UC Davis

Ca_V1.2 channels are critical players in cardiac excitation-contraction coupling, yet we do not understand how they are affected by an important therapeutic target of heart failure drugs and regulator of blood pressure, angiotensin II. Signaling through G_q-coupled AT1 receptors, angiotensin II triggers a decrease in PIP₂, a phosphoinositide component of the plasma membrane (PM) and known regulator of many ion channels. PIP₂ depletion suppresses Ca_V1.2 currents in heterologous expression systems but the mechanism of this regulation and whether a similar phenomenon occurs in cardiomyocytes is unknown. Previous studies have shown that Ca_V1.2 currents are also suppressed by angiotensin II. We hypothesized that these two observations are linked and that PIP₂ stabilizes Ca_V1.2 expression at the PM and angiotensin II depresses cardiac excitability by stimulating PIP₂ depletion and destabilization of Ca_V1.2 expression. We tested this hypothesis and report that Ca_V1.2 channels in tsA201 cells are destabilized after AT1 receptor-triggered PIP₂ depletion, leading to their dynamin-dependent endocytosis. Likewise, in cardiomyocytes, angiotensin II decreased t-tubular Ca_V1.2 expression and cluster size by inducing their dynamic removal from the sarcolemma. These effects were abrogated by PIP₂ supplementation. Functional data revealed acute angiotensin II reduced Ca_V1.2 currents and Ca²⁺ transient amplitudes thus diminishing excitation-contraction coupling. Finally, mass spectrometry results indicated whole-heart levels of PIP₂ are decreased by acute angiotensin II treatment. Based on these observations, we propose a model wherein PIP₂ stabilizes Ca_V1.2 membrane lifetimes, and angiotensin II-induced PIP₂ depletion destabilizes sarcolemmal Ca_V1.2, triggering their removal, and the acute reduction of Ca_V1.2 currents and contractility.