BioMed Research International: Developmental Biology The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Synchronization by Food Access Modifies the Daily Variations in Expression and Activity of Liver GABA Transaminase Mon, 07 Apr 2014 11:46:16 +0000 Daytime restricted feeding (DRF) is an experimental protocol that influences the circadian timing system and underlies the expression of a biological clock known as the food entrained oscillator (FEO). Liver is the organ that reacts most rapidly to food restriction by adjusting the functional relationship between the molecular circadian clock and the metabolic networks. -Aminobutyric acid (GABA) is a signaling molecule in the liver, and able to modulate the cell cycle and apoptosis. This study was aimed at characterizing the expression and activity of the mostly mitochondrial enzyme GABA transaminase (GABA-T) during DRF/FEO expression. We found that DRF promotes a sustained increase of GABA-T in the liver homogenate and mitochondrial fraction throughout the entire day-night cycle. The higher amount of GABA-T promoted by DRF was not associated to changes in GABA-T mRNA or GABA-T activity. The GABA-T activity in the mitochondrial fraction even tended to decrease during the light period. We concluded that DRF influences the daily variations of GABA-T mRNA levels, stability, and catalytic activity of GABA-T. These data suggest that the liver GABAergic system responds to a metabolic challenge such as DRF and the concomitant appearance of the FEO. Dalia De Ita-Pérez, Isabel Méndez, Olivia Vázquez-Martínez, Mónica Villalobos-Leal, and Mauricio Díaz-Muñoz Copyright © 2014 Dalia De Ita-Pérez et al. All rights reserved. Developmental Stage-Specific Regulation of the Circadian Clock by Temperature in Zebrafish Thu, 27 Mar 2014 00:00:00 +0000 The circadian clock enables animals to adapt their physiology and behaviour in anticipation of the day-night cycle. Light and temperature represent two key environmental timing cues (zeitgebers) able to reset this mechanism and so maintain its synchronization with the environmental cycle. One key challenge is to unravel how the regulation of the clock by zeitgebers matures during early development. The zebrafish is an ideal model for studying circadian clock ontogeny since the process of development occurs ex utero in an optically transparent chorion and many tools are available for genetic analysis. However, the role played by temperature in regulating the clock during zebrafish development is poorly understood. Here, we have established a clock-regulated luciferase reporter transgenic zebrafish line (Tg (−3.1) per1b::luc) to study the effects of temperature on clock entrainment. We reveal that under complete darkness, from an early developmental stage onwards (48 to 72 hpf), exposure to temperature cycles is a prerequisite for the establishment of self-sustaining rhythms of zfper1b, zfaanat2, and zfirbp expression and also for circadian cell cycle rhythms. Furthermore, we show that following the 5–9 somite stage, the expression of zfper1b is regulated by acute temperature shifts. Kajori Lahiri, Nadine Froehlich, Andreas Heyd, Nicholas S. Foulkes, and Daniela Vallone Copyright © 2014 Kajori Lahiri et al. All rights reserved. Neuropeptide Y in the Adult and Fetal Human Pineal Gland Mon, 17 Mar 2014 10:06:59 +0000 Neuropeptide Y was isolated from the porcine brain in 1982 and shown to be colocalized with noradrenaline in sympathetic nerve terminals. The peptide has been demonstrated to be present in sympathetic nerve fibers innervating the pineal gland in many mammalian species. In this investigation, we show by use of immunohistochemistry that neuropeptide Y is present in nerve fibers of the adult human pineal gland. The fibers are classical neuropeptidergic fibers endowed with large boutons en passage and primarily located in a perifollicular position with some fibers entering the pineal parenchyma inside the follicle. The distance from the immunoreactive terminals to the pinealocytes indicates a modulatory function of neuropeptide Y for pineal physiology. Some of the immunoreactive fibers might originate from neurons located in the brain and be a part of the central innervation of the pineal gland. In a series of human fetuses, neuropeptide Y-containing nerve fibers was present and could be detected as early as in the pineal of four- to five-month-old fetuses. This early innervation of the human pineal is different from most rodents, where the innervation starts postnatally. Morten Møller, Pansiri Phansuwan-Pujito, and Corin Badiu Copyright © 2014 Morten Møller et al. All rights reserved. An Immunocytochemical Study of Interchromatin Granule Clusters in Early Mouse Embryos Wed, 11 Sep 2013 13:48:30 +0000 Interchromatin granule clusters (IGCs) are universal nuclear domains. Their molecular composition and functions were studied in detail in somatic cells. Here, we studied IGCs in the nuclei of early mouse embryos during zygotic gene activation (ZGA). We found that the size of IGCs gradually increases during realization of ZGA events. Using immunocytochemical approaches, we showed that the molecular composition of IGCs is also modified in mouse embryos. The hyperphosphorylated form of RNA polymerase II and the transcription factor TFIID have been revealed in IGCs before the end of ZGA. Association of these factors with IGCs became more noticeable during ZGA realization. Our data suggest that IGCs in early mouse embryos have some functional peculiarities connected most probably with IGC formation de novo. We believe that IGCs in early mouse embryos not only are storage sites of splicing factors but also may be involved in mRNA metabolism and represent the multifunctional nuclear domains. Irina Bogolyubova and Dmitry Bogolyubov Copyright © 2013 Irina Bogolyubova and Dmitry Bogolyubov. All rights reserved. Antioxidant, Antiproliferative, and Antiangiogenesis Effects of Polyphenol-Rich Seaweed (Sargassum muticum) Tue, 03 Sep 2013 16:21:23 +0000 In the present study, we evaluated the effect of brown seaweeds Sargassum muticum methanolic extract (SMME), against MCF-7 and MDA-MB-231 breast cancer cell lines proliferation. This algae extract was also evaluated for reducing activity and total polyphenol content. The MTT assay results indicated that the extracts were cytotoxic against breast cancer cell lines in a dose-dependent manner, with IC50 of 22 μg/ml for MCF-7 and 55 μg/ml for MDA-MB-231 cell lines. The percentages of apoptotic MCF-7-treated cells increased from 13% to 67% by increasing the concentration of the SMME. The antiproliferative efficacy of this algal extract was positively correlated with the total polyphenol contents, suggesting a causal link related to extract content of phenolic acids. Cell cycle analysis showed a significant increase in the accumulation of SMME-treated cells at sub-G1 phase, indicating the induction of apoptosis by SMME. Further apoptosis induction was confirmed by Hoechst 33342 and AO/PI staining. Also SMME implanted in vivo into fertilized chicken eggs induced dose-related antiangiogenic activity in the chorioallantoic membrane (CAM). Our results imply a new insight on the novel function of Sargassum muticum polyphenol-rich seaweed in cancer research by induction of apoptosis, antioxidant, and antiangiogenesis effects. Farideh Namvar, Rosfarizan Mohamad, Javad Baharara, Saeedeh Zafar-Balanejad, Fahimeh Fargahi, and Heshu Sulaiman Rahman Copyright © 2013 Farideh Namvar et al. All rights reserved. Regulatory Interactions between Androgens, Hoxb5, and TGFβ Signaling in Murine Lung Development Tue, 03 Sep 2013 11:09:37 +0000 Androgens enhance airway branching but delay alveolar maturation contributing to increased respiratory morbidity in prematurely born male infants. Hoxb5 protein positively regulates airway branching in developing lung. In other organs, androgen regulation intersects with Hox proteins and TGFβ-SMAD signaling, but these interactions have not been studied in the lung. We hypothesized that androgen alteration of airway branching early in lung development requires Hoxb5 expression and that these androgen-Hoxb5 interactions occur partially through regional changes in TGFβ signaling. To evaluate acute effects of androgen and TGFβ on Hoxb5, E11 whole fetal mouse lungs were cultured with dihydrotestosterone (DHT) with/without Hoxb5 siRNA or TGFβ inhibitory antibody. Chronic in utero DHT exposure was accomplished by exposing pregnant mice to DHT (subcutaneous pellet) from E11 to E18. DHT’s ability to enhance airway branching and alter phosphorylated SMAD2 cellular localization was partially dependent on Hoxb5. Hoxb5 inhibition also changed the cellular distribution of SMAD7 protein. Chronic in utero DHT increased Hoxb5 and altered SMAD7 mesenchymal localization. TGFβ inhibition enhanced airway branching, and Hoxb5 protein cellular localization was more diffuse. We conclude that DHT controls lung airway development partially through modulation of Hoxb5 protein expression and that this level of regulation involves interactions with TGFβ signaling. MaryAnn V. Volpe, Sujatha M. Ramadurai, Sana Mujahid, Thanhxuan Vong, Marcia Brandao, Karen T. Wang, Lucia D. Pham, and Heber C. Nielsen Copyright © 2013 MaryAnn V. Volpe et al. All rights reserved. Expression and Cellular Distribution of INHA and INHB before and after In Vitro Cultivation of Porcine Oocytes Isolated from Follicles of Different Size Tue, 20 Nov 2012 15:00:20 +0000 Cumulus-oocyte-complexes (COCs) were collected from small (<3 mm), medium (3–5 mm), and large (>5 mm) porcine follicles, and the INHA and INHB expression and cellular localization were studied. Developmentally competent (BCB+) COCs were cultured for 44 h. Samples of mRNA were isolated before and after in vitro maturation (IVM) from oocytes collected from follicles of different size for RQ-PCR assay. The INHA and INHB protein distribution within the oocytes was observed by confocal microscopy. INHA mRNA expression was increased in oocytes from large compared to medium and small follicles before IVM (), and to oocytes of small follicles after IVM (). The INHB expression was not different before IVM, but the IHNB mRNA level was gradually higher in oocytes from large follicles after IVM (). INHA was not differently expressed before IVM; however, in large follicle oocytes the protein was distributed in the peripheral area of the cytoplasm; in oocytes from small follicles it was in the entire cytoplasm. After IVM, INHA was strongly expressed in oocytes from small follicles and distributed particularly in the zona pellucida (ZP). Similarly and both before and after IVM, INHB protein was highly expressed in small follicle oocytes and within the cytoplasm. In summary, INHs can be recognized as a marker of porcine oocyte quality. Bartosz Kempisty, Marta Jackowska, Magdalena Woźna, Paweł Antosik, Hanna Piotrowska, Piotr Zawierucha, Dorota Bukowska, Jędrzej M. Jaśkowski, Michał Nowicki, and Klaus P. Brüssow Copyright © 2012 Bartosz Kempisty et al. All rights reserved. Selection of Ovine Oocytes by Brilliant Cresyl Blue Staining Wed, 23 May 2012 10:40:09 +0000 Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB−) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye. Liqin Wang, Jiapeng Lin, Juncheng Huang, Jing Wang, Yuncheng Zhao, and Tong Chen Copyright © 2012 Liqin Wang et al. All rights reserved. A Symphony of Regulations Centered on p63 to Control Development of Ectoderm-Derived Structures Sun, 22 May 2011 16:15:45 +0000 The p53-related transcription factor p63 is critically important for basic cellular functions during development of the ectoderm and derived structure and tissues, including skin, limb, palate, and hair. On the one side, p63 is required to sustain the proliferation of keratinocyte progenitors, while on the other side it is required for cell stratification, commitment to differentiate, cell adhesion, and epithelial-mesenchymal signaling. Molecules that are components or regulators of the p63 pathway(s) are rapidly being identified, and it comes with no surprise that alterations in the p63 pathway lead to congenital conditions in which the skin and other ectoderm-derived structures are affected. In this paper, we summarize the current knowledge of the molecular and cellular regulations centered on p63, derived from the comprehension of p63-linked human diseases and the corresponding animal models, as well as from cellular models and high-throughput molecular approaches. We point out common themes and features, that allow to speculate on the possible role of p63 downstream events and their potential exploitation in future attempts to correct the congenital defect in preclinical studies. Luisa Guerrini, Antonio Costanzo, and Giorgio R. Merlo Copyright © 2011 Luisa Guerrini et al. All rights reserved. Immunohistochemical Localisation of PDE5 in Rat Lung during Pre- and Postnatal Development Thu, 20 Aug 2009 16:13:26 +0000 In mammalian lung, at the transition to extrauterine life, NO/cGMP signal transduction system is known to play crucial roles in the regulation of vascular resistance and is supposed to act in angiogenesis. PDE5, which is the most abundant cGMP metabolizing enzyme within the lung, is highly expressed in the perinatal period, but its localisation in the different pulmonary cells is still poorly known. In our research, PDE5 immunohistochemical distribution was investigated in foetal and neonatal rat lung. The highest expression of PDE5 was found in cells randomly located in the stroma; in newborns, in particular, many cells in the intersaccular walls were heavily labelled, while much lower staining levels were shown by smooth myocytes belonging to vessels and airways. On the basis of their immunoreactivity for 𝛼-SM actin and/or desmin, most of the heavily PDE5-positive cells were identified as interstitial myofibroblasts and transitional pericytes, while only a few were interpreted as interstitial lipofibroblasts. Angela Scipioni, Mauro Giorgi, Valeria Nuccetelli, and Stefania Stefanini Copyright © 2009 Angela Scipioni et al. All rights reserved.