Duffy Diane M
Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, Virginia 23507, USA.
Biol Reprod. 2003 Aug;69(2):725-32. doi: 10.1095/biolreprod.103.015891. Epub 2003 Apr 16.
Expression of growth differentiation factor 9 (GDF-9), an apparent regulator of follicular development, reportedly differs between compartments of the rodent (oocytes) and human (oocytes and granulosa cells) ovary. To further characterize GDF-9 expression and action in the primate periovulatory follicle, adult female rhesus monkeys received recombinant human gonadotropins to promote multiple follicular development. Whole ovaries or follicular aspirates were obtained before and at various times after administration of an ovulatory dose of hCG; time points for tissue collection spanned the 40-h periovulatory interval. GDF-9 mRNA was detected by reverse transcription polymerase chain reaction assay in each oocyte and every granulosa cell sample examined, but granulosa cell GDF-9 mRNA levels did not change across the periovulatory interval. GDF-9 was also detected in follicular fluid using Western blotting; GDF-9 protein concentration in follicular fluid did not change across the periovulatory interval. Immunocytochemical staining for GDF-9 indicated that oocytes of both small and large antral follicles were positive for GDF-9. GDF-9 immunoreactivity was also present in cumulus granulosa cells and mural granulosa cells near the cumulus stalk. When granulosa cells from preovulatory follicles were exposed to recombinant GDF-9 in culture, GDF-9 increased vascular endothelial growth factor levels in culture medium. These data demonstrate that the cells of the primate periovulatory follicle both produce and respond to GDF-9. However, GDF-9 expression and action differ between rodent and primate follicles, suggesting a possible regulatory role for GDF-9 that is unique to the primate follicle.
生长分化因子9（GDF - 9）是卵泡发育的一个明显调节因子，据报道，其在啮齿动物（卵母细胞）和人类（卵母细胞和颗粒细胞）卵巢的不同区域表达存在差异。为了进一步阐明GDF - 9在灵长类动物排卵前卵泡中的表达及作用，成年雌性恒河猴接受重组人促性腺激素以促进多个卵泡发育。在给予排卵剂量的hCG之前及之后的不同时间点获取整个卵巢或卵泡抽吸物；组织采集的时间点涵盖了40小时的排卵前间隔期。通过逆转录聚合酶链反应检测每个检测的卵母细胞和颗粒细胞样本中的GDF - 9 mRNA，但颗粒细胞GDF - 9 mRNA水平在排卵前间隔期内没有变化。使用蛋白质印迹法也在卵泡液中检测到了GDF - 9；卵泡液中GDF - 9蛋白浓度在排卵前间隔期内没有变化。GDF - 9的免疫细胞化学染色表明，大小不同的窦状卵泡的卵母细胞GDF - 9均呈阳性。GDF - 9免疫反应性也存在于卵丘颗粒细胞和卵丘柄附近的壁层颗粒细胞中。当将排卵前卵泡的颗粒细胞在培养中暴露于重组GDF - 9时，GDF - 9增加了培养基中血管内皮生长因子的水平。这些数据表明，灵长类动物排卵前卵泡的细胞既能产生GDF - 9，又能对其作出反应。然而，GDF - 9在啮齿动物和灵长类动物卵泡中的表达及作用存在差异，这表明GDF - 9可能具有灵长类动物卵泡特有的调节作用。
