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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
E-mail: ynzhang@yzu.edu.cn; 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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| Arbouzova N I, Zeidler M P. 2006. JAK/STAT signaling inDrosophila: Insights into conserved regulatory and cellularfunctions. Development, 133, 2605-2616Caires K C, de Avila J, McLean D J. 2009. Vascularendothelial growth factor regulates germ cell survival duringestablishment of spermatogenesis in the bovine testis.Reproduction, 138, 667-677Chen W, Jia W, Wang K, Zhou Q, Leng Y, Duan T, KangJ. 2012. Retinoic acid regulates germ cell differentiationin mouse embryonic stem cells through a Smaddependentpathway. Biochemical and Biophysical ResearchCommunications, 418, 571-577Chuang C Y, Lin K I, Hsiao M, Stone L, Chen H F, Huang Y H,Kuo H C. 2012. Meiotic competent human germ cell-likecells derived from human embryonic stem cells inducedby BMP4/WNT3A signaling and OCT4/EpCAM (epithelialcell adhesion molecule) selection. Journal of BiologicalChemistry, 287, 14389-14401Hamburger V, Hamilton H L. 1951. A series of normal stagesin the development of the chick embryo. Journal ofMorphology, 88, 49-92Hou S X, Zheng Z, Chen X, Perrimon N. 2002. The JAK/STAT pathway in model organisms: emerging roles in cellmovement. Developmental Cell, 3, 765-778Kiger A A, Jones D L, Schulz C, Rogers M B, Fuller M T. 2001.Stem cell self-renewal specified by JAK-STAT activation inresponse to a support cell cue. Science, 294, 2542-2545Kisseleva T, Bhattacharya S, Braunstein J, Schindler C W.2002. Signaling through the JAK/STAT pathway, recentadvances and future challenges. Gene, 285, 1-24Levy O, Ruvinov E, Reem T, Granot Y, Cohen S. 2010. Highlyefficient osteogenic differentiation of human mesenchymalstem cells by eradication of STAT3 signaling. TheInternational Journal of Biochemistry & Cell Biology, 42,1823-1830Oatley J M, Kaucher A V, Avarbock M R, Brinster R L. 2010.Regulation of mouse spermatogonial stem cell differentiationby STAT3 signaling. Biology of Reproduction, 83, 427-433Oatley J M, Brinster R L. 2008. Regulation of spermatogonialstem cell self-renewal in mammals. Annual Review of Celland Developmental Biology, 24, 263-286Pareek T K, Joshi A R, Sanyal A, Dighe R R. 2007. Insights intomale germ cell apoptosis due to depletion of gonadotropinscaused by GnRH antagonists. Apoptosis, 12, 1085-1100Rawlings J S, Rosler K M, Harrison D A. 2004. The JAK/STAT signaling pathway. Journal of Cell Science, 117,1281-1283Sheng X R, Posenau T, Gumulak J J, Matunis E, van DorenM, Wawersik M. 2009. JAK-STAT regulation of malegermline stem cell establishment during Drosophilaembryogenesis. Developmental Biology, 334, 335-344Shi Q Q, Zhang Z T, Li P C, Wang D, Huang X M, Zhang Y N,Li B C. 2013. Study on differentiation of chicken embryonicstem cells to male germ cells by BMP4. Chinese Journalof Animal and Veterinary Sciences, 44, 1749-1757 (inChinese)Shi Q Q, Sun M, Zhang Z T, Zhang Y N, Elsayed A K, ZhangL, Li B C. 2014. A screen of suitable inducers for germlinedifferentiation of chicken embryonic stem cells. AnimalReproduction Science, 147, 74-85Singh S R, Xiu C, Steven X. 2005. JAK/STAT signalingregulates tissue outgrowth and male germline stem cellfate in Drosophila. Cell Research, 15, 1-5Sun M. 2012. Study on male germ cell derived from chickenembryonic stem cell and the generation of transgenicchicken. Ph D thesis, Yangzhou University. (in Chinese)Sun M, Shi Q Q, Fu D Z, Yin Y H, Zhang Y N, Li B C. 2011. Studyof the expression of gene cell differentiation associatedgenes on chicken ESC and SSCs. Biotechnology, 21,16-19 (in Chinese)Tang F, Barbacioru C, Bao S, Lee C, Nordman E, Wang X, LaoK Q, Surani M A. 2010a. Tracing the derivation of embryonicstem cells from the inner cell mass by single-cell RNA-Seqanalysis. Cell Stem Cell, 6, 468-478Tang F, Barbacioru C, Nordman E, Li B, Xu N, BashkirovV I, Lao K Q, Surani M A. 2010b. RNA-Seq analysis tocapture the transcriptome landscape of a single cell. NatureProtocols, 5, 516-535Trapnell C, Williams B A, Pertea G, Mortazavi A, Kwan G,Baren M J, Salzber S L, World B J, Pachter L. 2010.Transcript assembly and quantification by RNA-Seq revealsunannotated transcripts and isoform switching during celldifferentiation. Nature Biotechnology, 28, 511-515Tulina N, Matunis E. 2001. Control of stem cell self-renewalin Drosophila spermatogenesis by JAK-STAT signaling.Science, 294, 2546-2549Wawersik M, Milutinovich A, Casper A L, Matunis E, WilliamsB, Van Doren M. 2005. Somatic control of germlinesexual development is mediated by the JAK/STATpathway. Nature, 436, 563-567Yang H, Sun M, Tian Z, Qin Y, Ren L, Xu F, Li B. 2010.Preliminary comparison of dispersed culture and wholeembryo culture of chicken blastodermal cells on Xperiod. China Poultry, 32, 14-17 (in Chinese)Zhang Z, Lv X, Jiang J, Zhang L, Zhao Y. 2013. Dual rolesof Hh signaling in the regulation of somatic stem cell selfrenewaland germline stem cell maintenance in Drosophilatestis. Cell Research, 23, 573-576 |
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