Aberrant Splicing Induced by Dysregulated Rbfox2 Produces Enhanced Function of Ca _V 1.2 Calcium Channel and Vascular Myogenic Tone in Hypertension

Abstract

Calcium influx from activated voltage-gated calcium channel Ca1.2 in vascular smooth muscle cells is indispensable for maintaining myogenic tone and blood pressure. The function of Ca1.2 channel can be optimized by alternative splicing, one of post-transcriptional modification mechanisms. The splicing factor Rbfox2 is known to regulate the Ca1.2 pre-mRNA alternative splicing events during neuronal development. However, Rbfox2's roles in modulating the key function of vascular Ca1.2 channel and in the pathogenesis of hypertension remain elusive. Here, we report that the proportion of Ca1.2 channels with alternative exon 9* is increased by 10.3%, whereas that with alternative exon 33 is decreased by 10.5% in hypertensive arteries. Surprisingly, the expression level of Rbfox2 is increased ≈3-folds, presumably because of the upregulation of a dominant-negative isoform of Rbfox2. In vascular smooth muscle cells, we find that knockdown of Rbfox2 dynamically increases alternative exon 9*, whereas decreases exon 33 inclusion of Ca1.2 channels. By patch-clamp studies, we show that diminished Rbfox2-induced alternative splicing shifts the steady-state activation and inactivation curves of vascular Ca1.2 calcium channel to hyperpolarization, which makes the window current potential to more negative. Moreover, siRNA-mediated knockdown of Rbfox2 increases the pressure-induced vascular myogenic tone of rat mesenteric artery. Taken together, our data indicate that Rbfox2 modulates the functions of vascular Ca1.2 calcium channel by dynamically regulating the expressions of alternative exons 9* and 33, which in turn affects the vascular myogenic tone. Therefore, our work suggests a key role for Rbfox2 in hypertension, which provides a rational basis for designing antihypertensive therapies.

Identification of putative biomarkers for Infantile Hemangiomas and Propranolol treatment via data integration

“…The activation of the Cav1.2 is a response to changes in intramural pressure due to blood flow changes. In an animal model, Zhou and colleagues 80 found that knocking down Rbfox2 up-regulates Cav1.2 channel activity and leads to vasoconstriction. Further, Murphy et al 81 found that Rbfox2 is an important regulator of endothelial cells exposed to an acute reduction in blood flow.…”

Diversification of the muscle proteome through alternative splicing

“…In the case of Ca V 1.1 ( CACNA1S ) alternative splicing of exon 29 generates a calcium conducting Ca V 1.1e that enhances calcium influx and generates spontaneous calcium sparklets in muscle fiber during EC coupling, resulting in reduced force and enhanced endurance as observed in myotonic dystrophies DM1 and DM2 [ 138 , 139 ]. Similarly, in the vascular smooth muscle and heart muscle cells, Ca V 1.2 is alternatively spliced by RBFOX2 generating a variant with exon 9 inclusion (Ca v 1.2SM) and exon 33 (Ca v 1.2CM) skipping during hypertension [ 140 ]. Functionally, these Ca v 1.2 splice variants differ in their Ca 2+ window current, where the Ca v 1.2SM isoform can stimulate Ca 2+ influx at a much-reduced action potential for basal contractility during blood flow [ 141 ].…”

Mutations in voltage-gated L-type calcium channel: implications in cardiac arrhythmia

“…Recently, multiply alternative splicing events have been found in CACNA1C , which optimize the functions of Ca V 1.2 channel [13,14]. Moreover, alternative splicing in Ca V 1.2 channels make some roles in several cardiovascular diseases, including cardiac arrhythmia [15–17]. Ca V 1.3 α 1D ( CACNA1D ) is highly expressed in both SANCs and cochlear inner hair cells [18–21].…”