生物技术通报 ›› 2016, Vol. 32 ›› Issue (9): 149-155.doi: 10.13560/j.cnki.biotech.bull.1985.2016.09.020

• 研究报告 • 上一篇    下一篇

雷州黑鸭胚胎期骨骼肌发育表达与功能分析

许冲, 陈琦, 苏瑛, 黄汉光, 崔红艳, 黄骏腾, 常羽   

  1. 广东海洋大学农学院,湛江 524088
  • 收稿日期:2016-03-10 出版日期:2016-09-25 发布日期:2016-10-10
  • 作者简介:许冲,女,硕士研究生,研究方向:动物遗传资源利用与品种选育,E-mail:810649753@qq.com;陈琦为本文并列第一作者
  • 基金资助:
    广东海洋大学广东省优秀学位论文培育项目(201412),广东海洋大学研究生示范课程建设项目(201411),广东省科技计划项目(2014A020208123),湛江市现代农业重点项目(2015A03003),广东星火计划项目(2012A020602104),湛江市科技计划项目(2012C0204)

Expression and Function of Embryonic Skeletal Muscle Development of Leizhou Black Duck

XU Chong, CHEN Qi, SU Ying, HUANG Han-guang, CUI Hong-yan, HUANG Jun-teng, CHANG Yu   

  1. College of Agricultural,Guangdong Ocean University,Zhanjiang 524088
  • Received:2016-03-10 Published:2016-09-25 Online:2016-10-10

摘要: 为保护雷州黑鸭优异的肉用性能,并开发利用这一珍贵的种质资源,研究选取雷州黑鸭为实验素材,对其胚胎期胸腿肌进行组织切片观察,并采用qRT-PCR与Western blot试验方法检测了Pax3、DCN和MSTN等基因在8、13、18、23和28胚龄胸腿肌中的发育性表达规律。同时用qRT-PCR法检测了雷州黑鸭28胚龄时Pax3、DCN和MSTN基因在各组织中mRNA的差异性表达量。胸腿肌组织切片观察雷州黑鸭胚胎期腿肌的发育明显早于胸肌;荧光定量PCR结果显示,雷州黑鸭Pax3、DCN和MSTN基因在各阶段胸腿肌中的相对表达量在23和28胚龄期最高,显著高于其它胚龄(P<0.05);雷州黑鸭28胚龄时各组织中Pax3、DCN和MSTN基因在胸肌和腿肌中的表达显著高于其他组织(P<0.05);Western blot结果表明,在胸腿肌中Pax3、DCN和MSTN蛋白的表达量在8胚龄期最低,在23胚龄时表达量显著高于其它胚龄(P<0.05),达到高峰。结合切片图对比表达模式的分析表明在肌纤维的形成以及生长发育过程中,Pax3、DCN和MSTN基因对骨骼肌的生理特性起到决定性作用,在雷州黑鸭胚胎期骨骼肌发育过程中的不同阶段调控着早期胚胎中前体肌细胞的分化、增殖、促进肌纤维的形成。

关键词: 雷州黑鸭, 骨骼肌, 切片, 时空表达

Abstract: To protect the excellent meat performance of Leizhou black duck,develop and utilize this precious germplasm,we selected Leizhou black duck as the experimental material and observed the tissue section of breast thigh meat in its fetal period,and used qRT-PCR and Western blot to detect the developmental expression patterns of gene Pax3,DCN and MSTN in breast thigh meat at 8th,13th,18th,23th,and 28th fetal period. Concurrently,qRT-PCR was used to test the mRNA difference expressions of genePax3,DCN and MSTN in all tissues of 28th fetal period. The observation of tissue section of breast thigh meat showed that in fetal period of Leizhou black duck,the development of thigh muscle was obviously earlier than the chest muscle. Fluorescence quantitative PCR showed that,in each stage of breast thigh meat,the relative expressions of gene Pax3,DCN and MSTN were the highest at 23th and 28th fetal period,and significantly higher than other fetal periods(P<0.05). In all tissues of Leizhou black duck,the expressions of Pax3,DCN and MSTN in thigh muscle and chest muscle was higher than other tissues in 28th fetal period(P<0.05);The results of Western blot showed that the expressions of Pax3,DCN and MSTN protein were the lowest in 8th fetal period and its expression in 23th fetal period was significantly higher than that in other fetal period(P< 0.05),and reached a peak. Combined the profiles of sections,comparing them with the expression pattern analysis showed that the gene Pax3,DCN and MSTN play a determining role for the physiological characteristics of skeletal muscles in the formation and growth of muscle fibers,and regulate the differentiation,proliferation and the formation of muscle cells of Leizhou black duck early embryo at different development stages of skeletal muscle in embryonic stage.

Key words: Leizhou black duck, skeletal muscle, sliced, temporal and spatial expression