Ghrelin对小鼠不同来源胚胎体外发育的影响及其受体的表达模式

doi:10.7668/hbnxb.2015.04.014

华北农学报 ›› 2015, Vol. 30 ›› Issue (4): 78-83. doi: 10.7668/hbnxb.2015.04.014

Ghrelin对小鼠不同来源胚胎体外发育的影响及其受体的表达模式

李运生^1,2, 杨宏星^1,2, 刘晓蕊³, 习海涛⁴, 方富贵^1,2, 刘亚^1,2

1. 安徽农业大学动物科技学院, 安徽合肥 230036;
2. 安徽农业大学安徽地方畜禽遗传资源保护与生物育种省级实验室, 安徽合肥 230036;
3. 南京医科大学代谢疾病研究中心, 江苏南京 210029;
4. 温州医学院附属第二医院生殖医学中心, 浙江温州 325000

收稿日期:2015-04-22 出版日期:2015-08-28
通讯作者: 刘亚(1971-),男,安徽亳州人,副教授,博士,主要从事动物胚胎工程和生殖生理研究。
作者简介:李运生(1980-),男,安徽凤台人,博士,主要从事动物胚胎工程和生殖生理研究。
基金资助:
高等学校博士学科点专项科研基金博导类资助课题(20123418110004);安徽农业大学学科骨干培育项目(2014XKPY-23)

Effects of Ghrelin on in vitro Development of Mouse Embryos and the Expression of Ghsr-1a in Different Embryos

LI Yun-sheng^1,2, YANG Hong-xing^1,2, LIU Xiao-rui³, XI Hai-tao⁴, FANG Fu-gui^1,2, LIU Ya^1,2

1. College of Animal Science and Technology, Anhui Agricutural University, Hefei 230036, China;
2. Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricutural University, Hefei 230036, China;
3. Metabolic Disease Research Center, Nanjing Medical University, Nanjing 210029, China;
4. Center for Reproductive Medicine, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325000, China

Received:2015-04-22 Published:2015-08-28

摘要/Abstract

摘要： 为探明Ghrelin及其受体Ghsr-1a在早期胚胎发育中的作用,以小鼠体内受精、体外受精和孤雌激活胚胎为研究对象,在其体外发育过程中添加不同浓度Ghrelin,观察胚胎发育效率;通过免疫荧光染色和荧光定量PCR,观察不同时期胚胎中Ghsr-1a的变化。结果表明,体内受精胚体外发育中添加0.1 ng/mL Ghrelin显著降低了囊胚率(P <0.05),添加1 ng/mL的Ghrelin 显著降低了小鼠体外受精胚的囊胚率 (P <0.05)。在体内受精胎中, Ghsr-1a 的mRNA表达水平在4细胞显著高于其他时期(P <0.05);体外受精胚中, Ghsr-1a 的mRNA水平在8细胞前无明显差异,桑椹胚和囊胚显著下降(P <0.05)。在孤雌激活胚胎中, Ghsr-1a 的mRNA表达水平从2细胞到8细胞显著降低,之后到囊胚期开始显著升高,其蛋白表达模式与mRNA表达模式相反。Ghsr-1a在体内受精和体外受精胚胎中的表达相似,囊胚期表达量最低。结果提示,Ghrelin能够抑制体内和体外受精胚的体外发育,不同来源胚胎 Ghsr-1a mRNA和蛋白表达模式均不相同。研究结果为进一步探索Ghrelin在胚胎发育中的作用机制提供了试验基础。

关键词: Ghrelin, Ghsr-1a, 小鼠, 胚胎

Abstract: The objective of the present study was to investigate the effect of addition of ghrelin to in vitro culture medium on preimplatation development quality of different sources(in vivo, in vitro fertilization,parthenogenetic) mouse embryos.The receptor of Ghrelin, Ghsr-1a which mRNA and protein expression also been checked in preimplatation embryos.The results showed that blastocyst rate of in vivo embryos was significantly decreased when 0.1 ng/mL Ghrelin added into in vitro culture medium (P <0.05).Blastocyst rate of in vitro fertilization embryos cultured in KSOM supplemented with 1 ng/mL ghrelin was significantly decline than control group (P <0.05).The mRNA levels and protein expression of Ghsr-1a were observed at different early stage of mouse.The transcription level of Ghsr-1a of 4-cell embryo was signification higher than other stages of embryos in in vivo embryos(P <0.05). Ghsr-1a levels was decreased from 8-cell to blastocyst in in vitro fertilization embryos,The mRNA levels of Ghsr-1a was declined from 2-cell to 8-cell and increased from 8-cell to blastocyt in parthenogenetic embryos and protein expression contrary.The protein expression of Ghsr-1a were decreased in blastocyst stage and same in vivo and in vitro fertilization embryos.Thus,ghrelin may inhibit embryo development quality through its receptors Ghsr-1a and give a reference for further researchers in function of ghrelin in mouse embryos.

Key words: Ghrelin, GHSR-1a, Mouse, Embryos

中图分类号:

S814.8

李运生, 杨宏星, 刘晓蕊, 习海涛, 方富贵, 刘亚. Ghrelin对小鼠不同来源胚胎体外发育的影响及其受体的表达模式[J]. 华北农学报, 2015, 30(4): 78-83. doi: 10.7668/hbnxb.2015.04.014.

LI Yun-sheng, YANG Hong-xing, LIU Xiao-rui, XI Hai-tao, FANG Fu-gui, LIU Ya. Effects of Ghrelin on in vitro Development of Mouse Embryos and the Expression of Ghsr-1a in Different Embryos[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(4): 78-83. doi: 10.7668/hbnxb.2015.04.014.

http://www.hbnxb.net/CN/Y2015/V30/I4/78

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Ghrelin对小鼠不同来源胚胎体外发育的影响及其受体的表达模式

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