A Comprehensive Sequencing Analysis of Testis-Born miRNAs in Immature and Mature Indigenous Wandong Cattle (Bos taurus)

Hongyu Liu; Ibrar Muhammad Khan; Yong Liu; Nazir Muhammad Khan; Kaiyuan Ji; Huiqun Yin; Wenliang Wang; Xinqi Zhou; Yunhai Zhang

doi:10.3390/genes13122185

A Comprehensive Sequencing Analysis of Testis-Born miRNAs in Immature and Mature Indigenous Wandong Cattle (Bos taurus)

Genes (Basel). 2022 Nov 23;13(12):2185. doi: 10.3390/genes13122185.

Authors

Hongyu Liu¹, Ibrar Muhammad Khan^{1

2}, Yong Liu², Nazir Muhammad Khan³, Kaiyuan Ji¹, Huiqun Yin⁴, Wenliang Wang¹, Xinqi Zhou¹, Yunhai Zhang¹

Affiliations

¹ Anhui Provincial Laboratory of Local Livestock and Poultry Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.
² Anhui Province Key Laboratory of Embryo Development and Reproduction Regulation, Anhui Province Key Laboratory of Environmental Hormone and Reproduction, School of Biological and Food Engineering, Fuyang Normal University, Fuyang 236037, China.
³ Department of Zoology, University of Science and Technology, Bannu 28100, Pakistan.
⁴ Reproductive Medicine Center, the 901st Hospital, Hefei 230031, China.

Abstract

Micro RNAs (miRNAs) have been recognized as important regulators that are indispensable for testicular development and spermatogenesis. miRNAs are endogenous transcriptomic elements and mainly regulate the gene expression at post-transcriptional levels; however, the key role of miRNA in bovine testicular growth is not clearly understood. Thus, supposing to unveil the transcriptomics expression changes in the developmental processes of bovine testes, we selected three immature calves and three sexually mature bulls of the local Wandong breed for testicular-tissue sample collection. The cDNA libraries of experimental animals were established for RNA-sequencing analysis. We detected the miRNA expression in testes by using high-throughput sequencing technology, and bioinformatics analysis followed. The differentially expressed (DE) data showed that 151 miRNAs linked genes were significantly DE between immature and mature bull testes. Further, in detail, 64 were significantly up-regulated and 87 were down-regulated in the immature vs. mature testes (p-value < 0.05). Pathway analyses for miRNA-linked genes were performed and identified JAG2, BCL6, CFAP157, PHC2, TYRO3, SEPTIN6, and BSP3; these genes were involved in biological pathways such as TNF signaling, T cell receptor, PI3KAkt signaling, and functions affecting testes development and spermatogenesis. The DE miRNAs including MIR425, MIR98, MIR34C, MIR184, MIR18A, MIR136, MIR15A, MIR1388 and MIR210 were associated with cattle-bull sexual maturation and sperm production. RT-qPCR validation analysis showed a consistent correlation to the sequencing data findings. The current study provides a good framework for understanding the mechanism of miRNAs in the development of testes and spermatogenesis.

Keywords: bovine; miRNA; spermatogenesis; testicular growth; total RNA-Seq.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cattle / genetics
Male
MicroRNAs* / genetics
MicroRNAs* / metabolism
Semen / metabolism
Spermatogenesis / genetics
Spermatozoa / metabolism
Testis* / metabolism

Substances

MicroRNAs

Grants and funding

This research was funded by National Natural Science Foundation of China (31101696); Science and Technology Major Project of Anhui Province (202103a06020016); Anhui Provincial Livestock Genebank and Anhui Province Modern Agriculture Industry (Cattle, Goat and Sheep) Technology System (AHCYJSTX-07).