Wnt/βcatenin信号通路相关基因及MyHCs在猪骨骼肌发育中的表达规律

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (12): 1686-1695.

Wnt/βcatenin信号通路相关基因及MyHCs在猪骨骼肌发育中的表达规律

杨秋梅，史新娥，沈清武，刘月光，高晓娟，陈宗正，杨公社*

西北农林科技大学动物科技学院动物脂肪沉积与肌肉发育实验室，杨凌 712100

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-12-15 发布日期:2011-12-15
通讯作者: 杨公社

The Expression Pattern of Wnt/βcatenin Signal Pathway Related Genes and MyHCs during Porcine Skeletal Muscle Development

YANG Qiumei，SHI Xine，SHEN Qingwu，LIU Yueguang，GAO Xiaojuan，CHEN Zongzheng，YANG Gongshe*

Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-12-15 Published:2011-12-15
Contact: YANG Gongshe

摘要/Abstract

摘要： 为研究Wnt/βcatenin信号通路对猪骨骼肌纤维类型发育的影响，并探讨其潜在的作用机制。本研究以1日龄、1周龄、2周龄、1月龄、2月龄和6月龄大白猪背最长肌组织为研究材料，利用Realtime PCR、Western blot、冰冻切片免疫组化等方法，研究Wnt/βcatenin信号通路相关基因及MyHCs在猪骨骼肌发育过程中的表达规律。Realtime PCR结果显示：MyHCⅠ、GSK3β、βcatenin和Fz3 mRNA随着个体日龄增加表达量下降，MyHCⅡb mRNA表达逐渐升高，MyHCⅡa mRNA表达量在1日龄～2周龄阶段升高后又降低，MyHCⅡx mRNA表达量变化不显著；免疫组化结果显示：快肌蛋白呈棕色，慢肌蛋白呈灰白色，在出生1日龄～1周龄时，快、慢肌呈弥散交叉分布，到2周龄时慢肌簇被快肌包被在肌束中间，随着年龄增加，慢肌簇逐渐减少。Western blot检测结果显示：慢肌蛋白从出生到2周龄显著下降，在2周龄～2月龄阶段下降不明显，生长到6月龄时又显著降低；快肌蛋白随日龄增加而增加，在2周龄～6月龄期间增加极显著；βcatenin、GSK3β蛋白表达量与其mRNA表达一致，而pβcatenin在2周龄时显著高于1日龄，且随后持续上调；Tyr216pGSK3β蛋白表达量早期生长下调极显著；Ser9pGSK3β蛋白表达量则是在后期生长显著降低。上述基因表达变化规律揭示，Wnt/βcatenin信号通路相关基因βcatenin、Fz3、GSK3β与MyHCⅠ 表达呈正相关，与MyHCⅡ表达呈负相关，其在骨骼肌生长过程中对肌纤维类型组成具有重要影响。

关键词: Wnt/βcatenin信号通路, 肌纤维类型, 猪, 背最长肌, 发育

Abstract: The aim of this study was to investigate the mechanism of effect of Wnt/βcatenin signal pathway on the porcine skeletal muscle fiber types development. The longissimus doris from Large White pigs at the age of 1 day, 1 week, 2 weeks, 1 month, 2 months and 6 months were used as research materials. Realtime PCR, Western blot and immunohistochemical methods were used to identify the morphologic and histologic change of porcine skeletal muscle during development process, and research the expression of Wnt/βcatenin signal pathway related genes and MyHCs during longissimus doris development. Realtime PCR results showed that MyHCⅠ, GSK3β, βcatenin and Fz3 mRNA expression decreased, MyHCⅡb mRNA increased gradually, MyHCⅡa mRNA increased and then decreased from 1 day to 2 weeks old, and MyHCⅡx mRNA did not change significantly during the development process of porcine longissimus doris. The results of immunohistochemical showed that the slow mucle protein was gray; fast muscle protein was brown. The fast and slow muscle fibers were crossdispersion distribution from 1 day to 1 week old, then slow fibers were coated by fast muscle fibers after 2 weeks old and then the slow muscle fibers decreased gradually. Western bolt results showed that the slow muscle protein decreased significantly from 1 day to 2 weeks old, its change was not significant from 2 weeks to 2 months old, then decreased at 6 months old. The fast muscle protein increased with development, it was extremely increased from 2 weeks to 6 months old. The protein expression of βcatenin and GSK3β were consistent with their mRNA. The protein of pβcatenin was much higher at 2 weeks old than that at 1 day old, and then increased continuously, Tyr216pGSK3β was reduced significantly during early growth, ser9pGSK3β was decreased during later growth. These results suggest that the expression of Wnt/βcatenin signaling pathway related genes βcatenin, Fz3 and GSK3β were positively correlated with MyHCⅠ, but negatively correlated with MyHCⅡ. It played an important role on composition of muscle fiber type during skeletal muscle development.

Key words: Wnt/βcatenin signaling pathway, muscle fiber types, porcine, longissimus doris, developme

杨秋梅;史新娥;沈清武;刘月光;高晓娟;陈宗正;杨公社. Wnt/βcatenin信号通路相关基因及MyHCs在猪骨骼肌发育中的表达规律[J]. 畜牧兽医学报, 2011, 42(12): 1686-1695.

YANG Qiumei;SHI Xine;SHEN Qingwu;LIU Yueguang;GAO Xiaojuan;CHEN Zongzheng;YANG Gongshe . The Expression Pattern of Wnt/βcatenin Signal Pathway Related Genes and MyHCs during Porcine Skeletal Muscle Development[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(12): 1686-1695.

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