Skip to main content
SpringerLink
Log in

Inhibin A regulates follicular development via hormone secretion and granulosa cell behaviors in laying hens

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Inhibin A regulates follicular development, and its expression level is related to physiological activities, such as the recruitment, selection, and predominance during follicular development. Therefore, examining inhibin A and its regulatory effects on the reproductive performance of poultry is crucial. In this study, we measured the mRNA and protein abundances of INHA and INHBA in the chicken reproductive system and determined the hormone secretion and apoptosis of follicular granulosa cells (GCs) after being treated with inhibin A protein, and flow cytometry was performed to analyze GC apoptosis in INHA-specific small RNA interference (siRNA). We detected that INHA and INHBA were mainly expressed in chicken follicles. The highest INHA mRNA abundance was found in the fifth largest preovulatory follicle (F5) (P < 0.05). INHBA mRNA expression in the largest preovulatory follicle (F1) was significantly higher than those in other follicles (P < 0.05). Similar results were found for INHA and INHBA protein expression in those follicles (P < 0.05). Treatment with inhibin A protein increased the activity of GCs in a dose-dependent manner (P < 0.05), which was characterized by decreased gene expression of pro-apoptotic factors Bax and Caspase-3 (P < 0.05) and increased expression of proliferation genes Bcl-2 and PCNA (P < 0.05). Additionally, inhibin A significantly increased the secretion of progesterone and estradiol (P < 0.05). RNAi-mediated knockdown of INHA increased apoptosis in GCs via a Caspase-3-dependent mitochondrial pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14
Fig. 15
Fig. 16
Fig. 17
Fig. 18
Fig. 19

Similar content being viewed by others

References

  • Anasti JN (1998) Premature ovarian failure: an update. Fertil Steril 70(1):1–15

    CAS  PubMed  Google Scholar 

  • Bartosz K, Hanna P, Marta R (2015) Expression of INHÎ2A and INHÎ2B proteins in porcine oocytes cultured in vitro is dependent on the follicle size. Zygote 23:205–211

    Google Scholar 

  • Bernard DJ, Chapman SC, Woodruff TK (2001) Mechanisms of inhibin signal transduction. Recent Prog Horm Res 56:417–450

    CAS  PubMed  Google Scholar 

  • Bristol-Gould SK, Kreeger PK, Selkirk CG (2006) Postnatal regulation of germ cells by activin: the establishment of the initial follicle pool. Dev Biol 298:132–148

    CAS  PubMed  Google Scholar 

  • Cai K, Hua G, Ahmad S (2011) Action mechanism of inhibin α-subunit on the development of Sertoli cells and first wave of spermatogenesis in mice. PLoS One 6:e25585

    CAS  PubMed  PubMed Central  Google Scholar 

  • Campbell BK, Baird DT (2001) Inhibin A is a follicle stimulating hormone-responsive marker of granulosa cell differentiation, which has both autocrine and paracrine actions in sheep. J Endocrinol 169:333–345

    CAS  PubMed  Google Scholar 

  • Chandrasekar B et al (2004) 17beta-estradiol and restenosis: a novel vaso-protective role for estrogen. J Invasive Cardiol 16:719–722

    PubMed  Google Scholar 

  • Chang HM, Cheng JC, Klausen C (2014) Effects of recombinant activins on steroidogenesis in human granulosa-lutein cells. J Clin Endocrinol Metab 99:E1922–E1932

    CAS  PubMed  Google Scholar 

  • Chen F, Jiang X, Chen X (2007) Effects of downregulation of inhibin alpha gene expression on apoptosis and proliferation of goose granulosa cells. J Genet GenoMics 34:1106–1113

    CAS  PubMed  Google Scholar 

  • Denkova R, Bourneva V, Staneva-Dobrovski L (2004) In vitro effects of inhibin on apoptosis and apoptosis related proteins in human ovarian granulosa cells. Endocr Regul 38:51–55

    CAS  PubMed  Google Scholar 

  • Fan TJ, Han LH, Cong RS (2005) Caspase family proteases and apoptosis. Acta Bioch Bioph Sin 37:719–727

    CAS  Google Scholar 

  • Findlay J, Drummond A, Britt K (2000) The roles of activins, inhibins and estrogen in early committed follicles. Mol Cell Endocrinol 163:81–87

    CAS  PubMed  Google Scholar 

  • Fortune JE et al (1994) Ovarian follicular growth and development in mammals. Biol Reprod 50:225–232

    CAS  PubMed  Google Scholar 

  • Gago LA, Ginsburg KA (2004) Premature ovarian failure. Encyclopedia Endocr Dis 18(4):65–72

    Google Scholar 

  • Gilbert AB, Evans AJ, Perry MM, Davidson MH (1977) A method for separating the granulosa cells, the basal lamina and the theca of the preovulatory ovarian follicle of the domestic fowl (Gallus domesticus). J Reprod Fertil 50:179–181

    CAS  PubMed  Google Scholar 

  • Hall PA, Levison DA, Woods AL (1990) Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some, neoplasms. J Pathol 162:285–294

    CAS  PubMed  Google Scholar 

  • Han L, Wu C, Riaz H (2013) Characterization of the mechanism of inhibin α-subunit gene in mouse anterior pituitary cells by RNA interference. PLoS One 8:e74596

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hillier SG et al (1991) Regulatory functions for inhibin and activin in human ovaries. J Endocrinol 131:171–175

    CAS  PubMed  Google Scholar 

  • Hillier SG et al (1994) Current concepts of the roles of follicle stimulating hormone and luteinizing hormone in folliculogenesis. Hum Reprod 9:188–191

    CAS  PubMed  Google Scholar 

  • Hrabia A, Paczoska-Eliasiewicz H, Rząsa J (2004) Effect of prolactin on estradiol and progesterone secretion by isolated chicken ovarian follicles. Folid Biol-Prague 52:197–203

    CAS  Google Scholar 

  • Hrabia A, Wilk M, Rząsa J (2008) Expression of α and β estrogen receptors in the chicken ovary. Folid Biol-Prague 56:187–191

    CAS  Google Scholar 

  • Hsueh AJ, Dahl KD, Vaughan J (1987) Heterodimers and homodimers of inhibin subunits have different paracrine action in the modulation of luteinizing hormone-stimulated androgen biosynthesis. P Natl Acad Sci USA 84:5082–5086

    CAS  Google Scholar 

  • Hulshof SC, Figueiredo JR, Beckers JF (1997) Bovine preantral follicles and activin: immunohistochemistry for activin and activin receptor and the effect of bovine activin A in vitro. Theriogenology 48:133–142

    CAS  PubMed  Google Scholar 

  • Ito M, Park Y, Weck J (2000) Synergistic activation of the inhibin alpha-promoter by steroidogenic factor-1 and cyclic adenosine 3′, 5′-monophosphate. Mol Endocrinol 14:66–81

    CAS  PubMed  Google Scholar 

  • Jiang XP, Yang LG, Liu GQ, Mao DG (2002) Effects of inhibin α (1-32) gene immunization on mouse reproductive traits. Chin J Vet Sci 22:368–370

    CAS  Google Scholar 

  • Jimenez-Krassel F, Winn ME, Burns D (2003) Evidence for a negative intrafollicular role for inhibin in regulation of estradiol production by granulosa cells. Endocrinology 144:1876–1886

    CAS  PubMed  Google Scholar 

  • Johnson PA, Brooks CF, Davis AJ (2005) Pattern of secretion of immunoreactive inhibin/activin subunits by avian granulosa cells. GenComp Endocrinol 141:233–239

    CAS  Google Scholar 

  • Kadariya I, Wang J, Ur RZ (2015) RNAi-mediated knockdown of inhibin α subunit increased apoptosis in granulosa cells and decreased fertility in mice. J Steroid Biochem 152:161–170

    CAS  Google Scholar 

  • Knight PG, Glister C (2001) Potential local regulatory functions of inhibins, activins and follistatin in the ovary. Reproduction 121:503–512

    CAS  PubMed  Google Scholar 

  • Li M, Yin Y, Hua H (2010) The reciprocal regulation of gamma-synuclein and insulin-like growth factor-I (IGF-I) receptor expression creates a circuit that modulates IGF-I signaling. J Biol Chem 285:30480

    CAS  PubMed  PubMed Central  Google Scholar 

  • Li T, Ma AT, Zhang YJ (2017) Effects of silencing INHα gene by RNAi on the mRNA expressions of StAR, FST, INHβB, and FSHR genes in cultured sheep granulosa cells. Small Ruminant Res 157:23–26

    Google Scholar 

  • Lu C, Yang W, Chen M (2009) Inhibin A inhibits follicle-stimulating hormone (FSH) action by suppressing its receptor expression in cultured rat granulosa cells. Mol Cell Endocrinol 298:48–56

    CAS  PubMed  Google Scholar 

  • Manabe N, Goto Y, Matsuda-Minehata F (2004) Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia. J Reprod Develop 50:493–514

    Google Scholar 

  • Matzuk MM, Finegold MJ, Mather JP (1994) Development of cancer cachexia-like syndrome and adrenal tumors in inhibin-deficient mice. P Natl A Sci 91:8817–8821

    CAS  Google Scholar 

  • Messinis IE, Messini CI, Dafopoulos K (2014) Novel aspects of the endocrinology of the menstrual cycle. Reprod BioMed Online 28:714–722

    CAS  PubMed  Google Scholar 

  • Meunier H, Rivier C, Evans RM (1988) Gonadal and extragonadal expression of inhibin alpha, beta a, and beta B subunits in various tissues predicts diverse functions. P Natl Acad Sci USA 85:247–251

    CAS  Google Scholar 

  • Miyazaki T, Liu ZJ, Kawahara A (1995) Three distinct IL-2 signaling pathways mediated by bcl-2, c-myc, and lck cooperate in hematopoietic cell proliferation. Cell 81:223–231

    CAS  PubMed  Google Scholar 

  • Mohamed SM, Toshiro T, Yoshito A, Masato K (2006) The effect of active immunizationagainst inhibin on gonadotropin secretions and follicular dynamics during the estrous cycle in cows. J Reprod Dev 52:107–113

    Google Scholar 

  • Nishimori K, Matzuk MM (1996) Transgenic mice in the analysis of reproductive development and function. Rev Peprod 1:203–212

    CAS  Google Scholar 

  • Onagbesan O, Bruggeman V, Decuypere E (2009) Intra-ovarian growth factors regulating ovarian function in avian species: a review. Anim Reprod Sci 111:121–140

    CAS  PubMed  Google Scholar 

  • Rabinovici J, Spencer SJ, Doldi N (1992) Activin-A as an intraovarian modulator: actions, ocalization, and regulation of the intact dimer in human ovarian cells. J Clin Invest 89:1528–1536

    CAS  PubMed  PubMed Central  Google Scholar 

  • Roberts VJ, Barth S, el-Roeiy A (1993) Expression of inhibin/activin subunits and follistatin messenger ribonucleic acids and proteins in ovarian follicles and the corpus luteum during the human menstrual cycle. J Clin Endocrinol Metab 77:1402–1410

    CAS  PubMed  Google Scholar 

  • Sadovsky Y, Dorn C (2000) Function of steroidogenic factor 1 during development and differentiation of the reproductive system. Rev Reprod 5:136–142

    CAS  PubMed  Google Scholar 

  • Safi M, Buys N, Onagbesan OM (1998) Quantification of inhibin/activin alpha and beta a subunit messenger ribonucleic acid by competitive reverse transcription-polymerase chain reaction in chicken granulosa cells during follicular development. Biol Reprod 59:1047–1054

    CAS  PubMed  Google Scholar 

  • Shi F, Mochida K, Suzuki O, Matsuda J (2000) Development of embryos in superovulated Guinea pigs following active immunization against the inhibin alpha-subunit. J Endocrinol 47:451–459

    CAS  Google Scholar 

  • Silva JR, Tharasanit T, Taverne MA (2006) The activin-follistatin system and in vitro early follicle development in goats. J Endocrinol 189:113–125

    CAS  PubMed  Google Scholar 

  • Sirotkin AV et al (2011) Cytokines: signalling molecules controlling ovarian functions. Int J Biochem Cell B 43:857–861

    CAS  Google Scholar 

  • Sisco B, Pfeffer PL (2007) Expression of activin pathway genes in granulosa cells of dominant and subordinate bovine follicles. Theriogenology 68:29–37

    CAS  PubMed  Google Scholar 

  • Tannetta DS, Feist SA, Bleach EC (1998) Effects of active immunization of sheep against an amino terminal peptide of the inhibin alpha C subunit on intrafollicular levels of activin A, inhibin A and follistatin. J Endocrinol 157:157–168

    CAS  PubMed  Google Scholar 

  • Tilly JL et al (1996) Apoptosis and ovarian function. Rev Peprod 1:162–172

    CAS  Google Scholar 

  • Van den Hurk R, Van de Pavert SA (2001) Localization of an activin/activin receptor system in the porcine ovary. Mol Reprod Dev 60:463–471

    PubMed  Google Scholar 

  • Wang C, Li C, Li H (2015) Downregulation of the expression of inhibin α subunit and betaglycan in porcine cystic follicles. J Vet Med Sci 77:1419–1425

    CAS  PubMed  PubMed Central  Google Scholar 

  • Webley GE, Marsden PL, Knight PG (1994) Differential control of immunoreactive alpha-inhibin and progesterone production by marmoset luteal cells in vitro: evidence for a paracrine action of alpha-inhibin on basal and gonadotropin-stimulated progesterone production. Biol Reprod 50:1394–1402

    CAS  PubMed  Google Scholar 

  • Xu J, Wang B, Xu Y (2012) Epigenetic regulation of HIF-1α in renal cancer cells involves HIF-1α/2α binding to a reverse hypoxia-response element. Oncogene 31:1065

    CAS  PubMed  Google Scholar 

  • Yamoto M, Minami S, Nakano R (1992) Immunohistochemical localization of inhibin/activin subunits in human ovarian follicles during the menstrual cycle. J Clin Endocrinol Metab 74:989–993

    CAS  PubMed  Google Scholar 

  • Ying SY et al (1987) Inhibins and activins: chemical properties and biological activity. P Soc Exp Biol Med 186:253–264

    CAS  Google Scholar 

  • Zhao J, Taverne MA, van der Weijden GC (2001) Effect of activing A on in vitro development of rat preantral follicles and localization of activin A and activin receptor II. Biol Reprod 65:967–977

    CAS  PubMed  Google Scholar 

Download references

Funding

This research was supported the Modern Agricultural Industry Technology System (CARS-41), the National Natural Science Foundation of China (Grant No. 31872347), and the Sichuan Provincial Improving Poverty Founding (Grant No. 2017NFP0211). We deeply appreciate Dr. Wei Zhang from Department of Animal Science, the University of Maryland for giving great comments on this manuscript.

Author information

Author notes

  1. Zhifu Cui and Lingbin Liu contributed equally to this work.

Authors and Affiliations

  1. Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, 611130, China

    Zhifu Cui, Qing Zhu, Yan Wang, Huadong Yin, Diyan Li, Yaofu Tian & Xiaoling Zhao

  2. College of Animal Science and Technology, Southwest University, Chongqing, China

    Lingbin Liu

  3. Department of Pharmacy, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China

    Gang Shu

  4. Department of Animal Science, Sichuan Agricultural University, Apt 211, Huimin Road, Wenjiang District, Chengdu, 611130, Sichuan Province, China

    Xiaoling Zhao

Authors
  1. Zhifu Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lingbin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qing Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Huadong Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Diyan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yaofu Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Gang Shu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xiaoling Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XZ and ZC conceived and designed the experiments; XZ, ZC, and LL performed the experiments; ZC, QZ, YW, HY, DL, YT, and GS analyzed the data; ZC and XZ wrote the paper.

Corresponding author

Correspondence to Xiaoling Zhao.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All experimental procedures in the present study complied with the recommendations of the Animal Care and Use Committee of Sichuan Agricultural University, and the study was approved by the Animal Care and Use Committee of Sichuan Agricultural University (Sichuan, China) under permit no. YCS-B2018102013.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Cui, Z., Liu, L., Zhu, Q. et al. Inhibin A regulates follicular development via hormone secretion and granulosa cell behaviors in laying hens. Cell Tissue Res 381, 337–350 (2020). https://doi.org/10.1007/s00441-020-03207-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-020-03207-8

Keywords

Search

Navigation