OBJECTIVE:Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored.METHODS:In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2'-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative polymerase chain reaction and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression.RESULTS:We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1).CONCLUSION:MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1, and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.

The Duroc pig has high adaptability and feeding efficiency, making it one of the most popular pig breeds worldwide. Over long periods of natural and artificial selection, genetic footprints, i.e., selective signatures, were left in the genome. In this study, a Duroc pig population (n=715) was genotyped with the Porcine SNP60K Bead Chip and the GeneSeek Genomic Profiler (GGP) Porcine Chip. The relative extended haplotype homozygosity (REHH) method was used for selective signature detection in a subset of the population (n=368), selected to represent a balanced family structure. In total, 154 significant core regions were detected as selective signatures (P<0.01), some of which overlap with previously reported quantitative trait loci associated with several economically important traits, including average daily gain and backfat thickness. Genome annotation for these significant core regions revealed a variety of interesting candidate genes including GATA3, TAF3, ATP5C1, and FGF1. These genes were functionally related to anterior/posterior pattern specification, phosphatidylinositol 3-kinase signaling, embryonic skeletal system morphogenesis, and oxidation-reduction processes. This research provides knowledge for the study of selection mechanisms and breeding practices in Duroc and other pigs.

In mammalian ovaries, many studies demonstrated that the proliferation and apoptosis of granulosa cells are involved in folliculogenesis. Previous evidence suggests that miR-126-3p might get involved in the proliferation and apoptosis of granulosa cells, and tuberous sclerosis complex 1 (TSC1) gene was predicted as one target of miR-126-3p, and moreover, granulosa cell-specific TSC1 knockout stimulated folliculogenesis in mice. However, the molecular regulation of miR-126-3p on TSC1 and its effects on cell proliferation and apoptosis remain virtually unexplored in granulosa cells. Using porcine granulosa cells as a model, the luciferase report assay, mutation, deletion, Annexin-V/PI staining, and EdU assays were applied to investigate the molecular mechanism for miR-126-3p regulating the expression of TSC1 and their effects on the cell proliferation and apoptosis. We found that miR-126-3p showed a positive effect on cell proliferation and a negative effect on cell apoptosis in porcine granulosa cells, and knockdown of TSC1 significantly promoted cell proliferation and significantly inhibited cell apoptosis in porcine granulosa cells. Furthermore, miR-126-3p might target and repress the expressions of TSC1 at the post-transcriptional level, thereby promoting cell proliferation and inhibiting cell apoptosis of granulosa cells. These findings would provide of great insight in further exploring the molecular regulation of miR-126-3p and TSC1 on the functions of granulosa cells during the folliculogenesis in mammals.

Reduced representation bisulfite sequencing (RRBS) has been widely used to profile genome-scale DNA methylation in mammalian genomes. However, the applications and technical performances of RRBS with different fragment sizes have not been systematically reported in pigs, which serve as one of the important biomedical models for humans. The aims of this study were to evaluate capacities of RRBS libraries with different fragment sizes to characterize the porcine genome.

对猪全基因组高密度SNP基因型数据及生长性状表型数据进行全基因组关联分析,以期找到影响这些性状的候选基因,更准确地了解这些生长性状的遗传基础.利用Illumina猪60K SNP芯片对191头杜洛克猪进行基因型检测,使用R语言环境下GenABEL软件包提供的单标记回归分析模型,对体重达100 kg日龄(D100)、活体背膘厚(BFT)和活体眼肌面积(LMA)3个生长性状的表型分别进行全基因组关联分析.在D100和LMA2个性状中分别检测到1个基因组水平和6个染色体水平显著关联的SNP,均位于5号染色体;没有检测到与BFT显著相关的SNP.生物信息学分析表明,BTG1和EFCA B6可能是影响生长性状的重要候选基因,但其功能有待进一步研究确认.