Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (18): 3716-3724.doi: 10.3864/j.issn.0578-1752.2014.18.020

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles    

Regulation of GnIH on Reproduction of Female Pig

WANG Yao1, LI Xun2, LI Min-jie1, GUO Meng-jie1, LEI Zhi-hai1   

  1. 1 College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095
    2 College of Animal Science and Technology, Guangxi University, Nanning 530004
  • Received:2013-09-23 Revised:2014-06-16 Online:2014-09-16 Published:2014-09-16

RichHTML

3

PDF

638

Abstract: 【Objective】The expression of GnIH mRNA in female pig, the effect of GnIH on steroid hormone secretion and cell proliferation proteins expression in granulosa cells cultured in vitro, and morphological relationship between GPR147 and GnRH were investigated in this study. The purpose of all the studies was to explore the regulatory mechanism of GnIH to pig reproduction in vitro, at molecular and cell levels, and from cell morphology images.【Method】Twenty-four tissue and organ samples were collected from healthy female piglets aged 30 d, expression levels of GnIH mRNA were detected by semi-quantitative RT-PCR. The effect of GnIH on steroid hormone secretion and cell proliferation proteins expression in granulosa cells cultured in vitro was analyzed by the method of radioimmunoassay and Western blotting. Hypothalamus tissues were also collected from healthy female piglets aged 30 d and used for preparation of histological sections. The morphological relationship between GnIH receptor and GnRH was revealed by immunohistochemistry and examined with confocal microscopy.【Result】The result showed that GnIH mRNA mainly expressed in the central nervous system, especially in the diencephalon. After in vitro cultured, the granulosa cells were treated with 10-6, 10-8, 10-10 and 10-12 mol•L-1 GnIH. Compared with the control cells, 10-6 and 10-10 mol•L-1 doses of GnIH significantly reduced (P＜0.01) E2 secretion. All concentrations of GnIH could attenuate P4 secretion. GnIH significantly inhibited (P＜0.05) PCNA and cyclin B1 expression in cultured granulosa cells. Moreover, immunohistochemistry analysis revealed the close contact between immunoreactive fibers or cell bodies of GPR147 and GnRH in pig hypothalamus tissue. 【Conclusion】These results demonstrated that GnIH not only plays a central regulatory role on GnRH secretion in the hypothalamus, but also peripherally regulates pig reproduction by affecting steroid hormone secretion and cell proliferation proteins expression in the ovary.

Key words: female pig, GnIH, semi-quantitative RT-PCR, reproductive regulation

[1] 雷治海, 李月. 促性腺激素抑制激素的研究进展. 畜牧与兽医, 2008, 40(9): 96-99.
Lei Z H, Li Y. Advances in gonadotropin-inhibitory hormone. Animal husbandry and Veterinary medicine, 2008, 40(9): 96-99. (in Chinese)
[2] Tsutsui K, Saigoh E, Ukena K, Teranishi H, Fujisawa Y, Kikuchi M, Ishii S, Sharp P J. A novel avian hypothalamic peptide inhibiting gonadotropin release. Biochemical and Biophysical Research Communications, 2000, 275(2): 661-667.
[3] Tsutsui K. A new key neurohormone controlling reproduction, gonadotropin-inhibitory hormone (GnIH): Biosynthesis, mode of action and functional significance. Progress in Neurobiology, 2009, 88(1): 76-88.
[4] Clarke I J, Qi Y, Puspita Sari I, Smith J T. Evidence that RF-amide related peptides are inhibitors of reproduction in mammals. Frontiers in Neuroendocrinology, 2009, 30(3): 371-378.
[5] Ikemoto T, Park M K. Chicken RFamide-related peptide (GnIH) and two distinct receptor subtypes: identi?cation, molecular characterization, and evolutionary considerations. Journal of Reproduction and Development, 2005, 51(3): 359-377.
[6] Hinuma S, Shintani Y, Fukusumi S, Iijima N, Matsumoto Y, Hosoya M, Fujii R, Watanabe T, Kikuchi K, Terao Y, Yano T, Yamamoto T, Kawamata Y, Habata Y, Asada M, Kitada C, Kurokawa T, Onda H, Nishimura O, Tanaka M, Ibata Y, Fujino M. New neuropeptides containing carboxy-terminal RFamide and their receptor in mammals. Nature Cell Biology, 2000, 2(10): 703-708.
[7] Ubuka T, Morgan K, Pawson A J, Osugi T, Chowdhury V S, Minakata H, Tsutsui K, Millar R P, Bentley G E. Identification of human GnIH homologs, RFRP-1 and RFRP-3, and the cognate receptor, GPR147 in the human hypothalamic pituitary axis. PloS ONE, 2009, 4(12): 1-7/e8400.
[8] Bentley G E, Tsutsui K, Kriegsfeld L J. Recent studies of gonadotropin-inhibitory hormone (GnIH) in the mammalian hypothalamus, pituitary and gonads. Brain Research, 2010, 1364: 62-71.
[9] Zhang Y, Li S, Liu Y, Lu D, Chen H, Huang X, Liu X, Meng Z, Lin H, Cheng C H. structural diversity of the GnIH/GnIH receptor system in teleost: its involvement in early development and the negative control of LH release. Peptides, 2010, 31(6): 1034-1043.
[10] Maddineni S, Ocón-Grove O M, Krzysik-Walker S M, Hendricks G L 3rd, Proudman J A, Ramachandran R. Gonadotrophin-inhibitory hormone receptor expression in the chicken pituitary gland: potential influence of sexual maturation and ovarian steroids. Journal of Neuroendocrinology, 2008, 20(9): 1078-1088.
[11] Tsutsui K, Bentley G E, Bedecarrats G, Osugi T, Ubuka T, Kriegsfeld L J. Gonadotropin-inhibitory hormone (GnIH) and its control of central and peripheral reproductive function. Front Neuroendocrinolgy, 2010, 31(3): 284-295.
[12] Shimizu M, Bedecarrats G Y. Activation of the chicken gonadotropin- inhibitory hormone receptor reduces gonadotropin releasing hormone receptor signaling. General and Comparative Endocrinology, 2010, 167(2): 331-337.
[13] Su J, Lei Z, Zhang W, Ning H, Ping J. Distribution of orexin B and its relationship with GnRH in the pig hypothalamus. Research in Veterinary Science, 2008, 85(2): 315-323.
[14] Hatey F, Langlois I, Mulsant P, Bonnet A, Benne F, Gasser F. Gonadotropins induce accumulation of insulin-like growth factor I mRNA in pig granulosa cells in vitro. Molecular and Cellular Endocrinology, 1992, 86(3): 205-211.
[15] Qi Y, Oldfield B J, Clarke I J. Projections of RFamide-related peptide-3 neurones in the ovine hypothalamus, with special reference to regions regulating energy balance and reproduction. Journal of  Neuroendocrinology, 2009, 21(8): 690-697.
[16] Wu M, Dumalska I, Morozova E, van den Pol AN, Alreja M. Gonadotropin inhibitory hormone inhibits basal forebrain vGluT2-世纪gonadotropin-releasing hormone neurons via a direct postsynaptic mechanism. Journal of Physiology, 2009, 587(7): 1401-1411.
[17] Ducret E, Anderson G M, Herbison A E. RFamide-related peptide-3, a mammalian gonadotropin-inhibitory hormone ortholog, regulates gonadotropin-releasing hormone neuron firing in the mouse. Endocrinology, 2009, 150(6): 2799-2804.
[18] Smith J T. The role of kisspeptin and gonadotropin inhibitory hormone in the seasonal regulation of reproduction in sheep. Domest Animal Endocrinology, 2012, 43(2): 75-84.
[19] Rizwan M Z, Poling M C, Corr M, Cornes P A, Augustine R A, Quennell J H, Kauffman A S, Anderson G M. RFamide-related peptide-3 receptor gene expression in GnRH and kisspeptin neurons and GnRH-dependent mechanism of action. Endocrinology, 2012, 153(8): 3770-3779.
[20] Khan A R, Kauffman A S. The role of kisspeptin and RFamide- related peptide-3 neurones in the circadian-timed preovulatory luteinising hormone surge. Journal of Neuroendocrinology, 2012, 24(1): 131-143.
[21] Li X, Su J, Lei Z, Zhao Y, Jin M, Fang R, Zheng L, Jiao Y. Gonadotropin-inhibitory hormone (GnIH) and its receptor in the female pig: cDNA cloning, expression in tissues and expression pattern in the reproductive axis during the estrous cycle. Peptides, 2012, 36(2): 176-185.
[22] Singh P, Krishna A and Tsutsui K. Effects of gonadotropin-inhibitory hormone on folliculogenesis, steroidogenesis of cyclic mice. Fertile Sterile, 2011, 95(4): 1397-1404.
[23] Maddineni S R, Ocón-Grove O M, Krzysik-Walker S M, Hendricks G L 3rd, Ramachandran R. Gonadotrophin-inhibitory hormone(GnIH) receptor gene is expressed in the chicken ovary: potential role of GnIH in follicular maturation. Reproduction, 2008, 135(2): 267-274.
[24] Bonini J A, Jones KA, Adham N, Forray C, Artymyshyn R, Durkin M M, Smith K E, Tamm J A, Boteju L W, Lakhlani P P, Raddatz R, Yao W J, Ogozalek K L, Boyle N, Kouranova EV, Quan Y, Vaysse P J, Wetzel J M, Branchek T A, Gerald C, Borowsky B. Identification and characterization of two G protein-coupled receptors for neuropeptide FF. Journal of Biological Chemistry, 2000, 275(50): 39324-39331.
[25] Oishi H, Klausen C, Bentley G E, Osugi T, Tsutsui K, Gilks C B, Yano T, Leung P C. The human gonadotropin-inhibitory hormone ortholog RFamide-related peptide-3 suppresses gonadotropin-induced progesterone poduction in human granulosa cells. Endocrinology, 2012, 153(7): 3435-3445.
[26] Meloche S, Pouyssegur J. The ERK1/2 mitogen-activated protein kinase pathway as a master regulator of the G1-to S-phase transition. Oncogene, 2007, 26(22): 3227-3239.
[27] Chaffin C L, Schwinof K M, Stouffer R L. Gonadotropin and steroid control of granulosa cell proliferation during the periovulatory interval in rhesus monkeys. Biology of Reproduction, 2001, 65(3): 755-762.
[28] Bentley G E, Ubuka T, McGuire N L, Chowdhury V S, Morita Y, Yano T, Hasunuma I, Binns M, Wingfield J C, Tsutsui K. Gonadotrophin-inhibitory hormone and its receptor in the avain reproductive system. General and Comparative Endocrinology, 2008, 156(1): 34-43.
[29] Ubuka T, Kim S, Huang Y C, Reid J, Jiang J, Osugi T, Chowdhury V S, Tsutsui K, Bentley G E. Gonadotrophin-inhibitory hormone neurons interact directly with gonadotrophin-releasing hormone-I and -II neurons in European starling brain. Endocrinology, 2008, 149(1): 268-278.
[1] Xin ZHANG,KongLin HUO,XingXing SONG,DuoNi ZHANG,Wen HU,ChuanHuo HU,Xun LI. Effects of GnIH on Autophagy and Apoptosis of Porcine Ovarian Granulosa Cells via p38MAPK Signaling Pathway [J]. Scientia Agricultura Sinica, 2020, 53(9): 1904-1912.
[2] ZHAO Wei,WANG Qian,HAO Zhi-min,WANG Qing,SONG Wen-jing,HAN Jian-min,DONG Jin-gao
. Cloning and Expression of the Gene Encoding the Protein Kinase C in Setosphaeria turcica
 [J]. Scientia Agricultura Sinica, 2011, 44(5): 933-938 .
[3] JIANG Jin, CAO Shao-Xian, MAO Da-Gan, HUI Feng-Ming. Expression of Uterine LHR mRNA During Estrous Cycle in Guinea Pig [J]. Scientia Agricultura Sinica, 2011, 44(16): 3420-3426.
[4] JIANG Jun-bing,YU Xiu-ju,HE Jun-ping,DONG Chang-sheng
. Study on the Expression and Localization of Tyrosinase in Skin of Alpacas with Different Coat Colors#br# [J]. Scientia Agricultura Sinica, 2010, 43(12): 2555-2560 .
[5] . Cloning of MSTN Gene of Landes Goose and The dfference of MSTN Gene Expression by Different Energe [J]. Scientia Agricultura Sinica, 2008, 41(1): 221-228 .
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd