ABSTRACT
Background: Polycystic ovarian syndrome [PCOS] is a metabolic and endocrine disorder which is characterized by hyperandrogenism, anovulation or oligomenor-rhea and polycystic ovarian morphology. It is believed that modulation in metabo-lism of granulosa cells of PCOS patients may lead to infertility. One of the metabolic modulators is FNDC5 and its cleaved form, irisin. The axis of PGC1 Alpha - FNDC5 pathway is one of the main factors affecting cellular energy balance the purpose of this study was to evaluate this pathway in granulosa cells derived from PCOS mice model in comparison with control group
Methods: In the present study, PCOS mouse model was developed by injection of dehydroepiandrosterone [DHEA] hormone in 20 mice for a period of 20 days. Also, 20 uninjected mice were used as the control. Meanwhile, a vehicle group consisted of mice which received daily subcutaneous sesame oil injection [n=20]. Relative ex-pressions of PGC1á and FNDC5 in granulosa cells were evaluated by RT-qPCR. Analysis of gene expressions was calculated by the Delta Delta CT method and the relative levels of mRNA were normalized to GAPDH transcript levels. Differences in genes expression among three groups were assessed using one-way ANOVA, Tukey's Post Hoc test
Results: Our results showed that expression of FNDC5 was significantly reduced in granulosa cells of DHEA-induced PCOS mice compared with control and vehicle groups [p<0.05], while there was no significant differences in PGC1 Alpha expression among different groups
Conclusion: Down regulation of FNDC5 transcript level may contribute in metabol-ic disturbance of granulosa cells derived from PCOS ovary apart from PGC1 Alpha levels which remained unchanged
ABSTRACT
In vitro simulation of developmental processes is an invaluable tool to shed light on the intrinsic mechanism of developmental biosystems such as central nervous system in mammals. Chick somites have been used to simulate the neural differentiation of human neural progenitor cells. In the present study, we aimed to indicate whether somites have the ability to express required neural differentiation factors at mRNA level. Chick embryos were isolated from the yolk surface of the fertilized eggs and somites were subsequently isolated from embryos under a dissecting microscope. Total RNA of the somites was extracted and RT-PCR carried out with specific primers of cerberus, chordin, FGF8, follistatin and noggin. Data showed that five aforementioned factors were co-expressed after 7 days in vitro by somites. We concluded that neural induction property of somites appeared by production of required neural differentiation factors including cerberus, chordin, FGF8, follistatin and noggin
ABSTRACT
Peroxisome Proliferator Activated Receptor gamma [PPAR[gamma]], a member of nuclear receptor superfamily, comprises two isoforms in mouse. These two isoforms are encoded by different mRNAs, which are arisen by alternative promoter usage. There are two promoter regions upstream of PPAR[gamma] gene. A 3 kb fragment, containing several transcription factor binding sites, acts as PPAR[gamma]1 promoter region. Thus, expression pattern of PPAR[gamma]1 isoform is due to the potential transcription factors that could influence its promoter activity. PPAR[gamma], Retinoid X Receptor [RXR] and Vitamin D Receptor [VDR], as nuclear receptors could influence PPAR[gamma] gene expression pattern during several differentiation processes. During neural differentiation, PPAR[gamma]1 isoform expression reaches to maximal level at neural precursor cell formation. A vast computational analysis was carried out to reveal the PPAR[gamma]1 promoter region. The putative promoter region was then subcloned upstream of an EGFP reporter gene. Then the functionality of PPAR[gamma]1 promoter was assessed in different cell lines. Results indicated that Rosiglitazone increased PPAR[gamma]1 promoter regulated EGFP expression of neural precursor cells during Embryoid Body [EB] formation. Furthermore vitamin D reduced PPAR[gamma]1 promoter regulated EGFP expression of neural precursor cells during EB formation through binding to its receptor. This study suggests that there are potential response elements for PPAR/RXR and VDR/RXR heterodimers in PPAR[gamma]1 isoform promoter. Also VDR/RXR heterodimers may decrease PPAR[gamma] expression through binding to its promoter
ABSTRACT
Peroxisomes are ubiquitous organelles in nearly all eukaryotes that participate in the metabolism of fatty acids and other metabolites. In order to investigate peroxisome biogenesis and gene profile expression analysis during neurogenesis, we have performed to set a semi-quantitative analysis for two peroxisomal genes expression [Catalase and PEX3] during neurogenesis comparing with stem cell marker genes [Oct4 and Nanog] in P19 cells. In this project we have used P19 cells which are suitable progenitor cells for the study of neurogenesis. Moreover, two different peroxisomal genes were selected to chase both kinetics of peroxisomal matrix protein synthesis [Catalase] and peroxisomal membrane protein biogenesis [Pex3p]. Total RNA extracted from P19 cells, and was used for cDNA synthesis at the next step. Specific primers of Oct4, Nanog, PEX3 and Catalase cDNAs were amplified by RT-PCR for quantitative analysis. Various parameters were changed to optimize the PCR profiles for each gene for further assessments. Here we are going to describe the conditions which we used, e.g.: PEX3 band was detected sharply after 35 cycles and at annealing temperature of 52.5-55.1°C. For PCR of Catalase we observed a sharp band after 35 cycles and annealing temperature of 52.7-55°C. Nanog cDNA was amplified after 36 cycles and between 60.1-64.1°C. Oct4 band was observed sharply after 28 cycles of amplification and annealing temperature in range of 53.4-57°C