Recent reports have demonstrated that follicular atresia is initiated or caused by granulosa cell apoptosis followed by theca cell degeneration in mammalian ovaries, but the mechanism of follicular atresia is still to be elucidated. Therefore, our present study was designed to examine our hypothesis that the changes of follicular microenvironment induce the granulosa cell apoptosis during pocrine follicular atresia in vivo. We firstly isolated intact porcine antral follicles and identified them into three groups, healthy follicles (HF), early atretic follicles (EAF) and progressed atretic follicles (PAF) through morphology and histology. To further confirm their status, we detected hormone levels in follicular fluids and the expression level of apoptosis gene Bax in granulosa cells. The rate of progesterone (P) and estradiol (E2) was increased with the expression of Bax, indicating hormone can be used as a marker of granulosa cell apoptosis or follicular atresia. Finally, we analyzed the expression level of hormone receptor genes in granulosa cells and their relationship with follicular atresia. In PAF, the expression of Progesterone receptor (PGR) was increased significantly while estradiol receptor (ER) had no notable changes, which suggesting the increased-PGR accelerated the effect of P-stimulated granulosa cell apoptosis. The dramatic increasing of androgen receptor (AR) expression in PAF and the obvious increase of tumor necrosis factor-α receptor (TNFR) in EAF indicated that there are different pathways regulating granulosa cell apoptosis during follicular atresia. Together, our results suggested that different pathways of granulosa cell apoptosis was induced by changing the follicular microenvironment during follicular atresia.

MicroRNAs (miRNAs), which post-transcriptionally regulate gene expression by binding to the 3´ untranslated region of mRNAs to either inhibit or enhance translation, are involved in diverse biological processes. The use of high-throughput Solexa sequencing plays important roles in the discovery of miRNAs. In this study, we used high-throughput Solexa sequencing to identify novel duck miRNAs and compare the miRNA expression profiles in laying and non-laying duck ovaries. Using a bioinformatic analysis, we discovered 86 potential duck miRNAs similar to known chicken miRNAs and 43 unique sequences that matched known miRNAs of other species. We also found that miRNA variations and isoforms were widespread in our two RNA libraries, with most of the variations occurring in the 3´ region of the miRNAs. Furthermore, we detected 55 miRNAs that exhibited significant expression differences between laying and non-laying ducks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses of the potential targets of the differentially expressed miRNAs indicated these miRNAs may play key roles in the egg laying process. Finally, we confirmed the differential expression of 5 miRNAs in the laying and non-laying samples by qRT-PCR. Cumulatively, our work provides the first look at the miRNA expression profile of the duck ovary and provides novel insight into the roles of miRNAs in egg laying and reproduction.