DiZerega G S, Richardson C L, Davies T F, Hodgen G D, Catt K J
Fertil Steril. 1980 Oct;34(4):379-85. doi: 10.1016/s0015-0282(16)45008-9.
A fluorescent derivative of human chorionic gonadotropin (hCG) for binding studies was prepared by conjugation of hightly purified hCG with fluorescein isothiocynate (FITC-hCG) in a molar ratio of 3:1. The resulting FITC-hCG conjugate retained full cross-reactivity with antibodies directed against intact hCG, the hCG beta-subunit, the carboxyl-terminal peptide sequence of beta-hCG. The fluorescent hormone retained binding activity similar to that of native hCG, and the capacity to induce progesterone production from primate corpora luteal cells in vitro. After intravenous administration to female rhesus monkeys, FITC-hCG was localized within the theca cell layer of certain antral follicles, including the dominant follicle, but no fluorescence was noted around preantral follicles. Areas of fluorescence were also localized to discrete groups of interstitial gland cells in the stroma. The degree of fluorescence associated with these cells was indistinguishable from that observed in the thecal cells. The development of a fluorescent derivative that is biologically and immunologically similar to native hCG has permitted differential microscopic localization of luteinizing hormone/hCG uptake in the preovulatory ovary.
一种用于结合研究的人绒毛膜促性腺激素(hCG)荧光衍生物,是通过将高度纯化的hCG与异硫氰酸荧光素以3:1的摩尔比偶联制备而成(FITC-hCG)。所得的FITC-hCG偶联物与针对完整hCG、hCGβ亚基、β-hCG羧基末端肽序列的抗体保持完全交叉反应性。这种荧光激素保留了与天然hCG相似的结合活性,以及在体外诱导灵长类动物黄体细胞产生孕酮的能力。给雌性恒河猴静脉注射后,FITC-hCG定位于某些窦状卵泡的卵泡膜细胞层内,包括优势卵泡,但在窦前卵泡周围未观察到荧光。荧光区域也定位于基质中离散的间质腺细胞群。与这些细胞相关的荧光程度与在卵泡膜细胞中观察到的无法区分。一种在生物学和免疫学上与天然hCG相似的荧光衍生物的开发,使得在排卵前卵巢中对促黄体生成素/hCG摄取进行差异显微镜定位成为可能。