The Induction Effect of Am80 and TSA on ESC Differentiation via Regulation of Stra8 in Chicken

Yani Zhang; Qisheng Zuo; Zhiyong Liu; Dong Li; Beibei Tang; Tian-Rong Xiao; Chao Lian; Yingjie Wang; Kai Jin; Yilin Wang; Wenhui Zhang; Bichun Li

doi:10.1371/journal.pone.0140262

The Induction Effect of Am80 and TSA on ESC Differentiation via Regulation of Stra8 in Chicken

PLoS One. 2015 Nov 25;10(11):e0140262. doi: 10.1371/journal.pone.0140262. eCollection 2015.

Authors

Yani Zhang^{1

2}, Qisheng Zuo^{1

2}, Zhiyong Liu^{1

2}, Dong Li^{1

2}, Beibei Tang^{1

2}, Tian-Rong Xiao^{1

2}, Chao Lian^{1

2}, Yingjie Wang^{1

2}, Kai Jin^{1

2}, Yilin Wang^{1

2}, Wenhui Zhang¹, Bichun Li^{1

2}

Affiliations

¹ College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
² Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, Yangzhou 225009, China.

Abstract

Stra8 encodes stimulated by retinoic acid gene 8, a protein that is important for initiation of meiosis in mammals and birds. This study was aimed at identifying the active control area of chicken STRA8 gene core promoter, to screen optimum inducers of the STRA8 gene, thus to enhance the differentiation of embryonic stem cells (ESCs) into spermatogonial stem cells. Fragments of chicken STRA8 gene promoter were cloned into fluorescent reporter plasmids and transfected into DF-1 cells. Then Dual-Luciferase® Reporter Assay System was used to identify the activity of the STRA8 gene under different inducers. Our studies showed that the promoter fragment -1055 bp to +54 bp of Suqin chicken Stra8 revealed the strongest activity. The dual-luciferase® reporter showed that Tamibarotene (Am80) and TrichostatinA (TSA) could significantly enhance STRA8 transcription. The in vitro inductive culture of chicken ESCs demonstrated that spermatogonial stem cells (SSC)-like cells appeared and Integrinβ1 protein was expressed on day 10, indicating that Am80 and TSA can promote ESCs differentiation into SSCs via regulation of Stra8.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Signal Transducing / genetics
Adaptor Proteins, Signal Transducing / metabolism*
Animals
Avian Proteins / genetics
Avian Proteins / metabolism*
Benzoates / pharmacology*
Cell Differentiation
Chick Embryo
Chickens
Embryonic Stem Cells / physiology*
Histone Deacetylase Inhibitors / pharmacology*
Hydroxamic Acids / pharmacology*
Promoter Regions, Genetic
Tetrahydronaphthalenes / pharmacology*
Tissue Culture Techniques
Transcriptional Activation / drug effects

Substances

Adaptor Proteins, Signal Transducing
Avian Proteins
Benzoates
Histone Deacetylase Inhibitors
Hydroxamic Acids
Tetrahydronaphthalenes
tamibarotene
trichostatin A

Grants and funding

This work was supported by the National Natural Science Foundation of China (31301959, 1272429, 31472087); Research Fund for the Doctoral Program of Higher Education (20123250120009), China Postdoctoral Science Foundation funded project (2012M511326, 2014T70550); A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There are not any conflicts for the above grant numbers.