绵羊胚胎发育中后期骨骼肌BTG2/3的表达动态及其与Myostatin的关系

doi:10.11843/j.issn.0366-6964.2015.11.002

摘要/Abstract

摘要：

旨在探究绵羊胚胎中后期骨骼肌BTG2/3（B cell translocation gene 2/3）的动态表达情况，探索肌肉生长抑制素(Myostatin，MSTN)基因对其表达的影响。以特克赛尔（Texel）和乌珠穆沁（Ujimqin)绵羊85、100、120和135 d 4个不同发育阶段的胎儿背最长肌为研究对象，利用Real-time PCR技术分别检测MSTN及BTG2/3的表达差异；选取100 d各胚胎半膜肌、背最长肌、半腱肌和股四头肌检测不同组织BTG2/3的表达差异；构建稳定慢病毒干扰载体pFU-GW-myostatin，分别感染处于增殖和分化阶段的绵羊成肌细胞，检测BTG2/3表达水平变化。结果显示，随着胚胎生长，特克赛尔羊胎儿MSTN表达量持续下降，而乌珠穆沁羊胎儿先下降后上升，特克赛尔羊胎儿BTG2表达量呈先升高后降低趋势，而乌珠穆沁羊表现出先降低后升高再降低的趋势；BTG3的表达量在特克赛尔羊与BTG2表达趋势相同，而乌珠穆沁羊则与BTG2相反；100 d胎儿4种不同骨骼肌组织中，特克赛尔羊与乌珠穆沁羊相比，背最长肌和半腱肌中BTG2表达量极显著偏高(P<0.01)，而半膜肌和背最长肌中BTG3表达量极显著偏低(P<0.01)；慢病毒干扰载体对成肌细胞具有较高的感染和干扰效率，增殖和分化阶段的成肌细胞被MSTN干扰后，BTG2和BTG3表达量极显著降低(P<0.01)。综上表明，BTG2和BTG3对绵羊胎儿中后期骨骼肌发育有重要作用，可能参与myostatin调节通路。本研究对绵羊胚胎发育过程的分子机制研究具有重要意义。

Abstract:

The aim of this study was to analyze the expression of the sheep BTG2/3 genes in the mid to late embryonic skeletal muscle and to investigate the relationship between MSTN and BTG2/3.Longissimus dorsi of 85 dpc(day postconception)，100 dpc，120 dpc and 135 dpc in Texel and Ujimqin sheep were collected as tested subjects.Real-time PCR was used to detect the expression of MSTN and BTG2/3 in the longissimus dorsi.Semimembranosus，longissimus dorsi，semitendinosus and quadriceps femoris of 100 dpc in the Texel and Ujimqin sheep were selected to detect the expression of BTG2/3.Lentiviral vectors pFU-GW-myostatin were constructed to infect sheep myoblasts at the proliferation and differentiation stage stably and the expression of BTG2/3 were detected at different stages.The results showed that a downward expression trend was emerged of MSTN in Texel fetuses，whereas the expression level in Ujimqin increased after decline at 100 dpc.The expression of BTG2 and BTG3 in Texel had downward trend after rising on 100 dpc，but BTG2 represented decrease-increase-decrease trend and BTG3 represented opposite trend to BTG2 in Ujimqin.Expression levels in different muscle on 100 dpc in the Texel and Ujimqin showed that，BTG2 gene expression was significantly higher(P<0.01) in Texel longissimus dorsi and semitendinosus than that in Ujimqin，and BTG3 gene expression was significantly lower(P<0.01) in Texel semimembranosus and longissimus dorsi than that in Ujimqin.Transfection and interference efficiency of lentiviral vector were very high.After transfected myoblast at proliferation and differentiation stage，respectively，expression levels of BTG2 and BTG3 also were decreased significantly(P<0.01) at both stages.The results indicate that BTG2 and BTG3 play an important role in sheep skeletal muscle growth from the mid to late embryonic stage and may be involved in myostatin regulatory pathway.Our study provide an insight into the molecular regulation mechanism of embryonic development in sheep.

中图分类号:

S826
S813.3

刘瑞凿，刘真，吴明明，王慧华，朱才业，张莉，赵福平，魏彩虹，杜立新. 绵羊胚胎发育中后期骨骼肌BTG2/3的表达动态及其与Myostatin的关系[J]. 畜牧兽医学报, 2015, 46(11): 1916-1923.

LIU Rui-zao，LIU Zhen，WU Ming-ming，WANG Hui-hua，ZHU Cai-ye，ZHANG Li，ZHAO Fu-ping，WEI Cai-hong，DU Li-xin. Expression Trend of BTG2/3 in the Mid to Late Embryonic Skeletal Muscle of Sheep and the Relationship between BTG2/3 and MSTN[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(11): 1916-1923.

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