Skip to main content
Log in

A functional polymorphism of inhibin alpha subunit at miR-181b-1-3p-binding site regulates proliferation and apoptosis of chicken ovarian granular cells

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

Abstract

INHA, the gene encoding the inhibin alpha subunit, was involved in folliculogenesis in mammals, but no study was reported for its working pathway in birds. Here we hypothesize that gene polymorphism in INHA 3′UTR might influence miRNAs binding efficiency and further affect the function of this gene. Thus, we investigated the association between the 3′UTR single-nucleotide polymorphisms (SNPs) in INHA and the laying performance in chickens and further explore their possible molecular cascades in granulosa cells (GC). Five SNPs were detected in Tianfu green-shell layers and g. 22,178,975 G > A was significantly associated with total egg numbers at the age of 300 days (EN, n = 286). Birds carrying the AA genotype laid more EN than those with GG (P < 0.05). The allele transition from G to A in the 3′UTR of INHA gene destroyed a binding site which was targeted by miR-181b-1-3p. The expression abundances of INHA mRNA increased firstly and then decreased with follicle growing, and reached the top in the sixth largest pre-ovulation follicle, whereas miR-181b-1-3p levels in chicken pre-hierarchical follicles had the contrary tendency. Further studies indicated that high levels of miR-181b-1-3p increased apoptosis and reduced GC proliferation while miR-181b-1-3p inhibitors decreased apoptosis and promoted GC proliferation. Additionally, depression of INHA increased apoptosis and reduced GC proliferation via a caspase-3-dependent mitochondrial pathway. Generally, the mutation in INHA 3ʹUTR was tightly correlated with egg production in chickens, and blocked a binding site of miR-181b-1-3p. miR-181b-1-3p inhibited GC proliferation and promoted apoptosis by targeting INHA.

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

Access this article

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

Similar content being viewed by others

Data availability statement

The data used to support the findings of this study are available from the corresponding author upon request.

References

  • Akbas Y, Takma C (2018) Canonical correlation analysis for studying the relationship between egg production traits and body weight, egg weight and age at sexual maturity in layers. Czech J Anim Sci 50:163–168

    Article  Google Scholar 

  • Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bino GD, Lassota P, Darzynkiewicz Z (1991) The s-phase cytotoxicity of camptothecin. Exp Cell Res 193:27–35

    Article  PubMed  Google Scholar 

  • Bras M, Queenan B, Susin SA (2005) Programmed cell death via mitochondria: different modes of dying. Biochemistry 70:231–239

    CAS  PubMed  Google Scholar 

  • Budihardjo I, Oliver H, Lutter M, Luo X, Wang X (1999) Biochemical Pathways of Caspase Activation During Apoptosis. Annu Rev Cell Dev Bi 15:269–290

    Article  CAS  Google Scholar 

  • Cannon JD, Cherianshaw M, Chaffin CL (2005) Proliferation of rat granulosa cells during the periovulatory interval. Endocrinology 146:414–422

    Article  CAS  PubMed  Google Scholar 

  • Chen F, Jiang X, Chen X, Liu G, Ding J (2008) Effects of Downregulation of Inhibin α Gene Expression on Apoptosis and Proliferation of Goose Granulosa Cells. J Genet Genomics 34:1106–1113

    Article  Google Scholar 

  • Cipolla GA, Park JK, Oliveira L, Loboalves SC, De Almeida RC, Farias TDJ, Lemos DS, Malheiros D, Lavker RM, Petzlerler ML (2016) A 3’UTR polymorphism marks differential KLRG1 mRNA levels through disruption of a miR-584-5p binding site and associates with pemphigus foliaceus susceptibility. BBA-Gene Regul Mech 1859:1306–1313

    CAS  Google Scholar 

  • Clop A, Marcq F, Takeda H, Pirottin D, Tordoir X, Bibe B, Bouix J, Caiment F, Elsen J, Eychenne F (2006) A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep. Nat Genet 38:813–818

    Article  CAS  PubMed  Google Scholar 

  • Cui Z, Liu L, Zhao X, Ran J, Wang Y, Yin H, Li D, Zhu Q (2019) Analysis of Expression and Single Nucleotide Polymorphisms of INHA Gene Associated with Reproductive Traits in Chickens. BioMed Res Int 2019:8572837

    Article  PubMed  PubMed Central  Google Scholar 

  • Den Hurk RV, De Pavert SAV (2001) Localization of an activin/activin receptor system in the porcine ovary. Mol Reprod Dev 60:463–471

    Article  PubMed  Google Scholar 

  • Fleury C, Mignotte B, Vayssiere JL (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84:131–141

    Article  CAS  PubMed  Google Scholar 

  • Francesch A, Estany J, Alfonso L, Iglesias M (1997) Genetic parameters for egg number, egg weight, and eggshell color in three Catalan poultry breeds. Poultry Sci 76:1627–1631

    Article  CAS  Google Scholar 

  • Garrido C, Galluzzi L, Brunet M, Puig P, Didelot C, Kroemer G (2006) Mechanisms of cytochrome c release from mitochondria. Cell Death Differ 13:1423–1433

    Article  CAS  PubMed  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). Reproduction 50:179–181

    Article  CAS  Google Scholar 

  • Goswami D, Conway GS (2005) Premature ovarian failure. Hum Reprod Update 11:391–410

    Article  CAS  PubMed  Google Scholar 

  • Green DR, Reed JC (1998) Mitochondria and Apoptosis. Science 281:1309–1312

    Article  CAS  PubMed  Google Scholar 

  • Hall PA, Levison DA, Woods AL, Yu CCW, Kellock DB, Watkins JA, Barnes DM, Gillett C, Camplejohn RS, Dover R (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

    Article  CAS  PubMed  Google Scholar 

  • Hulshof SCJ, Figueiredo JR, Beckers JF, Bevers MM, Vanderstichele H, Den Hurk RV (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

    Article  CAS  PubMed  Google Scholar 

  • Jiang L, Huang J, Li L, Chen Y, Chen X, Zhao X, Yang D (2015) MicroRNA-93 Promotes Ovarian Granulosa Cells Proliferation Through Targeting CDKN1A in Polycystic Ovarian Syndrome. J Clin Endocr and Metab 100:E729

    Article  Google Scholar 

  • Kadariya I, Wang J, Rehman ZU, Ali H, Riaz H, He J, Bhattarai D, Liu J, Zhang SJ (2015) RNAi-mediated knockdown of inhibin α subunit increased apoptosis in granulosa cells and decreased fertility in mice. J Steroid Biochem 152:161–170

    Article  CAS  Google Scholar 

  • Kang L, Cui X, Zhang Y, Yang C, Jiang Y (2013) Identification of miRNAs associated with sexual maturity in chicken ovary by Illumina small RNA deep sequencing. BMC Genomics 14:352–352

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kowaltowski AJ, Souza-Pinto NCD, Castilho RF, Vercesi AE (2009) Mitochondria and reactive oxygen species. Free Radical Bio Med 47:333–343

    Article  CAS  Google Scholar 

  • Li DY, Zhang L, Yang M, Xu HL, Yin HD, Li Y, Zhu Q (2013) Effect of luteinizing hormone/choriogonadotropin receptor (LHCGR) gene on chicken reproductive traits. Mol Biol Rep 40:7111–7116

    Article  CAS  PubMed  Google Scholar 

  • Li W, Sun L, Chen S, Chen L, Liu Z, Hou X, Chen C, Han Y, Wang C, Li C (2015) Association of inhibin-α gene polymorphisms with follicular cysts in large white sows. Theriogenology 84:839–845

    Article  CAS  PubMed  Google Scholar 

  • Liu L, Li D, Zhao X, Liu Y, Wang Y, Zhu Q (2012) Polymorphism of Prolactin Receptor Gene and its Association with Egg Production Traits in Erlang Mountainous Chicken. Asian J Anim Vet Adv 7:1183–1190

    Article  CAS  Google Scholar 

  • Liu X, Kim CN, Yang J, Jemmerson R, Wang X (1996) Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c. Cell 86:147–157

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Martin SJ, Green DR (1995) Protease activation during apoptosis: Death by a thousand cuts? Cell 82:349–352

    Article  CAS  PubMed  Google Scholar 

  • Matsuda F, Inoue N, Manabe N, Ohkura S (2012) Follicular Growth and Atresia in Mammalian Ovaries: Regulation by Survival and Death of Granulosa Cells. J Reprod Develop 58:44–50

    Article  CAS  Google Scholar 

  • Matsudaminehata F, Inoue N, Goto Y, Manabe N (2006) The Regulation of Ovarian Granulosa Cell Death by Pro- and Anti-apoptotic Molecules. J Reprod Develop 52:695–705

    Article  CAS  Google Scholar 

  • Mcstay GP, Salvesen GS, Green DR (2008) Overlapping cleavage motif selectivity of caspases: implications for analysis of apoptotic pathways. Cell Death Differ 15:322–331

    Article  CAS  PubMed  Google Scholar 

  • Meunier H, Rivier C, Evans RM, Vale W (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

    Article  CAS  Google Scholar 

  • Pilsworth JA, Cochrane DR, Xia Z, Aubert G, Farkkila A, Horlings HM, Yanagida S, Yang W, Lim JLP, Wang YK (2018) TERT promoter mutation in adult granulosa cell tumor of the ovary. Modern Pathol 31:1107–1115

    Article  CAS  Google Scholar 

  • Pirooz HJ, Jafari N, Rastegari M, Fathiroudsari M, Tasharrofi N, Shokri G, Tamadon M, Sazegar H, Kouhkan F (2018) Functional SNP in microRNA-491-5p binding site of MMP9 3’-UTR affects cancer susceptibility. J Cell Biochem 119:5126–5134

    Article  CAS  PubMed  Google Scholar 

  • Rabinovici J, Schwall R, Jaffe RB (1992) Activin-A as an intraovarian modulator: actions, localization, and regulation of the intact dimer in human ovarian cells. J Clin Invest 89:1528–1536

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rah H, Jeon YJ, Ko JJ, Kim JH, Kim Y, Cha SH, Choi Y, Lee WS, Kim NK (2014) Association of inhibin α gene promoter polymorphisms with risk of idiopathic primary ovarian insufficiency in Korean women. Maturitas 77:163–167

    Article  CAS  PubMed  Google Scholar 

  • Ro S, Park C, Young DL, Sanders KM, Yan W (2007) Tissue-dependent paired expression of miRNAs. Nucleic Acids Res 35:5944–5953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Silva JRV, Tharasanit T, Taverne MAM, Der Weijden GCV, Santos RR, Figueiredo JR, Den Hurk RV (2006) The activin-follistatin system and in vitro early follicle development in goats. J Endocrinol 189:113–125

    Article  CAS  PubMed  Google Scholar 

  • Song C, Yang J, Jiang R, Yang Z, Li H, Huang Y, Lan X, Lei C, Ma Y, Qi X (2019) miR-148a-3p regulates proliferation and apoptosis of bovine muscle cells by targeting KLF6. J Cell Physiol 234:15742–15750

    Article  CAS  Google Scholar 

  • Sun Y, Lin Y, Li H, Liu J, Sheng X, Zhang W (2012) 2,5-Hexanedione induces human ovarian granulosa cell apoptosis through BCL-2, BAX, and CASPASE-3 signaling pathways. Arch Toxicol 86:205–215

    Article  CAS  PubMed  Google Scholar 

  • Wang C, Youle RJ (2009) The Role of Mitochondria in Apoptosis. Annu Rev Genetics 43:95–118

    Article  CAS  Google Scholar 

  • Welt CK, Sidis Y, Keutmann HT, Schneyer AL (2002) Activins, Inhibins, and Follistatins: From Endocrinology to Signaling. A Paradigm for the New Millennium. Exp Biol Med 227:724–752

    Article  CAS  Google Scholar 

  • Wen R, Hu S, Xiao Q, Han C, Gan C, Gou H, Liu H, Li L, Xu H, He H (2015) Leptin exerts proliferative and anti-apoptotic effects on goose granulosa cells through the PI3K/Akt/mTOR signaling pathway. J Steroid Biochem 149:70–79

    Article  CAS  Google Scholar 

  • Wu N, Zhu Q, Chen B, Gao J, Xu Z, Li D (2017) High-throughput sequencing of pituitary and hypothalamic microRNA transcriptome associated with high rate of egg production. BMC Genomics 18:255–255

    Article  PubMed  PubMed Central  Google Scholar 

  • Wu W, Hua G, Yang L, Wen Q, Zhang C, Zoheir KMA, Chen S (2009) Association analysis of the INHA gene with litter size in Boer goats. Small Ruminant Res 82:139–143

    Article  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Yang X, Zhou Y, Peng S, Wu L, Lin H, Wang S, Wang H (2012) Differentially expressed plasma microRNAs in premature ovarian failure patients and the potential regulatory function of mir-23a in granulosa cell apoptosis. Reproduction 144:235–244

    Article  CAS  PubMed  Google Scholar 

  • Ying S (1987) Inhibins and Activins: Chemical Properties and Biological Activity. Exp Biol Med 186:253–264

    Article  CAS  Google Scholar 

  • Yuan M, Zhan Q, Duan X, Song B, Zeng S, Chen X, Yang Q, Xia J (2013) A functional polymorphism at miR-491-5p binding site in the 3’-UTR of MMP-9 gene confers increased risk for atherosclerotic cerebral infarction in a Chinese population. Atherosclerosis 226:447–452

    Article  CAS  PubMed  Google Scholar 

  • Zhao J, Taverne MAM, Der Weijden GCV, Bevers MM, Den Hurk RV (2001) Effect of Activin A on In Vitro Development of Rat Preantral Follicles and Localization of Activin A and Activin Receptor II. Biol Reprod 65:967–977

    Article  CAS  PubMed  Google Scholar 

  • Zhou X, Xian D, Xia J, Tang Y, Li W, Chen X, Zhou Z, Lu D, Feng X (2017) MicroRNA-34c is regulated by p53 and is involved in sevoflurane-induced apoptosis in the developing rat brain potentially via the mitochondrial pathway. Mol Med Rep 15:2204–2212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou Y, Tian W, Zhang M, Ren T, Sun G, Jiang R, Han R, Kang X, Yan F (2019) Transcriptom analysis revealed regulation of dexamethasone induced microRNAs in chicken thymus. J Cell Biochem 120:6570–6579

    Article  CAS  PubMed  Google Scholar 

  • Zhu L, Yuan H, Guo C, Lu Y, Deng S, Yang Y, Wei Q, Wen L, He Z (2012) Zearalenone induces apoptosis and necrosis in porcine granulosa cells via a caspase-3- and caspase-9-dependent mitochondrial signaling pathway. J Cell Physiol 227:1814–1820

    Article  CAS  PubMed  Google Scholar 

  • Zi X, Xu H, Wang Y (2012) Variation in sequences and mRNA expression levels of inhibin subunits α (INHA) and βA (INHBA) genes between prolific and nonprolific goat breeds. Mol Reprod Dev 79:238–238

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

We would like to thank the China Agriculture Research System of Ministry of Agriculture and Rural Areas (Grant No. CARS-41), and National Natural Science Foundation of China (Grant No. 31872347) for their financial supports.

Author information

Authors and Affiliations

Authors

Contributions

ZC and XS conceived and designed the experiments; ZC, XXZ and FL performed the experiments; ZC, LL, FA, QZ, YW, HY, DL, and YT analyzed the data; ZC and XLZ wrote the paper.

Corresponding author

Correspondence to Xiaoling Zhao.

Ethics declarations

Conflicts of interest

The authors declare that there are no conflicts of interest.

Ethical approval

Due diligence was followed bearing in mind the guidelines of the Sichuan Agricultural University (SAU) Laboratory Animal Welfare and Ethics. All experiments on animals carried out in this study, was endorsed by the Institutional Animal Care and Use committee of Sichuan Agricultural University with the permit number: YYS130125.

Additional information

Publisher’s Note

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

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (XLSX 42 KB)

Supplementary file2 (DOCX 18 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Cui, Z., Shen, X., Zhang, X. et al. A functional polymorphism of inhibin alpha subunit at miR-181b-1-3p-binding site regulates proliferation and apoptosis of chicken ovarian granular cells. Cell Tissue Res 384, 545–560 (2021). https://doi.org/10.1007/s00441-020-03356-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-020-03356-w

Keywords

Navigation