Seshagiri P B, Terasawa E, Hearn J P
Wisconsin Regional Primate Research Center, University of Wisconsin, Madison 53715-1299.
Hum Reprod. 1994 Jul;9(7):1300-7. doi: 10.1093/oxfordjournals.humrep.a138699.
Chorionic gonadotrophin (CG) is the first clear embryonic signal during early pregnancy in primates. CG has close structural and functional similarities to pituitary luteinizing hormone (LH) which is regulated by gonadotrophin releasing hormone (GnRH). To study the regulatory mechanism of CG secretion in primate embryos, we examined the production and timing of secretion of GnRH in peri-implantation embryos of the rhesus monkey. In-vivo fertilized/developed morulae and early blastocysts, recovered from non-superovulated, naturally-bred rhesus monkeys by non-surgical uterine flushing, were cultured in vitro to hatched, attached and post-attached blastocyst stages using a well-established culture system. We measured GnRH and CG in media samples from cultured embryos with a sensitive radioimmunoassay and bioassay, respectively. The secretion of GnRH (pg/ml; mean +/- SEM) by embryos (n = 20) commenced from low levels (0.32 +/- 0.05) during the pre-hatching blastocyst stage to 0.70 +/- 0.08 at 6-12 days and 1.30 +/- 0.23 at > or = 13 days of hatched blastocyst attachment and proliferation of trophoblast cells. GnRH concentrations in culture media obtained from embryos (n = 5) that failed to hatch and attach were mostly undetectable (< or = 0.1). Samples that did not contain detectable GnRH failed to show detectable CG. Immunocytochemical studies, using a specific monoclonal anti-GnRH antibody (HU4H) as well as polyclonal antisera (LR-1), revealed that immunopositive GnRH cells were localized in pre-hatching blastocysts (n = 4), in blastocysts (n = 2) after 5-10 days of attachment and in monolayer cultures (n = 4) of well-established embryonic trophoblast cells.(ABSTRACT TRUNCATED AT 250 WORDS)
绒毛膜促性腺激素(CG)是灵长类动物早期妊娠期间第一个明确的胚胎信号。CG与垂体促黄体生成素(LH)在结构和功能上有密切相似性,而LH受促性腺激素释放激素(GnRH)调节。为了研究灵长类胚胎中CG分泌的调节机制,我们检测了恒河猴着床前胚胎中GnRH的产生及分泌时间。通过非手术子宫冲洗从未超排、自然受孕的恒河猴中获取体内受精发育的桑葚胚和早期囊胚,利用成熟的培养系统将其体外培养至孵化、着床及着床后囊胚阶段。我们分别用灵敏的放射免疫分析法和生物分析法检测培养胚胎培养基样本中的GnRH和CG。胚胎(n = 20)分泌的GnRH(pg/ml;均值±标准误)在孵化前囊胚阶段从低水平(0.32±0.05)开始,在6 - 12天升至0.70±0.08,在孵化后囊胚着床及滋养层细胞增殖≥13天时升至1.30±0.23。未孵化和未着床胚胎(n = 5)培养基中的GnRH浓度大多检测不到(≤0.1)。不含可检测到的GnRH的样本未显示可检测到的CG。使用特异性单克隆抗GnRH抗体(HU4H)以及多克隆抗血清(LR - 1)进行的免疫细胞化学研究表明,免疫阳性的GnRH细胞定位于孵化前囊胚(n = 4)、着床5 - 10天后的囊胚(n = 2)以及成熟胚胎滋养层细胞的单层培养物(n = 4)中。(摘要截断于250字)