Cargando…

Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells

The present study investigated the dynamic expression and regulatory mechanism of transforming growth factor β (TGF-β) signaling involved in embryonic stem cells (ESCs) differentiation into male germ cells. Candidate genes involved in TGF-β signaling pathway were screened from RNA-sequencing (RNA-se...

Descripción completa

Detalles Bibliográficos
Autores principales: Zuo, Qisheng, Jin, Kai, Zhang, Yani, Song, Jiuzhou, Li, Bichun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434085/
https://www.ncbi.nlm.nih.gov/pubmed/28495881
http://dx.doi.org/10.1042/BSR20170179
_version_ 1783406410664509440
author Zuo, Qisheng
Jin, Kai
Zhang, Yani
Song, Jiuzhou
Li, Bichun
author_facet Zuo, Qisheng
Jin, Kai
Zhang, Yani
Song, Jiuzhou
Li, Bichun
author_sort Zuo, Qisheng
collection PubMed
description The present study investigated the dynamic expression and regulatory mechanism of transforming growth factor β (TGF-β) signaling involved in embryonic stem cells (ESCs) differentiation into male germ cells. Candidate genes involved in TGF-β signaling pathway were screened from RNA-sequencing (RNA-seq), which were further validated by quantitative real-time PCR (qRT-PCR). Bone morphogenetic protein 4 (BMP4) was used to induce differentiation of ESCs in vitro. Inhibition of TGF-β signaling pathway was reflected by Western blot of SMAD2 and SMAD5 expression. Differentiating efficiency of germ cells was evaluated by immunofluorescence and fluorescence-activated cell sorting (FACS). Germ cell marker genes were assessed by qRT-PCR in the differentiation process, with activation or inhibition of TGF-β signaling pathway. In the process of in vitro induction, SMAD2 and SMAD5 were found to significantly up-regulated in BMP4 group versus the control and inhibition groups after 4 and 14 days. Expression of CKIT, CVH, DAZL, STRA8, and INTEGRIN α6 were significantly increased in the BMP4 group compared with the control group, while down-regulated in the inhibition groups. The proportion of germ cell-like cells was decreased from 17.9% to 2.2% after 4 days induction, and further decreased from 14.1% to 2.1% after 14 days induction. Correspondingly, expression of marker genes in germ cells was significantly lower. In vivo inhibition of TGF-β signaling pathway reduced germ cells formation from 5.5% to 1.6%, and down-regulated the expression of CKIT, CVH, DAZL, STRA8, and INTEGRIN α6. In conclusion, our study reveals the mechanism regulating spermatogonial stem cells (SSCs) and lays the basis for further understanding of the regulatory network.
format Online
Article
Text
id pubmed-6434085
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Portland Press Ltd.
record_format MEDLINE/PubMed
spelling pubmed-64340852019-04-12 Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells Zuo, Qisheng Jin, Kai Zhang, Yani Song, Jiuzhou Li, Bichun Biosci Rep Research Articles The present study investigated the dynamic expression and regulatory mechanism of transforming growth factor β (TGF-β) signaling involved in embryonic stem cells (ESCs) differentiation into male germ cells. Candidate genes involved in TGF-β signaling pathway were screened from RNA-sequencing (RNA-seq), which were further validated by quantitative real-time PCR (qRT-PCR). Bone morphogenetic protein 4 (BMP4) was used to induce differentiation of ESCs in vitro. Inhibition of TGF-β signaling pathway was reflected by Western blot of SMAD2 and SMAD5 expression. Differentiating efficiency of germ cells was evaluated by immunofluorescence and fluorescence-activated cell sorting (FACS). Germ cell marker genes were assessed by qRT-PCR in the differentiation process, with activation or inhibition of TGF-β signaling pathway. In the process of in vitro induction, SMAD2 and SMAD5 were found to significantly up-regulated in BMP4 group versus the control and inhibition groups after 4 and 14 days. Expression of CKIT, CVH, DAZL, STRA8, and INTEGRIN α6 were significantly increased in the BMP4 group compared with the control group, while down-regulated in the inhibition groups. The proportion of germ cell-like cells was decreased from 17.9% to 2.2% after 4 days induction, and further decreased from 14.1% to 2.1% after 14 days induction. Correspondingly, expression of marker genes in germ cells was significantly lower. In vivo inhibition of TGF-β signaling pathway reduced germ cells formation from 5.5% to 1.6%, and down-regulated the expression of CKIT, CVH, DAZL, STRA8, and INTEGRIN α6. In conclusion, our study reveals the mechanism regulating spermatogonial stem cells (SSCs) and lays the basis for further understanding of the regulatory network. Portland Press Ltd. 2017-07-07 /pmc/articles/PMC6434085/ /pubmed/28495881 http://dx.doi.org/10.1042/BSR20170179 Text en © 2017 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Articles
Zuo, Qisheng
Jin, Kai
Zhang, Yani
Song, Jiuzhou
Li, Bichun
Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title_full Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title_fullStr Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title_full_unstemmed Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title_short Dynamic expression and regulatory mechanism of TGF-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
title_sort dynamic expression and regulatory mechanism of tgf-β signaling in chicken embryonic stem cells differentiating into spermatogonial stem cells
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434085/
https://www.ncbi.nlm.nih.gov/pubmed/28495881
http://dx.doi.org/10.1042/BSR20170179
work_keys_str_mv AT zuoqisheng dynamicexpressionandregulatorymechanismoftgfbsignalinginchickenembryonicstemcellsdifferentiatingintospermatogonialstemcells
AT jinkai dynamicexpressionandregulatorymechanismoftgfbsignalinginchickenembryonicstemcellsdifferentiatingintospermatogonialstemcells
AT zhangyani dynamicexpressionandregulatorymechanismoftgfbsignalinginchickenembryonicstemcellsdifferentiatingintospermatogonialstemcells
AT songjiuzhou dynamicexpressionandregulatorymechanismoftgfbsignalinginchickenembryonicstemcellsdifferentiatingintospermatogonialstemcells
AT libichun dynamicexpressionandregulatorymechanismoftgfbsignalinginchickenembryonicstemcellsdifferentiatingintospermatogonialstemcells