Expression of growth differentiation factor 9 in the oocytes is essential for the development of primordial follicles in the hamster ovary.
Expression of growth differentiation factor 9 in the oocytes is essential for the development of primordial follicles in the hamster ovary.
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DOI：
10.1210/en.2005-1208
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发表时间：
2006-04
期刊：
影响因子：
4.8
通讯作者：
Cheng Wang;S. Roy
中科院分区：
文献类型：
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作者：
Cheng Wang;S. Roy
出生后生长分化因子9(GDF-9)在金黄地鼠卵母细胞中的表达先于原始卵泡的形成。我们通过RNA干扰抑制GDF-9的mRNA和蛋白表达，研究了GDF-9在金黄地鼠卵巢原始卵泡发生中的功能意义。取15天龄的胎儿卵巢，分别加或不加1 ng FSH、1微l Metafectane、100 nM对照非靶向小干扰RNA(SiRNA)、GDF-9 siRNA或GDF-9 siRNA+FSH培养9d，观察原始卵泡发育情况。在器官培养中，用Siglo(一种标记有Cy3的非靶向对照siRNA)培养卵巢细胞，以测试siRNA转染卵巢细胞的效率。卵巢中90%以上的细胞呈Siglo阳性，Metafectane和siRNA均未诱导细胞凋亡。对照siRNA对GDF-9mRNA的基础水平没有影响，但GDF-9siRNA略有降低，但显著降低。FSH显著上调GDF-9的mRNA和蛋白水平，GDF-9 siRNA可完全抑制FSH上调GDF-9的作用。然而，GDF-9 siRNA不影响骨形态发生蛋白受体IA和β-肌动蛋白mRNA的表达。单独使用GDF-9siRNA也会降低GDF-9蛋白的表达。在表达GDF-9的同时，FSH显著促进原始卵泡的形成，但这种作用可被GDF-9 siRNA消除。这些结果表明，内源性GDF-9在体细胞分化和原始卵泡的形成中起重要作用。此外，FSH通过调节GDF-9的表达，调节卵母细胞对原始卵泡形成的调节。
Postnatal growth differentiation factor 9 (GDF-9) expression in the hamster oocytes precedes the formation of primordial follicles. We examined the functional significance of GDF-9 in primordial folliculogenesis in the hamster ovary using RNA interference knockdown of GDF-9 mRNA and protein expression. Fifteen-day-old fetal ovaries were cultured for 9 d with or without 1 ng FSH, 1 microl Metafectane, 100 nM control nontargeting small interfering RNA (siRNA), GDF-9 siRNA, or GDF-9 siRNA + FSH, and the development of primordial follicles examined. The efficiency of siRNA transfecting ovarian cells in the organ culture was tested by culturing ovaries with siGlo, a nontargeting control siRNA labeled with Cy3. More than 90% of cells in the ovary were siGlo positive, and neither the Metafectane nor the siRNA-induced cellular apoptosis. Control siRNA did not affect the basal levels of GDF-9 mRNA, but GDF-9 siRNA slightly but significantly reduced the level. FSH markedly up-regulated the levels of GDF-9 mRNA and protein, and the effect was completely suppressed by GDF-9 siRNA. However, GDF-9 siRNA did not affect the levels of bone morphogenetic protein receptor IA or beta-actin mRNA. GDF-9 siRNA alone also reduced GDF-9 protein expression. Concurrent with GDF-9 expression, FSH significantly augmented primordial follicle formation, but the effect was abolished by GDF-9 siRNA. These results suggest that endogenous GDF-9 plays an important role in somatic cell differentiation and the formation of primordial follicles. Furthermore, FSH, by virtue of regulating GDF-9 expression, modulates oocyte regulation of primordial follicles formation.