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αKAP剪接变体在兔快肌和慢肌骨骼肌中的产后发育表达

Post-natal developmental expression of alphaKAP splice variants in rabbit fast-twitch and slow-twitch skeletal muscle.

作者信息

Sacchetto Roberta, Salviati Leonardo, Damiani Ernesto, Margreth Alfredo

机构信息

Department of Experimental Biomedical Sciences, University of Padova, Padova, Italy.

出版信息

J Muscle Res Cell Motil. 2004;25(4-5):309-14. doi: 10.1007/s10974-004-1685-0.

Abstract

AlphaKAP is a non-kinase product of a gene within alpha CaMKII gene, that anchors catalytic subunits to skeletal muscle sarcoplasmic reticulum. alphaKAP gene expresses two distinct mRNA transcripts by alternative splicing in rabbit fast muscle, rather than one, as an apparently fiber-type specific and species-specific property. We report on the expression profile of alphaKAP gene in fast and slow muscles of adult rats and in rabbit post-natal developing muscles, by RT-PCR analysis of muscle RNA, together with Western blot analysis of expressed alphaKAP protein isoforms. We show that alphaKAP isoform predominantly expressed in muscle of newborn rabbits, corresponds to the fast-adult isoform. In fast muscle, the transition from neonatal isoform to the common adult isoform takes place early after birth, and is never complete, unlike in slow muscle. Relevant to these findings, is the observation that neonatal/embryonic alphaKAP mRNA is highly expressed in rat myotubes grown in vitro. Together, our findings suggest that alphaKAP splicing pathways are primarily modulated according to muscle developmental stage.

摘要

αKAP是α钙调蛋白激酶II基因内一个基因的非激酶产物,它将催化亚基锚定到骨骼肌肌浆网。αKAP基因在兔快肌中通过可变剪接表达两种不同的mRNA转录本,而不是一种,这显然是一种纤维类型特异性和物种特异性的特性。我们通过对肌肉RNA进行逆转录聚合酶链反应(RT-PCR)分析以及对表达的αKAP蛋白异构体进行蛋白质免疫印迹分析,报告了αKAP基因在成年大鼠的快肌和慢肌以及兔出生后发育肌肉中的表达谱。我们发现,在新生兔肌肉中主要表达的αKAP异构体与成年快肌异构体相对应。在快肌中,从新生儿异构体向常见成年异构体的转变在出生后早期发生,并且与慢肌不同,这种转变从未完全完成。与这些发现相关的是,观察到新生/胚胎αKAP mRNA在体外培养的大鼠肌管中高度表达。总之,我们的研究结果表明,αKAP剪接途径主要根据肌肉发育阶段进行调节。

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