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bioRxiv · 10.1101/2023.08.31.553598

Na+/K+ATPase - Cav1.2 nanodomain differentially regulates intracellular and local adrenergic signaling in cardiac myocytes

Karbowski, M.·Boyman, L.·Garber, L.·Joca, H.·Verhoeven, N.·Coleman, A. K.·Ward, C.·Lederer, W. J.·Greiser, M.
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Abstract

BackgroundThe intracellular Na+ concentration ([Na+]i) is a crucial but understudied regulator of cardiac myocyte function. The Na+/K+ ATPase (NKA) controls the steady-state [Na+]i and thereby determines the set-point for intracellular Ca2+. Here, we investigate the nanoscopic organization and local adrenergic regulation of the NKA macromolecular complex and how it differentially regulates the intracellular Na+ and Ca2+ homeostases in atrial and ventricular myocytes. MethodsMulticolor STORM super-resolution microscopy, Western Blot analyses, and in vivo examination of adrenergic regulation are employed to examine the organization and function of Na+ nanodomains in cardiac myocytes. Quantitative fluorescence microscopy at high spatiotemporal resolution is used in conjunction with cellular electrophysiology to investigate intracellular Na+ homeostasis in atrial and ventricular myocytes. ResultsThe NKA1 (NKA1) and the L-type Ca2+-channel (Cav1.2) form a nanodomain with a center-to center distance of [~]65 nm in both ventricular and atrial myocytes. NKA1 protein expression levels are [~]3 fold higher in atria compared to ventricle. 100% higher atrial INKA, produced by large NKA "superclusters", underlies the substantially lower Na+concentration in atrial myocytes compared to the benchmark values set in ventricular myocytes. The NKAs regulatory protein phospholemman (PLM) has similar expression levels across atria and ventricle resulting in a much lower PLM/NKA1 ratio for atrial compared to ventricular tissue. In addition, a huge PLM phosphorylation reserve in atrial tissue produces a high {beta}-adrenergic sensitivity of INKA in atrial myocytes. {beta}-adrenergic regulation of INKA is locally mediated in the NKA1-Cav1.2 nanodomain via A-kinase anchoring proteins. ConclusionsNKA1, Cav1.2 and their accessory proteins form a structural and regulatory nanodomain at the cardiac dyad. The tissue-specific composition and local adrenergic regulation of this "signaling cloud" is a main regulator of the distinct global intracellular Na+ and Ca2+ concentrations in atrial and ventricular myocytes.

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Karbowski, M., Boyman, L., Garber, L., Joca, H., Verhoeven, N., Coleman, A. K., Ward, C., Lederer, W. J., Greiser, M.. 2023-09-02. Na+/K+ATPase - Cav1.2 nanodomain differentially regulates intracellular and local adrenergic signaling in cardiac myocytes. https://doi.org/10.1101/2023.08.31.553598

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