Construction and expression of vectors encoding biologically active rodent gonadotropins

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  • OHTA Akihiko
    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 Department of Life Science, Meiji University, Kanagawa 214-8571, Japan
  • TSUNODA Yuichiro
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • TAMURA Yoshihiko
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • IINO Kayoko
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • NISHIMURA Naoto
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • NISHIHARA Hiroto
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • TAKANASHI Haruka
    Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
  • YOSHIDA Saishu
    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
  • KATO Takako
    Institute for Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
  • KATO Yukio
    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 Department of Life Science, Meiji University, Kanagawa 214-8571, Japan

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Abstract

<p> 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.</p>

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