Gene expression profiles of pro- and anti-apoptotic factors in porcine ovarian granulosa cells (single follicular level analyses)

抄録

In mammalian ovaries, more than 99% of follicles undergo atresia, and only a few follicles grow to ovulate during ovarian follicular growth and development. From many recent studies, it is suggested that the apoptosis in ovarian granulosa cells plays a crucial role in follicular atresia, but the molecular mechanism regulateing selective follicular atresia is still largely unknown. In the present study, we examined the precise gene expression levels of pro- and anti-apoptotic factors in porcine ovarian granulosa cells isolated from each single follicle. We obtained porcine ovaries from slaughterhouse, and isolated the granulosa cells from each follicle. The states of each follicle were determined by TUNEL staining on the smeared samples of granulosa cells, and categorized into healthy-, early atretic- and progressed atretic follicles. Then the RNA and cDNA samples were prepared from the single follicle derived granulosa cells, and the gene expression levels of pro- and anti-apoptotic factors were quantified by real time PCR. Here we report the gene expression profiles of death ligands and receptors [Fas ligand (FasL)-Fas, tumor necrosis factor (TNF)_ -TNF receptor (TNFR), and TNF_-related apoptosis-inducing ligand (TRAIL)-TRAIL receptor (TRAILR)] and intracellular death-signal mediators [Fas-associated death domain protein (FADD), TNF receptor 1-associated death domain protein (TRADD), caspases, apoptotic protease-activating factor 1 (Apaf1), TNFR-associated factor 2 (TRAF2), and cellular FLICE-like inhibitory protein (cFLIP) etc.] in granulosa cells of each follicle.