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| Study on the role of JAK/STAT signaling pathway during chicken spermatogonial stem cells generation based on RNA-Seq |
| ZHANG Lei, ZUO Qi-sheng, LI Dong, LIAN Chao, Kamel E Ahmed, TANG Bei-bei, SONG Jiu-zhou, ZHANG Ya-ni, LI Bi-chun |
1、Jiangsu Province Key Laboratory of Animal Breeding and Molecular Design, College of Animal Science and Technology,Yangzhou University, Yangzhou 225009, P.R.China
2、Animal & Avian Sciences, University of Maryland, Washington, D.C. MD20741, USA
3、Department of Anatomy and Embryology, College of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt |
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摘要 Spermatogonial stem cells (SSCs) form the foundation for spermatogenesis and sustain male fertility. To explore the regulatory mechanisms of chicken SSCs generation, we obtained highly purified chicken embryonic stem cells (ESCs), primordial germ cells (PGCs) and SSCs by fluorescence-activated cell sorting (FACS). High-throughput analysis methods (RNA-Seq) were used to sequence the transcriptome level of these cells. Gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were used to analyze RNA-Seq results. BMP4 was used to induce chicken ESCs differentiation to SSCs-like cells in vitro. The quantitative real-time (qRT)-PCR was used to detect the expression changes of the key genes. The results showed that 22 relevant critical pathways were found by RNA-Seq, one of them was the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Total of 103 related genes were detected in this pathway. Protein-protein interactions analysis found that 87 proteins were significantly related to 19 key proteins in this pathway. These 87 proteins were enriched in 21 biological processes and 18 signaling pathways. Moreover, during the differentiation of chicken ESCs to SSCs-like cells induced by BMP4 in vitro, JAK2 and STAT3 were activated. The qRT-PCR results showed that the expression trends of JAK2 and STAT3 were basically the same as in vivo. We concluded that JAK/STAT signaling pathway plays an important role in the process of chicken SSCs generation both in vivo and in vitro; it may achieve its function through multiple biological processes and other related pathways.
Abstract Spermatogonial stem cells (SSCs) form the foundation for spermatogenesis and sustain male fertility. To explore the regulatory mechanisms of chicken SSCs generation, we obtained highly purified chicken embryonic stem cells (ESCs), primordial germ cells (PGCs) and SSCs by fluorescence-activated cell sorting (FACS). High-throughput analysis methods (RNA-Seq) were used to sequence the transcriptome level of these cells. Gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were used to analyze RNA-Seq results. BMP4 was used to induce chicken ESCs differentiation to SSCs-like cells in vitro. The quantitative real-time (qRT)-PCR was used to detect the expression changes of the key genes. The results showed that 22 relevant critical pathways were found by RNA-Seq, one of them was the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Total of 103 related genes were detected in this pathway. Protein-protein interactions analysis found that 87 proteins were significantly related to 19 key proteins in this pathway. These 87 proteins were enriched in 21 biological processes and 18 signaling pathways. Moreover, during the differentiation of chicken ESCs to SSCs-like cells induced by BMP4 in vitro, JAK2 and STAT3 were activated. The qRT-PCR results showed that the expression trends of JAK2 and STAT3 were basically the same as in vivo. We concluded that JAK/STAT signaling pathway plays an important role in the process of chicken SSCs generation both in vivo and in vitro; it may achieve its function through multiple biological processes and other related pathways.
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Received: 20 October 2014
Accepted:
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| Fund: This work was supported by the National Natural Science Foundation of China (31272429, 31472087), the Specialized Research Fund for the Doctoral Program of Higher Education, China (20123250120009), the China Postdoctoral Science Foundation Funded Project (2012M511326, 2014T70550), the Research and Innovation Program for Graduate Cultivation of Jiangsu Province, China (CXZZ13_0909), and the project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China. |
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Corresponding Authors:
ZHANG Ya-ni, E-mail: ynzhang@yzu.edu.cn;LI Bi-chun, Tel: +86-514-87997207, E-mail: yubcli@yzu.edu.cn
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| About author: ZHANG Lei, E-mail: leizhang17@sina.com; |
Cite this article:
ZHANG Lei, ZUO Qi-sheng, LI Dong, LIAN Chao, Kamel E Ahmed, TANG Bei-bei, SONG Jiu-zhou, ZHANG Ya-ni, LI Bi-chun.
2015.
Study on the role of JAK/STAT signaling pathway during chicken spermatogonial stem cells generation based on RNA-Seq. Journal of Integrative Agriculture, 14(5): 939-948.
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