视黄酸及Stra8基因联合作用对雄性鸡胚胎干细胞分化的影响

doi:10.11843/j.issn.0366-6964.2014.01.006

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (1): 38-45.doi: 10.11843/j.issn.0366-6964.2014.01.006

视黄酸及Stra8基因联合作用对雄性鸡胚胎干细胞分化的影响

张振韬¹，王丹¹，施青青¹，ELSAYED A K^1,2，张亚妮¹，韦光辉¹，朱睿¹，左其生¹，李东¹，刘堃¹，李碧春^1*

（1.扬州大学动物科学与技术学院，扬州 225009；2.College of Veterinary Medicine，Suez Canal University，Ismailia 41522，Egypt）

收稿日期:2013-08-14 出版日期:2014-01-23 发布日期:2014-01-23
通讯作者: 李碧春，教授，E-mail:yubcli@yzu.edu.cn
作者简介:张振韬，(1986-)，男，甘肃兰州人，博士生，主要从事动物胚胎工程研究，E-mail：yzuztzhang@gmail.com
基金资助:
国家自然科学基金（31272429）；江苏省研究生培养创新工程(CXLX12_0934)

Effect of Combination of Retinoic Acid and Stra8 Gene on Male cESCs Differentiation

ZHANG Zhen-tao¹，WANG Dan¹，SHI Qing-qing¹，ELSAYED A K^1,2，ZHANG Ya-ni¹，WEI Guang-hui¹，ZHU Rui¹，ZUO Qi-sheng¹，LI Dong¹，LIU Kun¹，LI Bi-chun^1*

(1.College of Animal Science and Technology，Yangzhou University，Yangzhou 225009，China；2.College of Veterinary Medicine，Suez Canal University，Ismailia 41522，Egypt)

Received:2013-08-14 Online:2014-01-23 Published:2014-01-23

摘要/Abstract

摘要：

旨在研究视黄酸（All-trans retinoic acid,RA）及Stra8（Stimulated by retinoic acid gene 8）基因联合作用对雄性鸡胚胎干细胞（Chicken embryonic stem cells，cESCs）分化的影响。选用PCR方法鉴定雄性cESCs为研究对象，转染Stra 8基因，对Stra8基因阳性cESCs使用RA进行12 d连续诱导，同时设立Stra8基因转染组、RA诱导组及空白组对照，各组均进行3次重复试验。细胞形态学、qRT-PCR检测RA及Stra8基因对cESCs分化的影响。结果表明，转染Stra8基因进行RA诱导，在诱导12 d后，出现精原干细胞样的圆形小克隆，其他组cESCs出现类胚体或解体；qRT-PCR鉴定发现各组中干细胞标记基因表达量随着培养时间的延长逐渐下降，同时检测到生殖细胞相关基因表达，转染Stra8基因并使用RA诱导组中表达量均高于对照组（P<0.01），6 d表达量达到最高，然后逐渐下降。Stra8基因和RA的联合作用能够促进cESCs向着生殖细胞方向的分化，不仅改变了原有细胞形态，同时出现了生殖相关基因的表达。通过该方法可以在体外建立一个ESCs诱导分化模型，将为生殖细胞的发生、增殖、分化及调控机理的研究和人类不孕不育症的治疗奠定良好的基础。

Abstract:

This experiment was conducted to explore the effect of combination of retinoic acid and Stra8 gene on male cESCs
differentiation.The sex of cESCs was determined by PCR to select only the male cESCs which divided into four groups，(1) both Stra8
transfected and RA-induced group，(2) only Stra8 transfected group，(3) only RA-induced group and (4) control group.Each group was repeated for three times.Cell morphology changes observations and qRT-PCR were performed to detect the effect of RA，Stra8 gene，and their combination on male cESCs differentiation.The results showed that there were small and round clones in the Stra8 transfected and RA-induced group in contrast to the other groups in which the cESCs were disintegrated.The expression quantity of stem cell marker genes was gradually declined in each group with extension of the incubation time.In the Transected Stra8 and RA induced group，the germ line related genes expressions were higher than that of the other groups (P<0.05) with the maximum amount at 6 d post induction and then gradually decreased.These results indicated that RA and Stra8 gene combination can promote the male cESCs differentiation to germ cell direction synactic that was proved by both significant cellular morphological changes and also the highly expressed germ cell marker genes.Through this induction system，it will not only establish a good foundation for germ cell induction and differentiation，but also provide basic idea for the late stage cESCs differentiation mechanisms.

中图分类号:

S831
S813.3

张振韬，王丹，施青青，ELSAYED A K，张亚妮，韦光辉，朱睿，左其生，李东，刘堃，李碧春. 视黄酸及Stra8基因联合作用对雄性鸡胚胎干细胞分化的影响[J]. 畜牧兽医学报, 2014, 45(1): 38-45.

ZHANG Zhen-tao，WANG Dan，SHI Qing-qing，ELSAYED A K，ZHANG Ya-ni，WEI Guang-hui，ZHU Rui，ZUO Qi-sheng，LI Dong，LIU Kun，LI Bi-chun. Effect of Combination of Retinoic Acid and Stra8 Gene on Male cESCs Differentiation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(1): 38-45.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2014.01.006

https://www.xmsyxb.com/CN/Y2014/V45/I1/38

参考文献

［1］KISSEL H，TIMOKHINA I，HARDY M P，et al.Point mutation in Kit receptor tyrosine kinase reveals essential roles for Kit signaling in spermatogenesis and oogenesis without affecting other Kit responses ［J］. EMBO J，2000，19：1312-1326．

［2］ROSSI P，DOLCI S，ALBANESI C，et al.Follicle- stimulating hormone induction of steel factor ( SLF) mRNA in mouse Sertoli cells and stimulation of DNA synthesis in spermatogonia by soluble SLF［J］. Dev Biol，1993，155:68-74．

［3］BLUME J P，JIANG G，HYMAN R，et al.Kit / stem cell factor receptor- induced activation of phosphatidylinositol 3’- kinase is essential for male fertility［J］.Nat Genet，2000，24:157-162．

［4］ZHOU Q，LI Y，NIE R，et al.Expression of stimulated by retinoic acid gene 8 (Stra8) and maturation of murine gonocytes and spermatogonia induced by retinoic acid in vitro［J］.Biol Reprod，2008，78:537-545．

［5］MIYAMOTO T，SENGOKU K，TAKUMA N，et al.Isolation and expression analysis of the testis-specific gene，stra，stimulated by retinoic acid gene 8［J］.Assist Reprod Genet，2002，19:531-535.

［6］HUBNER K，FUHRMANN G，CHRISTENSON L K，et al.Derivation of oocytes from mouse embryonic stem cells［J］.Science，2003，300:1251-1256.

［7］NAYERNIA K，NOLTE J，MICHELMANN H W，et al. In vitro differentiated embryonic stem cell give rise to male gametes that can generate offspring mice［J］.Dev Cell，2006,11:125-132.

［8］KERKIS A，FONSECA S A，SERAFIM R C，et al.In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes［J］. Cloning Stem Cell，2007，9:535-548.

［9］常卓，侯玲玲，周雅琼，等.哺乳动物精子体外诱导的研究进展［J］.中国生物工程杂志，2013，33(3)：135-142.

［10］王丹，张振韬，施青青，等.鸡Stra8基因真核表达载体构建及亚细胞定位的研究［J］.中国家禽，2013，35(8)：10-13.

［11］HAMBURGER V，HAMILTON H L.A series of normal stages in the development of the chicken［J］.Morphol，1951,88:49-92.

［12］李碧春，陈国宏，赵东伟，等.鸡胚PGCs迁移与性腺发育关系的研究［J］.扬州大学学报，2002，23(1)：18-26.

［13］王嘉力，贾存灵，苏利红，等.家鸡CHD1基因PCR快速性别鉴定方法的建立及其在早期鸡胚中的应用［J］.家禽科学，2009，12：3-7.

［14］戴建明，何先红，赵文明，等.鸡胚胎干细胞的分离和培养［J］.中国家禽，2007，43(19)： 15-19.

［15］GUBBAY J，COLLIGNON J，KOOPMAN P，et al.A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes［J］.Nature，1990,7:245-250.

［16］AVILION A A，NICOLIS S K，PEVNY L H，et al.Multipotent cell lineages in early mouse development depend on Sox2 function ［J］. Genes Dev，2003,17:126-140.

［17］YUAN H，CORBI N，BASILICO C，et al.Development-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3［J］.Genes Dev，1995,9:2635-2645.

［18］VAN P A，ROEPERS G H，GADEMAN I S，et al.Establishment of cell lines with rat spermatogonial stem cell characteristics ［J］.Endocrinology，2002,l43:l845-1850.

［19］SHINPHAAR T，AVARBOCK M R，BRINSTER R L.Betal-and alpha6-integrin are surface markers on mouse spermatogonial stem cells［J］. Prco Natl Acad Sci USA，1999,96:504-509.

[1]	牟湘钰, 徐云若, 胡静怡, 周欣妍, 朱勇文. 家禽支链氨基酸营养需要研究进展[J]. 畜牧兽医学报, 2024, 55(1): 31-38.
[2]	郭妍婷, 周建民, 王晶, 齐广海. 产蛋鸡子宫部钙离子转运及调控的研究进展[J]. 畜牧兽医学报, 2024, 55(1): 39-50.
[3]	唐鑫鑫, 郑炬梅, 骆娜, 营凡, 朱丹, 李森, 刘大伟, 安炳星, 文杰, 赵桂苹, 李和刚. 基于全基因组关联分析揭示肉鸡腿病发生的遗传机制[J]. 畜牧兽医学报, 2024, 55(1): 99-109.
[4]	卢建, 张欣, 江栋材, 马猛, 王强, 王星果, 李永峰, 郭军, 窦套存, 胡玉萍, 李尚民, 邵丹, 曲亮. 甘氨酸锰对产蛋后期蛋鸡产蛋性能和肠道微生物的影响[J]. 畜牧兽医学报, 2024, 55(1): 218-231.
[5]	徐小静, 董瑞玲, 魏立民, 赵少猛, 赵桂苹, 张敏红, 冯京海. 文昌鸡母鸡生长模型的研究[J]. 畜牧兽医学报, 2023, 54(12): 4952-4961.
[6]	齐丽娜, 陆雪林, 杨凯旋, 周瑾, 李先玉, 刘珂, 张文刚, 顾欣, 章伟建. 基于SNP芯片分析新浦东鸡的遗传多样性和遗传结构[J]. 畜牧兽医学报, 2023, 54(12): 4962-4971.
[7]	王燕星, 张雨时, 姬海港, 刘阳, 牛玉芳, 韩瑞丽, 刘小军, 田亚东, 康相涛, 李转见. 鸡骨骼肌卫星细胞系的建立及分析[J]. 畜牧兽医学报, 2023, 54(12): 4972-4981.
[8]	范晨宇, 单艳菊, 章明, 姬改革, 巨晓军, 屠云洁, 贺喜, 束婧婷, 刘一帆, 张海涵. 立华麻黄鸡体重和肉品质性状全基因组关联分析[J]. 畜牧兽医学报, 2023, 54(12): 4982-4992.
[9]	李贞明, 余苗, 唐延天, 李袁飞, 刘志昌, 容庭, 马现永. 腐殖酸钠对白羽肉鸡营养物质表观消化率、粪便微生物及其代谢产物的影响[J]. 畜牧兽医学报, 2023, 54(12): 5091-5100.
[10]	毕睿晨, 刘相泽, 胡泽琼, 杨美雪, 乔嘉宁, 黄嘉, 郭芳申, 孔令华, 王忠. 植物多酚在家禽领域应用研究进展[J]. 畜牧兽医学报, 2023, 54(11): 4488-4501.
[11]	陈诚, 乔西波, 孙亿, 康丽, 姜运良. 琅琊鸡FSHR基因-868位点的多态性及对产蛋性能的遗传效应研究[J]. 畜牧兽医学报, 2023, 54(11): 4560-4568.
[12]	金四华, 贾富民, 何培莉, 贾羽晴, 税斐, 刘旭玲, 刘星, 王新, 徐嫚, 耿照玉. IHH过表达对鸡原代软骨细胞增殖和分化的影响[J]. 畜牧兽医学报, 2023, 54(11): 4569-4576.
[13]	秦士贞, 杨敏敏, 任志雄, 李金录, 唐德富, 史兆国. 枯草芽孢杆菌对脂多糖应激肉仔鸡肠道免疫、肠道组织形态以及肠道屏障的影响[J]. 畜牧兽医学报, 2023, 54(11): 4676-4690.
[14]	徐自强, 刘金松, 孙耀威, 史永浩, 吴艳萍, 张瑞强. 月桂酸对肉鸡屠宰性能、肌肉品质和抗氧化功能的影响[J]. 畜牧兽医学报, 2023, 54(11): 4691-4701.
[15]	苏丹, 文晓宾, 马腾, 钟儒清, 王阳, 陈亮. 羟基酪醇对肉仔鸡生长性能、抗氧化能力和肠道炎性因子的影响[J]. 畜牧兽医学报, 2023, 54(11): 4851-4859.

视黄酸及Stra8基因联合作用对雄性鸡胚胎干细胞分化的影响

Effect of Combination of Retinoic Acid and Stra8 Gene on Male cESCs Differentiation

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价