miR-204 Regulates the Proliferation of Dairy Goat Spermatogonial Stem Cells via Targeting to Sirt1

Abstract

The regulation of spermatogonial stem cell (SSC) proliferation and self-renewal is a complex process. Several studies on the microRNA regulation of mammalian spermatogenesis have been reported. Here, we predicted miRNA targeting of Sirt1, and a dual luciferase experiment confirmed that miR-204 interacted with the Sirt1 3'-untranslated region (3'-UTR). The expression of miR-204 and Sirt1 in dairy goat testicles was investigated, and the results showed that the expression pattern of Sirt1 was similar to that of miR-204 in the temporal-spatial distribution. The over-expression of Sirt1 in goat SSCs can promote SSCs' self-renewal gene expression and cell proliferation. Furthermore, miRNA sequencing results showed that Sirt1 had a higher expression level in dairy goat CD49f(+) and CD90(+) SSCs, but the expression level of miR-204 was lower. In an in vitro assay, Sirt1 was significantly down-regulated in dairy goat SSCs when transfected with miR-204 mimics, indicating that Sirt1 was a target of miR-204 in the dairy goat. On the basis of the results of RT-qPCR, fluorescence-activated cell sorting (FACS), and western blotting, we found that the over-expression of Sirt1 in goat SSCs can promote cellular proliferation and change self-renewal and pluripotent gene expression. Thus, miR-204 was involved in the regulation of dairy goat SSCs proliferation via Sirt1.

Role of bta‐miR‐204 in the regulation of adipocyte proliferation, differentiation, and apoptosis

“…However, no significantly alteration was observed in cell proliferation. In addition, previous studies have demonstrated that miR‐204 inhibits the proliferation of dairy goat spermatogonial stem cells and osteosarcoma cells via targeting to SIRT1 (Niu et al, ; Shi et al, ). In contrast, miR‐204 enhances cell proliferation in MCF‐7 and MDA‐MB‐231 breast cancer cells by downregulating tumor suppressor genes (Lee, Lee, Bae, Kang, & Kim, ).…”

SIRT1 regulates macrophage self‐renewal

“…Reversely, overexpression of SIRT1 increased the low proliferative capacities of bone marrow‐derived macrophages. Interestingly, SIRT1 has also been shown to positively affect the self‐renewal capacity of several different types of stem cells (Rathbone et al , ; Yuan et al , ; Niu et al , ; Zhang et al , ), including ES and iPS cells (Calvanese et al , ; Saunders et al , ). These data suggest that high levels of SIRT1 are important for active self‐renewal potential in macrophages and stem cells.…”

MicroRNA‐10b regulates the renewal of spermatogonial stem cells through Kruppel‐like factor 4

“…Many miRs participate in regulating developmental stages of the mouse testis and controlling spermatogenesis . For example, miR‐20 and miR‐106 maintain SSC self‐renewal through the inhibition of signal transducer and activator of transcription 3 and cyclin‐D1 genes, which are specifically expressed in mouse SSCs, miR‐184 inhibits SSC differentiation through the inhibition of the nuclear receptor corepressor 2 gene, enabling functional sperms to be expressed in the testes of mice after birth, miR‐204 regulates the proliferation of goat SSCs through the target gene sirtuin‐1, and miR‐21 regulates the transcription factor ETV5 to maintain SSC self‐renewal . In the present study, we used qRT‐PCR analysis to show that miR‐10b was expressed in mouse testis at different ages.…”