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Journal of Integrative Agriculture  2021, Vol. 20 Issue (10): 2749-2757    DOI: 10.1016/S2095-3119(21)63606-7
Special Issue: 动物科学合辑Animal Science
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Follicle-stimulating hormone is expressed in ovarian follicles of chickens and promotes ovarian granulosa cell proliferation
BI Yu-lin1, 2*, YANG Shu-yan3*, WANG Hai-yan4*, CHANG Guo-bin1, CHEN Guo-hong1 
1 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P.R.China
2 Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education, Yangzhou University, Yangzhou 225009, P.R.China
3 Department of Biochemistry, Capital Institute of Pediatrics, Beijing 100020, P.R.China
4 Aquacultural College, Rizhao Polytechnic, Rizhao 276826, P.R.China
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摘要  

卵泡刺激素(FSH)是一种重要的下丘脑-垂体-性腺轴(HPG)激素,由垂体分泌。本研究证实FSH在鸡卵泡不同时期均有表达,颗粒细胞中FSHβ mRNA表达信号强于卵母细胞(P<0.05)。卵巢中FSHβ 369 bp的编码序列与垂体中FSHβ的编码序列完全相同。体外实验表明卵巢具有FSH分泌功能。此外,卵泡中FSHβ mRNA表达显著上调(P<0.05),且随着卵泡发育,FSHβ mRNA表达水平显著高于垂体(P<0.05),约为垂体的2~23倍。siRNA处理后卵巢颗粒细胞数量明显减少(P<0.05),说明卵巢FSH可促进颗粒细胞增殖。这一观点得到了细胞周期分析和CCND2CCNE2表达的支持。进一步研究表明,经FSHβ siRNA处理的颗粒细胞数与外源性FSH处理的颗粒细胞数无显著差异(P>0.05),而未经FSHβ siRNA转染的颗粒细胞数明显高于外源性FSH处理的颗粒细胞数(P<0.05)。提示外源性FSH对卵巢颗粒细胞的增殖作用依赖于内源性FSH。本研究表明FSH基因在鸡卵泡中表达,促进卵巢颗粒细胞增殖。本研究可以丰富HPG轴的理论。




Abstract  
Follicle-stimulating hormone (FSH), an important hypothalamic-pituitary-gonadal axis (HPG) hormone, is secreted by the pituitary gland.  This study confirms that FSH is expressed in chicken follicles at different stages, and positive FSHβ mRNA signals were stronger (P<0.05) in granulosa cells than in oocytes.  The 369 bp coding sequence of FSHβ in ovaries is 100% identical to that in the pituitary gland.  The experiment in vitro revealed that the ovary possessed FSH secretory capacity.  Further, FSHβ mRNA was significantly upregulated (P<0.05) in follicles and significantly higher (P<0.05) than that in the pituitary gland by approximately 2–23 times with the development.  The number of granulosa cells decreased significantly (P<0.05) in the cells with siRNA treatment, confirming that the ovarian FSH could promote granulosa cell proliferation.  This view was supported by cell cycle analysis and CCND2 and CCNE2 expression.  Further research indicated that no difference (P>0.05) was observed between the number of granulosa cells treated with FSHβ siRNA and in exogenous FSH. However, the number of granulosa cells without FSHβ siRNA transfection was significantly higher (P<0.05) for exogenous FSH.  This finding suggests that the proliferative effect of exogenous FSH on ovarian granulosa cells depend on endogenous FSH.  This study demonstrated that the FSH gene was expressed in chicken follicles and promoted ovarian granulosa cell proliferation, which enriched the theory on HPG axis.
Keywords:  FSH        expression        follicle        granulosa cell proliferation        chicken  
Received: 04 February 2020   Accepted:
Fund: This work was funded by the National Natural Science Foundation of China (31802057), the Second Batch of Special Grant from China Postdoctoral Science Foundation (2020TQ0252), the Postdoctoral Research Funding Project of Jiangsu Province, China in 2020 (2020Z213), and the Open Project Program of Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China (JILAR-KF202017).
Corresponding Authors:  Correspondence CHEN Guo-hong, Tel: +86-514-87971833, E-mail: ghchen@yzu.edu.cn; CHANG Guo-bin, Tel: +86-514-87971833, E-mail: passioncgb@163.com    
About author:  BI Yu-lin, Tel: +86-514-87971833, E-mail: ylbi@yzu.edu.cn; * These authors contributed equally to this study.

Cite this article: 

BI Yu-lin, YANG Shu-yan, WANG Hai-yan, CHANG Guo-bin, CHEN Guo-hong. 2021. Follicle-stimulating hormone is expressed in ovarian follicles of chickens and promotes ovarian granulosa cell proliferation. Journal of Integrative Agriculture, 20(10): 2749-2757.

Bergandi L, Canosa S, Pittatore G, Silvagno F, Doublier S, Gennarelli G, Benedetto C, Revelli A. 2019. Human recombinant FSH induces chemoresistance in human breast cancer cells via hif-1α activation. Biology of Reproduction, 100, 1521–1535.
Campbell B K, Scaramuzzi R J, Webb R. 1995. Control of antral follicle development and selection in sheep and cattle. Journal of Reproduction and Fertility, 49, 335–350.
Casarini L, Crépieux P. 2019. Molecular mechanisms of action of FSH. Front Endocrinol (Lausanne), 10, 305.
Chrusciel M, Ponikwicka-Tyszko D, Wolczynski S, Huhtaniemi I, Rahman N A. 2019. Extragonadal FSHR expression and function - is it real? Frontiers in Endocrinology (Lausanne), 10, 32.
Cui H X, Zhao G P, Liu R R, Zheng M Q, Chen J L, Wen J. 2012. FSH stimulates lipid biosynthesis in chicken adipose tissue by upregulating the expression of its receptor FSHR. Journal of Lipid Research, 53, 909–917.
Deng M Q, Huang X Y, Tang T S, Sun F Z. 1998. Spontaneous and fertilization induced Ca2+ oscillations in mouse immaturegerminal vesicle-stage oocytes. Biology of Reproduction, 58, 807–813.
Downs S M, Hunzicker-Dunn M. 1995. Differential regulation of oocyte maturation and cumulus expansion in the mouse oocyte-cumulus cell complex by site-selective analogs of cyclic adenosine monophosphate. Developmental Biology, 172, 72–85.
Eppig J J. 2001. Oocyte control of ovarian follicular development and function in mammals. Reproduction, 122, 829–838.
Escamilla-Hernandez R, Little-Ihrig L, Orwig K E, Yue J, Chandran U, Zeleznik A J. 2008. Constitutively active protein kinase A qualitatively mimics the effects follicle- stimulating hormone on granulosa cell differentiation. Molecular Endocrinology, 22, 1842–1852.
Gao P, Wang H, Yu J, Zhang J, Yang Z, Liu M, Niu Y, Wei X, Wang W, Li H, Wang Y, Sun G. 2018. miR-3607-3p suppresses non-small cell lung cancer (NSCLC) by targeting TGFBR1 and CCNE2. PLoS Genetic, 14, e1007790.
Gjerdrum L M, Lielpetere I, Rasmussen L M, Bendix K, Hamilton-Dutoit S. 2001. Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis. Journal of Molecular Diagnostics, 3, 105–110.
Hernández-Coronado C G, Guzmán A, Rodríguez A, Mondragón J A, Romano M C, Gutiérrez C G, Rosales-Torres A M. 2016. Sphingosine-1-phosphate, regulated by FSH and VEGF, stimulates granulosa cell proliferation. General and Comparative Endocrinology, 236, 1–8.
Huang Z, Li X, Li Y, Liu R, Chen Y, Wu N, Wang M, Song Y, Yuan X, Lan L, Xu Q, Chen G, Zhao W. 2015. Molecular cloning and functional analysis of the goose FSHβ gene. British Poultry Science, 56, 284–292.
Ji Y, Liu P, Yuen T, Haider S, He J, Romero R, Chen H, Bloch M, Kim S M, Lizneva D, Munshi L, Zhou C, Lu P, Iqbal J, Cheng Z, New M I, Hsueh A J, Bian Z, Rosen C J, Sun L, et al. 2018. Epitope-specific monoclonal antibodies to FSHβ increase bone mass. Proceedings of the National Academy of Sciences of the United States of America, 115, 2192–2197.
Kayampilly P P, Menon K M. 2009. Follicle-stimulating hormone inhibits adenosine 5´-monophosphate-activated protein kinase activation and promotes cell proliferation of primary granulosa cells in culture through an Akt-dependent pathway. Endocrinology, 150, 929–935.
Kong D, Guan Q, Li G, Xin W, Qi X, Guo Y, Zhao J, Xu J, Sun S, Gao L. 2018. Expression of FSHR in chondrocytes and the effect of FSH on chondrocytes. Biochemical and Biophysical Research Communications, 495, 587–593.
Leoni G G, Naitana S. 2018. Ovine granulosa cells isolation and culture to improve oocyte quality. Methods in Molecular Biology, 1817, 95–106.
Li C H, Simpson M E, Evans H M. 1949. Isolation of pituitary follicle-stimulating hormone (FSH). Science, 109, 445–446.
Liu P, Ji Y, Yuen T, Rendina-Ruedy E, DeMambro V E, Dhawan S, Abu-Amer W, Izadmehr S, Zhou B, Shin A C, Latif R, Thangeswaran P, Gupta A, Li J, Shnayder V, Robinson S T, Yu Y E, Zhang X, Yang F, Lu P, et al. 2017. Blocking FSH induces thermogenic adipose tissue and reduces body fat. Nature, 546, 107–112.
Liu X M, Chan H C, Ding G L, Cai J, Song Y, Wang T T, Zhang D, Chen H, Yu M K, Wu Y T, Qu F, Liu Y, Lu Y C, Adashi E Y, Sheng J Z, Huang H F. 2015. FSH regulates fat accumulation and redistribution in aging through the Gαi/Ca2+/CREB pathway. Aging Cell, 14, 409–420.
Lizneva D, Rahimova A, Kim S M, Atabiekov I, Javaid S, Alamoush B, Taneja C, Khan A, Sun L, Azziz R, Yuen T, Zaidi M. 2019. FSH beyond fertility. Frontiers in Endocrinology (Lausanne), 10, 136.
Meinsohn M C, Morin F, Bertolin K, Duggavathi R, Schoonjans K, Murphy B D. 2017. The orphan nuclear receptor liver homolog receptor-1 (Nr5a2) regulates ovarian granulosa cell proliferation. Journal of Endocrine Society, 2, 24–41.
Nemer A, Azab A N, Rimon G, Lamprecht S, Ben-Menahem D. 2018. Different roles of cAMP/PKA and PKC signaling in regulating progesterone and PGE2 levels in immortalized rat granulosa cell cultures. General and Comparative Endocrinology, 269, 88–95.
Olejar T, Vetvicka D, Boucek J, Zabrodsky M, Benes J, Kabesova M, Pouckova P. 2020. The FSHR expression in head and neck squamous cell cancer, a pilot immunohistochemical study. Anticancer Research, 40, 349–356.
Orisaka M, Orisaka S, Jiang J Y, Craig J, Wang Y, Kotsuji F, Tsang B K. 2006. Growth differentiation factor 9 is antiapoptotic during follicular development from preantral to early antral stage. Molecular Endocrinology, 20, 2456–2468.
Semeshin V F, Andreyeva E N, Shloma V V, Saumweber H, Zhimulev I F. 2002. Immunogold electron microscope localization of proteins in Drosophila polytene chromosomes: applications and limitations of the method. Chromosome Research, 10, 429–433.
Shi X, Qiu S, Zhuang W, Wang C, Zhang S, Yuan N, Yuan F, Qiao Y. 2018. Follicle-stimulating hormone inhibits cervical cancer via NF-κB pathway. OncoTargets and Therapy, 11, 8107–8115.
Sirotkin A, Alexa R, Kádasi A, Adamcová E, Alwasel S, Harrath A H. 2019. Resveratrol directly affects ovarian cell sirtuin, proliferation, apoptosis, hormone release and response to follicle-stimulating hormone (FSH) and insulin-like growth factor I (IGF-I). Reproduction Fertility and Development, doi: 10.1071/RD18425.
Su S, Sun X, Zhou X, Fang F, Li Y. 2015. Effects of GnRH immunization on the reproductive axis and thymulin. Journal of Endocrinology, 226, 93–102.
Su Y Q, Xia G L, Byskov A G, Fu G D, Yang C R. 1999. Protein kinase C and intracellular calcium are involved in follicle-stimulating hormone-mediated meiotic resumption of cumulus cell-enclosed porcine oocytes in hypoxanthine-supplemented medium. Molecular Reproduction and Development, 53, 51–58.
Sturkie P D. 2012. Avian Physiology. 6th ed. Springer Science & Business Media, New York, NY, USA.
Ulloa-Aguirre A, Reiter E, Crépieux P. 2018. FSH receptor signaling: Complexity of interactions and signal diversity. Endocrinology, 159, 3020–3035.
Volonteri M C , Regueira E, Scaia MF, Ceballos N R. 2013. Characterization and seasonal changes in LHβ and FSHβ mRNA of Rhinella arenarum (Amphibia, Anura). General and Comparative Endocrinology, 187, 95–103.
Xia G L, Byskov A G, Andersen C Y. 1994. Cumulus cells secrete a meiosis-inducing substance by stimulation with forskolin and dibutyric cyclic adenosine monophosphate. Molecular Reproduction and Development, 39, 17–24.
Yang P X, Roy S K. 2006. A novel mechanism of FSH regulation of DNA synthesis in the granulosa cells of hamster preantral follicles. Biology of Reproduction, 75, 149–157.
Yivgi-Ohana N, Sher N, Melamed-Book N, Eimerl S, Koler M, Manna P R, Stocco D M, Orly J. 2009. Transcription of steroidogenic acute regulatory protein in the rodent ovary and placenta: Alternative modes of cyclic adenosine 3´, 5’-monophosphate dependent and independent regulation. Endocrinology, 150, 977–989.
Zhang L, Zhang X, Zhang X, Lu Y, Li L, Cui S. 2017. MiRNA-143 mediates the prolifeative signaling pathway of FSH and regulates estradiol production. Journal of Endocrinology, 234, 1–14.
Zheng J, Mao J, Cui M, Liu Z, Wang X, Xiong S, Nie M, Wu X. 2017. Novel FSHβ mutation in a male patient with isolated FSH deficiency and infertility. European Journal of Medical Genetics, 60, 335–339. 
Zhou J, Ju W Q, Yuan X P, Zhu X F, Wang D P, He X S. 2016. miR-26a regulates mouse hepatocyte proliferation via directly targeting the 3´ untranslated region of CCND2 and CCNE2. Hepatobiliary & Pancreatic Diseases International, 15, 65–72.
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