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鉴定分离的大鼠心室肌细胞中不同通道的 RNA 读取,以及细胞拉伸对 L 型钙电流的影响。

Identification of RNA reads encoding different channels in isolated rat ventricular myocytes and the effect of cell stretching on L-type Cacurrent.

机构信息

Department of Physiology, Pirogov Russian National Research Medical University, Moscow, Russian Federation.

Faculty of Natural Sciences and Mathematics, Institute of Biology, "Ss. Cyril and Methodius" University, Skopje, North, Macedonia.

出版信息

Biol Direct. 2023 Oct 30;18(1):70. doi: 10.1186/s13062-023-00427-0.

Abstract

BACKGROUND

The study aimed to identify transcripts of specific ion channels in rat ventricular cardiomyocytes and determine their potential role in the regulation of ionic currents in response to mechanical stimulation. The gene expression levels of various ion channels in freshly isolated rat ventricular cardiomyocytes were investigated using the RNA-seq technique. We also measured changes in current through Ca1.2 channels under cell stretching using the whole-cell patch-clamp method.

RESULTS

Among channels that showed mechanosensitivity, significant amounts of TRPM7, TRPC1, and TRPM4 transcripts were found. We suppose that the recorded L-type Ca current is probably expressed through Ca1.2. Furthermore, stretching cells by 6, 8, and 10 μm, which increases I through the TRPM7, TRPC1, and TRPM4 channels, also decreased I through the Ca1.2 channels in K /K , Cs /K , K /Cs , and Cs /Cs  solutions. The application of a nonspecific I blocker, Gd, during cell stretching eliminated I through nonselective cation channels and I through Ca1.2 channels. Since the response to Gd was maintained in Cs /Cs  solutions, we suggest that voltage-gated Ca1.2 channels in the ventricular myocytes of adult rats also exhibit mechanosensitive properties.

CONCLUSIONS

Our findings suggest that TRPM7, TRPC1, and TRPM4 channels represent stretch-activated nonselective cation channels in rat ventricular myocytes. Probably the Ca1.2 channels in these cells exhibit mechanosensitive properties. Our results provide insight into the molecular mechanisms underlying stretch-induced responses in rat ventricular myocytes, which may have implications for understanding cardiac physiology and pathophysiology.

摘要

背景

本研究旨在鉴定大鼠心室心肌细胞中特定离子通道的转录本,并确定它们在响应机械刺激调节离子电流中的潜在作用。使用 RNA-seq 技术研究了新鲜分离的大鼠心室心肌细胞中各种离子通道的基因表达水平。我们还使用全细胞膜片钳技术测量了细胞拉伸时 Ca1.2 通道电流的变化。

结果

在表现出机械敏感性的通道中,发现了大量的 TRPM7、TRPC1 和 TRPM4 转录本。我们推测记录的 L 型 Ca 电流可能通过 Ca1.2 表达。此外,将细胞拉伸 6、8 和 10 μm,这会增加通过 TRPM7、TRPC1 和 TRPM4 通道的 I,也会降低在 K/K、Cs/K、K/Cs 和 Cs/Cs 溶液中通过 Ca1.2 通道的 I。在细胞拉伸时应用非特异性 I 阻断剂 Gd 消除了非选择性阳离子通道和 Ca1.2 通道的 I。由于在 Cs/Cs 溶液中对 Gd 的反应得以维持,我们推测成年大鼠心室心肌细胞中的电压门控 Ca1.2 通道也表现出机械敏感性。

结论

我们的研究结果表明,TRPM7、TRPC1 和 TRPM4 通道代表大鼠心室心肌细胞中的拉伸激活非选择性阳离子通道。可能这些细胞中的 Ca1.2 通道具有机械敏感性。我们的研究结果为理解大鼠心室心肌细胞中拉伸诱导反应的分子机制提供了线索,这可能对理解心脏生理学和病理生理学具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ea6/10614344/7e15b6f1b404/13062_2023_427_Fig1_HTML.jpg

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