Boudes Mathieu, Sar Chamroeun, Menigoz Aurélie, Hilaire Cécile, Péquignot Marie O, Kozlenkov Alexei, Marmorstein Alan, Carroll Patrick, Valmier Jean, Scamps Frédérique
INSERM, Montpellier, France.
J Neurosci. 2009 Aug 12;29(32):10063-71. doi: 10.1523/JNEUROSCI.1312-09.2009.
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.
我们研究了神经损伤后成年感觉神经元中表达的钙激活氯电流（CaCC）的分子决定因素。背根神经节表达已知在异源系统中诱导CaCC的三个基因家族的转录本：贝斯特rophin、tweety和TMEM16。我们通过定量转录分析和原位杂交发现，神经损伤仅诱导感觉神经元中贝斯特rophin-1转录本上调。在单个神经元中用RNA干扰进行基因筛选表明，小鼠Best1是损伤感觉神经元中CaCC表达所必需的。用贝斯特rophin-1突变体转染损伤的感觉神经元可抑制内源性CaCC。在未损伤的神经元中外源表达融合蛋白绿色荧光蛋白-贝斯特rophin-1表明该蛋白定位于质膜，产生具有与内源性CaCC相似的生物物理和药理学特性的CaCC。我们的数据表明，Best1属于一组因神经损伤而上调的基因，并支持损伤感觉神经元中功能性CaCC的表达。
