Down-regulation of endogenous KLHL1 decreases voltage-gated calcium current density Journal Article

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Authors: Perissinotti, P. P.; Ethington, E. G.; Cribbs, L.; Koob, M. D.; Martin, J.; Piedras-Renteria, E. S.
Article Title: Down-regulation of endogenous KLHL1 decreases voltage-gated calcium current density
Abstract: The actin-binding protein Kelch-like 1 (KLHL1) can modulate voltage-gated calcium channels in vitro. KLHL1 interacts with actin and with the pore-forming subunits of Cav2.1 and CaV3.2 calcium channels, resulting in up-regulation of P/Q and T-type current density. Here we tested whether endogenous KLHL1 modulates voltage gated calcium currents in cultured hippocampal neurons by down-regulating the expression of KLHL1 via adenoviral delivery of shRNA targeted against KLHL1 (shKLHL1). Control adenoviruses did not affect any of the neuronal properties measured, yet down-regulation of KLHL1 resulted in HVA current densities approximately 68% smaller and LVA current densities 44% smaller than uninfected controls, with a concomitant reduction in alpha1A and alpha1H protein levels. Biophysical analysis and western blot experiments suggest CaV3.1 and 3.3 currents are also present in shKLHL1-infected neurons. Synapsin I levels, miniature postsynaptic current frequency, and excitatory and inhibitory synapse number were reduced in KLHL1 knockdown. This study corroborates the physiological role of KLHL1 as a calcium channel modulator and demonstrates a novel, presynaptic role.
Journal Title: Cell calcium
Volume: 55
Issue: 5
ISSN: 1532-1991; 0143-4160
Publisher: Elsevier Inc  
Date Published: 2014
Start Page: 269
End Page: 280
Language: ENG
DOI/URL:

10.1016/j.ceca.2014.03.002

10.1016/j.ceca.2014.03.002
Notes: CI: Copyright (c) 2014; JID: 8006226; 0 (Calcium Channels, N-Type); 0 (Calcium Channels, T-Type); 0 (KLHL1 protein, human); 0 (Microfilament Proteins); 0 (Quinoxalines); 0 (RNA, Small Interfering); 0 (voltage-dependent calcium channel (P-Q type)); 118876-58-7 (2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline); SY7Q814VUP (Calcium); Y37615DVKC (Bicuculline); OTO: NOTNLM; 2013/07/27 [received]; 2014/03/04 [revised]; 2014/03/09 [accepted]; 2014/03/21 [aheadofprint]; ppublish