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

Paula P. Perissinotti, Elizabeth G. Ethington, Leanne Cribbs, Michael D. Koob, Jody Martin, Erika S. Piedras-Rentería

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

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 ~68% smaller and LVA current densities 44% smaller than uninfected controls, with a concomitant reduction in α1A and α1H 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.

Original languageEnglish (US)
Pages (from-to)269-280
Number of pages12
JournalCell Calcium
Volume55
Issue number5
DOIs
StatePublished - May 2014

Bibliographical note

Funding Information:
We thank all members of the Piedras laboratory for helpful comments and discussions. This paper is based upon work supported by the National Science Foundation under Grant no. 1022075 (EPR).

Keywords

  • ATXN8OS
  • Actin-binding protein
  • HVA calcium current
  • LVA calcium currents
  • P/Q-type channel
  • Spinocerebellar ataxia type 8
  • Synaptic
  • T-type channel

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