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. 2022 Nov 25;13(12):2212.
doi: 10.3390/genes13122212.

Transcriptomic Study of Spermatogenesis in the Testis of Hu Sheep and Tibetan Sheep

Xiaoyu Fu  1 , Yanan Yang  1 , Zunqiang Yan  1 , Miaomiao Liu  1 , Xinrong Wang  1
Affiliations

Transcriptomic Study of Spermatogenesis in the Testis of Hu Sheep and Tibetan Sheep

Xiaoyu Fu et al. Genes (Basel). .

Abstract

Numerous genes involved in male reproduction regulate testis development and spermatogenesis. In this study, the testis tissue transcriptome was used to identify candidate genes and key pathways associated with fecundity in sheep. Histological analysis of testis tissue using hematoxylin-eosin (HE) routine staining was performed for two sheep breeds. Overall, 466 differentially expressed genes (DEGs) were identified between Hu sheep (HS) and Tibetan sheep (TS) through RNA sequencing technology (RNA-Seq), including 226 upregulated and 240 downregulated genes. Functional analysis showed that several terms and pathways, such as "protein digestion and absorption", "cAMP signaling pathway", "focal adhesion", and "p53 signaling pathway" were closely related to testis development and spermatogenesis. Several genes (including COL1A1, COL1A2, COL3A1, SOX9, BCL2, HDC, and GGT5) were significantly enriched in these terms and pathways and might affect the reproduction of sheep by regulating the migration of spermatogenic cells, apoptosis of spermatogenic cells, and secretion of sterol hormones via testicular interstitial cells. Our results provide a theoretical basis for better understanding the molecular mechanisms of reproduction in sheep.

Keywords: Hu sheep; RNA-Seq; Tibetan sheep; histological observation; testis tissue.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Determination of morphologic parameters of testes in Hu sheep and Tibetan sheep. The unit of weight is g, the unit of scrotal circumference is cm, and the unit of the major axis and minor axis is mm. Phenotypic values are shown as Mean ± SD. n = 3 sheep, * p < 0.05, ** p < 0.01.
Figure 2
Figure 2
Histological observation of testicular tissue of sheep with different fecundities. (A,B): Hu sheep testes. (C,D): Tibetan sheep testes. (A,C) represent ×200 magnification. (B,D) represent ×400 magnification. ST: seminiferous tubules; BC: blood capillary; BM: basement membrane; MC: myoid cells; S: spermatogonia; PS: primary spermatocytes; SZ: spermatozoa; Sp: sperm cell; Sc: Sertoli cells.
Figure 3
Figure 3
Transcriptomic analysis of the testis in sheep. (A) Distribution of gene expression levels. (B) PCA analysis of all mapped genes in HS and TS groups. Triangles represent the mean of each group of samples.
Figure 4
Figure 4
Clusters of orthologous groups (COG) function classification of transcriptome unigenes.
Figure 5
Figure 5
Differential expression of genes between Hu and Tibetan sheep groups. (A) Volcanic plot of the differentially expressed genes. (B) Heat map of the differentially expressed gene clustering analysis. TS: Tibetan sheep group; HS: Hu sheep group.
Figure 6
Figure 6
The expression levels of BCL2 in all samples and its relationship with spermatogenic cells and Sertoli cells. (A) The number of spermatogenic cells and Sertoli cells in all samples. (B) The expression levels of BCL2 in all samples.
Figure 7
Figure 7
GO annotation of the differentially expressed genes.
Figure 8
Figure 8
KEGG enrichment analysis of the differentially expressed genes.
Figure 9
Figure 9
(A) Analysis diagram of protein–protein interaction network. Red indicates upregulated genes and blue indicates downregulated genes. (B) Validation of differentially expressed genes by RT-qPCR.
See this image and copyright information in PMC

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