Inhibition of CaV2.3 channels by NK1 receptors is sensitive to membrane cholesterol but insensitive to caveolin-1
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Voltage-gated, CaV2.3 calcium channels and neurokinin-1 (NK1) receptors are both present in nuclei of the central nervous system. When transiently coexpressed in human embryonic kidney (HEK) 293 cells, CaV2.3 is primarily inhibited during strong, agonist-dependent activation of NK1 receptors. NK1 receptors localize to plasma membrane rafts, and their modulation by Gq/11 protein-coupled signaling is sensitive to plasma membrane cholesterol. Here, we show that inhibition of CaV2.3 by NK1 receptors is attenuated following methyl-β-cyclodextrin (MBCD)-mediated depletion of membrane cholesterol. By contrast, inhibition of CaV2.3 was unaffected by intracellular diffusion of caveolin-1 scaffolding peptide or by overexpression of caveolin-1. Interestingly, MΒCD treatment had no effect on the macroscopic biophysical properties of CaV2.3, though it significantly decreased whole-cell membrane capacitance. Our data indicate that (1) cholesterol supports at least one component of the NK1 receptor-linked signaling pathway that inhibits CaV2.3 and (2) caveolin-1 is dispensable within this pathway. Our findings suggest that NK1 receptors reside within non-caveolar membrane rafts and that CaV2.3 resides nearby but outside the rafts. Raft-dependent modulation of CaV2.3 could be important in the physiological and pathophysiological processes in which these channels participate, including neuronal excitability, synaptic plasticity, epilepsy, and chronic pain. © 2014, Springer-Verlag Berlin Heidelberg.
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Caveolin-1; Membrane capacitance; Membrane rafts; Methyl-β-cyclodextrin; NK1 receptors; R-type channels beta 2 adrenergic receptor; calcium channel R type; caveolin 1; cholesterol; methyl beta cyclodextrin; neurokinin 1 receptor; beta cyclodextrin derivative; calcium channel R type; Cav1 protein, rat; caveolin 1; cholesterol; methyl-beta-cyclodextrin; neurokinin 1 receptor; Article; cell membrane potential; concentration (parameters); conductance; controlled study; diffusion; electric potential; embryo; enzyme activation; enzyme inactivation; enzyme inhibition; gene overexpression; HEK293 cell line; human; human cell; membrane formation; nonhuman; priority journal; protein protein interaction; receptor sensitivity; signal transduction; steady state; systematic review; animal; cell membrane; drug effects; genetic transfection; genetics; membrane potential; metabolism; rabbit; rat; signal transduction; Animals; beta-Cyclodextrins; Calcium Channels, R-Type; Caveolin 1; Cholesterol; HEK293 Cells; Humans; Membrane Microdomains; Membrane Potentials; Rabbits; Rats; Receptor Cross-Talk; Receptors, Neurokinin-1; Transfection
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