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烟草天蛾(Manduca sexta)的钙激活钾通道:分子特征与表达分析

Calcium-activated potassium channel of the tobacco hornworm, Manduca sexta: molecular characterization and expression analysis.

作者信息

Keyser Matthew R, Witten Jane L

机构信息

Department of Biological Sciences, PO Box 413, University of Wisconsin-Milwaukee, Milwaukee, WI 53201, USA.

出版信息

J Exp Biol. 2005 Nov;208(Pt 21):4167-79. doi: 10.1242/jeb.01857.

Abstract

Large-conductance calcium- and voltage-gated potassium channels (BK or Slowpoke) serve as dynamic integrators linking electrical signaling and intracellular activity. These channels can mediate many different Ca2+-dependent physiological processes including the regulation of neuronal and neuroendocrine cell excitability and muscle contraction. To gain insights into the function of BK channels in vivo, we isolated a full-length cDNA encoding the alpha subunit of a Slowpoke channel from the tobacco hornworm, Manduca sexta (msslo). Amino acid sequence comparison of the deduced Manduca protein revealed at least 80% identity to the insect Slo channels. The five C-terminal alternative splice regions are conserved, but the cloned cDNA fragments contained some unique combinations of exons E, G and I. Our spatial profile revealed that transcript levels were highest in skeletal muscle when compared with the central nervous system (CNS) and visceral muscle. The temporal profile suggested that msslo expression is regulated developmentally in a tissue- and regional-specific pattern. The levels of msslo transcripts remain relatively constant throughout metamorphosis in the CNS, transiently decline in the heart and are barely detectable in the gut except in adults. A dramatic upregulation of msslo transcript levels occurs in thoracic but not abdominal dorsal longitudinal body wall muscles (DLM), suggesting that the msSlo current plays an important role in the excitation or contractile properties of the phasic flight muscle. Our developmental profile of msslo expression suggests that msSlo currents may contribute to the changes in neural circuits and muscle properties that produce stage-specific functions and behaviors.

摘要

大电导钙激活钾通道(BK或Slowpoke通道)作为连接电信号与细胞内活动的动态整合器。这些通道可介导许多不同的钙依赖性生理过程,包括调节神经元和神经内分泌细胞的兴奋性以及肌肉收缩。为了深入了解BK通道在体内的功能,我们从烟草天蛾(Manduca sexta,简称msslo)中分离出了一个编码Slowpoke通道α亚基的全长cDNA。推导的烟草天蛾蛋白的氨基酸序列比较显示,与昆虫Slo通道至少有80%的同一性。五个C末端可变剪接区域是保守的,但克隆的cDNA片段包含外显子E、G和I的一些独特组合。我们的空间分布显示,与中枢神经系统(CNS)和内脏肌肉相比,骨骼肌中的转录水平最高。时间分布表明,msslo的表达在发育过程中以组织和区域特异性模式受到调控。在中枢神经系统中,msslo转录本水平在整个变态过程中保持相对恒定,在心脏中短暂下降,在肠道中除成虫外几乎检测不到。msslo转录本水平在胸部而非腹部背纵体壁肌肉(DLM)中显著上调,这表明msSlo电流在相飞行肌的兴奋或收缩特性中起重要作用。我们对msslo表达的发育分析表明,msSlo电流可能有助于神经回路和肌肉特性的变化,从而产生特定阶段的功能和行为。

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