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.
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Data availability statement
The data used to support the findings of this study are available from the corresponding author upon request.
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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.
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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.
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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.
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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
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DOI: https://doi.org/10.1007/s00441-020-03356-w