Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep 5;13(18):2815.
doi: 10.3390/ani13182815.

Transcriptome Studies Reveal the N6-Methyladenosine Differences in Testis of Yaks at Juvenile and Sexual Maturity Stages

Shaoke Guo  1   2 , Jie Pei  1   2 , Xingdong Wang  1   2 , Mengli Cao  1   2 , Lin Xiong  1   2 , Yandong Kang  1   2 , Ziqiang Ding  1   2 , Yongfu La  1   2 , Min Chu  1   2 , Pengjia Bao  1   2 , Xian Guo  1   2
Affiliations

Transcriptome Studies Reveal the N6-Methyladenosine Differences in Testis of Yaks at Juvenile and Sexual Maturity Stages

Shaoke Guo et al. Animals (Basel). .

Abstract

Studying the mechanism of spermatogenesis is key to exploring the reproductive characteristics of male yaks. Although N6-methyladenosine (m6A) RNA modification has been reported to regulate spermatogenesis and reproductive function in mammals, the molecular mechanism of m6A in yak testis development and spermatogenesis remains largely unknown. Therefore, we collected testicular tissue from juvenile and adult yaks and found that the m6A level significantly increased after sexual maturity in yaks. In MeRIP-seq, 1702 hypermethylated peaks and 724 hypomethylated peaks were identified. The hypermethylated differentially methylated RNAs (DMRs) (CIB2, AK1, FOXJ2, PKDREJ, SLC9A3, and TOPAZ1) mainly regulated spermatogenesis. Functional enrichment analysis showed that DMRs were significantly enriched in the adherens junction, gap junction, and Wnt, PI3K, and mTOR signaling pathways, regulating cell development, spermatogenesis, and testicular endocrine function. The functional analysis of differentially expressed genes showed that they were involved in the biological processes of mitosis, meiosis, and flagellated sperm motility during the sexual maturity of yak testis. We also screened the key regulatory factors of testis development and spermatogenesis by combined analysis, which included BRCA1, CREBBP, STAT3, and SMAD4. This study indexed the m6A characteristics of yak testicles at different developmental stages, providing basic data for further research of m6A modification regulating yak testicular development.

Keywords: N6-methyladenosine; mRNA; reproduction; spermatogenesis.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Tissue structure and relative m6A level of yak testis at different stages. (a) Tissue structure of yak testis at different stages. SPG—spermatogonia; SC—Sertoli cell; LC—Leydig cell; SPC—spermatocyte; RS—round sperm; SPT—spermatid. (b) Relative m6A levels of yak testis at different stages. (c) Expression of writers in yak testis at different stages. (d) Expression of erasers in yak testis at different stages. (e) Expression of readers in yak testis at different stages (* represents a significant difference in gene expression between the two groups (p < 0.05), ** represents a very significant difference in gene expression between the two groups (p < 0.01), and ns represents no significant difference in gene expression between the two groups (p > 0.05)).
Figure 2
Figure 2
Characteristics of m6A in yak testis before and after sexual maturity. (a) Distribution of reference genome alignment. (b) Violin plot of m6A peak length distribution. (c) Annotation classification of the peak in the gene functional element region. (d) Distribution circle of reads on the reference genome. (e) Statistics of the concentration location of the m6A peak in yak testis. (f) Data visualization analysis of differential m6A peaks in the selected mRNAs (RBM11 and EURL). (g) Motif sequence enriched by the m6A peak.
Figure 3
Figure 3
Results of differentially methylated peaks (DMPs) before and after sexual maturity of yak testis. (a) Statistical histogram of DMPs. (b) Volcano plot of DMP distribution. (c) GO enrichment analysis of DMPs. (d) KEGG enrichment analysis of DMPs.
Figure 4
Figure 4
Results of differentially expressed genes (DEGs) before and after sexual maturity of yak testis. (a) Box diagram of gene FPKM value of each testicular sample. (b) Density distribution curve of gene FPKM value of each testicular sample. (c) Statistical histogram of DEGs. (d) Volcano plot of DEG distribution. (e) Cluster heatmap of DEGs. (f) GO enrichment analysis of DEGs. (g) KEGG enrichment analysis of DEGs.
Figure 5
Figure 5
Combined analysis of m6A-seq and RNA-seq data. (a) Combined four-quadrant map of differentially methylated peaks and differentially expressed genes. (b) Network diagram of 30 differentially co-expressed genes. (c) qRT-PCR results of DEGs. (d) Transcriptome sequencing expression of DEGs (** represents a very significant difference in gene expression between the two groups (p < 0.01)).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Qiu Q., Zhang G., Ma T., Qian W., Wang J., Ye Z., Cao C., Hu Q., Kim J., Larkin D.M., et al. The yak genome and adaptation to life at high altitude. Nat. Genet. 2012;44:946–949. doi: 10.1038/ng.2343. - DOI - PubMed
    1. Ding L., Wang Y., Kreuzer M., Guo X., Mi J., Gou Y., Shang Z., Zhang Y., Zhou J., Wang H., et al. Seasonal variations in the fatty acid profile of milk from yaks grazing on the Qinghai-Tibetan plateau. J. Dairy Res. 2013;80:410–417. doi: 10.1017/S0022029913000496. - DOI - PubMed
    1. Shah M.A., Xu C., Wu S., Zhao W., Luo H., Yi C., Liu W., Cai X. Isolation and characterization of spermatogenic cells from cattle, yak and cattleyak. Anim. Reprod. Sci. 2018;193:182–190. doi: 10.1016/j.anireprosci.2018.04.067. - DOI - PubMed
    1. Cannarella R., Condorelli R.A., Duca Y., La Vignera S., Calogero A.E. New insights into the genetics of spermatogenic failure: A review of the literature. Hum. Genet. 2019;138:125–140. doi: 10.1007/s00439-019-01974-1. - DOI - PubMed
    1. Mäkelä J.A., Koskenniemi J.J., Virtanen H.E., Toppari J. Testis Development. Endocr. Rev. 2019;40:857–905. doi: 10.1210/er.2018-00140. - DOI - PubMed

LinkOut - more resources