Unité Mixte de Recherche 1272 Physiologie de l'Insecte, Signalisation et Communication, Institut National de la Recherche Agronomique, F-78000 Versailles, France.
J Neurosci. 2010 May 5;30(18):6323-33. doi: 10.1523/JNEUROSCI.0261-10.2010.
The response of insect olfactory receptor neurons (ORNs) to odorants involves the opening of Ca(2+)-permeable channels, generating an increase in intracellular Ca(2+) concentration. Here, we studied the downstream effect of this Ca(2+) rise in cultured ORNs of the moth Spodoptera littoralis. Intracellular dialysis of Ca(2+) from the patch pipette in whole-cell patch-clamp configuration activated a conductance with a K(1/2) of 2.8 microm. Intracellular and extracellular anionic and cationic substitutions demonstrated that Cl(-) carries this current. The anion permeability sequence I(-) > NO(3)(-) > Br(-) > Cl(-) > CH(3)SO(3)(-) >> gluconate(-) of the Ca(2+)-activated Cl(-) channel suggests a weak electrical field pore of the channel. The Ca(2+)-activated current partly inactivated over time and did not depend on protein kinase C (PKC) and CaMKII activity or on calmodulin. Application of Cl(-) channel blockers, flufenamic acid, 5-nitro-2-(3-phenylpropylamino) benzoic acid, or niflumic acid reversibly blocked the Ca(2+)-activated current. In addition, lowering Cl(-) concentration in the sensillar lymph bathing the ORN outer dendrites caused a significant delay in pheromone response termination in vivo. The present work identifies a new Cl(-) conductance activated by Ca(2+) in insect ORNs presumably required for ORN repolarization.
昆虫嗅觉受体神经元 (ORNs) 对气味的反应涉及钙 (Ca(2+))-通透通道的打开,导致细胞内 Ca(2+)浓度增加。在这里,我们研究了鳞翅目昆虫斜纹夜蛾 ORNs 培养物中这种 Ca(2+)上升的下游效应。在全细胞膜片钳配置中,通过贴壁式细胞内灌流从膜片钳电极内液中提取 Ca(2+),激活了一种 K(1/2)为 2.8 μm 的电导。细胞内和细胞外的阴离子和阳离子取代实验表明,Cl(-)携带这种电流。Ca(2+)-激活 Cl(-)通道的阴离子通透性序列 I(-) > NO(3)(-) > Br(-) > Cl(-) > CH(3)SO(3)(-) >> gluconate(-) 表明通道具有较弱的电场孔。Ca(2+)-激活电流随时间部分失活,不依赖于蛋白激酶 C (PKC) 和 CaMKII 活性或钙调蛋白。Cl(-)通道阻滞剂、氟芬那酸、5-硝基-2-(3-苯丙基氨基)苯甲酸或尼氟灭酸的应用可可逆地阻断 Ca(2+)-激活的电流。此外,降低 ORN 外树突感觉淋巴液中的 Cl(-)浓度会导致体内信息素反应终止的显著延迟。本工作鉴定了一种新的 Ca(2+)-激活的 Cl(-)电导,它可能是昆虫 ORNs 复极化所必需的。