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The Journal of Neuroscience, August 12, 2009, 29(32):10063-10071; doi:10.1523/JNEUROSCI.1312-09.2009

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Cellular/Molecular
Best1 Is a Gene Regulated by Nerve Injury and Required for Ca2+-Activated Cl– Current Expression in Axotomized Sensory Neurons

Mathieu Boudes,1,2 Chamroeun Sar,1 Aurélie Menigoz,1,2 Cécile Hilaire,1 Marie O. Péquignot,1 Alexei Kozlenkov,1 Alan Marmorstein,3 Patrick Carroll,1 Jean Valmier,1,2 and Frédérique Scamps1

1INSERM, U-583, F-34000 Montpellier, France, 2University Montpellier II, F-34000 Montpellier, France, and 3Department of Ophthalmology and Vision Science, University of Arizona, Tucson, Arizona 85724

Correspondence should be addressed to Dr. Frédérique Scamps, INM-Hôpital St. Eloi, INSERM U-583, 80, rue Augustin Fliche, 34091 Montpellier Cedex 5, France. Email: scamps{at}univ-montp2.fr

We investigated the molecular determinants of Ca2+-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.

Received March 18, 2009; revised April 28, 2009; accepted May 11, 2009.

Correspondence should be addressed to Dr. Frédérique Scamps, INM-Hôpital St. Eloi, INSERM U-583, 80, rue Augustin Fliche, 34091 Montpellier Cedex 5, France. Email: scamps{at}univ-montp2.fr






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