Gene Expression In Corpus Luteum Research Articles

Effect of neonatal exposure to endocrine-active compounds on epigenetic regulation of gene expression in corpus luteum of gilts

Recently, we have demonstrated that neonatal exposure to environmental endocrine-active compounds (EACs) with androgenic/antiandrogenic and estrogenic/antiestrogenic activities led to morphological and functional changes in the porcine corpus luteum (CL). To gain insight into the regulatory mechanisms of the long-term effects of EACs, we analyzed the impact of neonatal exposure of such compounds on global DNA methylation and the expression of miRNA biogenesis components in the porcine CL. Piglets were injected subcutaneously with testosterone propionate (TP, an androgen), flutamide (FLU, an antiandrogen), 4-tert-octylphenol (OP, an estrogenic compound), ICI 182,780 (ICI, an antiestrogen), methoxychlor (MXC, a compound with mixed activities) or corn oil (control) between postnatal days 1 and 10 (n = 5/group). The CLs from sexually mature gilts were examined for global DNA methylation and for the abundance of proteins related to DNA methylation (DNMT1, DNMT3A and DNMT3B) and miRNA biogenesis (DROSHA, XPO5, DICER1, AGO2) using an enzyme-linked immunosorbent assay, Western blotting and immunohistochemical staining. ICI and MXC increased the global DNA methylation levels and DNMT1 protein abundance in the luteal tissue. OP treatment led to a lower DROSHA protein abundance, while ICI treatment resulted in a greater DROSHA protein abundance. Both FLU and ICI increased DICER1 protein abundance in the luteal tissue. In addition, XPO5 showed immunolocalization exclusively in small luteal cells in the OP-treated pigs, in contrast to localization in both small and large luteal cells in the controls. In conclusion, the changes in DNA methylation, as well as the altered miRNA biogenesis components, seem to be a part of the regulatory network that mediates the long-term effects of EACs on CL function in pigs.

Tissue-specific promoter methylation and histone modification regulate CYP19 gene expression during folliculogenesis and luteinization in buffalo ovary

Aromatase, the key enzyme of estrogen biosynthesis, is encoded by the CYP19 gene. The expression of CYP19 gene is regulated in species- and tissue-specific manner by alternate use of different promoters. We have previously, cloned and characterized the tissue-specific promoter and tissue-specific transcripts in preovulatory (granulosa cells) and postovulatory (corpus luteum) structure of buffalo ovary. The present study was aimed to understand if epigenetic gene regulation through DNA methylation and histone modifications is involved in tissue-specific CYP19 gene regulation during folliculogenesis and luteinization in buffalo ovary. Methylation analysis of five CpG dinucleotides of ovary specific proximal promoter II showed hypo-methylation in large follicle while hyper-methylation in corpus luteum. However, PI.1, the exclusive promoter responsible for residual CYP19 gene expression in corpus luteum, was found to be hypermethylated. Analysis of histone modifications using ChIP assay revealed that the distal promoter (PI.1) of CYP19 gene is ∼40-fold more enriched with acetylated Histone H3 in corpus luteum than in the large follicle. This indicates that PI.1 chromatin was more accessible for transcription in corpus luteum as compared to large follicles. The chromatin accessibility for the proximal promoter (PII) in the preovulatory stage tends to be higher than the luteal tissue. However, the difference was not found to be significant. In vitro experiments showed the similar results. In conclusion, results of the present study suggests that tissue-specific methylation status of PII and chromatin remodeling through histone modifications of PI.1, coincides with the changes in expression of CYP19 gene and thus are the regulatory mechanism controlling its tissue-specific expression and promoter activity during folliculogenesis and luteinization.

Progesterone receptor membrane component 1 (PGRMC1) gene expression in corpus luteum during the estrous cycle in cows

The aim of the study was to investigate progesterone receptor membrane component 1 (PGRMC1) mRNA expression in bovine corpus luteum (CL) obtained from heifers or non-pregnant cows on the following days of the estrous cycle: 1-5, 6-10, 11-16 and 17-21 (n=4/each time period). The expression of PGRMC1 mRNA, analyzed by semiquantitative RT-PCR, was the highest on days 6-10 (p<0.01) and then it declined (p<0.05). The lowest expression was found on days 1-5 (p<0.05). A significant correlation (p<0.05) was also observed between luteal progesterone (P(4)) concentration and PGRMC1 mRNA expression. These data indicate that PGRMC1 mRNA is expressed in bovine CL and this expression varies throughout the luteal phase. It is assumed that PGRMC1 may be involved in a non-genomic effect of P(4) on luteal cells.