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Bta-miR-6531 Regulates Calcium Influx in Bovine Leydig Cells and Is Associated with Sperm Motility

文献类型： 外文期刊

作者： Ding, Qiang 1 ; Ding, Xiuhu 3 ; Xia, Shuwen 1 ; Zhao, Fang 1 ; Chen, Kunlin 1 ; Qian, Yong 1 ; Cao, Shaoxian 1 ; Lin, Zhiping 4 ; Gao, Yundong 5 ; Wang, Huili 1 ; Zhong, Jifeng 2 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Anim Sci, Nanjing 210014, Peoples R China

2.Jiangsu Acad Agr Sci, Key Lab Crop & Anim Integrated Farming, Minist Agr & Rural Affairs, Nanjing 210014, Peoples R China

3.Nanjing Agr Univ, Coll Anim Sci & Technol, Nanjing 210095, Peoples R China

4.Jiangsu Youyuan Dairy Res Inst Co Ltd, Nanjing 211106, Peoples R China

5.Shandong OX Livestock Breeding Co Ltd, Jinan 250131, Peoples R China

关键词： sperm motility; bta-miR-6531; calcium signaling pathway; SNP

期刊名称：GENES （ 影响因子：4.141； 五年影响因子：4.474 ）

ISSN：

年卷期： 2022 年 13 卷 10 期

页码：

收录情况： SCI

摘要： MicroRNAs (miRNAs) play key roles in sperm as the regulatory factors involved in fertility and subsequent early embryonic development. Bta-miR-6531 is specifically a highly enriched miRNA in low-motility sperms in previous study. To investigate the mechanism of bta-miR-6531, 508 shared target genes of bta-miR-6531 were predicted using two miRNA target databases (TargetScan7 and miRWalk). According to the Kyoto Encyclopedia of Genes and Genomes (KEGG), the calcium and cAMP signaling pathways were the most enriched of the target genes. A dual-luciferase assay indicated that bta-miR-6531 targeted ATP2A2 mRNA by binding to the coding sequence region. In bovine Leydig cells, bta-miR-6531 overexpression affected the intracellular calcium concentration by restraining ATP2A2 expression. Moreover, we observed high calcium concentrations and high ATP2A2 protein levels in high-motility sperm compared with those in low-motility sperms. Furthermore, high-linkage single-nucleotide polymorphisms (SNPs) of the pre-bta-miR-6531 sequence were identified that related to sperm traits. Genotype TCTC of bta-miR-6531 showed high sperm motility and density and low deformity rate in Holstein bulls. However, the mutation in pre-miR-6531 did not significantly affect mature bta-miR-6531 expression in the sperm or cell models. Our results demonstrate that bta-miR-6531 might involve in sperm motility regulation by targeting ATP2A2 of the calcium signaling pathway in bovine spermatozoa.

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