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. 2022 Jan;101(1):101558.
doi: 10.1016/j.psj.2021.101558. Epub 2021 Oct 21.

Epididymal mRNA and miRNA transcriptome analyses reveal important genes and miRNAs related to sperm motility in roosters

Kai Xing  1 , Yu Chen  2 , Liang Wang  2 , Xueze Lv  2 , Zheng Li  3 , Xiaolong Qi  1 , Xiangguo Wang  1 , Longfei Xiao  1 , Hemin Ni  1 , Yong Guo  1 , Xihui Sheng  4
Affiliations

Epididymal mRNA and miRNA transcriptome analyses reveal important genes and miRNAs related to sperm motility in roosters

Kai Xing et al. Poult Sci. 2022 Jan.

Abstract

Sperm motility is a crucial trait in chicken production, and the epididymis is an essential organ in the reproductive system. Currently, the molecular mechanisms underlying sperm motility in the epididymis are unclear. In this study, 8 cDNA libraries and eight miRNA libraries were constructed from roosters (4 chickens per group) with diverse sperm motility. After a comparative analysis of epididymal transcriptomes, we detected 84 differentially expressed genes (DEGs) using the edgeR package. Integrated interpretation of DEGs indicated that MMP9, SLN, WT1, PLIN1, and LRRIQ1 are the most promising candidate genes affecting sperm motility in the epididymis of roosters. MiR-146a, mir-135b, and mir-205 could play important regulatory roles in sperm maturation, capacitation, and motility. Additionally, a comprehensive analysis of the mRNA and miRNAs transcriptomes in silico identified a promising gene-miRNA pair miR-135b-HPS5, which may be a vital regulator of sperm motility in the epididymis. Our findings provide novel integrated information of miRNAs and genes that shed light on the regulatory mechanisms of fertility in roosters.

Keywords: epididymis; miRNA; rooster; sperm motility; transcriptome.

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Figures

Figure 1
Figure 1
Gene expression and hierarchical clustering of all samples. (A) Gene expression level across all samples. (B) Hierarchical clustering of all samples.
Figure 2
Figure 2
Gene expression in the epididymis of roosters with high sperm motility (HSM) and low sperm motility (LSM). (A) Venn diagrams showing the total number of shared and unique expressed genes in each group. (B) Volcano plot of differentially expressed genes; significantly upregulated genes are represented as red dots and significantly downregulated genes are represented as green dots. (C) Number of significantly upregulated genes and downregulated genes.
Figure 3
Figure 3
Heatmap of DEGs and their enriched GO terms. Abbreviations: DEGs, differentially expressed genes; GO, Gene Ontology.
Figure 4
Figure 4
Expression scatter plot of the DEMs between HSM and LSM groups. Red dot represents differently expressed miRNAs with fold change ≥ 2 and P-value ≤ 0.05. X-axis values are average log2 (counts per million) and y-axis values are log2 (fold change). Abbreviations: DEMs, Differentially expressed miRNAs; HSM, high sperm motility; LSM, low sperm motility.
Figure 5
Figure 5
Validation of DEGs and DEMs expression level using next generation sequencing (NGS) and qPCR. Abbreviations: DEGs, differentially expressed genes; DEMs, Differentially expressed miRNAs.
Figure 6
Figure 6
Putative regulatory relationships between differentially expressed genes (DEGs) and miRNAs (DEMs). (A) Potential network of genes and miRNAs differentially expressed between HSM and LSM roosters. Squares represent miRNAs, and ellipses represent DEGs, respectively. Upregulated genes or miRNAs are shown in red, and downregulated genes or miRNAs are shown in green to LSM. The network diagram was generated using Cytoscape. (B) The correlation coefficient and P-value of each miRNA-gene pair. R means pearson correlation value. Abbreviations: HSM, high sperm motility; LSM, low sperm motility.
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