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

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...

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Bibliographic Details
Published in:Genes 2022-10, Vol.13 (10), p.1788
Main Authors: Ding, Qiang, Ding, Xiuhu, Xia, Shuwen, Zhao, Fang, Chen, Kunlin, Qian, Yong, Cao, Shaoxian, Lin, Zhiping, Gao, Yundong, Wang, Huili, Zhong, Jifeng
Format: Article
Language:English
Subjects:
Biotechnology
Calcium (intracellular)
Calcium influx
Calcium signalling
Calcium, Dietary
Cattle
Cell culture
Dehydrogenases
Embryogenesis
Fertility
Genes
Genetic aspects
Genetic regulation
Genomes
Health aspects
Leydig cells
MicroRNA
MicroRNAs
miRNA
Motility
mRNA
Mutagenesis
Mutation
Proteins
Signal transduction
Single-nucleotide polymorphism
Sperm
Spermatozoa
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Summary: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.
ISSN:2073-4425
2073-4425
DOI:10.3390/genes13101788