Bocca Silvina M, Billiar Reinhart B, Albrecht Eugene D, Pepe Gerald J
Department of Obstetrics,, The Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA 23501, USA.
Endocrine. 2008 Jun;33(3):254-60. doi: 10.1007/s12020-008-9081-y.
In fetal ovaries of estrogen-suppressed baboons, we have previously shown that follicle numbers were 50% lower than in estrogen-replete animals and contained oocytes with a reduced number of microvilli. In the baboon fetal ovary, although estrogen receptor (ER)alpha and beta have been detected by immunocytochemistry in granulosa cells, it is not known whether oocytes express ER. Because the actions of estrogen are mediated by interaction with cell-specific receptors, the current study determined whether ERalpha/beta mRNA were expressed in oocytes of baboon fetal ovaries obtained on day 165 (term = day 184) of gestation. Oocyte nuclei and cytoplasm from primordial follicles were isolated by laser capture microdissection and ERalpha, ERbeta, GATA-4 (granulosa cell specific marker) mRNAs, and 18S rRNA determined by RT-PCR and products verified by sequencing. ERbeta mRNA was expressed in oocytes of 5 of 5 fetuses. In contrast, fetal oocytes did not express ERalpha mRNA. Although 18S rRNA was expressed in all oocytes, GATA-4 mRNA was not detected in oocytes and only detected in granulosa cells confirming purity of oocytes sampled. We conclude that oocytes of the fetal baboon ovary express ERbeta mRNA, thereby providing a mechanism by which estrogen regulates oocyte function, e.g. microvillus development.
在雌激素抑制的狒狒胎儿卵巢中,我们之前已经表明,卵泡数量比雌激素充足的动物低50%,并且所含卵母细胞的微绒毛数量减少。在狒狒胎儿卵巢中,尽管通过免疫细胞化学在颗粒细胞中检测到了雌激素受体(ER)α和β,但尚不清楚卵母细胞是否表达ER。由于雌激素的作用是通过与细胞特异性受体相互作用介导的,因此本研究确定了在妊娠第165天(足月为第184天)获得的狒狒胎儿卵巢卵母细胞中是否表达ERα/β mRNA。通过激光捕获显微切割分离原始卵泡的卵母细胞核和细胞质,并通过逆转录聚合酶链反应(RT-PCR)测定ERα、ERβ、GATA-4(颗粒细胞特异性标志物)mRNA和18S rRNA,并通过测序验证产物。5只胎儿的卵母细胞中有5只表达ERβ mRNA。相比之下,胎儿卵母细胞不表达ERα mRNA。尽管所有卵母细胞中都表达18S rRNA,但在卵母细胞中未检测到GATA-4 mRNA,仅在颗粒细胞中检测到,这证实了所采集卵母细胞的纯度。我们得出结论,狒狒胎儿卵巢的卵母细胞表达ERβ mRNA,从而提供了一种雌激素调节卵母细胞功能(如微绒毛发育)的机制。