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Identification of miR-34c-5p as a potential regulatory molecule associated with testicular development of black goat

ABSTRACT: The growth and mature of Sertoli cells (SCs) are important part of testicular development and miRNAs are considered as novel regulatory molecules involved in it. Here we systematically screened and analyzed the key miRNAs participating in testicular development of black goat, as well as identifying underlying mechanism of candidate mRNAs contributes to spermatogenesis. The miRNA transcriptional profiles of testicular tissues at 3 days old (D3), 30 days old (D30), 90 days old (D90) and 180 days old (D180) were constructed and comparative analyzed. Results showed that the most differentially expressed (DE) miRNAs were found between the testicular tissues of D30 and D90 black goats. Moreover, the predicted target genes of DE miRNAs are mainly enriched in biological processes including cell proliferation, cell composition and enriched in signaling pathways including p53, FoxO and PI3K-Akt. The top one DE miRNA (miR-34c-5p), which significantly highly expressed in the testis of D90 when compared to that of D30, was mainly located within SCs and targeted to IGF1R. By performing miR-34c-5p overexpression and RNA interference, we found that the function of miR-34c-5p in SCs was inhibited cell proliferation, increased apoptosis rate, and significantly reduced the expression of growth factors (CXCL12, BMP4, GDNF and IGF1). In summary, our study suggests that miR-34c-5p can serve as a potential target for regulation of testicular development and a reference marker of sex maturation applicated in black goat genetic breeding.

ORGANISM(S): Capra hircus

PROVIDER: GSE279238 | GEO | 2025/10/01

REPOSITORIES: GEO

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Project description:Sertoli cells (SCs), the only somatic constituent of the testicular seminiferous epithelium, are vital to spermatogenesis. We previously found that vitamin C (ascorbic acid, AA) can reprogram the transcriptome, and promote the proliferation and reproductive function of pig immature SCs (iSCs). However, the global change of microRNAs (miRNA) expression and its effect on pig iSCs as induced by vitamin C treatment is still unknown. Here, we performed small RNA sequencing on pig iSCs after 250μM AA2P (L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate) treatment for 36h. Total number of detected miRNAs ranged from 326-335 known, and 400-570 novel miRNAs. Of the top ten highly expressed miRNAs, we found that 8 were common (miR-21-5p, let-7i-5p, miR-30a-5p, let-7f-5p, let-7g, miR-100, miR-10a-5p and miR-30d), which were predicted to target mRNAs involved in cell development and differentiation. We identified 78 differentially expressed (DE) miRNAs (|log2 foldchange|≥1; Padj.<0.05), including 7 known and 71 novel miRNAs. We further selected 13 highly and stably expressed DE miRNAs (4 up-regulated: miR-184, novel-miR-610, novel-miR-316 and novel-miR-1274; 9 down-regulated: miR-222, miR-221-5p, miR-221-3p, miR-210, miR-146b, miR-146a-5p, novel-miR-182, novel-miR-1088 and novel-miR-1016), and performed integrated analysis on the miRNA-mRNA regulatory network. DE mRNAs negatively targeted by these 13 DE miRNAs were enriched in multiple GO and KEGG signaling pathways (e.g. pyruvate and steroid metabolic processes, developmental process in reproduction, response to oxidative stress, Glycolysis/Gluconeogenesis and HIF-1). We validated 8 DE miRNAs and their 12 DE mRNA targets, with expression patterns consistent with (mi)RNA-seq results. Taken together, our findings demonstrate that vitamin C could induce the global change of miRNAs, which possibly regulate cell proliferation, energy metabolism and male reproduction as induced by vitamin C treatment on pig iSCs.

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Identification of miR-34c-5p as a potential regulatory molecule associated with testicular development of black goat

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