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编码具有生物活性的啮齿动物促性腺激素的载体的构建与表达。

Construction and expression of vectors encoding biologically active rodent gonadotropins.

作者信息

Ohta Akihiko, Tsunoda Yuichiro, Tamura Yoshihiko, Iino Kayoko, Nishimura Naoto, Nishihara Hiroto, Takanashi Haruka, Yoshida Saishu, Kato Takako, Kato Yukio

机构信息

Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan.

Institute for Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan.

出版信息

J Reprod Dev. 2017 Dec 15;63(6):605-609. doi: 10.1262/jrd.2017-091. Epub 2017 Oct 13.

Abstract

The gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are important hormones in vertebrate reproduction. The isolation of gonadotropins from the pituitary gland is sub-optimal, as the cross-contamination of one hormone with another is common and often results in the variation in the measured activity of LH and FSH. The production of recombinant hormones is, therefore, a viable approach to solve this problem. This study aimed to express recombinant rat, mouse, and mastomys FSH and LH in Chinese hamster ovary (CHO) cells. Their common α-subunits along with their hormone-specific β-subunits were encoded in a single mammalian expression vector. FSH from all three species was expressed, whereas expression was achieved only for the mouse LH. Immunohistochemistry for rat alpha subunit of glycoprotein hormone (αGSU) and LHβ and FSHβ subunits confirmed the production of the dimeric hormone in CHO cells. The recombinant rodent gonadotropins were confirmed to be biologically active; estradiol production was increased by recombinant FSH in granulosa cells, while recombinant LH increased testosterone production in Leydig cells.

摘要

促性腺激素,即黄体生成素(LH)和卵泡刺激素(FSH),是脊椎动物繁殖中的重要激素。从垂体中分离促性腺激素并不理想,因为一种激素与另一种激素的交叉污染很常见,并且常常导致所测得的LH和FSH活性出现差异。因此,生产重组激素是解决这一问题的可行方法。本研究旨在在中国仓鼠卵巢(CHO)细胞中表达重组大鼠、小鼠和非洲沼鼠的FSH和LH。它们的共同α亚基及其激素特异性β亚基在单个哺乳动物表达载体中编码。所有三个物种的FSH均得以表达,而仅小鼠LH实现了表达。针对糖蛋白激素大鼠α亚基(αGSU)以及LHβ和FSHβ亚基的免疫组织化学证实了CHO细胞中产生了二聚体激素。重组啮齿动物促性腺激素被证实具有生物活性;重组FSH可增加颗粒细胞中的雌二醇生成,而重组LH可增加睾丸间质细胞中的睾酮生成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6383/5735272/1f1751e83fad/jrd-63-605-g001.jpg

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