Departments of Experimental Cardiology, Amsterdam UMC, location AMC, 1105 AZ, Amsterdam, The Netherlands.
Clinical Epidemiology, Biostatistics and Bioinformatics, Amsterdam UMC, location AMC, 1105 AZ, Amsterdam, The Netherlands.
J Cell Sci. 2022 Jan 1;135(1). doi: 10.1242/jcs.259191. Epub 2022 Jan 7.
Eukaryotic genomes contain a tiny subset of 'minor class' introns with unique sequence elements that require their own splicing machinery. These minor introns are present in certain gene families with specific functions, such as voltage-gated Na+ and voltage-gated Ca2+ channels. Removal of minor introns by the minor spliceosome has been proposed as a post-transcriptional regulatory layer, which remains unexplored in the heart. Here, we investigate whether the minor spliceosome regulates electrophysiological properties of cardiomyocytes by knocking down the essential minor spliceosome small nuclear snRNA component U6atac in neonatal rat ventricular myocytes. Loss of U6atac led to robust minor intron retention within Scn5a and Cacna1c, resulting in reduced protein levels of Nav1.5 and Cav1.2 channels. Functional consequences were studied through patch-clamp analysis, and revealed reduced Na+ and L-type Ca2+ currents after loss of U6atac. In conclusion, minor intron splicing modulates voltage-dependent ion channel expression and function in cardiomyocytes. This may be of particular relevance in situations in which minor splicing activity changes, such as in genetic diseases affecting minor spliceosome components, or in acquired diseases in which minor spliceosome components are dysregulated, such as heart failure.
真核基因组包含一小部分具有独特序列元件的“次要类”内含子,这些内含子需要自己的剪接机制。这些次要内含子存在于某些具有特定功能的基因家族中,如电压门控 Na+和电压门控 Ca2+通道。通过次要剪接体去除次要内含子被认为是一种转录后调控层,这在心脏中尚未得到探索。在这里,我们通过在新生大鼠心室肌细胞中敲低必需的次要剪接体小核 snRNA 成分 U6atac 来研究次要剪接体是否通过调节心肌细胞的电生理特性。U6atac 的缺失导致 Scn5a 和 Cacna1c 中的次要内含子大量保留,从而导致 Nav1.5 和 Cav1.2 通道的蛋白水平降低。通过膜片钳分析研究了功能后果,结果表明 U6atac 缺失后 Na+和 L 型 Ca2+电流减少。总之,次要内含子剪接调节心肌细胞中电压依赖性离子通道的表达和功能。在影响次要剪接体成分的遗传疾病或次要剪接体成分失调的获得性疾病(如心力衰竭)中,这种情况可能特别相关。