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. 2022 Sep;57(9):1082-1087.
doi: 10.1111/rda.14159. Epub 2022 May 26.

Localization and in silico-based functional analysis of miR-202 in bull testis

Affiliations

Localization and in silico-based functional analysis of miR-202 in bull testis

Bushra T Mohammed et al. Reprod Domest Anim. 2022 Sep.

Abstract

Bull fertility is pivotal to the prosperity of the cattle industry worldwide. miR-202 has been shown to be gonad specific and to have key roles in gonad function in different species. To further understand the involvement of miR-202 in bull reproduction, this study aimed to establish its localization in bovine testicular tissue and to identify putative biological functions using bioinformatics approaches. We assessed the miR-202 expression in paraffin-embedded tissue samples collected form an abattoir using in situ hybridization. miR-202 was present in Sertoli cells and in germ cells at different stages of development. Using available databases, a total of 466 predicted gene targets of miR-202 were identified. Functional annotation revealed that miR-202 target genes were mainly associated with protein modification and phosphorylation processes as well as longevity regulating pathway. Moreover, genes in the longevity regulating pathway mapped to PI3K/Akt/mTOR pathway which is involved in promoting proliferation of testicular cells and spermatogenesis. These findings suggest that miR-202 plays important roles in regulating proliferation and viability of testicular cells including somatic and germ cells.

Keywords: bull; in situ hybridization; miR-202; testicular cells.

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Conflict of interest statement

The authors declare that they have no conflict of interests.

Figures

FIGURE 1
FIGURE 1
Representative images of in situ hybridization detection of miR‐202 in sections of bull testes (n = 3 animals). Sertoli cells (St), spermatogonia (sg), primary spermatocytes (Ps), secondary spermatocytes (ss) and spermatids (S) are indicated by white arrows. Blood vessels (Bv) and interstitial cells (ins) are also shown. Sections hybridized with probes against miR‐202 (a, b, c), U6B (d, e) and scrambled sequence controls (f) are shown. Original magnification 200x,400x and 1,000x. Scale bar, 100 μm
FIGURE 2
FIGURE 2
Functional GO terms enriched for target genes of miR‐202 identified using ClueGO and CluePedia (p ≤ .01). Different functional groups are represented by colours. Each node represents a GO term, and node size represents level of significance for term enrichment. Terms are connected based on shared genes. Enriched biological processes (a), cellular components (b) and molecular functions (c) of miR‐202 targets
FIGURE 3
FIGURE 3
Functional KEGG pathways enriched for target genes of miR‐202 identified using ClueGO and CluePedia (p ≤ .05). Different functional groups are represented by colours. Each node represents a GO term, and node size represents level of significance for term enrichment. Pathways are connected based on shared genes

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