Determination of the Excitatory Effects of MicroRNA-30 in the Self-Renewal and Differentiation Process of Neonatal Mouse Spermatogonial Stem Cells

Abstract

Background: Spermatogonial stem cells (SSCs) are considered as special stem cells since they have the ability of self-renewal, differentiation, and transferring genetic information to the next generation. Also, they considered as vital players in initiating and preserving spermatogenesis. The fate decisions of SSCs are mediated by intrinsic and extrinsic factors, among which microRNAs (miRNAs) are one of the most essential factors in spermatogenesis among endogenous regulators. However, the mechanisms by which individual miRNAs regulate self-renewal and differentiation of SSCs are unclear. The present study aimed to evaluate the impact of miRNA-30 mimic on fate determinations of SSCs. Materials and Methods: The obtained SSCs from neonatal mice (3-6 days old) were purified by MACS and flow cytometry with a promyelocytic leukemia zinc-finger marker. Then, the cultured cells were transfected with miRNA- 30 mimic, and finally, the changes in expressing ID4 and c-kit proteins were assessed by western blot analysis. Results: According to flow cytometry findings, the percentage of SSC purity was about 98.32. The expression of ID4 protein and colonization increased significantly through the transfection of miRNA-30 mimic (P<0.05). Conclusion: The miRNA-30 controls spermatogonial stem cell self-renewal and differentiation, which may have significant implications for treating male infertility. [GMJ.2020;9:e1829]

MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs)

“…A considerable point in this regard is that although these markers have been introduced by other researchers as possible markers for SSC identity, no exclusive marker has yet been recommended for purification of SSCs. Other markers such as Plzf, α6, β1, and GFR α1 have also been used for purification of SSCs in the literature [ 31 , 32 , 38 ]. In this study, the high percentage of expression for these markers confirmed that the collected cells were undifferentiated since SSCs first appear at 3–6 days postpartum in mice.…”

“…Melting curve analysis was used to identify the non-specific real-time PCR products. U6 snRNA was selected as an internal control gene and the miR-30a-5p expression was calculated through 2-∆∆CT method [ 31 , 32 ]. Quantitative real-time PCR was done in triplicate for each sample.…”

MicroRNAs in spermatogenesis dysfunction and male infertility: clinical phenotypes, mechanisms and potential diagnostic biomarkers

“…In a previous study, the proliferation or undifferentiation marker ID4 was considerably upregulated in SSCs treated with an miR-30 mimic, suggesting that miR-30 regulates the self-renewal capability of SSCs ( 83 ). miR-31-5p inhibits proliferation and DNA synthesis and increases early and late apoptosis of human SSCs by targeting JAZF1 and cyclin A2 ( 84 ).…”

Updated Detailed Review of Trachyspermum Ammi: Composition, Applications and Pharmacological Profile

“…All the inventors of different countries representing the particular invention, along with the patent number, title of invention, granting country, and authority, have been listed in this table 1. The chemical composition of Ajwain oil with different major constituents including thymol 28 , γ-terpinene, and p-cymene 2,29 carvone, limonene, and dillapiole and carvacrol 2 . Fruit extract showed antispasmodic and antihypertensive effects by preventing contraction caused by K+.…”