelial PPAR- activation has proven to be protective against ischemia-induced cell death through inhibition of the miR-15a microRNA, thus strengthening the therapeutic concept based on activation of PPARs for the treatment of stroke-related microvascular dysfunction [180].

5.3. Inhibition of JNK Activation and NVU Protection

The c-Jun N-terminal kinases (JNKs) belong to the mitogen activated protein kinase (MAPK) family; the two other members being p38 and ERK [181, 182]. The isoforms JNK1 and JNK2 are ubiquitously distributed, while JNK3 is primarily expressed in the heart, brain, endocrine pancreas and testis [183]. JNKs are activated by phosphorylation, which is catalyzed by upstream kinases—MKK 4 and 7 [182184]. JNK activation is essential for normal brain development and organogenesis during embryonic development [185]. However, the activation of JNKs plays several roles ranging from regulation of cell survival and apoptosis to cell proliferation [183, 185187]. They are activated under pathological conditions both in the brain [188, 189] and in the periphery [190, 191]. In fact, JNK phosphorylation initially decreases after stroke and then starts to increase at 1.5 hours with a maximum at 9 hours after onset [192]. Phosphorylation of c-Jun, a JNK substrate, follows the same temporal pattern, peaking at 8 hours post-stroke [192, 193].

The development of the peptide named DJNKi, a competitive inhibitor of the JNK signaling pathway, has been shown to reduce lesion volume of mice with transient MCAO by 90% even when induced 6 hours after injury. This lesion volume decrease was accompanied by behavior improvements as well [193], suggesting an increase of the therapeutic time window almost 2 times longer than tPA. This positive outcome was also observed in a more severe model with a permanent occlusion model [194]. Moreover, DJNKi has been shown to be compatible for treatment of ischemic stroke even in the presence of rtPA and was shown to decrease lesion volume [195]. DJNKi also improved neurobehavior scores and decreased hemispheric swelling after a model of intra-cerebral hemorrhage [196]. Thus, DJNKi could possibly attenuate the highly probable side effect of hemorrhagic transformation caused by rtPA. Interestingly, in this model of intracerebral hemorrhage, DJNKi administration significantly increased AQP4 expression 48 hours after injury. This increase in AQP4 expression negatively correlated with decreased hemispheric swelling, thus pointing towards a possible role of DJNKi controlling edema as well. In fact, activation of the JNK pathway is present not only in the neurons but also in glial cells [197] and brain endothelial cells [198]. Such activation in nonneuronal cells may negatively impact neuronal cell death and function [197]. In the context of broad effects of this drug, Benakis et al. [199] showed that DJNKI-1, injected peripherally, is able to modulate some nonneuronal inflammatory processes. As discussed previously, the development of a drug targeting several cells such as in the NVU may help to move towards success in the clinic.

6. Summary and Perspectives in Stroke Research

In summary, the data found in the literature suggest that the failure of agents in protecting the brain against stroke may come from the fact that each developed compound targeted only one mechanism and one cell type of stroke pathophysiology. Ischemic preconditioning appears to be an attractive experimental strategy that would identify endogenous mechanisms of protection and regeneration. Recent evidence of such protective mechanisms supports a complex action on cells of the NVU, underlining the importance of the interactions between endothelial cells and astrocytes in the pathophysiology after stroke. As our knowledge of the NVU increases, molecules with pleiotropic activity will become increasing useful in the development of post-ischemic treatments in the clinics.

Conflict of Interest

The authors declare that they have no conflict of interests.

Acknowledgments

The authors thank Jacqueline Coats (Loma Linda University) for text revision. They also thank Françoise Dieterlen and the French Société de Biologie” for permitting the translation of some topics previously published in the French journal “Biologie Aujourd’hui. This paper was supported in part by the NIH R01HD061946 (to J. Badaut), the Swiss Science Foundation (FN 31003A-122166 to J. Badaut), and the European Union's Seventh Framework Programme (FP7/2007-2013) under Grant agreements nos. 201024 and 202213 (European Stroke Network).

References

  1. V. L. Roger, A. S. Go, D. M. Lloyd-Jones et al., “Heart disease and stroke statistics-2011 update: a report from the American Heart Association,” Circulation, vol. 123, no. 4, pp. e18–e19, 2011. View at Publisher · View at Google Scholar · View at Scopus
  2. W. Rosamond, K. Flegal, G. Friday et al., “Heart disease and stroke statistics—2007 Update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee,” Circulation, vol. 115, no. 5, pp. e69–e171, 2007. View at Publisher · View at Google Scholar · View at Scopus
  3. J. L. Saver, “Time is brain—quantified,” Stroke, vol. 37, no. 1, pp. 263–266, 2006. View at Publisher · View at Google Scholar · View at Scopus
  4. J. R. Marler, “Tissue plasminogen activator for acute ischemic stroke. The National Institute of Neurological Disorders and Stroke rt-PA Stroke study group,” New England Journal of Medicine, vol. 333, no. 24, pp. 1581–1587, 1995. View at Publisher · View at Google Scholar · View at Scopus
  5. W. Hacke, M. Kaste, E. Bluhmki et al., “Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke,” New England Journal of Medicine, vol. 359, no. 13, pp. 1317–1329, 2008. View at Publisher · View at Google Scholar · View at Scopus
  6. J. L. Saver, J. Gornbein, J. Grotta et al., “Number needed to treat to benefit and to harm for intravenous tissue plasminogen activator therapy in the 3- to 4.5-hour window Joint outcome table analysis of the ECASS 3 trial,” Stroke, vol. 40, no. 7, pp. 2433–2437, 2009. View at Publisher · View at Google Scholar · View at Scopus
  7. J. P. Broderick, “Intracerebral hemorrhage after intravenous t-PA therapy for ischemic stroke,” Stroke, vol. 28, no. 11, pp. 2109–2118, 1997. View at Scopus
  8. G. J. Del Zoppo, “The neurovascular unit in the setting of stroke,” Journal of Internal Medicine, vol. 267, no. 2, pp. 156–171, 2010. View at Publisher · View at Google Scholar · View at Scopus
  9. C. Iadecola, “Neurovascular regulation in the normal brain and in Alzheimer's disease,” Nature Reviews Neuroscience, vol. 5, no. 5, pp. 347–360, 2004. View at Scopus
  10. C. Iadecola and M. Nedergaard, “Glial regulation of the cerebral microvasculature,” Nature Neuroscience, vol. 10, no. 11, pp. 1369–1376, 2007. View at Publisher · View at Google Scholar · View at Scopus
  11. E. A. Neuwelt, B. Bauer, C. Fahlke et al., “Engaging neuroscience to advance translational research in brain barrier biology,” Nature Reviews Neuroscience, vol. 12, no. 3, pp. 169–182, 2011. View at Publisher · View at Google Scholar · View at Scopus
  12. B. Engelhardt and C. Coisne, “Fluids and barriers of the CNS establish immune privilege by confining immune surveillance to a two-walled castle moat surrounding the CNS castle,” Fluids and Barriers of the CNS, vol. 8, no. 1, Article ID 4, 2011. View at Publisher · View at Google Scholar · View at Scopus
  13. N. J. Abbott, L. Rönnbäck, and E. Hansson, “Astrocyte-endothelial interactions at the blood-brain barrier,” Nature Reviews Neuroscience, vol. 7, no. 1, pp. 41–53, 2006. View at Publisher · View at Google Scholar · View at Scopus
  14. E. Hamel, “Perivascular nerves and the regulation of cerebrovascular tone,” Journal of Applied Physiology, vol. 100, no. 3, pp. 1059–1064, 2006. View at Publisher · View at Google Scholar · View at Scopus
  15. L. Pellerin, “Food for thought: the importance of glucose and other energy substrates for sustaining brain function under varying levels of activity,” Diabetes and Metabolism, vol. 36, no. 3, pp. S59–S63, 2010. View at Publisher · View at Google Scholar · View at Scopus
  16. N. J. Abbott, A. A. K. Patabendige, D. E. M. Dolman, S. R. Yusof, and D. J. Begley, “Structure and function of the blood-brain barrier,” Neurobiology of Disease, vol. 37, no. 1, pp. 13–25, 2010. View at Publisher · View at Google Scholar · View at Scopus
  17. T. Owens, I. Bechmann, and B. Engelhardt, “Perivascular spaces and the two steps to neuroinflammation,” Journal of Neuropathology and Experimental Neurology, vol. 67, no. 12, pp. 1113–1121, 2008. View at Publisher · View at Google Scholar · View at Scopus
  18. A. W. Unterberg, J. Stover, B. Kress, and K. L. Kiening, “Edema and brain trauma,” Neuroscience, vol. 129, no. 4, pp. 1021–1029, 2004. View at Publisher · View at Google Scholar · View at Scopus
  19. J. M. Simard, T. A. Kent, M. Chen, K. V. Tarasov, and V. Gerzanich, “Brain oedema in focal ischaemia: molecular pathophysiology and theoretical implications,” The Lancet Neurology, vol. 6, no. 3, pp. 258–268, 2007. View at Publisher · View at Google Scholar · View at Scopus
  20. J. Badaut, S. Ashwal, and A. Obenaus, “Aquaporins in cerebrovascular disease: a target for treatment of brain edema?” Cerebrovascular Diseases, vol. 31, no. 6, pp. 521–531, 2011. View at Publisher · View at Google Scholar · View at Scopus
  21. Z. Zador, G. T. Manley, S. Stiver, and V. Wang, “Role of aquaporin-4 in cerebral edema and stroke,” Handbook of Experimental Pharmacology, vol. 190, pp. 159–170, 2009. View at Publisher · View at Google Scholar · View at Scopus
  22. W. C. Risher, R. D. Andrew, and S. A. Kirov, “Real-time passive volume responses of astrocytes to acute osmotic and ischemic stress in cortical slices and in vivo revealed by two-photon microscopy,” GLIA, vol. 57, no. 2, pp. 207–221, 2009. View at Publisher · View at Google Scholar · View at Scopus
  23. M. E. O'Donnell, T. I. Lam, L. Tran, and S. E. Anderson, “The role of the blood-brain barrier Na-K-2Cl cotransporter in stroke,” Advances in Experimental Medicine and Biology, vol. 559, pp. 67–75, 2005. View at Scopus
  24. S. P. Duckles and D. N. Krause, “Mechanisms of cerebrovascular protection: oestrogen, inflammation and mitochondria,” Acta Physiologica, vol. 203, no. 1, pp. 149–154, 2011. View at Publisher · View at Google Scholar · View at Scopus
  25. S. P. Duckles, D. N. Krause, C. Stirone, and V. Procaccio, “Estrogen and mitochondria: a new paradigm for vascular protection?” Molecular Interventions, vol. 6, no. 1, pp. 26–35, 2006. View at Publisher · View at Google Scholar · View at Scopus
  26. M. De Castro Ribeiro, L. Hirt, J. Bogousslavsky, L. Regli, and J. Badaut, “Time course of aquaporin expression after transient focal cerebral ischemia in mice,” Journal of Neuroscience Research, vol. 83, no. 7, pp. 1231–1240, 2006. View at Publisher · View at Google Scholar · View at Scopus
  27. L. Hirt, B. Ternon, M. Price, N. Mastour, J. F. Brunet, and J. Badaut, “Protective role of early Aquaporin 4 induction against postischemic edema formation,” Journal of Cerebral Blood Flow and Metabolism, vol. 29, no. 2, pp. 423–433, 2009. View at Publisher · View at Google Scholar · View at Scopus
  28. D. Strbian, A. Durukan, M. Pitkonen et al., “The blood-brain barrier is continuously open for several weeks following transient focal cerebral ischemia,” Neuroscience, vol. 153, no. 1, pp. 175–181, 2008. View at Publisher · View at Google Scholar · View at Scopus
  29. E. C. Henning, L. L. Latour, and S. Warach, “Verification of enhancement of the CSF space, not parenchyma, in acute stroke patients with early blood-brain barrier disruption,” Journal of Cerebral Blood Flow and Metabolism, vol. 28, no. 5, pp. 882–886, 2008. View at Publisher · View at Google Scholar · View at Scopus
  30. A. M. Romanic, R. F. White, A. J. Arleth, E. H. Ohlstein, and F. C. Barone, “Matrix metalloproteinase expression increases after cerebral focal ischemia in rats: inhibition of matrix metalloproteinase-9 reduces infarct size,” Stroke, vol. 29, no. 5, pp. 1020–1030, 1998. View at Scopus
  31. A. Kastrup, T. Engelhorn, C. Beaulieu, A. De Crespigny, and M. E. Moseley, “Dynamics of cerebral injury, perfusion, and blood-brain barrier changes after temporary and permanent middle cerebral artery occlusion in the rat,” Journal of the Neurological Sciences, vol. 166, no. 2, pp. 91–99, 1999. View at Publisher · View at Google Scholar · View at Scopus
  32. S. Nagahiro, S. Goto, K. Kogo, M. Sumi, M. Takahashi, and Y. Ushio, “Sequential changes in ischemic edema following transient focal cerebral ischemia in rats: magnetic resonance imaging study,” Neurologia Medico-Chirurgica, vol. 34, no. 7, pp. 412–417, 1994. View at Scopus
  33. M. Plateel, E. Teissier, and R. Cecchelli, “Hypoxia dramatically increases the nonspecific transport of blood-borne proteins to the brain,” Journal of Neurochemistry, vol. 68, no. 2, pp. 874–877, 1997. View at Scopus
  34. A. Rosell, A. Ortega-Aznar, J. Alvarez-Sabín et al., “Increased brain expression of matrix metalloproteinase-9 after ischemic and hemorrhagic human stroke,” Stroke, vol. 37, no. 6, pp. 1399–1406, 2006. View at Publisher · View at Google Scholar · View at Scopus
  35. G. A. Rosenberg, E. Y. Estrada, and J. E. Dencoff, “Matrix metalloproteinases and TIMPs are associated with blood-brain barrier opening after reperfusion in rat brain,” Stroke, vol. 29, no. 10, pp. 2189–2195, 1998. View at Scopus
  36. E. Candelario-Jalil, Y. Yang, and G. A. Rosenberg, “Diverse roles of matrix metalloproteinases and tissue inhibitors of metalloproteinases in neuroinflammation and cerebral ischemia,” Neuroscience, vol. 158, no. 3, pp. 983–994, 2009. View at Publisher · View at Google Scholar · View at Scopus
  37. A. Obenaus and S. Ashwal, “Magnetic resonance imaging in cerebral ischemia: focus on neonates,” Neuropharmacology, vol. 55, no. 3, pp. 271–280, 2008. View at Publisher · View at Google Scholar · View at Scopus
  38. C. A. Chastain, U. E. Oyoyo, M. Zipperman et al., “Predicting outcomes of traumatic brain injury by imaging modality and injury distribution,” Journal of Neurotrauma, vol. 26, no. 8, pp. 1183–1196, 2009. View at Publisher · View at Google Scholar · View at Scopus
  39. J. Badaut, S. Ashwal, B. Tone, L. Regli, H. R. Tian, and A. Obenaus, “Temporal and regional evolution of aquaporin-4 expression and magnetic resonance imaging in a rat pup model of neonatal stroke,” Pediatric Research, vol. 62, no. 3, pp. 248–254, 2007. View at Publisher · View at Google Scholar · View at Scopus
  40. D. R. Pillai, M. S. Dittmar, D. Baldaranov et al., “Cerebral ischemia-reperfusion injury in rats—A 3 T MRI study on biphasic blood-brain barrier opening and the dynamics of edema formation,” Journal of Cerebral Blood Flow and Metabolism, vol. 29, no. 11, pp. 1846–1855, 2009. View at Publisher · View at Google Scholar · View at Scopus
  41. C. S. Ábrahám, N. Harada, M. A. Deli, and M. Niwa, “Transient forebrain ischemia increases the blood-brain barrier permeability for albumin in stroke-prone spontaneously hypertensive rats,” Cellular and Molecular Neurobiology, vol. 22, no. 4, pp. 455–462, 2002. View at Publisher · View at Google Scholar · View at Scopus
  42. R. S. Bourke, H. K. Kimelberg, L. R. Nelson et al., “Biology of glial swelling in experimental brain edema,” Advances in Neurology, vol. 28, pp. 99–109, 1980. View at Scopus
  43. H. K. Kimelberg, “Astrocytic swelling in cerebral ischemia as a possible cause of injury and target for therapy,” GLIA, vol. 50, no. 4, pp. 389–397, 2005. View at Publisher · View at Google Scholar · View at Scopus
  44. J. M. Rutkowsky, B. K. Wallace, P. M. Wise, and M. E. O'Donnell, “Effects of estradiol on ischemic factor-induced astrocyte swelling and AQP4 protein abundance,” American Journal of Physiology, vol. 301, no. 1, pp. C204–C212, 2011. View at Publisher · View at Google Scholar · View at Scopus
  45. M. J. Tait, S. Saadoun, B. A. Bell, and M. C. Papadopoulos, “Water movements in the brain: role of aquaporins,” Trends in Neurosciences, vol. 31, no. 1, pp. 37–43, 2008. View at Publisher · View at Google Scholar · View at Scopus
  46. J. Badaut, A. Nehlig, J. M. Verbavatz, M. E. Stoeckel, M. J. Freund-Mercier, and F. Lasbennes, “Hypervascularization in the magnocellular nuclei of the rat hypothalamus: relationship with the distribution of aquaporin-4 and markers of energy metabolism,” Journal of Neuroendocrinology, vol. 12, no. 10, pp. 960–969, 2000. View at Publisher · View at Google Scholar · View at Scopus
  47. J. Badaut, J. M. Verbavatz, M. J. Freund-Mercier, and F. Lasbennes, “Presence of aquaporin-4 and muscarinic receptors in astrocytes and ependymal cells in rat brain: a clue to a common function?” Neuroscience Letters, vol. 292, no. 2, pp. 75–78, 2000. View at Publisher · View at Google Scholar · View at Scopus
  48. I. I. Arciénega, J. F. Brunet, J. Bloch, and J. Badaut, “Cell locations for AQP1, AQP4 and 9 in the non-human primate brain,” Neuroscience, vol. 167, no. 4, pp. 1103–1114, 2010. View at Publisher · View at Google Scholar · View at Scopus
  49. D. Dolman, S. Drndarski, N. J. Abbott, and M. Rattray, “Induction of aquaporin 1 but not aquaporin 4 messenger RNA in rat primary brain microvessel endothelial cells in culture,” Journal of Neurochemistry, vol. 93, no. 4, pp. 825–833, 2005. View at Publisher · View at Google Scholar · View at Scopus
  50. M. Amiry-Moghaddam, R. Xue, F. M. Haug et al., “Alpha-syntrophin deletion removes the perivascular but not endothelial pool of aquaporin-4 at the blood-brain barrier and delays the development of brain edema in an experimental model of acute hyponatremia,” The FASEB Journal, vol. 18, no. 3, pp. 542–544, 2004. View at Scopus
  51. S. Nielsen, E. A. Nagelhus, M. Amiry-Moghaddam, C. Bourque, P. Agre, and O. R. Ottersen, “Specialized membrane domains for water transport in glial cells: high- resolution immunogold cytochemistry of aquaporin-4 in rat brain,” Journal of Neuroscience, vol. 17, no. 1, pp. 171–180, 1997. View at Scopus
  52. G. P. Nicchia, B. Nico, L. M. A. Camassa et al., “The role of aquaporin-4 in the blood-brain barrier development and integrity: studies in animal and cell culture models,” Neuroscience, vol. 129, no. 4, pp. 935–945, 2004. View at Publisher · View at Google Scholar · View at Scopus
  53. H. Wolburg, S. Noell, A. Mack, K. Wolburg-Buchholz, and P. Fallier-Becker, “Brain endothelial cells and the glio-vascular complex,” Cell and Tissue Research, vol. 335, no. 1, pp. 75–96, 2009. View at Publisher · View at Google Scholar · View at Scopus
  54. H. Wolburg, S. Noell, K. Wolburg-Buchholz, A. MacK, and P. Fallier-Becker, “Agrin, aquaporin-4, and astrocyte polarity as an important feature of the blood-brain barrier,” Neuroscientist, vol. 15, no. 2, pp. 180–193, 2009. View at Publisher · View at Google Scholar · View at Scopus
  55. J. Badaut, S. Ashwal, A. Adami et al., “Brain water mobility decreases after astrocytic aquaporin-4 inhibition using RNA interference,” Journal of Cerebral Blood Flow and Metabolism, vol. 31, no. 3, pp. 819–831, 2011. View at Publisher · View at Google Scholar · View at Scopus
  56. J. Yu, A. J. Yool, K. Schulten, and E. Tajkhorshid, “Mechanism of gating and ion conductivity of a possible tetrameric pore in aquaporin-1,” Structure, vol. 14, no. 9, pp. 1411–1423, 2006. View at Publisher · View at Google Scholar · View at Scopus
  57. J. E. Rash, K. G. V. Davidson, T. Yasumura, and C. S. Furman, “Freeze-fracture and immunogold analysis of aquaporin-4 (AQP4) square arrays, with models of AQP4 lattice assembly,” Neuroscience, vol. 129, no. 4, pp. 915–934, 2004. View at Publisher · View at Google Scholar · View at Scopus
  58. C. S. Furman, D. A. Gorelick-Feldman, K. G. V. Davidson et al., “Aquaporin-4 square array assembly: opposing actions of M1 and M23 isoforms,” Proceedings of the National Academy of Sciences of the United States of America, vol. 100, no. 23, pp. 13609–13614, 2003. View at Publisher · View at Google Scholar · View at Scopus
  59. M. Suzuki, Y. Iwasaki, and T. Yamamoto, “Disintegration of orthogonal arrays in perivascular astrocytic processes as an early event in acute global ischemia,” Brain Research, vol. 300, no. 1, pp. 141–145, 1984. View at Publisher · View at Google Scholar · View at Scopus
  60. R. Musa-Aziz, L. M. Chen, M. F. Pelletier, and W. F. Boron, “Relative CO2/NH3 selectivities of AQP1, AQP4, AQP5, AmtB, and RhAG,” Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 13, pp. 5406–5411, 2009. View at Publisher · View at Google Scholar · View at Scopus
  61. J. Zhou, H. Kong, X. Hua, M. Xiao, J. Ding, and G. Hu, “Altered blood-brain barrier integrity in adult aquaporin-4 knockout mice,” NeuroReport, vol. 19, no. 1, pp. 1–5, 2008. View at Publisher · View at Google Scholar · View at Scopus
  62. B. Nico, A. Frigeri, G. P. Nicchia et al., “Role of aquaporin-4 water channel in the development and integrity of the blood-brain barrier,” Journal of Cell Science, vol. 114, no. 7, pp. 1297–1307, 2001. View at Scopus
  63. Y. Hiroaki, K. Tani, A. Kamegawa et al., “Implications of the aquaporin-4 structure on array formation and cell adhesion,” Journal of Molecular Biology, vol. 355, no. 4, pp. 628–639, 2006. View at Publisher · View at Google Scholar · View at Scopus
  64. T. Gonen, P. Silz, J. Kistler, Y. Cheng, and T. Walz, “Aquaporin-0 membrane junctions reveal the structure of a closed water pore,” Nature, vol. 429, no. 6988, pp. 193–197, 2004. View at Publisher · View at Google Scholar · View at Scopus
  65. A. Warth, S. Kröger, and H. Wolburg, “Redistribution of aquaporin-4 in human glioblastoma correlates with loss of agrin immunoreactivity from brain capillary basal laminae,” Acta Neuropathologica, vol. 107, no. 4, pp. 311–318, 2004. View at Publisher · View at Google Scholar · View at Scopus
  66. E. Guadagno and H. Moukhles, “Laminin-induced aggregation of the inwardly rectifying potassium channel, Kir4.1, and the water-permeable channel, AQP4, via a dystroglycan-containing complex in astrocytes,” GLIA, vol. 47, no. 2, pp. 138–149, 2004. View at Publisher · View at Google Scholar · View at Scopus
  67. J. D. Neely, M. Amiry-Moghaddam, O. P. Ottersen, S. C. Froehner, P. Agre, and M. E. Adams, “Syntrophin-dependent expression and localization of Aquaporin-4 water channel protein,” Proceedings of the National Academy of Sciences of the United States of America, vol. 98, no. 24, pp. 14108–14113, 2001. View at Publisher · View at Google Scholar · View at Scopus
  68. M. Amiry-Moghaddam, T. Otsuka, P. D. Hurn et al., “An α-syntrophin-dependent pool of AQP4 in astroglial end-feet confers bidirectional water flow between blood and brain,” Proceedings of the National Academy of Sciences of the United States of America, vol. 100, no. 4, pp. 2106–2111, 2003. View at Publisher · View at Google Scholar · View at Scopus
  69. S. Saadoun, M. J. Tait, A. Reza et al., “AQP4 gene deletion in mice does not alter blood-brain barrier integrity or brain morphology,” Neuroscience, vol. 161, no. 3, pp. 764–772, 2009. View at Publisher · View at Google Scholar · View at Scopus
  70. G. Bix and R. V. Iozzo, “Novel interactions of perlecan: unraveling Perlecan's role in angiogenesis,” Microscopy Research and Technique, vol. 71, no. 5, pp. 339–348, 2008. View at Publisher · View at Google Scholar · View at Scopus
  71. S. Noell, P. Fallier-Becker, U. Deutsch, A. F. MacK, and H. Wolburg, “Agrin defines polarized distribution of orthogonal arrays of particles in astrocytes,” Cell and Tissue Research, vol. 337, no. 2, pp. 185–195, 2009. View at Publisher · View at Google Scholar · View at Scopus
  72. M. Ramos-Fernandez, M. F. Bellolio, and L. G. Stead, “Matrix metalloproteinase-9 as a marker for acute ischemic stroke: a systematic review,” Journal of Stroke and Cerebrovascular Diseases, vol. 20, no. 1, pp. 47–54, 2011. View at Publisher · View at Google Scholar · View at Scopus
  73. B. Lee, D. Clarke, A. Al Ahmad et al., “Perlecan domain V is neuroprotective and proangiogenic following ischemic stroke in rodents,” Journal of Clinical Investigation, vol. 121, no. 8, pp. 3005–3023, 2011. View at Publisher · View at Google Scholar · View at Scopus
  74. Z. G. Zhang, L. Zhang, Q. Jiang et al., “VEGF enhances angiogenesis and promotes blood-brain barrier leakage in the ischemic brain,” Journal of Clinical Investigation, vol. 106, no. 7, pp. 829–838, 2000. View at Scopus
  75. A. J. Al-Ahmad, B. Lee, M. Saini, and G. J. Bix, “Perlecan domain V modulates astrogliosis In vitro and after focal cerebral ischemia through multiple receptors and increased nerve growth factor release,” GLIA, vol. 59, no. 12, pp. 1822–1840, 2011. View at Publisher · View at Google Scholar · View at Scopus
  76. D. K. Binder, X. Yao, Z. Zador, T. J. Sick, A. S. Verkman, and G. T. Manley, “Increased seizure duration and slowed potassium kinetics in mice lacking aquaporin-4 water channels,” GLIA, vol. 53, no. 6, pp. 631–636, 2006. View at Publisher · View at Google Scholar · View at Scopus
  77. V. Benfenati, G. P. Nicchia, M. Svelto, C. Rapisarda, A. Frigeri, and S. Ferroni, “Functional down-regulation of volume-regulated anion channels in AQP4 knockdown cultured rat cortical astrocytes,” Journal of Neurochemistry, vol. 100, no. 1, pp. 87–104, 2007. View at Publisher · View at Google Scholar · View at Scopus
  78. J. E. Rash, “Molecular disruptions of the panglial syncytium block potassium siphoning and axonal saltatory conduction: pertinence to neuromyelitis optica and other demyelinating diseases of the central nervous system,” Neuroscience, vol. 168, no. 4, pp. 982–1008, 2010. View at Publisher · View at Google Scholar · View at Scopus
  79. C. Giaume, A. Koulakoff, L. Roux, D. Holcman, and N. Rouach, “Astroglial networks: a step further in neuroglial and gliovascular interactions,” Nature Reviews Neuroscience, vol. 11, no. 2, pp. 87–99, 2010. View at Publisher · View at Google Scholar · View at Scopus
  80. A. Tabernero, J. M. Medina, and C. Giaume, “Glucose metabolism and proliferation in glia: role of astrocytic gap junctions,” Journal of Neurochemistry, vol. 99, no. 4, pp. 1049–1061, 2006. View at Publisher · View at Google Scholar · View at Scopus
  81. A. L. Harris, “Connexin channel permeability to cytoplasmic molecules,” Progress in Biophysics and Molecular Biology, vol. 94, no. 1-2, pp. 120–143, 2007. View at Publisher · View at Google Scholar · View at Scopus
  82. G. Zoidl and R. Dermietzel, “On the search for the electrical synapse: a glimpse at the future,” Cell and Tissue Research, vol. 310, no. 2, pp. 137–142, 2002. View at Publisher · View at Google Scholar · View at Scopus
  83. J. E. Rash, T. Yasumura, K. G. V. Davidson, C. S. Furman, F. E. Dudek, and J. I. Nagy, “Identification of cells expressing Cx43, Cx30, Cx26, Cx32 and Cx36 in gap junctions of rat brain and spinal cord,” Cell Communication and Adhesion, vol. 8, no. 4-6, pp. 315–320, 2001. View at Scopus
  84. J. I. Nagy, F. E. Dudek, and J. E. Rash, “Update on connexins and gap junctions in neurons and glia in the mammalian nervous system,” Brain Research Reviews, vol. 47, no. 1-3, pp. 191–215, 2004. View at Publisher · View at Google Scholar · View at Scopus
  85. J. E. Rash, T. Yasumura, F. E. Dudek, and J. I. Nagy, “Cell-specific expression of connexins and evidence of restricted gap junctional coupling between glial cells and between neurons,” Journal of Neuroscience, vol. 21, no. 6, pp. 1983–2000, 2001. View at Scopus
  86. K. Nagasawa, H. Chiba, H. Fujita et al., “Possible involvement of gap junctions in the barrier function of tight junctions of brain and lung endothelial cells,” Journal of Cellular Physiology, vol. 208, no. 1, pp. 123–132, 2006. View at Publisher · View at Google Scholar · View at Scopus
  87. M. A. Avila, S. L. Sell, B. E. Hawkins et al., “Cerebrovascular connexin expression: effects of traumatic brain injury,” Journal of Neurotrauma, vol. 28, no. 9, pp. 1803–1811, 2011. View at Publisher · View at Google Scholar · View at Scopus
  88. R. Farahani, M. H. Pina-Benabou, A. Kyrozis et al., “Alterations in metabolism and gap junction expression may determine the role of astrocytes as “good Samaritans” or executioners,” GLIA, vol. 50, no. 4, pp. 351–361, 2005. View at Publisher · View at Google Scholar · View at Scopus
  89. T. Nakase, Y. Yoshida, and K. Nagata, “Enhanced connexin 43 immunoreactivity in penumbral areas in the human brain following ischemia,” GLIA, vol. 54, no. 5, pp. 369–375, 2006. View at Publisher · View at Google Scholar · View at Scopus
  90. W. M. Armstead, “Superoxide generation links protein kinase C activation to impaired ATP- sensitive K+ channel function after brain injury,” Stroke, vol. 30, no. 1, pp. 153–159, 1999. View at Scopus
  91. W. M. Armstead, “Age-dependent impairment of K(ATP) channel function following brain injury,” Journal of Neurotrauma, vol. 16, no. 5, pp. 391–402, 1999. View at Scopus
  92. J. Badaut, J. F. Brunet, and L. Regli, “Aquaporins in the brain: from aqueduct to “multi-duct”,” Metabolic Brain Disease, vol. 22, no. 3-4, pp. 251–263, 2007. View at Publisher · View at Google Scholar · View at Scopus
  93. A. S. Verkman, D. K. Binder, O. Bloch, K. Auguste, and M. C. Papadopoulos, “Three distinct roles of aquaporin-4 in brain function revealed by knockout mice,” Biochimica et Biophysica Acta, vol. 1758, no. 8, pp. 1085–1093, 2006. View at Publisher · View at Google Scholar · View at Scopus
  94. G. T. Manley, M. Fujimura, T. Ma et al., “Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke,” Nature Medicine, vol. 6, no. 2, pp. 159–163, 2000. View at Publisher · View at Google Scholar · View at Scopus
  95. M. C. Papadopoulos, G. T. Manley, S. Krishna, and A. S. Verkman, “Aquaporin-4 facilitates reabsorption of excess fluid in vasogenic brain edema,” FASEB Journal, vol. 18, no. 11, pp. 1291–1293, 2004. View at Publisher · View at Google Scholar · View at Scopus
  96. Q. Guo, I. Sayeed, L. M. Baronne, S. W. Hoffman, R. Guennoun, and D. G. Stein, “Progesterone administration modulates AQP4 expression and edema after traumatic brain injury in male rats,” Experimental Neurology, vol. 198, no. 2, pp. 469–478, 2006. View at Publisher · View at Google Scholar · View at Scopus
  97. C. H. Chen, R. Xue, J. Zhang, X. Li, S. Mori, and A. Bhardwaj, “Effect of osmotherapy with hypertonic saline on regional cerebral edema following experimental stroke: a study utilizing magnetic resonance imaging,” Neurocritical Care, vol. 7, no. 1, pp. 92–100, 2007. View at Publisher · View at Google Scholar · View at Scopus
  98. D. F. Chapman, P. Lyden, P. A. Lapchak, S. Nunez, H. Thibodeaux, and J. Zivin, “Comparison of TNK with wild-type tissue plasminogen activator in a rabbit embolic stroke model,” Stroke, vol. 32, no. 3, pp. 748–752, 2001. View at Scopus
  99. P. A. Lapchak, D. M. Araujo, and J. A. Zivin, “Comparison of Tenecteplase with Alteplase on clinical rating scores following small clot embolic strokes in rabbits,” Experimental Neurology, vol. 185, no. 1, pp. 154–159, 2004. View at Publisher · View at Google Scholar · View at Scopus
  100. J. H. Gurwitz, J. M. Gore, R. J. Goldberg et al., “Risk for intracranial hemorrhage after tissue plasminogen activator treatment for acute myocardial infarction,” Annals of Internal Medicine, vol. 129, no. 8, pp. 597–604, 1998. View at Scopus
  101. O. Nicole, F. Docagne, C. Ali et al., “The proteolytic activity of tissue-plasminogen activator enhances NMDA receptor-mediated signaling,” Nature Medicine, vol. 7, no. 1, pp. 59–64, 2001. View at Publisher · View at Google Scholar · View at Scopus
  102. K. Benchenane, J. P. López-Atalaya, M. Fernández-Monreal, O. Touzani, and D. Vivien, “Equivocal roles of tissue-type plasminogen activator in stroke-induced injury,” Trends in Neurosciences, vol. 27, no. 3, pp. 155–160, 2004. View at Publisher · View at Google Scholar · View at Scopus
  103. A. L. Samson and R. L. Medcalf, “Tissue-type plasminogen activator: a multifaceted modulator of neurotransmission and synaptic plasticity,” Neuron, vol. 50, no. 5, pp. 673–678, 2006. View at Publisher · View at Google Scholar · View at Scopus
  104. M. Yepes, B. D. Roussel, C. Ali, and D. Vivien, “Tissue-type plasminogen activator in the ischemic brain: more than a thrombolytic,” Trends in Neurosciences, vol. 32, no. 1, pp. 48–55, 2009. View at Publisher · View at Google Scholar · View at Scopus
  105. C. J. Siao, S. R. Fernandez, and S. E. Tsirka, “Cell type-specific roles for tissue plasminogen activator released by neurons or microglia after excitotoxic injury,” Journal of Neuroscience, vol. 23, no. 8, pp. 3234–3242, 2003. View at Scopus
  106. M. Yepes, M. Sandkvist, E. G. Moore, T. H. Bugge, D. K. Strickland, and D. A. Lawrence, “Tissue-type plasminogen activator induces opening of the blood-brain barrier via the LDL receptor-related protein,” Journal of Clinical Investigation, vol. 112, no. 10, pp. 1533–1540, 2003. View at Publisher · View at Google Scholar · View at Scopus
  107. M. Fernández-Monreal, J. P. López-Atalaya, K. Benchenane et al., “Arginine 260 of the amino-terminal domain of NR1 subunit is critical for tissue-type plasminogen activator-mediated enhancement of N-methyl-D-aspartate receptor signaling,” Journal of Biological Chemistry, vol. 279, no. 49, pp. 50850–50856, 2004. View at Publisher · View at Google Scholar · View at Scopus
  108. M. Fernández-Monreal, J. P. López-Atalaya, K. Benchenane et al., “Is tissue-type plasminogen activator a neuromodulator?” Molecular and Cellular Neuroscience, vol. 25, no. 4, pp. 594–601, 2004. View at Publisher · View at Google Scholar · View at Scopus
  109. R. MacRez, P. Obiang, M. Gauberti et al., “Antibodies preventing the interaction of tissue-type plasminogen activator with N-methyl-D-aspartate receptors reduce stroke damages and extend the therapeutic window of thrombolysis,” Stroke, vol. 42, no. 8, pp. 2315–2322, 2011. View at Publisher · View at Google Scholar · View at Scopus
  110. K. Benchenane, V. Berezowski, C. Ali et al., “Tissue-type plasminogen activator crosses the intact blood-brain barrier by low-density lipoprotein receptor-related protein-mediated transcytosis,” Circulation, vol. 111, no. 17, pp. 2241–2249, 2005. View at Publisher · View at Google Scholar · View at Scopus
  111. K. Benchenane, V. Berezowski, M. Fernández-Monreal et al., “Oxygen glucose deprivation switches the transport of tPA across the blood-brain barrier from an LRP-dependent to an increased LRP-independent process,” Stroke, vol. 36, no. 5, pp. 1059–1064, 2005. View at Publisher · View at Google Scholar · View at Scopus
  112. C. S. Kidwell, L. Latour, J. L. Saver et al., “Thrombolytic toxicity: blood brain barrier disruption in human ischemic stroke,” Cerebrovascular Diseases, vol. 25, no. 4, pp. 338–343, 2008. View at Publisher · View at Google Scholar · View at Scopus
  113. T. Aoki, T. Sumii, T. Mori, X. Wang, and E. H. Lo, “Blood-brain barrier disruption and matrix metalloproteinase-9 expression during reperfusion injury mechanical versus embolic focal ischemia in spontaneously hypertensive rats,” Stroke, vol. 33, no. 11, pp. 2711–2717, 2002. View at Publisher · View at Google Scholar · View at Scopus
  114. M. A. Kelly, A. Shuaib, and K. G. Todd, “Matrix metalloproteinase activation and blood-brain barrier breakdown following thrombolysis,” Experimental Neurology, vol. 200, no. 1, pp. 38–49, 2006. View at Publisher · View at Google Scholar · View at Scopus
  115. S. R. Lee, S. Z. Guo, R. H. Scannevin et al., “Induction of matrix metalloproteinase, cytokines and chemokines in rat cortical astrocytes exposed to plasminogen activators,” Neuroscience Letters, vol. 417, no. 1, pp. 1–5, 2007. View at Publisher · View at Google Scholar · View at Scopus
  116. X. Wang, K. Tsuji, S. R. Lee et al., “Mechanisms of hemorrhagic transformation after tissue plasminogen activator reperfusion therapy for ischemic stroke,” Stroke, vol. 35, no. 11, pp. 2726–2730, 2004. View at Publisher · View at Google Scholar · View at Scopus
  117. J.-C. Copin, D. J. Bengualid, R. F. Da Silva, O. Kargiotis, K. Schaller, and Y. Gasche, “Recombinant tissue plasminogen activator induces blood-brain barrier breakdown by a matrix metalloproteinase-9-independent pathway after transient focal cerebral ischemia in mouse,” European Journal of Neuroscience, vol. 34, no. 7, pp. 1085–1092, 2011. View at Publisher · View at Google Scholar · View at Scopus
  118. S. Gautier, O. Petrault, P. Gele et al., “Involvement of thrombolysis in recombinant tissue plasminogen activator-induced cerebral hemorrhages and effect on infarct volume and postischemic endothelial function,” Stroke, vol. 34, no. 12, pp. 2975–2979, 2003. View at Publisher · View at Google Scholar · View at Scopus
  119. P. A. Lapchak, D. F. Chapman, and J. A. Zivin, “Metalloproteinase inhibition reduces thrombolytic (tissue plasminogen activator)-induced hemorrhage after thromboembolic stroke,” Stroke, vol. 31, no. 12, pp. 3034–3040, 2000. View at Scopus
  120. X. Wang, S. R. Lee, K. Arai et al., “Lipoprotein receptor-mediated induction of matrix metalloproteinase by tissue plasminogen activator,” Nature Medicine, vol. 9, no. 10, pp. 1313–1317, 2003. View at Publisher · View at Google Scholar · View at Scopus
  121. X. Zhang, R. Polavarapu, H. She, Z. Mao, and M. Yepes, “Tissue-type plasminogen activator and the low-density lipoprotein receptor-related protein mediate cerebral ischemia-induced nuclear factor-κB pathway activation,” American Journal of Pathology, vol. 171, no. 4, pp. 1281–1290, 2007. View at Publisher · View at Google Scholar · View at Scopus
  122. R. Polavarapu, J. An, C. Zhang, and M. Yepes, “Regulated intramembrane proteolysis of the low-density lipoprotein receptor-related protein mediates ischemic cell death,” American Journal of Pathology, vol. 172, no. 5, pp. 1355–1362, 2008. View at Publisher · View at Google Scholar · View at Scopus
  123. R. Polavarapu, M. C. Gongora, H. Yi et al., “Tissue-type plasminogen activator-mediated shedding of astrocytic low-density lipoprotein receptor-related protein increases the permeability of the neurovascular unit,” Blood, vol. 109, no. 8, pp. 3270–3278, 2007. View at Publisher · View at Google Scholar · View at Scopus
  124. J. O'Rourke, X. Jiang, Z. Hao, R. E. Cone, and A. R. Hand, “Distribution of sympathetic tissue plasminogen activator (tPA) to a distant microvasculature,” Journal of Neuroscience Research, vol. 79, no. 6, pp. 727–733, 2005. View at Publisher · View at Google Scholar · View at Scopus
  125. R. Wong, “NMDA receptors expressed in oligodendrocytes,” BioEssays, vol. 28, no. 5, pp. 460–464, 2006. View at Publisher · View at Google Scholar · View at Scopus
  126. A. Reijerkerk, G. Kooij, S. M. A. Van Der Pol et al., “The NR1 subunit of NMDA receptor regulates monocyte transmigration through the brain endothelial cell barrier,” Journal of Neurochemistry, vol. 113, no. 2, pp. 447–453, 2010. View at Publisher · View at Google Scholar · View at Scopus
  127. G. T. Liberatore, A. Samson, C. Bladin, W. D. Schleuning, and R. L. Medcalf, “Vampire bat salivary plasminogen activator (desmoteplase): a unique fibrinolytic enzyme that does not promote neurodegeneration,” Stroke, vol. 34, no. 2, pp. 537–543, 2003. View at Publisher · View at Google Scholar · View at Scopus
  128. J. P. López-Atalaya, B. D. Roussel, C. Ali et al., “Recombinant Desmodus rotundus salivary plasminogen activator crosses the blood-brain barrier through a low-density lipoprotein receptor-related protein-dependent mechanism without exerting neurotoxic effects,” Stroke, vol. 38, no. 3, pp. 1036–1043, 2007. View at Publisher · View at Google Scholar · View at Scopus
  129. W. Hacke, A. J. Furlan, Y. Al-Rawi et al., “Intravenous desmoteplase in patients with acute ischaemic stroke selected by MRI perfusion-diffusion weighted imaging or perfusion CT (DIAS-2): a prospective, randomised, double-blind, placebo-controlled study,” The Lancet Neurology, vol. 8, no. 2, pp. 141–150, 2009. View at Publisher · View at Google Scholar · View at Scopus
  130. E. J. Su, L. Fredriksson, M. Geyer et al., “Activation of PDGF-CC by tissue plasminogen activator impairs blood-brain barrier integrity during ischemic stroke,” Nature Medicine, vol. 14, no. 7, pp. 731–737, 2008. View at Publisher · View at Google Scholar · View at Scopus
  131. J. Alvarez-Sabín, P. Delgado, S. Abilleira et al., “Temporal profile of matrix metalloproteinases and their inhibitors after spontaneous intracerebral hemorrhage: relationship to clinical and radiological outcome,” Stroke, vol. 35, no. 6, pp. 1316–1322, 2004. View at Publisher · View at Google Scholar · View at Scopus
  132. S. Horstmann, P. Kalb, J. Koziol, H. Gardner, and S. Wagner, “Profiles of matrix metalloproteinases, their inhibitors, and laminin in stroke patients: influence of different therapies,” Stroke, vol. 34, no. 9, pp. 2165–2170, 2003. View at Publisher · View at Google Scholar · View at Scopus
  133. J. Montaner, “Stroke biomarkers: can they help us to guide stroke thrombolysis?” Drug News and Perspectives, vol. 19, no. 9, pp. 523–532, 2006. View at Publisher · View at Google Scholar · View at Scopus
  134. A. Rosell and E. H. Lo, “Multiphasic roles for matrix metalloproteinases after stroke,” Current Opinion in Pharmacology, vol. 8, no. 1, pp. 82–89, 2008. View at Publisher · View at Google Scholar · View at Scopus
  135. A. R. Green, “Pharmacological approaches to acute ischaemic stroke: reperfusion certainly, neuroprotection possibly,” British Journal of Pharmacology, vol. 153, no. 1, pp. S325–S338, 2008. View at Publisher · View at Google Scholar · View at Scopus
  136. A. F. Ducruet, B. T. Grobelny, B. E. Zacharia, Z. L. Hickman, M. L. Yeh, and E. S. Connolly Jr., “Pharmacotherapy of cerebral ischemia,” Expert Opinion on Pharmacotherapy, vol. 10, no. 12, pp. 1895–1906, 2009. View at Publisher · View at Google Scholar · View at Scopus
  137. U. Dirnagl, K. Becker, and A. Meisel, “Preconditioning and tolerance against cerebral ischaemia: from experimental strategies to clinical use,” The Lancet Neurology, vol. 8, no. 4, pp. 398–412, 2009. View at Publisher · View at Google Scholar · View at Scopus
  138. J. Chen and R. Simon, “Ischemic tolerance in the brain,” Neurology, vol. 48, no. 2, pp. 306–311, 1997. View at Scopus
  139. C. E. Murry, R. B. Jennings, and K. A. Reimer, “Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium,” Circulation, vol. 74, no. 5, pp. 1124–1136, 1986. View at Scopus
  140. T. Kirino, “Ischemic tolerance,” Journal of Cerebral Blood Flow and Metabolism, vol. 22, no. 11, pp. 1283–1296, 2002. View at Scopus
  141. M. P. Stenzel-Poore, S. L. Stevens, and R. P. Simon, “Genomics of preconditioning,” Stroke, vol. 35, no. 11, supplement 1, pp. 2683–2686, 2004. View at Publisher · View at Google Scholar · View at Scopus
  142. M. P. Stenzel-Poore, S. L. Stevens, Z. Xiong et al., “Effect of ischaemic preconditioning on genomic response to cerebral ischaemia: similarity to neuroprotective strategies in hibernation and hypoxia-tolerant states,” The Lancet, vol. 362, no. 9389, pp. 1028–1037, 2003. View at Publisher · View at Google Scholar · View at Scopus
  143. J. L. Cadet and I. N. Krasnova, “Cellular and molecular neurobiology of brain preconditioning,” Molecular Neurobiology, vol. 39, no. 1, pp. 50–61, 2009. View at Publisher · View at Google Scholar · View at Scopus
  144. A. Kunz, L. Park, T. Abe et al., “Neurovascular protection by ischemic tolerance: role of nitric oxide and reactive oxygen species,” Journal of Neuroscience, vol. 27, no. 27, pp. 7083–7093, 2007. View at Publisher · View at Google Scholar · View at Scopus
  145. T. Masada, Y. Hua, G. Xi, S. R. Ennis, and R. F. Keep, “Attenuation of ischemic brain edema and cerebrovascular injury after ischemic preconditioning in the rat,” Journal of Cerebral Blood Flow and Metabolism, vol. 21, no. 1, pp. 22–33, 2001. View at Scopus
  146. F. Y. Zhang, X. C. Chen, H. M. Ren, and W. M. Bao, “Effects of ischemic preconditioning on blood-brain barrier permeability and MMP-9 expression of ischemic brain,” Neurological Research, vol. 28, no. 1, pp. 21–24, 2006. View at Publisher · View at Google Scholar · View at Scopus
  147. S. Zahler, C. Kupatt, and B. F. Becker, “Endothelial preconditioning by transient oxidative stress reduces inflammatory responses of cultured endothelial cells to TNF-α,” FASEB Journal, vol. 14, no. 3, pp. 555–564, 2000. View at Scopus
  148. S. G. Zhou, X. Y. Lei, and D. F. Liao, “Effects of hypoxic preconditioning on the adhesion of neutrophils to vascular endothelial cells induced by hypoxia/reoxygenation,” Chinese Critical Care Medicine, vol. 15, no. 3, pp. 159–162, 2003. View at Scopus
  149. P. An and Y. X. Xue, “Effects of preconditioning on tight junction and cell adhesion of cerebral endothelial cells,” Brain Research, vol. 1272, no. C, pp. 81–88, 2009. View at Publisher · View at Google Scholar · View at Scopus
  150. A. V. Andjelkovic, S. M. Stamatovic, and R. F. Keep, “The protective effects of preconditioning on cerebral endothelial cells in vitro,” Journal of Cerebral Blood Flow and Metabolism, vol. 23, no. 11, pp. 1348–1355, 2003. View at Scopus
  151. R. Gesuete, F. Orsini, E. R. Zanier et al., “Glial cells drive preconditioning-induced blood-brain barrier protection,” Stroke, vol. 42, no. 5, pp. 1445–1453, 2011. View at Publisher · View at Google Scholar · View at Scopus
  152. T. Yamashita, K. Deguchi, S. Nagotani, and K. Abe, “Vascular protection and restorative therapy in ischemic stroke,” Cell Transplantation, vol. 20, no. 1, pp. 95–97, 2011. View at Publisher · View at Google Scholar · View at Scopus
  153. P. A. Lapchak, “A critical assessment of edaravone acute ischemic stroke efficacy trials: is edaravone an effective neuroprotective therapy?” Expert Opinion on Pharmacotherapy, vol. 11, no. 10, pp. 1753–1763, 2010. View at Publisher · View at Google Scholar · View at Scopus
  154. S. Feng, Q. Yang, M. Liu et al., “Edaravone for acute ischaemic stroke,” Cochrane Database of Systematic Reviews, vol. 12, p. CD007230, 2011. View at Scopus
  155. D. Deplanque and R. Bordet, “Physical activity: one of the easiest ways to protect the brain?” Journal of Neurology, Neurosurgery and Psychiatry, vol. 80, no. 9, p. 942, 2009. View at Publisher · View at Google Scholar · View at Scopus
  156. P. Amarenco, “Hypercholesterolemia, lipid-lowering agents, and the risk for brain infarction,” Neurology, vol. 57, no. 5, pp. S35–S44, 2001. View at Scopus
  157. H. B. Rubins, J. Davenport, V. Babikian et al., “Reduction in stroke with gemfibrozil in men with coronary heart disease and low HDL cholesterol the veterans affairs HDL intervention trial (VA-HIT),” Circulation, vol. 103, no. 23, pp. 2828–2833, 2001. View at Scopus
  158. D. Deplanque, I. Masse, C. Lefebvre, C. Libersa, D. Leys, and R. Bordet, “Prior TIA, lipid-lowering drug use, and physical activity decrease ischemic stroke severity,” Neurology, vol. 67, no. 8, pp. 1403–1410, 2006. View at Publisher · View at Google Scholar · View at Scopus
  159. B. Desvergne and W. Wahli, “Peroxisome proliferator-activated receptors: nuclear control of metabolism,” Endocrine Reviews, vol. 20, no. 5, pp. 649–688, 1999. View at Scopus
  160. P. Lefebvre, G. Chinetti, J. C. Fruchart, and B. Staels, “Sorting out the roles of PPARα in energy metabolism and vascular homeostasis,” Journal of Clinical Investigation, vol. 116, no. 3, pp. 571–580, 2006. View at Publisher · View at Google Scholar · View at Scopus
  161. S. Moreno, S. Farioli-vecchioli, and M. P. Cerù, “Immunolocalization of peroxisome proliferator-activated receptors and retinoid X receptors in the adult rat CNS,” Neuroscience, vol. 123, no. 1, pp. 131–145, 2004. View at Publisher · View at Google Scholar · View at Scopus
  162. M. Ricote and C. K. Glass, “PPARs and molecular mechanisms of transrepression,” Biochimica et Biophysica Acta, vol. 1771, no. 8, pp. 926–935, 2007. View at Publisher · View at Google Scholar · View at Scopus
  163. P. Delerive, K. De Bosscher, S. Besnard et al., “Peroxisome proliferator-activated receptor α negatively regulates the vascular inflammatory gene response by negative cross-talk with transcription factors NF-κB and AP-1,” Journal of Biological Chemistry, vol. 274, no. 45, pp. 32048–32054, 1999. View at Publisher · View at Google Scholar · View at Scopus
  164. M. Collino, M. Aragno, R. Mastrocola et al., “Oxidative stress and inflammatory response evoked by transient cerebral ischemia/reperfusion: effects of the PPAR-α agonist WY14643,” Free Radical Biology and Medicine, vol. 41, no. 4, pp. 579–589, 2006. View at Publisher · View at Google Scholar · View at Scopus
  165. D. Deplanque, P. Gelé, O. Pétrault et al., “Peroxisome proliferator-activated receptor-α activation as a mechanism of preventive neuroprotection induced by chronic fenofibrate treatment,” Journal of Neuroscience, vol. 23, no. 15, pp. 6264–6271, 2003. View at Scopus
  166. H. Inoue, X. F. Jiang, T. Katayama, S. Osada, K. Umesono, and S. Namura, “Brain protection by resveratrol and fenofibrate against stroke requires peroxisome proliferator-activated receptor α in mice,” Neuroscience Letters, vol. 352, no. 3, pp. 203–206, 2003. View at Publisher · View at Google Scholar · View at Scopus
  167. C. Mysiorek, M. Culot, L. Dehouck et al., “Peroxisome proliferator-activated receptor-α activation protects brain capillary endothelial cells from oxygen-glucose deprivation-induced hyperpermeability in the blood-brain barrier,” Current Neurovascular Research, vol. 6, no. 3, pp. 181–193, 2009. View at Publisher · View at Google Scholar · View at Scopus
  168. Y. Luo, W. Yin, A. P. Signore et al., “Neuroprotection against focal ischemic brain injury by the peroxisome proliferator-activated receptor-γ agonist rosiglitazone,” Journal of Neurochemistry, vol. 97, no. 2, pp. 435–448, 2006. View at Publisher · View at Google Scholar · View at Scopus
  169. T. Mabuchi, K. Kitagawa, T. Ohtsuki et al., “Contribution of microglia/macrophages to expansion of infarction and response of oligodendrocytes after focal cerebral ischemia in rats,” Stroke, vol. 31, no. 7, pp. 1735–1743, 2000. View at Scopus
  170. L. Pantoni, C. Sarti, and D. Inzitari, “Cytokines and cell adhesion molecules in cerebral ischemia: experimental bases and therapeutic perspectives,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 18, no. 4, pp. 503–513, 1998. View at Scopus
  171. T. Kielian and P. D. Drew, “Effects of peroxisome proliferator-activated receptor-γ agonists on central nervous system inflammation,” Journal of Neuroscience Research, vol. 71, no. 3, pp. 315–325, 2003. View at Publisher · View at Google Scholar · View at Scopus
  172. X. Zhao, Z. Ou, J. C. Grotta, N. Waxham, and J. Aronowski, “Peroxisome-proliferator-activated receptor-gamma (PPARγ) activation protects neurons from NMDA excitotoxicity,” Brain Research, vol. 1073-1074, no. 1, pp. 460–469, 2006. View at Publisher · View at Google Scholar · View at Scopus
  173. M. Endres, “Statins and stroke,” Journal of Cerebral Blood Flow and Metabolism, vol. 25, no. 9, pp. 1093–1110, 2005. View at Publisher · View at Google Scholar · View at Scopus
  174. A. Zacco, J. Togo, K. Spence et al., “3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitors protect cortical neurons from excitotoxicity,” Journal of Neuroscience, vol. 23, no. 35, pp. 11104–11111, 2003. View at Scopus
  175. C. R. W. Kuhlmann, M. Gerigk, B. Bender, D. Closhen, V. Lessmann, and H. J. Luhmann, “Fluvastatin prevents glutamate-induced blood-brain-barrier disruption in vitro,” Life Sciences, vol. 82, no. 25-26, pp. 1281–1287, 2008. View at Publisher · View at Google Scholar · View at Scopus
  176. S. Wang, S. R. Lee, S. Z. Guo et al., “Reduction of tissue plasminogen activator-induced matrix metalloproteinase-9 by simvastatin in astrocytes,” Stroke, vol. 37, no. 7, pp. 1910–1912, 2006. View at Publisher · View at Google Scholar · View at Scopus
  177. M. Jasińska, J. Owczarek, and D. Orszulak-Michalak, “Statins: a new insight into their mechanisms of action and consequent pleiotropic effects,” Pharmacological Reports, vol. 59, no. 5, pp. 483–499, 2007. View at Scopus
  178. G. Martin, H. Duez, C. Blanquart et al., “Statin-induced inhibition of the rho-signaling pathway activates PPARα and induces HDL apoA-I,” Journal of Clinical Investigation, vol. 107, no. 11, pp. 1423–1432, 2001. View at Scopus
  179. R. Paumelle and B. Staels, “Cross-talk between statins and PPARα in cardiovascular diseases: clinical evidence and basic mechanisms,” Trends in Cardiovascular Medicine, vol. 18, no. 3, pp. 73–78, 2008. View at Publisher · View at Google Scholar · View at Scopus
  180. K. J. Yin, Z. Deng, M. Hamblin, J. Zhang, and Y. E. Chen, “Vascular PPARδ protects against stroke-induced brain injury,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 31, no. 3, pp. 574–581, 2011. View at Publisher · View at Google Scholar · View at Scopus
  181. A. Plotnikov, E. Zehorai, S. Procaccia, and R. Seger, “The MAPK cascades: signaling components, nuclear roles and mechanisms of nuclear translocation,” Biochimica et Biophysica Acta, vol. 1813, no. 9, pp. 1619–1633, 2011. View at Publisher · View at Google Scholar · View at Scopus
  182. C. Widmann, S. Gibson, M. B. Jarpe, and G. L. Johnson, “Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human,” Physiological Reviews, vol. 79, no. 1, pp. 143–180, 1999. View at Scopus
  183. V. Waetzig, Y. Zhao, and T. Herdegen, “The bright side of JNKs-Multitalented mediators in neuronal sprouting, brain development and nerve fiber regeneration,” Progress in Neurobiology, vol. 80, no. 2, pp. 84–97, 2006. View at Publisher · View at Google Scholar · View at Scopus
  184. J. M. Kyriakis and J. Avruch, “Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation,” Physiological Reviews, vol. 81, no. 2, pp. 807–869, 2001. View at Scopus
  185. W. Haeusgen, R. Boehm, Y. Zhao, T. Herdegen, and V. Waetzig, “Specific activities of individual c-Jun N-terminal kinases in the brain,” Neuroscience, vol. 161, no. 4, pp. 951–959, 2009. View at Publisher · View at Google Scholar · View at Scopus
  186. H. Nishina, T. Wada, and T. Katada, “Physiological roles of SAPK/JNK signaling pathway,” Journal of Biochemistry, vol. 136, no. 2, pp. 123–126, 2004. View at Publisher · View at Google Scholar · View at Scopus
  187. V. Waetzig and T. Herdegen, “MEKK1 controls neurite regrowth after experimental injury by balancing ERK1/2 and JNK2 signaling,” Molecular and Cellular Neuroscience, vol. 30, no. 1, pp. 67–78, 2005. View at Publisher · View at Google Scholar · View at Scopus
  188. M. A. Bogoyevitch, “The isoform-specific functions of the c-Jun N-terminal kinases (JNKs): differences revealed by gene targeting,” BioEssays, vol. 28, no. 9, pp. 923–934, 2006. View at Publisher · View at Google Scholar · View at Scopus
  189. S. Brecht, R. Kirchhof, A. Chromik et al., “Specific pathophysiological functions of JNK isoforms in the brain,” European Journal of Neuroscience, vol. 21, no. 2, pp. 363–377, 2005. View at Publisher · View at Google Scholar · View at Scopus
  190. H. Chaudhury, M. Zakkar, J. Boyle et al., “C-Jun N-terminal kinase primes endothelial cells at atheroprone sites for apoptosis,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 30, no. 3, pp. 546–553, 2010. View at Publisher · View at Google Scholar · View at Scopus
  191. J. Cui, M. Zhang, Y. Q. Zhang, and Z. H. Xu, “JNK pathway: diseases and therapeutic potential,” Acta Pharmacologica Sinica, vol. 28, no. 5, pp. 601–608, 2007. View at Publisher · View at Google Scholar · View at Scopus
  192. Y. Gao, A. P. Signore, W. Yin et al., “Neuroprotection against focal ischemic brain injury by inhibition of c-Jun N-terminal kinase and attenuation of the mitochondrial apoptosis-signaling pathway,” Journal of Cerebral Blood Flow and Metabolism, vol. 25, no. 6, pp. 694–712, 2005. View at Publisher · View at Google Scholar · View at Scopus
  193. T. Borsellol, P. G. H. Clarkel, L. Hirt et al., “A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia,” Nature Medicine, vol. 9, no. 9, pp. 1180–1186, 2003. View at Publisher · View at Google Scholar · View at Scopus
  194. L. Hirt, J. Badaut, J. Thevenet et al., “D-JNKI1, a cell-penetrating c-Jun-N-terminal kinase inhibitor, protects against cell death in severe cerebral ischemia,” Stroke, vol. 35, no. 7, pp. 1738–1743, 2004. View at Publisher · View at Google Scholar · View at Scopus
  195. K. Wiegler, C. Bonny, D. Coquoz, and L. Hirt, “The JNK inhibitor XG-102 protects from ischemic damage with delayed intravenous administration also in the presence of recombinant tissue plasminogen activator,” Cerebrovascular Diseases, vol. 26, no. 4, pp. 360–366, 2008. View at Publisher · View at Google Scholar · View at Scopus
  196. D. Michel-Monigadon, C. Bonny, and L. Hirt, “C-Jun N-terminal kinase pathway inhibition in intracerebral hemorrhage,” Cerebrovascular Diseases, vol. 29, no. 6, pp. 564–570, 2010. View at Publisher · View at Google Scholar · View at Scopus
  197. Z. Xie, C. J. Smith, and L. J. Van Eldik, “Activated glia induce neuron death Via MAP kinase signaling pathways involving JNK and p38,” GLIA, vol. 45, no. 2, pp. 170–179, 2004. View at Publisher · View at Google Scholar · View at Scopus
  198. R. Kacimi, R. G. Giffard, and M. A. Yenari, “Endotoxin-activated microglia injure brain derived endothelial cells via NF-κB, JAK-STAT and JNK stress kinase pathways,” Journal of Inflammation, vol. 8, article 7, 2011. View at Publisher · View at Google Scholar · View at Scopus
  199. C. Benakis, C. Bonny, and L. Hirt, “JNK inhibition and inflammation after cerebral ischemia,” Brain, Behavior, and Immunity, vol. 24, no. 5, pp. 800–811, 2010. View at Publisher · View at Google Scholar · View at Scopus