Abstract
Testis is the primary organ of the male reproductive tract in mammals that plays a substantial role in spermatogenesis. Improvement of our knowledge regarding the molecular mechanisms in testicular development and spermatogenesis will be reflected in producing spermatozoa of superior fertility. Evidence showed that N6-Methyladenosine (m6A) plays a dynamic role in post-transcription gene expression regulation and is strongly associated with production traits. However, the role of m6A in bovine testis has not been investigated yet. In this study, we conducted MeRIP-Seq analysis to explore the expression profiles of the m6A and its potential mechanism underlying spermatogenesis in nine bovine testes at three developmental stages (prepuberty, puberty and postpuberty). The experimental animals with triplicate in each stage were chosen based on their semen volume and sperm motility except for the prepuberty bulls and used for testes collection. By applying MeRIP-Seq analysis, a total of 8,774 m6A peaks and 6,206 m6A genes among the studied groups were identified. All the detected peaks were found to be mainly enriched in the coding region and 3′- untranslated regions. The cross-analysis of m6A and mRNA expression exhibited 502 genes with concomitant changes in the mRNA expression and m6A modification. Notably, 30 candidate genes were located in the largest network of protein-protein interactions. Interestingly, four key node genes (PLK4, PTEN, EGR1, and PSME4) were associated with the regulation of mammal testis development and spermatogenesis. This study is the first to present a map of RNA m6A modification in bovine testes at distinct ages, and provides new insights into m6A topology and related molecular mechanisms underlying bovine spermatogenesis, and establishes a basis for further studies on spermatogenesis in mammals.
Highlights
Xia-Nan (XN) cattle, the first specialized beef breed produced by the crossbreeding of French Charolais and Nanyang cattle in China, has important features including fast growth rate and high meat production performance
These mRNA m6A sites were mainly concentrated around the CDS and 3′untranslated regions (3′UTRs) region (95.28%), in agreement with the distributional characteristics of the Pattern m6A level change mRNA level change
The results of qPCR supported that the expression level of the 4 selected DMGs exhibited marked differences between before and after puberty, which displayed a similar tendency with that of the RNA-Seq. These results indicated the putative role of these four genes in bovine testis development and spermatogenesis
Summary
Xia-Nan (XN) cattle, the first specialized beef breed produced by the crossbreeding of French Charolais (male) and Nanyang cattle (female) in China, has important features including fast growth rate and high meat production performance. Different methods and common practices are traditionally used to check male reproductive performance, including a physical assessment of the bull (e.g., volume of the testicles) and semen evaluation (e.g., sperm morphology, concentration, and motility). These procedures offered a considerable background on testicular and epididymal function and quantitative production of sperms, infertility due to major reasons was not well recognized. The testes at different developmental stages in XN cattle were taken as the experimental object in the present study, taking into consideration that identifying key regulators and signaling pathways related to testis development and spermatogenesis will provide valuable insights into improving semen quality
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