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鸡翅膀和腿部肌肉中快速骨骼肌肌钙蛋白T基因互斥外显子的差异表达。

Differential expression of mutually exclusive exons of the fast skeletal muscle troponin T gene in the chicken wing and leg muscles.

作者信息

Jozaki Miho, Hosoda Kouji, Miyazaki Jun-Ichi

机构信息

Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan.

出版信息

J Muscle Res Cell Motil. 2002;23(3):235-43. doi: 10.1023/a:1020956216423.

Abstract

To determine the physiological significance of developmentally regulated and muscle tissue-specific expression of troponin T (TnT) isoforms and also to elucidate mechanisms of alternative splicing, we investigated splicing patterns of fast skeletal muscle TnT by sequencing cDNAs obtained from the chicken wing and leg muscles. The chicken fast skeletal muscle TnT gene has 27 exons, and produces potentially 2(15) transcripts if alternative exons are randomly spliced. For the fourteen alternative exons in the 5'-region of the gene, we found 7 and 12 splice variants in the wing biceps brachii and leg gastrocnemius, respectively. Out of 19, 12 variants were newly found in this study. Twenty two different splice variants were reported by the previous studies, but this number has now increased to 34 in the chicken TnT gene, suggesting that alternative splicing is not random, but strictly regulated. We also investigated splicing patterns of mutually exclusive exons 16 and 17 in the 3'-region of the gene. Approximately equal amounts of cDNAs containing exons 16 and 17 were detected in proximal regions of adult biceps brachii and gastrocnemius, respectively, whereas cDNA with exon 17 dominated in distal regions. Only cDNA containing exon 17 was found in both proximal and distal regions of 5-day-old gastrocnemius. The inclusion of exon 16 into cDNAs was detected in the proximal region of 15-day-old gastrocnemius and increased during development.

摘要

为了确定肌钙蛋白T(TnT)亚型的发育调控和肌肉组织特异性表达的生理意义,并阐明可变剪接机制,我们通过对从鸡翼和腿部肌肉获得的cDNA进行测序,研究了快速骨骼肌TnT的剪接模式。鸡的快速骨骼肌TnT基因有27个外显子,如果外显子随机剪接,可能产生2(15)种转录本。对于该基因5'区域的14个可变外显子,我们分别在翼部肱二头肌和腿部腓肠肌中发现了7种和12种剪接变体。在这19种变体中,有12种是本研究中新发现的。先前的研究报道了22种不同的剪接变体,但现在鸡TnT基因中的这一数量已增加到34种,这表明可变剪接并非随机,而是受到严格调控。我们还研究了该基因3'区域中相互排斥的外显子16和17的剪接模式。在成年肱二头肌和腓肠肌的近端区域,分别检测到含有外显子16和17的cDNA数量大致相等,而在远端区域,含有外显子17的cDNA占主导。在5日龄腓肠肌的近端和远端区域,仅发现了含有外显子17的cDNA。在15日龄腓肠肌的近端区域检测到外显子16包含在cDNA中,且在发育过程中增加。

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