Furuya K, Mizumoto Y, Makimura N, Mitsui C, Murakami M, Tokuoka S, Ishikawa N, Nagata I, Kimura T, Ivell R
Department of Obstetrics and Gynecology, National Defense Medical College, Saitama, Japan.
Adv Exp Med Biol. 1995;395:523-8.
Recently, several authors have reported the existence of oxytocin (OT) in mammalian granulosa-luteal cells after ovulation. The purpose of this study was to examine the evidence for gene expression and localization of OT and OT receptor (OTR) in the cumulus cells surrounding the oocytes. Cumulus cells with mature oocytes were obtained from experimental and clinical in vitro fertilization-embryo transfer (IVF-ET) programs. OT and OTR gene expression was analyzed with reverse transcriptase-polymerase chain reaction (RT-PCR) and RT-PCR/single strand conformation polymorphism (SSCP). OT gene expression was detected in mouse and human cumulus cells. The results of RT-PCR/SSCP showed that the structure of OT mRNA in cumulus cells was similar to that in the hypothalamus. Furthermore, OTR gene expression was clearly demonstrated in human cumulus cells, and a weak positive signal was observed in human oocytes. Immunocytochemical staining of OTR was clearly detected in human cumulus cells. The rate of mouse blastocyst development was significantly higher in the group cultured with OT than that without OT. These results are the first observations of simultaneous OT and OTR gene expression in cumulus cells, suggesting that ovarian OT might have some physiological role in the early stage of embryo development.
最近,几位作者报道了排卵后哺乳动物颗粒黄体细胞中存在催产素(OT)。本研究的目的是检测卵母细胞周围卵丘细胞中OT和OT受体(OTR)的基因表达及定位证据。从实验性和临床体外受精-胚胎移植(IVF-ET)程序中获取带有成熟卵母细胞的卵丘细胞。采用逆转录-聚合酶链反应(RT-PCR)和RT-PCR/单链构象多态性(SSCP)分析OT和OTR基因表达。在小鼠和人类卵丘细胞中检测到OT基因表达。RT-PCR/SSCP结果显示,卵丘细胞中OT mRNA的结构与下丘脑相似。此外,在人类卵丘细胞中明确证实了OTR基因表达,在人类卵母细胞中观察到弱阳性信号。在人类卵丘细胞中清晰检测到OTR的免疫细胞化学染色。与未添加OT的组相比,添加OT培养的组中小鼠囊胚发育率显著更高。这些结果首次观察到卵丘细胞中同时存在OT和OTR基因表达,表明卵巢OT可能在胚胎发育早期具有某些生理作用。