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

Autor: Azim Hedayatpour, Seyed Mehdi Nourashrafeddin, Zahra Khosravizadeh, Farid Abolhassani, Elham Shiri, Soheila Madadi, Shokoofeh Kazemzadeh, Maryam Khanehzad, Parastoo Khanlari
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Male
0301 basic medicine
endocrine system
Glial Cell Line-Derived Neurotrophic Factor Receptors
QH471-489
Blotting
Western
Immunocytochemistry
Endogeny
Biology
Flow cytometry
Mice
03 medical and health sciences
0302 clinical medicine
Endocrinology
Western blot
microRNA
medicine
Animals
Spermatogonial stem cells
Promyelocytic Leukemia Zinc Finger Protein
Cell Self Renewal
Spermatogenesis
Adaptor Proteins
Signal Transducing
Adult Germline Stem Cells
medicine.diagnostic_test
Research
Reproduction
Obstetrics and Gynecology
Spermatogonial stem cell (SSC)
Transfection
Gynecology and obstetrics
Flow Cytometry
Immunohistochemistry
Cell biology
MicroRNAs
Proto-Oncogene Proteins c-kit
miRNA-30a-5p
030104 developmental biology
Animals
Newborn
Reproductive Medicine
030220 oncology & carcinogenesis
Differentiation
RG1-991
Thy-1 Antigens
Inhibitor of Differentiation Proteins
Developmental Biology
Zdroj: Reproductive Biology and Endocrinology, Vol 19, Iss 1, Pp 1-14 (2021)
Reproductive Biology and Endocrinology : RB&E
ISSN: 1477-7827
Popis: 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.
Databáze: OpenAIRE
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