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MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs)
BACKGROUND: The importance of spermatogonial stem cells (SSCs) in spermatogenesis is crucial and intrinsic factors and extrinsic signals mediate fate decisions of SSCs. Among endogenous regulators, microRNAs (miRNAs) play critical role in spermatogenesis. However, the mechanisms which individual miR...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8188658/ https://www.ncbi.nlm.nih.gov/pubmed/34108007 http://dx.doi.org/10.1186/s12958-021-00758-5 |
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author | Khanehzad, Maryam Nourashrafeddin, Seyed Mehdi Abolhassani, Farid Kazemzadeh, Shokoofeh Madadi, Soheila Shiri, Elham Khanlari, Parastoo Khosravizadeh, Zahra Hedayatpour, Azim |
author_facet | Khanehzad, Maryam Nourashrafeddin, Seyed Mehdi Abolhassani, Farid Kazemzadeh, Shokoofeh Madadi, Soheila Shiri, Elham Khanlari, Parastoo Khosravizadeh, Zahra Hedayatpour, Azim |
author_sort | Khanehzad, Maryam |
collection | PubMed |
description | BACKGROUND: The importance of spermatogonial stem cells (SSCs) in spermatogenesis is crucial and intrinsic factors and extrinsic signals mediate fate decisions of SSCs. Among endogenous regulators, microRNAs (miRNAs) play critical role in spermatogenesis. However, the mechanisms which individual miRNAs regulate self- renewal and differentiation of SSCs are unknown. The aim of this study was to investigate effects of miRNA-30a-5p inhibitor on fate determinations of SSCs. METHODS: SSCs were isolated from testes of neonate mice (3–6 days old) and their purities were performed by flow cytometry with ID4 and Thy1 markers. Cultured cells were transfected with miRNA- 30a-5p inhibitor. Evaluation of the proliferation (GFRA1, PLZF and ID4) and differentiation (C-Kit & STRA8) markers of SSCs were accomplished by immunocytochemistry and western blot 48 h after transfection. RESULTS: Based on the results of flow cytometry with ID4 and Thy1 markers, percentage of purity of SSCs was about 84.3 and 97.4 % respectively. It was found that expression of differentiation markers after transfection was significantly higher in miRNA-30a- 5p inhibitor group compared to other groups. The results of proliferation markers evaluation also showed decrease of GFRA1, PLZF and ID4 protein in SSCs transfected with miRNA-30a-5p inhibitor compared to the other groups. CONCLUSIONS: It can be concluded that inhibition of miRNA-30a-5p by overexpression of differentiation markers promotes differentiation of Spermatogonial Stem Cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12958-021-00758-5. |
format | Online Article Text |
id | pubmed-8188658 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-81886582021-06-10 MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) Khanehzad, Maryam Nourashrafeddin, Seyed Mehdi Abolhassani, Farid Kazemzadeh, Shokoofeh Madadi, Soheila Shiri, Elham Khanlari, Parastoo Khosravizadeh, Zahra Hedayatpour, Azim Reprod Biol Endocrinol Research BACKGROUND: The importance of spermatogonial stem cells (SSCs) in spermatogenesis is crucial and intrinsic factors and extrinsic signals mediate fate decisions of SSCs. Among endogenous regulators, microRNAs (miRNAs) play critical role in spermatogenesis. However, the mechanisms which individual miRNAs regulate self- renewal and differentiation of SSCs are unknown. The aim of this study was to investigate effects of miRNA-30a-5p inhibitor on fate determinations of SSCs. METHODS: SSCs were isolated from testes of neonate mice (3–6 days old) and their purities were performed by flow cytometry with ID4 and Thy1 markers. Cultured cells were transfected with miRNA- 30a-5p inhibitor. Evaluation of the proliferation (GFRA1, PLZF and ID4) and differentiation (C-Kit & STRA8) markers of SSCs were accomplished by immunocytochemistry and western blot 48 h after transfection. RESULTS: Based on the results of flow cytometry with ID4 and Thy1 markers, percentage of purity of SSCs was about 84.3 and 97.4 % respectively. It was found that expression of differentiation markers after transfection was significantly higher in miRNA-30a- 5p inhibitor group compared to other groups. The results of proliferation markers evaluation also showed decrease of GFRA1, PLZF and ID4 protein in SSCs transfected with miRNA-30a-5p inhibitor compared to the other groups. CONCLUSIONS: It can be concluded that inhibition of miRNA-30a-5p by overexpression of differentiation markers promotes differentiation of Spermatogonial Stem Cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12958-021-00758-5. BioMed Central 2021-06-09 /pmc/articles/PMC8188658/ /pubmed/34108007 http://dx.doi.org/10.1186/s12958-021-00758-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Khanehzad, Maryam Nourashrafeddin, Seyed Mehdi Abolhassani, Farid Kazemzadeh, Shokoofeh Madadi, Soheila Shiri, Elham Khanlari, Parastoo Khosravizadeh, Zahra Hedayatpour, Azim MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title | MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title_full | MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title_fullStr | MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title_full_unstemmed | MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title_short | MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs) |
title_sort | microrna-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (sscs) |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8188658/ https://www.ncbi.nlm.nih.gov/pubmed/34108007 http://dx.doi.org/10.1186/s12958-021-00758-5 |
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