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Experimental Diabetes Research
Volume 2012 (2012), Article ID 589589, 11 pages
ER Stress and Apoptosis: A New Mechanism for Retinal Cell Death
1Department of Medicine, Endocrinology and Diabetes, University of Oklahoma Health Sciences Center, 941 Stanton L. Young Blvd., Oklahoma City, OK 73104, USA
2Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, 941 Stanton L. Young Blvd., Oklahoma City, OK 73104, USA
3Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, 941 Stanton L. Young Blvd., Oklahoma City, OK 73104, USA
Received 1 June 2011; Accepted 9 September 2011
Academic Editor: Rangasamy Sampathkumar
Copyright © 2012 Guangjun Jing 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.
The endoplasmic reticulum (ER) is the primary subcellular organelle where proteins are synthesized and folded. When the homeostasis of the ER is disturbed, unfolded or misfolded proteins accumulate in the ER lumen, resulting in ER stress. In response to ER stress, cells activate a set of tightly controlled regulatory programs, known as the unfolded protein response (UPR), to restore the normal function of the ER. However, if ER stress is sustained and the adaptive UPR fails to eliminate unfolded/misfolded proteins, apoptosis will occur to remove the stressed cells. In recent years, a large body of studies has shown that ER stress-induced apoptosis is implicated in numerous human diseases, such as diabetes and neurogenerative diseases. Moreover, emerging evidence supports a role of ER stress in retinal apoptosis and cell death in blinding disorders such as age-related macular degeneration and diabetic retinopathy. In the present review, we summarize recent progress on ER stress and apoptosis in retinal diseases, focusing on various proapoptotic and antiapoptotic pathways that are activated by the UPR, and discuss how these pathways contribute to ER stress-induced apoptosis in retinal cells.
Retinal cell death has been widely held as a central event that leads to retinal neurodegeneration, vascular dysfunction, and eventually irreversible blindness in ocular diseases, such as glaucoma, retinal degeneration, diabetic retinopathy, and uveitis. Loss of retinal ganglion cells is considered to be a direct cause of vision loss in experimental glaucoma , correlating with elevated intraocular pressure (IOP) . Injury of retinal pigment epithelial (RPE) cells and photoreceptors leads to photoreceptor dysfunction and retinal degeneration , as seen in both inherited and acquired degenerative retinal diseases such as Stargardt’s Disease , retinitis pigmentosa , and age-related macular degeneration . In diabetic retinopathy, high glucose and other diabetic insults, such as oxidants, advanced glycation end products (AGEs), and inflammatory cytokines, result in neural and vascular cell death [7, 8]. In streptozotocin-induced diabetic rats, both retinal neurons and vascular cells become apoptotic soon after the onset of diabetes . Inflammation-driven neural and vascular cell death is also a hallmark characteristic of uveitis, a chronic eye disease that cause vision loss [10–13]. Together, these findings support a pivotal role of cell death in the pathogenesis of retinal diseases. Apoptosis, that is, programmed cell death, is the most common form of cell death in various cell types, including retinal cells. Apoptosis is tightly controlled by a variety of signaling pathways that either promote or inhibit the apoptotic cascades. Among the most extensively studied proapoptotic factors in retinal cells are oxidative stress, mitochondrial dysfunction, inflammation, ischemia, hyperglycemia, and excitotoxicity [14–18]. Intriguingly, recent evidence suggests that disturbed protein homeostasis and endoplasmic reticulum (ER) stress also contribute to apoptosis of retinal cells . Moreover, ER stress activates a large number of genes involved in the control of cell fate, including antiapoptotic and proapoptotic molecules such as Bax and Bcl-2 [20, 21]. Therefore, elucidating the role and mechanisms of ER stress in retinal cell apoptosis may provide important insight into the pathogenesis of retinal diseases and help in developing new drugs to protect retinal cells and to prevent vision loss. In the present review, we discuss the potential implication of ER stress in retinal cell apoptosis, with a primary focus on the signaling transduction pathways that link ER stress with apoptosis in general as well as specific for retinal cells.
2. ER Stress and the Unfolded Protein Response (UPR)
The endoplasmic reticulum (ER) is the primary intracellular organelle responsible for protein folding, maturation, and trafficking [22, 23]. The ER consists of a network of folded membranes in which secretory and most membrane proteins are synthesized, posttranslationally modified, and folded into their correct three-dimensional conformations. Only properly folded (mature) proteins can be transported to the Golgi apparatus for further processing. In addition, the ER also serves as a dynamic pool of calcium, governing the intracellular calcium homeostasis . Other major functions of the ER include lipid and steroid hormone synthesis, carbohydrate metabolism, and drug detoxification. Importantly, compelling evidence indicates that the ER is one of the major machinery that senses subtle environmental changes and cellular stresses, coordinates signaling pathways, and modulates cell function and cell survival. Various physiological and pathological circumstances, such as excessive mutant proteins, viral infection, energy or nutrient deprivation, as well as alteration in the redox status, can compromise the ER capacity in protein folding, resulting in the accumulation of unfolded or misfolded proteins in the ER lumen, or ER stress. In turn, misfolded proteins aggregate to form insoluble intracellular or extracellular deposit, which is toxic to the cell. It has been demonstrated that a number of age-related diseases, such as Alzheimer’s diseases; inflammatory disorders, such as diabetes; and neurodegenerative diseases, such as Parkinson’s disease, are associated with the build-up of misfolded or unfolded protein aggregates [25–28]. To eliminate the toxic protein components, cells activate an adaptive mechanism that consists of a number of intracellular signaling pathways, collectively known as unfolded protein response (UPR). The UPR relieves ER stress and restores the protein homeostasis through three complementary strategies: (1) halt the generation of more unfolded proteins by suppression of protein translation; (2) induce ER-related molecular chaperones to promote refolding of the unfolded proteins, and (3) activate the ER-associated protein degradation (ERAD) system to remove the unfolded proteins.
There are three branches of UPR that are initiated by distinct ER stress transducers located on the ER membrane: PKR-like endoplasmic reticulum kinase (PERK) , inositol-requiring enzyme 1 (IRE1) [30, 31], and activating transcription factor 6 (ATF6) . In nonstressed cells, all three ER stress transducers are kept in an inactive state through binding to the ER chaperon glucose-regulated protein 78 (Bip), which is also known as immunoglobulin binding protein (Bip) [32, 33]. Upon ER stress, excessive unfolded proteins accumulate in the ER lumen, resulting in the dissociation of GPR78 from the ER stress transducers , which triggers activation of the UPR branches. In eukaryotic cells, UPR is an adaptive cellular response to the disturbance of normal ER functions, which attenuate the aggregation of unfolded or misfolded proteins and promote cell survival . However, during prolonged or overwhelming ER stress, UPR fails to restore the normal function of the ER, and apoptotic cascade will be activated [36, 37] (Figure 1). The exact mechanism underlying the switch of the UPR from a prosurvival mechanism to a proapoptotic response is not clear.
2.1. The IRE1/XBP1 Pathway
IRE1 was firstly identified as an ER transmembrane protein kinase that is essential for signaling transduction from the ER to the nucleus  and was subsequently found to be involved in the initiation of the UPR . There are two different IRE1 proteins in mammalian cells, both of which participate in the ER stress response or UPR. IRE1α is ubiquitously expressed while IRE1β is tissue-specific [30, 40]. During ER stress, IRE1 dissociates with Bip/Bip and becomes activated. Activated IRE1 acquires the function as endogenous ribonuclease (RNase) and splices a 26-nucleotide intron from the mRNA of XBP1. The splicing results in a shift in the translational frame of the XBP1 gene, leading to the translation of a new protein, named spliced XBP1 [31, 41]. The newly generated spliced XBP1 is an active transcription factor, which in turn induces diverse downstream genes, such as ER chaperones  and proteins involved in ER-associated protein degradation (ERAD) . These proteins work together to restore the ER homeostasis and promote cell survival. Indeed, cells deficient of XBP1 are susceptible to oxidative stress- and inflammation-induced cell death [44, 45], suggesting that XBP1-mediated adaptive UPR is an important mechanism that protects the cell from apoptosis during ER stress. In addition, the IRE1/XBP1 pathway is also essential for embryonic development. Genetic deletion of IRE1 or XBP1 is lethal to mouse embryo due to fetal liver hyperplasia [46, 47]. In addition, a recent study shows that loss of IRE1 results in severe dysfunction of the placenta and also contributes to the embryonic lethality of IRE1 KO mice .
2.2. The PERK/eIF2α/ATF4 Pathway
PERK is a serine/threonine protein kinase located on the ER membrane. Like IRE1, PERK is activated by ER stress via dimerization and autophosphorylation upon the dissociation with Bip. Activated PERK phosphorylates its downstream target protein, eIF2α, resulting in the inhibition of global protein translation . However, some genes with upstream open reading frames (uORFs) in its 5′ untranslated region could escape from the eIF2α-initiated translational attenuation. A representative example is activating transcription factor 4 (ATF4)—human ATF4 gene contains multiple uORFs in its 5′UTR whereas the murine mRNA has two uORFs . These uORFs prevent the translation of ATF4 under normal conditions but enhance its expression when eIF2α is phosphorylated [51, 52]. ATF4 belongs to the superfamily of DNA-binding proteins that includes the activator protein-1 (AP-1) family, cAMP-response element binding proteins (CREBs), and CREB-like proteins. As a transcription factor, ATF4 binds to the CRE site in the promoter region of target genes, inducing a battery of stress response genes involved in oxidative stress, amino acid synthesis, and transportation. In addition, ATF4 is a major inducer of C/EBP homologous protein (CHOP), which has been considered as a central mediator of ER stress-induced apoptosis. The role of CHOP in coordinating the apoptotic pathways will be discussed in detail in the following chapters.
2.3. The ATF6 Pathway
Besides XBP-1 and ATF4, ATF6 has been identified as another basic leucine zipper- (bZIP-) containing transcription factor induced by ER stress. ATF6 is a type II ER transmembrane protein. Like IRE1 and PERK, ATF6 binds to Bip and remains in an inactive state in unstressed cells. In response to ER stress, the Bip/ATF6 complex is dissociated, resulting in the translocation of ATF6 from ER membrane to Golgi apparatus. In Golgi apparatus, ATF6 is cleaved by two proteases, serine protease site-1 protease (S1P) and the metalloprotease site-2 protease (S2P), to produce the active form of the transcription factor [53–55]. The active ATF6 then moves to the nucleus and activates the ER stress response element- (ERSE-) related genes through binding their promoters . ATF6 also regulates other URP genes, such as XBP-1 and CHOP .
3. ER Stress-Associated Apoptosis in Retina Cells
Previous studies suggest that the cell fate is dependent on the balance between the extent/severity of ER stress and the capacity of the ER to restore ER homeostasis through the UPR [57–59]. Temporal and mild ER stress can be overcome by the adaptive UPR, cell function maintained, and cells survive. However, if the stress condition is prolonged and the UPR fails to restore the ER homeostasis, the apoptotic signaling pathways will be initiated to remove the unhealthy cells. Recently, several independent studies have provided ample evidence that ER stress is a potential cause of retinal vascular and neuronal cell death in diseases such as glaucoma, diabetic retinopathy and age-related macular degeneration [19, 60–62]. ER stress has been observed in both cultured retinal cells (vascular endothelial cells, pericytes, ganglion cells, Muller cells, as well as RPE cells) and in the retina from animal models of various diseases. Not surprisingly, the role of ER stress has been extensively studied in the pathogenesis of retinitis pigmentosa (RP) with mutations of various retinal genes. In 2004, Rebello and associates reported that expression of a mutant (R14W) of carbonic anhydrase IV, a glycosylphosphatidylinositol-anchored protein that is highly expressed in the choriocapillaris of the human eye, induced upregulation of Bip, PERK, and CHOP, markers of ER stress and the unfolded protein response, accompanied by apoptosis . Similarly, enhanced ER stress was reported in RP induced by the rhodopsin mutation P23H in Xenopus laevis  and in rats [65, 66]. Further, stimulation of the UPR in the retina or cultured retinal cells by preexposure to mild ER stress protected photoreceptor neurons from oxidative damage and cell death . Moreover, overexpression of Bip, an ER chaperone that facilitates protein folding and reduces ER stress, attenuated retinal expression of CHOP and the activation of apoptotic cascade and restored retinal photoreceptor function in P23H rats . In addition to the genetic models of RP, ER stress was found remarkably enhanced in retinal photoreceptors, coincident with photoreceptor cell apoptosis, in a rodent model of light damage-induced retinal degeneration . These findings collectively support a causal role of ER stress in photoreceptor cell death and retinal degeneration.
Another well-studied area for ER stress-related retinal cell death is glaucoma. Increased ER stress markers were observed in retinal ganglion cells in animal models of ischemia-reperfusion and chronic glaucoma . Cultured retinal ganglion cells (RGC-5, a transformed rat ganglion cell line) treated with tunicamycin, a common ER stress inducer, undergo apoptosis, accompanied by increased production of ER stress-related proteins [19, 69]. In vivo, intravitreal injection of tunicamycin resulted in loss of retinal ganglion cells and reduced thickness of the inner retina. Moreover, raising IOP or intravitreal injection of N-methyl-D-aspartate (NMDA), an excitotoxin that binds to the NMDA receptor and induces neuron cell death, also increased the expression of ER stress markers in retinal ganglion cells, amacrine cells, and microglial cells . Pharmaceutical induction of Bip significantly attenuated tunicamycin- or NMDA-induced apoptosis in retinal ganglion cells, suggesting a pivotal role of ER stress in retinal neuron cell death .
Loss of retinal vascular cells and apoptosis of retinal neurons have been recognized as critical events and pathological features of diabetic retinopathy [70–72]. Although currently it remains to be investigated how ER stress signaling pathways contribute to retinal cell death induced by diabetes, recent studies by our group and others demonstrated that ER stress was induced in early stage of diabetic retinopathy and was implicated in retinal inflammation and vascular damage [73–76]. In 2009, we reported increased ER stress markers in the retina of diabetic Akita mice, in parallel with elevated expression of inflammatory genes . In cultured retinal endothelial cells, ER stress was induced by hypoxia, a potent stimulator of inflammation and angiogenesis, and prevented by chemical chaperones. Moreover, we showed that induction of ER stress in the retina was sufficient to trigger an upregulation of inflammatory genes. Conversely, inhibiting ER stress protected the retina and retinal endothelial cells from inflammatory damage. In addition, activation of the adaptive UPR by preconditioning with ER stress also successfully prevented inflammatory damage to retinal endothelial cells and vascular leakage induced by diabetic stimulus . These results suggest that ER stress is implicated in retinal cell damage caused by diabetes.
4. Signaling Pathways of ER Stress-Associated Apoptosis
It is currently unclear how ER stress induces apoptosis in various retinal cells. Generally, there are two major pathways for the initiation of apoptosis: extrinsic and intrinsic pathways . The extrinsic pathway is mediated by the cell membrane death receptors. Activation of the death receptor recruits adaptor molecules and activates caspase-8 or caspase-10, which cleaves the downstream substrates, that is, other caspases including caspase-3, resulting in apoptosis . The intrinsic pathway is closely related to factors anchored on the mitochondria. The insertion of these proapoptotic proteins changes the mitochondrial membrane permeability, resulting in the release of cytochrome c from mitochondria into the cytosol. Then cytochrome c binds to Apaf-1 and activates caspase-9 and then caspase-3, leading to the execution of cell death. In addition, accumulating evidence suggests that calcium release from the ER can also initiate the cell death signals, either by directly activating death receptors or by altering the sensitivity of mitochondria. Finally, these apoptotic pathways converge on caspase-3, resulting in the cleavage of other proteases and leading to apoptosis. In addition to caspase-dependent pathways, caspase-independent pathway is also implicated in retina apoptosis . In the following sections, we discuss the potential pathways that may play a role in ER stress-associated apoptosis in various retinal diseases, such as age-related macular degeneration, glaucoma, and diabetic retinopathy (Figure 2).
4.1. CHOP: A Key Mediator of ER Stress-Induced Apoptosis
CHOP, also named as growth-arrest and DNA-damage-inducible gene 153 (GADD153), is a major stress-inducible proapoptotic gene in ER stress-induced apoptosis . All three branches of the UPR regulate the activation of CHOP; however, ATF4 is considered as the major inducer of CHOP expression. CHOP is expressed at a very low level under physiological conditions but its expression level significantly increases in the presence of severe or persistent ER stress. Notably, the induction of CHOP well correlates with the onset of ER stress-associated apoptosis [81, 82], silencing CHOP expression protects cells against apoptosis induced by prolonged ER stress . As a transcription factor, CHOP has been shown to regulate numerous pro- and antiapoptotic genes, including Bcl-2, GADD34, and TRB3 . CHOP directly binds the promoter of TRB3 gene and upregulates its expression , which in turn inhibits AKT activation, resulting in apoptosis and cell death . Intriguingly, TRB3 also regulates CHOP expression through negative feedback. Overexpressing TRB3 inhibits the transcriptional induction of CHOP while silencing TRB3 results in upregulation of CHOP under both normal and stressed conditions . Treatment with PBA, a chemical chaperone that attenuates ER stress, restores AKT phosphorylation, reduces CHOP and TRB3 expression, and prevents apoptosis. These findings indicate that CHOP is a key mediator of ER stress-induced apoptosis and is tightly regulated by multiple factors, including UPR components such as ATF4 and its downstream genes such as TRB3.
4.2. Mitochondria and the Bcl-2 Family
Recent evidence suggests that mitochondrial dysfunction plays a role in ER stress-induced apoptosis [20, 87]. ER stress, via the UPR, also regulates a number of apoptosis-associated proteins that localize on the mitochondrial membrane, notably the members of the Bcl-2 family. These proteins are widely held as the central coordinators of mitochondria-mediated apoptotic pathways. The Bcl-2 family consists of antiapoptotic members, such as Bcl-2 and Bcl-xL, and proapoptotic proteins, such as Bax, Bak, and Bik . The balance between the anti- and proapoptotic proteins is important for maintaining normal mitochondrial function as well as cell survival. Cells overexpressing Bcl-2 or deficient of Bax and Bak are resistant to ER stress-induced apoptosis . Conversely, overexpressing Bax promotes cytochrome c release and activates apoptotic enzymes, leading to cell death . BH3-only proteins, such as Bim and Bax, are proapoptotic members of the Bcl-2 protein family, playing an essential role in the initiation of programmed cell death and stress-induced apoptosis . Recent studies show that both the antiapoptotic gene Bcl-2 and the proapoptotic proteins, for example, Bim and Bax, are regulated by CHOP during ER stress [87, 92]. CHOP downregulates Bcl-2 expression but upregulates Bim and promotes the translocation of Bax into the mitochondria . Another BH3-only protein, p53-upregulated modulator of apoptosis (PUMA), is induced by p53 during ER stress, and PUMA-deficient cells are resistant to ER stress-elicited apoptosis. These results imply an important role of p53 and PUMA in ER stress-associated cell death . In addition to mediating ER stress-driven apoptosis, the Bcl-2 family also regulates ER stress through physical interaction with ER stress sensors and UPR components. For example, both Bax and Bak have been reported to form a protein complex with IRE1α, which is essential for IRE1α activation . Double knockout mice that lack Bax and Bak exhibited decreased expression of XBP1, a substrate of IRE1, and developed extensive tissue damage in the liver in response to ER stress induced by tunicamycin . Thus, the mediators and pathways implicated in ER stress-related apoptosis are very complex. Nevertheless, the interdependent regulation of Bcl-2 proteins and the UPR appears to be a key event in the process of fine tuning of pro- and antiapoptotic system during ER stress.
4.3. Caspase-12: An ER-Resident Caspase
Caspase-12 is a member of the inflammatory group of the caspase family, localized to the ER. Moreover, it has been shown that caspase-12 is specifically activated by ER stress, including disruption of ER calcium homeostasis and accumulation of excess proteins in ER, but not by membrane- or mitochondrial-targeted apoptotic signals . Mice deficient of caspase-12 are resistant to ER stress-induced apoptosis, suggesting that caspase-12 plays a critical role in this process . However, the human caspase-12 gene has a single nucleotide polymorphism, which results in the production of either a truncated caspase-12 protein or a full-length protein with no enzymatic activity . In human, caspase-4, a member of caspase-1 subfamily that includes caspase-12, was found localized to the ER membrane and activated specifically by ER stress-inducers . Cleavage of caspase-4 was not affected by overexpression of Bcl-2, which prevents signal transduction on the mitochondria, suggesting that caspase-4 is primarily activated in ER stress-induced apoptosis . Furthermore, a reduction of caspase-4 expression by small interfering RNA decreased ER stress-induced apoptosis in some cell lines, but not other ER stress-independent apoptosis . Although the role of caspase-12 (caspase-4 in human) has been well established, it remains unclear how caspase-12 is activated during ER stress. Recent studies suggest that caspase-12 activation requires the IRE1 signal . Upon activation by ER stress, the cytosolic domain of IRE1 recruits TNF receptor-associated factor 2 (TRAF2), which interacts with caspase-12 and induces the cleavage and activation of the enzyme . In turn, activated caspase-12 cleaves procaspase-9 into active caspase-9, which further cleaves and activates caspase-3, resulting in apoptosis . In addition, caspase-12 can also be activated by its downstream executioner caspase-7, indicating a possible amplification loop in the apoptotic cascades though caspase-12 . Notably, in caspase-12-mediated apoptotic process, cytochrome c is not released from mitochondria, which suggests that cytochrome c is not involved in the caspase-12-dependent apoptosis .
4.4. The JNK Pathway in ER Stress-Mediated Apoptosis
The c-Jun N-terminal kinases/stress-activated protein kinase (JNK/SAPK) pathway is one of three members of the mitogen-activated protein kinase (MAPK) superfamily which also includes the ERK and the p38 MAPKinases . JNK is originally identified for specifically phosphorylating the transcription factor c-jun in its N-terminal transactivation domain . There are three different isoforms of JNK (JNK1, 2, and 3). Among these isoforms, JNK1 and JNK2 are ubiquitously expressed while the expression of JNK3 is tissue specific . It has been reported that JNK is activated by various stress factors and contributes to apoptosis and cell death [105, 106]. Recent evidence suggests that JNK activation is also involved in ER stress-initiated apoptotic cascades . For example, activation of IRE1 by ER stress recruits and activates tumor necrosis factor receptor-associated factor 2 (TRAF2), which further activates JNK , resulting in caspase-12 activation and apoptosis . IL-1β, a proinflammatory cytokine, stimulates JNK activation and enhances ER stress in pancreatic epithelial cells . Pretreatment with JNK inhibitor abrogates IL-1β-induced ER stress, indicated by phosphorylation of eIF2α, and increased expression of CHOP, GADD34, ATF4, and spliced XBP-1 while inhibition of ER stress does not affect JNK activation by IL-1β . This suggests that JNK activation is required for IL-1β-induced ER stress. In addition, recent studies demonstrate that inhibiting JNK resulted in reduced ATF4 expression during osteoblast differentiation. JNK inhibition also alleviated Bcl-2 antagonist-induced ER stress in a lymphoma cell line [109, 110]. These findings collectively indicate a pivotal role of JNK in induction of ER stress and in mediating ER stress-induced apoptosis, which is yet to be studied in retinal cell apoptosis and retinal diseases.
4.5. Fas-FasL-Induced Apoptosis
The Fas death receptor belongs to the TNF receptor superfamily and is known as important inducer of apoptosis. Fas, through binding to its ligand FasL, recruits and activates the zymogen (precursor) form of cysteine protease caspases, particularly procaspase-8 and -10, which in turn activate caspase-3 and the downstream apoptotic cascades [111, 112]. Previous studies reported that Fas-FasL system is activated in diabetic retinopathy and is implicated in retinal vascular cell death in diabetic animals . Treatment of retinal endothelial cells with neutrophils isolated from patients with diabetic retinopathy induced adhesion of neutrophils to endothelium and caused endothelial apoptosis . Blockade of the Fas-FasL interaction prevented retinal endothelial apoptosis . In an in vivo study, inhibiting FasL potently reduced retinal vascular endothelial cell injury, apoptosis, and blood-retinal barrier breakdown in diabetic animals . Increased immunoreactivity of Fas/FasL and Fas-associated death domain (FADD) was observed in retinal glial cells and ganglion cells in rats with experimental glaucoma . These findings suggest an important role of Fas-FasL system in retinal cell death. While it is widely held that binding and interaction of Fas and FasL are important for activation of the Fas signaling, recent studies suggest that Fas can also be regulated independently of FasL. Timmins and associates reported that Fas expression was induced by ER stress through a pathway involving calcium/calmodulin-dependent protein kinase IIgamma (CaMKIIgamma) and JNK . In addition, activation of CaMKII by ER stress also activated STAT1, a proapoptotic signal transducer, and induced mitochondrial-dependent apoptosis, including release of mitochondrial cytochrome c and loss of mitochondrial membrane potential . It was proposed that prolonged CHOP expression leads to the release of ER calcium stores, which increases cytosolic calcium concentration, resulting in CaMKII activation and apoptotsis. The role of CaMKII in retinal cell apoptosis remains to be elucidated.
Emerging evidence suggests that ER stress plays a pivotal role in retinal apoptosis and cell death. Studies in other fields have identified a number of signaling pathways that are implicated in ER stress-mediated apoptotic process. These include CHOP induction, caspase-12 activation, mitochondria dysfunction, JNK activation, Fas-FasL system and the STAT1 pathway. Blocking each of these pathways reduces or prevents ER stress-induced apoptosis to a certain extent; however, induction of an individual proapoptotic pathway may not be sufficient to induce apoptosis . This suggests that prolonged ER stress may activate multiple subthreshold proapoptotic pathways, and these pathways interact and regulate each other to execute apoptosis. In addition, prolonged ER stress may also suppress the compensatory cell survival pathways induced by the UPR, such as the IFN-β and Akt-p38α pathways . Recently, we demonstrated that enhancing the endogenous adaptive UPR system by preconditioning retinal cells with mild ER stress was able to reduce vascular inflammation and retinal vascular leakage . Moreover, inducing molecular chaperones, for example, heat shock protein 90 (Hsp90), by antibiotics  prevents protein aggregation and that protects photoreceptors against retinal degeneration in a murine model of autosomal dominant retinitis pigmentosa (ADRP) . In addition, overexpressing Bip/Bip in retinal photoreceptors alleviated ER stress, reduced CHOP expression, and mitigated photoreceptor apoptosis in P23H rhodopsin transgenic rats . These findings support an essential role of the adaptive URP system and the URP-activated survival pathways in protecting retinal cells against apoptosis and cell death. Therefore, identifying the key proapoptotic and antiapoptotic pathways implicated in ER stress-associated apoptosis and addressing how these pathways are involved in different pathological conditions of retinal cells may offer the opportunity for developing new drugs to treat retinal diseases.
This work was supported by the National Institutes of Health Grant EY019949 and research awards from the American Diabetes Association, Juvenile Diabetes Research Foundation, Oklahoma Center for the Advancement of Science and Technology, American Health Assistance Foundation, and Harold Hamm Diabetes Center (all to S. X. Zhang).
- E. Garcia-Valenzuela, S. Shareef, J. Walsh, and S. C. Sharma, “Programmed cell death of retinal ganglion cells during experimental glaucoma,” Experimental Eye Research, vol. 61, no. 1, pp. 33–44, 1995.
- L. Guo, S. E. Moss, R. A. Alexander, R. R. Ali, F. W. Fitzke, and M. F. Cordeiro, “Retinal ganglion cell apoptosis in glaucoma is related to intraocular pressure and IOP-induced effects on extracellular matrix,” Investigative Ophthalmology and Visual Science, vol. 46, no. 1, pp. 175–182, 2005.
- G. H. Travis, “Mechanisms of cell death in the inherited retinal degenerations,” American Journal of Human Genetics, vol. 62, no. 3, pp. 503–508, 1998.
- R. Allikmets, N. Singh, H. Sun et al., “A photoreceptor cell-specific ATP-binding transporter gene (ABCR) is mutated in recessive Stargardt macular dystrophy,” Nature Genetics, vol. 15, no. 3, pp. 236–246, 1997.
- C. Portera-Cailliau, C. H. Sung, J. Nathans, and R. Adler, “Apoptotic photoreceptor cell death in mouse models of retinitis pigmentosa,” Proceedings of the National Academy of Sciences of the United States of America, vol. 91, no. 3, pp. 974–978, 1994.
- P. T. V. M. De Jong, “Age-related macular degeneration,” New England Journal of Medicine, vol. 355, no. 14, pp. 1474–1485, 2006.
- J. R. Wolter, “Diabetic retinopathy,” American Journal of Ophthalmology, vol. 51, no. 5, pp. 1123–1141, 1961.
- A. M. Joussen, V. Poulaki, N. Mitsiades et al., “Nonsteroidal anti-inflammatory drugs prevent early diabetic retinopathy via TNF-alpha suppression,” The FASEB Journal, vol. 16, no. 3, pp. 438–440, 2002.
- A. J. Barber, E. Lieth, S. A. Khin, D. A. Antonetti, A. G. Buchanan, and T. W. Gardner, “Neural apoptosis in the retina during experimental and human diabetes: early onset and effect of insulin,” Journal of Clinical Investigation, vol. 102, no. 4, pp. 783–791, 1998.
- T. K. Tarrant, P. B. Silver, J. L. Wahlsten et al., “Interleukin 12 protects from a T helper type 1-mediated autoimmune disease, experimental autoimmune uveitis, through a mechanism involving interferon γ, nitric oxide, and apoptosis,” Journal of Experimental Medicine, vol. 189, no. 2, pp. 219–230, 1999.
- V. Poulaki, N. Mitsiades, G. Mastorakos, R. R. Caspi, G. P. Chrousos, and E. Bouzas, “Fas/Fas ligand-associated apoptosis in experimental autoimmune uveoretinitis in rodents: role of proinflammatory corticotropin-releasing hormone,” Experimental Eye Research, vol. 72, no. 6, pp. 623–629, 2001.
- J. Sueda, N. Hikita, M. Mochizuki, A. Jimi, and M. Kojiro, “Kinetics of apoptotic cells in experimental autoimmune uveoretinitis,” Investigative Ophthalmology and Visual Science, vol. 41, no. 3, pp. 799–804, 2000.
- P. Yang, N. H. Herzberg, H. Zhou, L. Broersma, M. De Smet, and A. Kijlstra, “Apoptosis of infiltrating cells in experimental autoimmune uveoretinitis,” Chinese Medical Journal, vol. 113, no. 7, pp. 643–646, 2000.
- S. Amano, S. I. Yamagishi, Y. Inagaki et al., “Pigment epithelium-derived factor inhibits oxidative stress-induced apoptosis and dysfunction of cultured retinal pericytes,” Microvascular Research, vol. 69, no. 1-2, pp. 45–55, 2005.
- Y. Behl, P. Krothapalli, T. Desta, A. DiPiazza, S. Roy, and D. T. Graves, “Diabetes-enhanced tumor necrosis factor-α production promotes apoptosis and the loss of retinal microvascular cells in type 1 and type 2 models of diabetic retinopathy,” American Journal of Pathology, vol. 172, no. 5, pp. 1411–1418, 2008.
- S. Choudhary, T. Xiao, S. Srivastava et al., “Toxicity and detoxification of lipid-derived aldehydes in cultured retinal pigmented epithelial cells,” Toxicology and Applied Pharmacology, vol. 204, no. 2, pp. 122–134, 2005.
- L. L. Kusner, V. P. Sarthy, and S. Mohr, “Nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase: a role in high glucose-induced apoptosis in retinal Müller cells,” Investigative Ophthalmology and Visual Science, vol. 45, no. 5, pp. 1553–1561, 2004.
- R. A. Kowluru, “Diabetic retinopathy: mitochondrial dysfunction and retinal capillary cell death,” Antioxidants and Redox Signaling, vol. 7, no. 11-12, pp. 1581–1587, 2005.
- M. Shimazawa, Y. Inokuchi, Y. Ito et al., “Involvement of ER stress in retinal cell death,” Molecular Vision, vol. 13, pp. 578–587, 2007.
- K. D. McCullough, J. L. Martindale, L. O. Klotz, T. Y. Aw, and N. J. Holbrook, “Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bc12 and perturbing the cellular redox state,” Molecular and Cellular Biology, vol. 21, no. 4, pp. 1249–1259, 2001.
- Z. Galehdar, P. Swan, B. Fuerth, S. M. Callaghan, D. S. Park, and S. P. Cregan, “Neuronal apoptosis induced by endoplasmic reticulum stress is regulated by ATF4-CHOP-mediated induction of the Bcl-2 homology 3-only member PUMA,” Journal of Neuroscience, vol. 30, no. 50, pp. 16938–16948, 2010.
- D. Ron and P. Walter, “Signal integration in the endoplasmic reticulum unfolded protein response,” Nature Reviews Molecular Cell Biology, vol. 8, no. 7, pp. 519–529, 2007.
- D. J. Todd, A. H. Lee, and L. H. Glimcher, “The endoplasmic reticulum stress response in immunity and autoimmunity,” Nature Reviews Immunology, vol. 8, no. 9, pp. 663–674, 2008.
- A. Görlach, P. Klappa, and T. Kietzmann, “The endoplasmic reticulum: folding, calcium homeostasis, signaling, and redox control,” Antioxidants and Redox Signaling, vol. 8, no. 9-10, pp. 1391–1418, 2006.
- L. Bouman, A. Schlierf, A. K. Lutz et al., “Parkin is transcriptionally regulated by ATF4: evidence for an interconnection between mitochondrial stress and ER stress,” Cell Death and Differentiation, vol. 18, pp. 769–782, 2011.
- S. S. Rajan, V. Srinivasan, M. Balasubramanyam, and U. Tatu, “Endoplasmic reticulum (ER) stress & diabetes,” Indian Journal of Medical Research, vol. 125, no. 3, pp. 411–424, 2007.
- D. Lindholm, H. Wootz, and L. Korhonen, “ER stress and neurodegenerative diseases,” Cell Death and Differentiation, vol. 13, no. 3, pp. 385–392, 2006.
- E. Szegezdi, A. Duffy, M. E. O'Mahoney et al., “ER stress contributes to ischemia-induced cardiomyocyte apoptosis,” Biochemical and Biophysical Research Communications, vol. 349, no. 4, pp. 1406–1411, 2006.
- H. P. Harding, Y. Zhang, and D. Ron, “Protein translation and folding are coupled by an endoplasmic- reticulum-resident kinase,” Nature, vol. 397, no. 6716, pp. 271–274, 1999.
- W. Tirasophon, A. A. Welihinda, and R. J. Kaufman, “A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells,” Genes and Development, vol. 12, no. 12, pp. 1812–1824, 1998.
- M. Calfon, H. Zeng, F. Urano et al., “IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA,” Nature, vol. 415, no. 6867, pp. 92–96, 2002.
- A. Bertolotti, Y. Zhang, L. M. Hendershot, H. P. Harding, and D. Ron, “Dynamic interaction of BiP and ER stress transducers in the unfolded-protein response,” Nature Cell Biology, vol. 2, no. 6, pp. 326–332, 2000.
- D. T. W. Ng, S. S. Watowich, and R. A. Lamb, “Analysis in vivo of GRP78-BiP/substrate interactions and their role in induction of the GRP78-BiP gene,” Molecular Biology of the Cell, vol. 3, no. 2, pp. 143–155, 1992.
- K. Haze, T. Okada, H. Yoshida et al., “Identification of the G13 (cAMP-response-element-binding protein-related protein) gene product related to activating transcription factor 6 as a transcriptional activator of the mammalian unfolded protein response,” Biochemical Journal, vol. 355, no. 1, pp. 19–28, 2001.
- M. Schröder and R. J. Kaufman, “The mammalian unfolded protein response,” Annual Review of Biochemistry, vol. 74, pp. 739–789, 2005.
- W. Paschen and A. Frandsen, “Endoplasmic reticulum dysfunction—a common denominator for cell injury in acute and degenerative diseases of the brain?” Journal of Neurochemistry, vol. 79, no. 4, pp. 719–725, 2001.
- R. V. Rao, H. M. Ellerby, and D. E. Bredesen, “Coupling endoplasmic reticulum stress to the cell death program,” Cell Death and Differentiation, vol. 11, no. 4, pp. 372–380, 2004.
- K. Mori, W. Ma, M. J. Gething, and J. Sambrook, “A transmembrane protein with a cdc2+/CDC28-related kinase activity is required for signaling from the ER to the nucleus,” Cell, vol. 74, no. 4, pp. 743–756, 1993.
- J. S. Cox and P. Walter, “A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response,” Cell, vol. 87, no. 3, pp. 391–404, 1996.
- X. Z. Wang, H. P. Harding, Y. Zhang, E. M. Jolicoeur, M. Kuroda, and D. Ron, “Cloning of mammalian Ire1 reveals diversity in the ER stress responses,” EMBO Journal, vol. 17, no. 19, pp. 5708–5717, 1998.
- H. Yoshida, T. Matsui, A. Yamamoto, T. Okada, and K. Mori, “XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor,” Cell, vol. 107, no. 7, pp. 881–891, 2001.
- A. H. Lee, N. N. Iwakoshi, and L. H. Glimcher, “XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response,” Molecular and Cellular Biology, vol. 23, no. 21, pp. 7448–7459, 2003.
- H. Yoshida, T. Matsui, N. Hosokawa, R. J. Kaufman, K. Nagata, and K. Mori, “A time-dependent phase shift in the mammalian unfolded protein response,” Developmental Cell, vol. 4, no. 2, pp. 265–271, 2003.
- Y. Liu, M. Adachi, S. Zhao et al., “Preventing oxidative stress: a new role for XBP1,” Cell Death and Differentiation, vol. 16, no. 6, pp. 847–857, 2009.
- A. Kaser, A. H. Lee, A. Franke et al., “XBP1 links ER stress to intestinal inflammation and confers genetic risk for human inflammatory bowel disease,” Cell, vol. 134, no. 5, pp. 743–756, 2008.
- K. Lee, W. Tirasophon, X. Shen et al., “IRE1-mediated unconventional mRNA splicing and S2P-mediated ATF6 cleavage merge to regulate XBP1 in signaling the unfolded protein response,” Genes and Development, vol. 16, no. 4, pp. 452–466, 2002.
- A. M. Reimold, A. Etkin, I. Clauss et al., “An essential role in liver development for transcription factor XBP-1,” Genes and Development, vol. 14, no. 2, pp. 152–157, 2000.
- T. Iwawaki, R. Akai, S. Yamanaka, and K. Kohno, “Function of IRE1 alpha in the placenta is essential for placental development and embryonic viability,” Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 39, pp. 16657–16662, 2009.
- H. P. Harding, Y. Zhang, A. Bertolotti, H. Zeng, and D. Ron, “Perk is essential for translational regulation and cell survival during the unfolded protein response,” Molecular Cell, vol. 5, no. 5, pp. 897–904, 2000.
- H. P. Harding, Y. Zhang, H. Zeng et al., “An integrated stress response regulates amino acid metabolism and resistance to oxidative stress,” Molecular Cell, vol. 11, no. 3, pp. 619–633, 2003.
- H. P. Harding, I. Novoa, Y. Zhang et al., “Regulated translation initiation controls stress-induced gene expression in mammalian cells,” Molecular Cell, vol. 6, no. 5, pp. 1099–1108, 2000.
- K. M. Vattem and R. C. Wek, “Reinitiation involving upstream ORFs regulates ATF4 mRNA translation in mammalian cells,” Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 31, pp. 11269–11274, 2004.
- K. Haze, H. Yoshida, H. Yanagi, T. Yura, and K. Mori, “Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress,” Molecular Biology of the Cell, vol. 10, no. 11, pp. 3787–3799, 1999.
- M. Q. Li, P. Baumeister, B. Roy et al., “ATF6 as a transcription activator of the endoplasmic reticulum stress element: thapsigargin stress-induced changes and synergistic interactions with NF-Y and YY1,” Molecular and Cellular Biology, vol. 20, no. 14, pp. 5096–5106, 2000.
- X. Chen, J. Shen, and R. Prywes, “The luminal domain of ATF6 senses endoplasmic reticulum (ER) stress and causes translocation of ATF6 from the er to the Golgi,” Journal of Biological Chemistry, vol. 277, no. 15, pp. 13045–13052, 2002.
- H. Yoshida, T. Okada, K. Haze et al., “ATF6 activated by proteolysis binds in the presence of NF-Y (CBF) directly to the cis-acting element responsible for the mammalian unfolded protein response,” Molecular and Cellular Biology, vol. 20, no. 18, pp. 6755–6767, 2000.
- H. Yoshida, “ER stress and diseases,” FEBS Journal, vol. 274, no. 3, pp. 630–658, 2007.
- N. Naidoo, M. Ferber, M. Master, Y. Zhu, and A. I. Pack, “Aging impairs the unfolded protein response to sleep deprivation and leads to proapoptotic signaling,” Journal of Neuroscience, vol. 28, no. 26, pp. 6539–6548, 2008.
- M. J. Kang and D. R. Hyung, “Suppression of retinal degeneration in Drosophila by stimulation of ER-associated degradation,” Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 40, pp. 17043–17048, 2009.
- H. D. Ryoo, P. M. Domingos, M. J. Kang, and H. Steller, “Unfolded protein response in a Drosophila model for retinal degeneration,” EMBO Journal, vol. 26, no. 1, pp. 242–252, 2007.
- M. S. Gorbatyuk, T. Knox, M. M. LaVail et al., “Restoration of visual function in P23H rhodopsin transgenic rats by gene delivery of BiP/Grp78,” Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 13, pp. 5961–5966, 2010.
- L. P. Yang, L. M. Wu, X. J. Guo, Y. Li, and M. O. M. Tso, “Endoplasmic reticulum stress is activated in light-induced retinal degeneration,” Journal of Neuroscience Research, vol. 86, no. 4, pp. 910–919, 2008.
- G. Rebello, R. Ramesar, A. Vorster et al., “Apoptosis-inducing signal sequence mutation in carbonic anhydrase IV identified in patients with the RP17 form of retinitis pigmentosa,” Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 17, pp. 6617–6622, 2004.
- B. M. Tam and O. L. Moritz, “Characterization of rhodopsin P23H-induced retinal degeneration in a Xenopus laevis model of retinitis pigmentosa,” Investigative Ophthalmology and Visual Science, vol. 47, no. 8, pp. 3234–3241, 2006.
- T. Shinohara, M. L. Mulhern, and C. J. Madson, “Silencing gene therapy for mutant membrane, secretory, and lipid proteins in retinitis pigmentosa (RP),” Medical Hypotheses, vol. 70, no. 2, pp. 378–380, 2008.
- J. H. Lin, H. Li, D. Yasumura et al., “IRE1 signaling affects cell fate during the unfolded protein response,” Science, vol. 318, no. 5852, pp. 944–949, 2007.
- C. S. Mendes, C. Levet, G. Chatelain et al., “ER stress protects from retinal degeneration,” EMBO Journal, vol. 28, no. 9, pp. 1296–1307, 2009.
- S. H. Doh, J. H. Kim, K. M. Lee, H. Y. Park, and C. K. Park, “Retinal ganglion cell death induced by endoplasmic reticulum stress in a chronic glaucoma model,” Brain Research, vol. 1308, pp. 158–166, 2010.
- Y. Inokuchi, Y. Nakajima, M. Shimazawa et al., “Effect of an inducer of BiP, a molecular chaperone, on endoplasmic reticulum (ER) stress-induced retinal cell death,” Investigative Ophthalmology and Visual Science, vol. 50, no. 1, pp. 334–344, 2009.
- P. M. Martin, P. Roon, T. K. Van Ells, V. Ganapathy, and S. B. Smith, “Death of retinal neurons in streptozotocin-induced diabetic mice,” Investigative Ophthalmology and Visual Science, vol. 45, no. 9, pp. 3330–3336, 2004.
- A. M. Joussen, V. Poulaki, A. Tsujikawa et al., “Suppression of diabetic retinopathy with angiopoietin-1,” American Journal of Pathology, vol. 160, no. 5, pp. 1683–1693, 2002.
- D. A. Antonetti, A. J. Barber, S. Khin, E. Lieth, J. M. Tarbell, and T. W. Gardner, “Vascular permeability in experimental diabetes is associated with reduced endothelial occludin content. Vascular endothelial growth factor decreases occludin in retinal endothelial cells,” Diabetes, vol. 47, no. 12, pp. 1953–1959, 1998.
- J. Li, J. J. Wang, Q. Yu, M. Wang, and S. X. Zhang, “Endoplasmic reticulum stress is implicated in retinal inflammation and diabetic retinopathy,” FEBS Letters, vol. 583, no. 9, pp. 1521–1527, 2009.
- J. Li, J. J. Wang, and S. X. Zhang, “Preconditioning with endoplasmic reticulum stress mitigates retinal endothelial inflammation via activation of X-box binding protein 1,” Journal of Biological Chemistry, vol. 286, no. 6, pp. 4912–4921, 2011.
- Y. Zhong, J. J. Wang, and S. X. Zhang, “Intermittent but not constant high glucose induces ER stress and inflammation in human retinal pericytes,” Advances in Experimental Medicine and Biology. In press.
- B. Li, D. Li, G. G. Li, H. W. Wang, and A. X. Yu, “P58IPK inhibition of endoplasmic reticulum stress in human retinal capillary endothelial cells in vitro,” Molecular Vision, vol. 14, pp. 1122–1128, 2008.
- C. E. Remé, C. Grimm, F. Hafezi, A. Wenzel, and T. P. Williams, “Apoptosis in the Retina: The Silent Death of Vision,” News in Physiological Sciences, vol. 15, no. 3, pp. 120–125, 2000.
- A. Degterev, M. Boyce, and J. Yuan, “A decade of caspases,” Oncogene, vol. 22, no. 53, pp. 8543–8567, 2003.
- R. J. Carmody and T. G. Cotter, “Oxidative stress induces caspase-independent retinal apoptosis in vitro,” Cell Death and Differentiation, vol. 7, no. 3, pp. 282–291, 2000.
- X. Wang and D. Ron, “Stress-induced phosphorylation and activation of the transcription factor CHOP (GADD153) by p38 MAP kinase,” Science, vol. 272, no. 5266, pp. 1347–1349, 1996.
- B. Eymin, L. Dubrez, M. Allouche, and E. Solary, “Increased gadd153 messenger RNA level is associated with apoptosis in human leukemic cells treated with etoposide,” Cancer Research, vol. 57, no. 4, pp. 686–695, 1997.
- A. D. Friedman, “GADD153/CHOP, a DNA damage-inducible protein, reduced CAAT/enhancer binding protein activities and increased apoptosis in 32d cl3 myeloid cells,” Cancer Research, vol. 56, no. 14, pp. 3250–3256, 1996.
- S. Oyadomari, A. Koizumi, K. Takeda et al., “Targeted disruption of the Chop gene delays endoplasmic reticulum stress-mediated diabetes,” Journal of Clinical Investigation, vol. 109, no. 4, pp. 525–532, 2002.
- C. R. Bromati, C. Lellis-Santos, T. S. Yamanaka et al., “UPR induces transient burst of apoptosis in islets of early lactating rats through reduced AKT phosphorylation via ATF4/CHOP stimulation of TRB3 expression,” American Journal of Physiology, vol. 300, no. 1, pp. R92–R100, 2011.
- N. Ohoka, S. Yoshii, T. Hattori, K. Onozaki, and H. Hayashi, “TRB3, a novel ER stress-inducible gene, is induced via ATF4-CHOP pathway and is involved in cell death,” EMBO Journal, vol. 24, no. 6, pp. 1243–1255, 2005.
- C. Jousse, C. Deval, A. C. Maurin et al., “TRB3 inhibits the transcriptional activation of stress-regulated genes by a negative feedback on the ATF4 pathway,” Journal of Biological Chemistry, vol. 282, no. 21, pp. 15851–15861, 2007.
- H. Puthalakath, L. A. O'Reilly, P. Gunn et al., “ER stress triggers apoptosis by activating BH3-only protein Bim,” Cell, vol. 129, no. 7, pp. 1337–1349, 2007.
- N. N. Danial and S. J. Korsmeyer, “Cell death: critical control points,” Cell, vol. 116, no. 2, pp. 205–219, 2004.
- M. C. Wei, W. X. Zong, E. H. Y. Cheng et al., “Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death,” Science, vol. 292, no. 5517, pp. 727–730, 2001.
- N. S. Wang, M. T. Unkila, E. Z. Reineks, and C. W. Distelhorst, “Transient expression of wild-type or mitochondrially targeted Bcl-2 induces apoptosis, whereas transient expression of endoplasmic reticulum-targeted Bcl-2 is protective against Bax-induced cell death,” Journal of Biological Chemistry, vol. 276, no. 47, pp. 44117–44128, 2001.
- D. C. S. Huang and A. Strasser, “BH3-only proteins—essential initiators of apoptotic cell death,” Cell, vol. 103, no. 6, pp. 839–842, 2000.
- K. D. McCullough, J. L. Martindale, L. O. Klotz, T. Y. Aw, and N. J. Holbrook, “Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bc12 and perturbing the cellular redox state,” Molecular and Cellular Biology, vol. 21, no. 4, pp. 1249–1259, 2001.
- E. Szegezdi, S. E. Logue, A. M. Gorman, and A. Samali, “Mediators of endoplasmic reticulum stress-induced apoptosis,” EMBO Reports, vol. 7, no. 9, pp. 880–885, 2006.
- J. Li, B. Lee, and A. S. Lee, “Endoplasmic reticulum stress-induced apoptosis: multiple pathways and activation of p53-UP-regulated modulator of apoptosis (PUMA) and NOXA by p53,” Journal of Biological Chemistry, vol. 281, no. 11, pp. 7260–7270, 2006.
- C. Hetz, P. Bernasconi, J. Fisher et al., “Proapoptotic BAX and BAK modulate the unfolded protein response by a direct interaction with IRE1α,” Science, vol. 312, no. 5773, pp. 572–576, 2006.
- T. Nakagawa, H. Zhu, N. Morishima et al., “Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-β,” Nature, vol. 403, no. 6765, pp. 98–103, 2000.
- M. Saleh, J. P. Vaillancourt, R. K. Graham et al., “Differential modulation of endotoxin responsiveness by human caspase-12 polymorphisms,” Nature, vol. 429, no. 6987, pp. 75–79, 2004.
- J. Hitomi, T. Katayama, Y. Eguchi et al., “Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis and Aβ-induced cell death,” Journal of Cell Biology, vol. 165, no. 3, pp. 347–356, 2004.
- F. Urano, X. Wang, A. Bertolotti et al., “Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1,” Science, vol. 287, no. 5453, pp. 664–666, 2000.
- T. Yoneda, K. Imaizumi, K. Oono et al., “Activation of caspase-12, an endoplastic reticulum (ER) resident caspase, through tumor necrosis factor receptor-associated factor 2-dependent mechanism in response to the ER stress,” Journal of Biological Chemistry, vol. 276, no. 17, pp. 13935–13940, 2001.
- N. Morishima, K. Nakanishi, H. Takenouchi, T. Shibata, and Y. Yasuhiko, “An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12,” Journal of Biological Chemistry, vol. 277, no. 37, pp. 34287–34294, 2002.
- R. V. Rao, S. Castro-Obregon, H. Frankowski et al., “Coupling endoplasmic reticulum stress to the cell death program. An Apaf-1-independent intrinsic pathway,” Journal of Biological Chemistry, vol. 277, no. 24, pp. 21836–21842, 2002.
- M. Hibi, A. Lin, T. Smeal, A. Minden, and M. Karin, “Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain,” Genes and Development, vol. 7, no. 11, pp. 2135–2148, 1993.
- 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.
- S. Leppä and D. Bohmann, “Diverse functions of JNK signaling and c-Jun in stress response and apoptosis,” Oncogene, vol. 18, no. 45, pp. 6158–6162, 1999.
- J. Liu, Y. Minemoto, and A. Lin, “c-Jun N-terminal protein kinase 1 (JNK1), but not JNK2, is essential for tumor necrosis factor alpha-induced c-Jun kinase activation and apoptosis,” Molecular and Cellular Biology, vol. 24, no. 24, pp. 10844–10856, 2004.
- D. Ron and P. Walter, “Signal integration in the endoplasmic reticulum unfolded protein response,” Nature Reviews Molecular Cell Biology, vol. 8, no. 7, pp. 519–529, 2007.
- G. Verma and M. Datta, “IL-1β induces ER stress in a JNK dependent manner that determines cell death in human pancreatic epithelial MIA PaCa-2 cells,” Apoptosis, vol. 15, no. 7, pp. 864–876, 2010.
- T. Matsuguchi, N. Chiba, K. Bandow, K. Kakimoto, A. Masuda, and T. Ohnishi, “JNK activity is essential for Atf4 expression and late-stage osteoblast differentiation,” Journal of Bone and Mineral Research, vol. 24, no. 3, pp. 398–410, 2009.
- G. Dasmahapatra, D. Lembersky, M. Rahmani et al., “Bcl-2 antagonists interact synergistically with bortezomib in DLBCL cells in association with JNK activation and induction of ER stress,” Cancer Biology and Therapy, vol. 8, no. 9, pp. 808–819, 2009.
- K. A. Koeplinger, A. M. Mildner, J. W. Leone, J. S. Wheeler, R. L. Heinrikson, and A. G. Tomasselli, “Caspase 8: an efficient method for large-scale autoactivation of recombinant procaspase 8 by matrix adsorption and characterization of the active enzyme,” Protein Expression and Purification, vol. 18, no. 3, pp. 378–387, 2000.
- P. A. Gregoli and M. C. Bondurant, “Function of caspases in regulating apoptosis caused by erythropoietin deprivation in erythroid progenitors,” Journal of Cellular Physiology, vol. 178, no. 2, pp. 133–143, 1999.
- K. Miyamoto, S. Khosrof, S. E. Bursell et al., “Prevention of leukostasis and vascular leakage in streptozotocin-induced diabetic retinopathy via intercellular adhesion molecule-1 inhibition,” Proceedings of the National Academy of Sciences of the United States of America, vol. 96, no. 19, pp. 10836–10841, 1999.
- R. Arita, Y. Hata, S. Nakao et al., “Rho kinase inhibition by fasudil ameliorates diabetes-induced microvascular damage,” Diabetes, vol. 58, no. 1, pp. 215–226, 2009.
- A. M. Joussen, V. Poulaki, N. Mitsiades et al., “Suppression of Fas-FasL-induced endothelial cell apoptosis prevents diabetic blood-retinal barrier breakdown in a model of streptozotocin-induced diabetes,” The FASEB Journal, vol. 17, no. 1, pp. 76–78, 2003.
- K. R. Ju, H. S. Kim, J. H. Kim, N. Y. Lee, and C. K. Park, “Retinal glial cell responses and Fas/FasL activation in rats with chronic ocular hypertension,” Brain Research, vol. 1122, no. 1, pp. 209–221, 2006.
- J. M. Timmins, L. Ozcan, T. A. Seimon et al., “Calcium/calmodulin-dependent protein kinase II links ER stress with Fas and mitochondrial apoptosis pathways,” Journal of Clinical Investigation, vol. 119, no. 10, pp. 2925–2941, 2009.
- A. Sittler, R. Lurz, G. Lueder et al., “Geldanamycin activates a heat shock response and inhibits huntingtin aggregation in a cell culture model of Huntington's disease,” Human Molecular Genetics, vol. 10, no. 12, pp. 1307–1315, 2001.
- L. C. Tam, A. S. Kiang, M. Campbell et al., “Prevention of autosomal dominant retinitis pigmentosa by systemic drug therapy targeting heat shock protein 90 (Hsp90),” Human Molecular Genetics, vol. 19, no. 22, pp. 4421–4436, 2010.