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雌激素受体基因在小鼠卵母细胞及胚胎发育过程中的表达。

Expression of estrogen receptor gene in mouse oocyte and during embryogenesis.

作者信息

Wu T C, Wang L, Wan Y J

机构信息

Department of Obstetrics and Gynecology, UCLA School of Medicine 90024.

出版信息

Mol Reprod Dev. 1992 Dec;33(4):407-12. doi: 10.1002/mrd.1080330406.

Abstract

Estrogen is required for oocyte maturation and embryonic development in vivo; however, the mechanism involved is not clear. Since the effect of estrogen is mediated through the estrogen receptor (ER), we examined the ontogeny and expression of the ER gene in mouse oocytes and embryos of various gestational stages using the highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Total RNA, extracted from 40 ovulated oocytes, 2-cell embryos, morulae, and blastocysts, was reverse transcribed into cDNA. A pair of primers flanking the 453-bp region encoding the hormone-binding domain of ER was used for 30 cycles of PCR. The identity of the amplified product was confirmed by sizing and Southern blot hybridization. The results indicated that ER gene is expressed in unfertilized oocytes and cumulus-oocyte complexes. The amount of ER mRNA decreases in 2-cell embryos, coincident with degradation of maternal mRNA. No ER transcript can be detected in the morulae or blastocyst stage when the embryonic genome has been activated. Postimplantation embryos do not contain detectable ER mRNA until gestation day 8. The levels of ER mRNA increase from day 10 to day 18 of gestation. These data suggest that estrogen, secreted by granulosa cells, may directly influence oocyte growth and maturation in vivo. Since estrogen is known to stimulate the production of growth factors in mouse uteri, the absence of ER mRNA in periimplantation embryos suggests that the effects of estrogen on early embryogenesis may be indirect, i.e., through estrogen-regulated growth-promoting factors produced by the reproductive tract. In mid- and late-post-implantation embryos which contain ER mRNA, estrogen may affect embryonic development through the receptor-mediated mechanisms.

摘要

雌激素是体内卵母细胞成熟和胚胎发育所必需的;然而,其涉及的机制尚不清楚。由于雌激素的作用是通过雌激素受体(ER)介导的,我们使用高灵敏度逆转录聚合酶链反应(RT-PCR)技术检测了小鼠不同妊娠阶段卵母细胞和胚胎中ER基因的个体发生及表达。从40个排卵后的卵母细胞、2细胞胚胎、桑椹胚和囊胚中提取总RNA,并逆转录成cDNA。一对位于编码ER激素结合域的453 bp区域两侧的引物用于30个循环的PCR。通过大小测定和Southern印迹杂交确认扩增产物的同一性。结果表明,ER基因在未受精的卵母细胞和卵丘-卵母细胞复合体中表达。ER mRNA的量在2细胞胚胎中减少,与母体mRNA的降解同时发生。当胚胎基因组被激活时,在桑椹胚或囊胚阶段检测不到ER转录本。植入后胚胎直到妊娠第8天才含有可检测到的ER mRNA。ER mRNA的水平在妊娠第10天到第18天增加。这些数据表明,颗粒细胞分泌的雌激素可能在体内直接影响卵母细胞的生长和成熟。由于已知雌激素可刺激小鼠子宫中生长因子的产生,植入前胚胎中缺乏ER mRNA表明雌激素对早期胚胎发生的影响可能是间接的,即通过生殖道产生的雌激素调节的生长促进因子。在含有ER mRNA的植入后中晚期胚胎中,雌激素可能通过受体介导的机制影响胚胎发育。

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