Effects of gonadotropin-releasing hormone agonist on human chorionic gonadotropin activity in granulosa cells of immature female rats

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  • TUNGMAHASUK Doungrut
    Laboratory of Veterinary Physiology, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan
  • FUNGBUN Numfa
    Laboratory of Veterinary Physiology, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan
  • LAOHARATCHATATHANIN Titaree
    Clinic for Small Domestic Animals and Radiology, Faculty of Veterinary Medicine, Mahanakorn University of Technology, Bangkok 10530, Thailand
  • TERASHIMA Ryota
    Laboratory of Veterinary Physiology, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan
  • KURUSU Shiro
    Laboratory of Veterinary Physiology, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan
  • KAWAMINAMI Mitsumori
    Laboratory of Veterinary Physiology, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan

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Abstract

<p> Although the expression of gonadotropin-releasing hormone (GnRH) in the ovaries is well established, its physiological role remains unknown. The aim of this study was to determine whether ovarian GnRH mediates the actions of human chorionic gonadotropin (hCG) in the granulosa cells of immature female rats. Follicular growth was induced by administration of pregnant mare serum gonadotropin (PMSG, 15 IU/0.15 ml) on day 25 after birth, and hCG (20 IU/0.2 ml) was administered on day 27 revealing the increase of plasma progesterone level. Primary cultures of granulosa cells were established from large follicles 2 days after PMSG treatment. Progesterone synthesis was augmented by hCG in a dose-dependent manner. Annexin A5 (ANXA5), a biomarker of GnRH, was expressed in the granulosa-luteal cells after hCG treatment, as shown by immunohistochemistry, suggesting that hCG treatment induced GnRH action. The GnRH mRNA level was increased by hCG, and treatment with GnRH agonist (GnRHa) increased ANXA5 mRNA levels in the primary cultures of granulosa cells. Concomitant incubation of GnRH (10–7 M) or GnRHa (fertirelin acetate, 10–8 M) with hCG suppressed progesterone synthesis during a 3 h incubation period. The mRNA expression of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) was synergistically stimulated and suppressed by hCG and GnRHa, respectively. GnRHa stimulated p21 expression, and GnRHa and hCG synergistically reduced the mRNA expression levels of p27 and FOXO1. These data suggest that GnRH induced by LH may have a role for the LH-mediated luteinization of granulosa cells. In addition, ANXA5 may be involved in GnRH action. GnRH-ANXA5 would be an important mechanism in cell differentiation.</p>

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