快速起搏降低l型Ca2+电流并改变原代培养大鼠左心室肌细胞Cacna1c异基因表达。

IF 2.3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Anne Ritzer, Tobias Roeschl, Sandra Nay, Elena Rudakova, Tilmann Volk
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引用次数: 0

摘要

l型钙电流(ICaL)是心脏兴奋-收缩-耦合的第一步,不仅在调节收缩力方面起着重要作用,而且在电重构和机械重构中也起着重要作用。心肌细胞原代培养是心脏离子通道研究中广泛使用的一种工具,它与大量的形态学、功能和电学变化有关,其中一些变化可能被电起搏阻止。因此,我们在大鼠左心室肌细胞分离后,以及在1和3hz频率下进行和不进行常规起搏的24小时原代培养后直接研究了ICaL。此外,我们分析了l型Ca2+通道的孔形成亚基(cacna1c)的总mRNA表达以及其外显子1的剪接变体的表达,这些变体有助于心肌细胞或平滑肌等不同组织中ICaL的特异性。无起搏24 h孵育仅使ICaL密度降低约10%。与这种减少一致,我们观察到总cacna1c和外显子1a(心肌细胞的优势变体)的表达减少,而外显子1b和1c的表达增加。在1和3hz频率下起搏24 h,导致ICaL密度大幅下降30%,略微减缓了ICaL失活,并将稳态失活转移到更多的负电位。起搏器显著降低了cacna1c mRNA的总表达,外显子1b和1c的表达也是如此。综上所述,与起搏24小时相比,电沉默对ICaL密度和cacna1c mRNA表达的影响更小,因此应该是心肌细胞原代培养的首选方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Rapid Pacing Decreases L-type Ca<sup>2+</sup> Current and Alters Cacna1c Isogene Expression in Primary Cultured Rat Left Ventricular Myocytes.

Rapid Pacing Decreases L-type Ca2+ Current and Alters Cacna1c Isogene Expression in Primary Cultured Rat Left Ventricular Myocytes.

The L-type calcium current (ICaL) is the first step in cardiac excitation-contraction-coupling and plays an important role in regulating contractility, but also in electrical and mechanical remodeling. Primary culture of cardiomyocytes, a widely used tool in cardiac ion channel research, is associated with substantial morphological, functional and electrical changes some of which may be prevented by electrical pacing. We therefore investigated ICaL directly after cell isolation and after 24 h of primary culture with and without regular pacing at 1 and 3 Hz in rat left ventricular myocytes. Moreover, we analyzed total mRNA expression of the pore forming subunit of the L-type Ca2+ channel (cacna1c) as well as the expression of splice variants of its exon 1 that contribute to specificity of ICaL in different tissue such as cardiac myocytes or smooth muscle. 24 h incubation without pacing decreased ICaL density by ~ 10% only. Consistent with this decrease we observed a decrease in the expression of total cacna1c and of exon 1a, the dominant variant of cardiomyocytes, while expression of exon 1b and 1c increased. Pacing for 24 h at 1 and 3 Hz led to a substantial decrease in ICaL density by 30%, mildly slowed ICaL inactivation and shifted steady-state inactivation to more negative potentials. Total cacna1c mRNA expression was substantially decreased by pacing, as was the expression of exon 1b and 1c. Taken together, electrical silence introduces fewer alterations in ICaL density and cacna1c mRNA expression than pacing for 24 h and should therefore be the preferred approach for primary culture of cardiomyocytes.

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来源期刊
Journal of Membrane Biology
Journal of Membrane Biology 生物-生化与分子生物学
CiteScore
4.80
自引率
4.20%
发文量
63
审稿时长
6-12 weeks
期刊介绍: The Journal of Membrane Biology is dedicated to publishing high-quality science related to membrane biology, biochemistry and biophysics. In particular, we welcome work that uses modern experimental or computational methods including but not limited to those with microscopy, diffraction, NMR, computer simulations, or biochemistry aimed at membrane associated or membrane embedded proteins or model membrane systems. These methods might be applied to study topics like membrane protein structure and function, membrane mediated or controlled signaling mechanisms, cell-cell communication via gap junctions, the behavior of proteins and lipids based on monolayer or bilayer systems, or genetic and regulatory mechanisms controlling membrane function. Research articles, short communications and reviews are all welcome. We also encourage authors to consider publishing ''negative'' results where experiments or simulations were well performed, but resulted in unusual or unexpected outcomes without obvious explanations. While we welcome connections to clinical studies, submissions that are primarily clinical in nature or that fail to make connections to the basic science issues of membrane structure, chemistry and function, are not appropriate for the journal. In a similar way, studies that are primarily descriptive and narratives of assays in a clinical or population study are best published in other journals. If you are not certain, it is entirely appropriate to write to us to inquire if your study is a good fit for the journal.
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