Please wait a minute...
Journal of Integrative Agriculture  2015, Vol. 14 Issue (5): 939-948    DOI: 10.1016/S2095-3119(14)60938-2
Animal Science · Veterinary Science Advanced Online Publication | Current Issue | Archive | Adv Search |
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
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
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.
Keywords:  spermatogonial stem cell (SSC)       JAK/STAT signaling pathway       RNA-Seq       induction       chicken  
Received: 20 October 2014   Accepted:
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.

Corresponding Authors:  ZHANG Ya-ni, E-mail: ynzhang@yzu.edu.cn;LI Bi-chun, Tel: +86-514-87997207, E-mail: yubcli@yzu.edu.cn   
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.

Arbouzova N I, Zeidler M P. 2006. JAK/STAT signaling inDrosophila: Insights into conserved regulatory and cellularfunctions. Development, 133, 2605-2616

Caires 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, 667-677

Chen 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, 571-577

Chuang 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, 14389-14401

Hamburger V, Hamilton H L. 1951. A series of normal stagesin the development of the chick embryo. Journal ofMorphology, 88, 49-92

Hou S X, Zheng Z, Chen X, Perrimon N. 2002. The JAK/STAT pathway in model organisms: emerging roles in cellmovement. Developmental Cell, 3, 765-778

Kiger 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, 2542-2545

Kisseleva T, Bhattacharya S, Braunstein J, Schindler C W.2002. Signaling through the JAK/STAT pathway, recentadvances and future challenges. Gene, 285, 1-24

Levy 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, 42,1823-1830

Oatley 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, 427-433

Oatley J M, Brinster R L. 2008. Regulation of spermatogonialstem cell self-renewal in mammals. Annual Review of Celland Developmental Biology, 24, 263-286

Pareek 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, 1085-1100

Rawlings J S, Rosler K M, Harrison D A. 2004. The JAK/STAT signaling pathway. Journal of Cell Science, 117,1281-1283

Sheng 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, 335-344

Shi 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, 74-85

Singh S R, Xiu C, Steven X. 2005. JAK/STAT signalingregulates tissue outgrowth and male germline stem cellfate in Drosophila. Cell Research, 15, 1-5

Sun 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, 468-478

Tang 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, 516-535

Trapnell 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, 511-515

Tulina N, Matunis E. 2001. Control of stem cell self-renewalin Drosophila spermatogenesis by JAK-STAT signaling.Science, 294, 2546-2549

Wawersik 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, 563-567

Yang 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
[1] Qing Li, Zhuangzhuang Sun, Zihan Jing, Xiao Wang, Chuan Zhong, Wenliang Wan, Maguje Masa Malko, Linfeng Xu, Zhaofeng Li, Qin Zhou, Jian Cai, Yingxin Zhong, Mei Huang, Dong Jiang. Time-course transcriptomic information reveals the mechanisms of improved drought tolerance by drought priming in wheat[J]. >Journal of Integrative Agriculture, 2025, 24(8): 2902-2919.
[2] Changning Wei, Hui Cao, Chenxu Li, Hongyu Song, Qing Liu, Xingquan Zhu, Wenbin Zheng. Differences in N6-methyladenosine (m6A) methylation among the three major clonal lineages of Toxoplasma gondii tachyzoites[J]. >Journal of Integrative Agriculture, 2025, 24(7): 2810-2825.
[3] Guangzheng Liu, Wenjie Ren, Kai Jin, Dan Zheng, Qisheng Zuo, Yani Zhang, Guohong Chen, Bichun Li, Yingjie Niu. PGC-mediated conservation strategies for germplasm resources of Rugao Yellow chicken and Shouguang chicken in China[J]. >Journal of Integrative Agriculture, 2025, 24(6): 2327-2341.
[4] Tengfei Wang, Changyong Fan, Yufei Xiao, Shan Lü, Guangyang Jiang, Mengyun Zou, Yingjie Wang, Qiao Guo, Zhenghao Che, Xiuli Peng. Protection of chickens from Mycoplasma gallisepticum through the MAPK/ERK/JNK pathway by a compound of ten Chinese medicine formulas[J]. >Journal of Integrative Agriculture, 2025, 24(6): 2356-2370.
[5] Qianqian Shi, Xue Han, Xinhao Zhang, Jie Zhang, Qi Fu, Chen Liang, Fangmeng Duan, Honghai Zhao, Wenwen Song. Transcriptome-wide N6-methyladenosine (m6A) profiling of compatible and incompatible responses reveals a nonhost resistance-specific m6A modification involved in soybean–soybean cyst nematode interaction[J]. >Journal of Integrative Agriculture, 2025, 24(5): 1875-1891.
[6] Xuemei Hou, Meimei Shi, Zhuohui Zhang, Yandong Yao, Yihua Li, Changxia Li, Wenjin Yu, Chunlei Wang, Weibiao Liao. DNA demethylation is involved in nitric oxide-induced flowering in tomato[J]. >Journal of Integrative Agriculture, 2025, 24(5): 1769-1785.
[7] Jiamao Gu, Pengkun Liu, Wenting Nie, Zhijun Wang, Xiaoyu Cui, Hongdan Fu, Feng Wang, Mingfang Qi, Zhouping Sun, Tianlai Li, Yufeng Liu. Abscisic acid alleviates photosynthetic damage in the tomato ABA-deficient mutant sitiens and protects photosystem II from damage via the WRKY22–PsbA complex under low-temperature stress[J]. >Journal of Integrative Agriculture, 2025, 24(2): 546-563.
[8] Jie Zhao, Sa Xiao, Wei Yao, Xudong Chang, Xinglong Wang, Zengqi Yang, Wenbin Wang. Velogenic Newcastle disease virus invades chicken brain by infecting brain microvascular endothelial cells to increase blood-brain barrier permeability[J]. >Journal of Integrative Agriculture, 2025, 24(2): 712-723.
[9] Yuanmei Wang, Jingwei Yuan, Yanyan Sun, Aixin Ni, Jinmeng Zhao, Yunlei Li, Panlin Wang, Lei Shi, Yunhe Zong, Pingzhuang Ge, Shixiong Bian, Hui Ma, Jilan Chen. Genome-wide circular RNAs signatures involved in sexual maturation and its heterosis in chicken[J]. >Journal of Integrative Agriculture, 2025, 24(2): 697-711.
[10] Yanxing Wang, Haigang Ji, Liyang He, Yufang Niu, Yushi Zhang, Yang Liu, Yadong Tian, Xiaojun Liu, Hong Li, Xiangtao Kang, Yanling Gao, Zhuanjian Li. Establishment and analysis of a chicken skeletal muscle satellite cell line using TERT[J]. >Journal of Integrative Agriculture, 2025, 24(11): 4370-4378.
[11] Yong Yang, Rong Fan, Xuejun Zhang, Meihua Li, Yongbing Zhang, Hongping Yi, Manrui Ma, Yun Yang, Bin Liu, Xingwang Liu, Huazhong Ren. Mutation in CmGhc1 confers the white hypocotyl phenotype in melon (Cucumis melo L.)[J]. >Journal of Integrative Agriculture, 2025, 24(11): 4242-4254.
[12] Qi Zeng, Qingguo Sun, Xinru Hou, Lin Chen, Ruixing Zhang, Xue Bai, Xifan Liu, Xiaowu Wang, Lugang Zhang, Baohua Li. Comparative transcriptomic analysis of Chinese cabbage’s defense responses to Alternaria brassicae[J]. >Journal of Integrative Agriculture, 2025, 24(10): 3895-3908.
[13] Wenya Li, Haoxiang Ma, Yanxing Wang, Yushi Zhang, Yang Liu, Ruili Han, Hong Li, Hanfang Cai, Xiaojun Liu, Xiangtao Kang, Ruirui Jiang, Zhuanjian Li. The VGLL2 gene participates in muscle development in Gushi chickens[J]. >Journal of Integrative Agriculture, 2025, 24(1): 246-260.
[14] Lei Shi, Yanyan Sun, Yunlei Li, Hao Bai, Jingwei Yuan, Hui Ma, Yuanmei Wang, Panlin Wang, Aixin Ni, Linlin Jiang, Pingzhuang Ge, Shixiong Bian, Yunhe Zong, Jinmeng Zhao, Adamu M. Isa, Hailai H. Tesfay, Jilan Chen. Asymmetric expression of CA2 and CA13 linked to calcification in the bilateral mandibular condyles cause crossed beaks in chickens[J]. >Journal of Integrative Agriculture, 2024, 23(7): 2379-2390.
[15] Yuan Gao, Fuxia Bai, Qi Zhang, Xiaoya An, Zhaofei Wang, Chuzhao Lei, Ruihua Dang. Dynamic transcriptome profiles and novel markers in bovine spermatogenesis revealed by single-cell sequencing[J]. >Journal of Integrative Agriculture, 2024, 23(7): 2362-2378.
No Suggested Reading articles found!