Best1 is a gene regulated by nerve injury and required for Ca2+-activated Cl- current expression in axotomized sensory neurons

J Neurosci. 2009 Aug 12;29(32):10063-71. doi: 10.1523/JNEUROSCI.1312-09.2009.

Abstract

We investigated the molecular determinants of Ca(2+)-activated chloride current (CaCC) expressed in adult sensory neurons after a nerve injury. Dorsal root ganglia express the transcripts of three gene families known to induce CaCCs in heterologous systems: bestrophin, tweety, and TMEM16. We found with quantitative transcriptional analysis and in situ hybridization that nerve injury induced upregulation of solely bestrophin-1 transcripts in sensory neurons. Gene screening with RNA interference in single neurons demonstrated that mouse Best1 is required for the expression of CaCC in injured sensory neurons. Transfecting injured sensory neurons with bestrophin-1 mutants inhibited endogenous CaCC. Exogenous expression of the fusion protein green fluorescent protein-Bestrophin-1 in naive neurons demonstrated a plasma membrane localization of the protein that generates a CaCC with biophysical and pharmacological properties similar to endogenous CaCC. Our data suggest that Best1 belongs to a group of genes upregulated by nerve injury and supports functional CaCC expression in injured sensory neurons.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bestrophins
  • Calcium / metabolism*
  • Cell Membrane / metabolism
  • Chlorides / metabolism*
  • Eye Proteins / genetics
  • Eye Proteins / metabolism*
  • Ganglia, Spinal / physiology*
  • Gene Knockdown Techniques
  • Green Fluorescent Proteins / genetics
  • Ion Channels
  • Membrane Potentials / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation
  • Patch-Clamp Techniques
  • RNA Interference
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sciatic Nerve / injuries*
  • Sensory Receptor Cells / physiology*
  • Transfection

Substances

  • Best1 protein, mouse
  • Bestrophins
  • Chlorides
  • Eye Proteins
  • Ion Channels
  • Vmd2L3 protein, mouse
  • Green Fluorescent Proteins
  • Calcium