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免疫细胞化学定位大鼠运动神经元中的 TASK-3 通道。

Immunocytochemical localization of TASK-3 channels in rat motor neurons.

机构信息

Institut für Integrative Neuroanatomie, Charité, Philippstr. 12, 10115 Berlin, Germany.

出版信息

Cell Mol Neurobiol. 2012 Mar;32(2):309-18. doi: 10.1007/s10571-011-9762-6. Epub 2011 Oct 20.

Abstract

Motor neurons are large cholinergic neurons located in the brain stem and spinal cord. In recent years, a functional role for TASK channels in cellular excitability and vulnerability to anesthetics of motor neurons has been described. Using a polyclonal monospecific antibody against the tandem pore domain K(+) channel (K2P channel) TWIK-related acid-sensitive K(+) channel (TASK-3), we analyzed the expression of the TASK-3 protein in motor systems of the rat CNS. Immunocytochemical staining showed strong TASK-3 expression in motor neurons of the facial, trigeminal, ambiguus, and hypoglossal nuclei. Oculomotor nuclei (including trochlear and abducens nucleus) were also strongly positive for TASK-3. The parasympathetic Edinger-Westphal nucleus and dorsal vagal nucleus showed significant, but weaker expression compared with somato- and branchiomotoric neurons. In addition, motor neurons in the anterior horn of the spinal cord were also strongly labeled for TASK-3 immunoreactivity. Based on morphological criteria, TASK-3 was found in the somatodendritic compartment of motor neurons. Cellular staining using methyl green and immunofluorescence double-labeling with anti-vesicular acetylcholine transporter (anti-vAChT) indicated ubiquitous TASK-3 expression in motor neurons, whereas in other brain regions TASK-3 showed a widespread but not ubiquitous expression. In situ hybridization using a TASK-3 specific riboprobe verified the expression of TASK-3 in motor neurons at the mRNA level.

摘要

运动神经元是位于脑干和脊髓中的大型胆碱能神经元。近年来,已经描述了 TASK 通道在运动神经元的细胞兴奋性和对麻醉剂的敏感性中的功能作用。使用针对串联孔域 K(+)通道(K2P 通道)TWIK 相关酸敏感 K(+)通道(TASK-3)的多克隆单特异性抗体,我们分析了 TASK-3 蛋白在大鼠中枢神经系统运动系统中的表达。免疫细胞化学染色显示 TASK-3 在面神经、三叉神经、疑核和舌下神经核的运动神经元中表达强烈。动眼神经核(包括滑车神经核和展神经核)也对 TASK-3 呈强阳性。副交感神经的 Edinger-Westphal 核和背侧迷走神经核的表达与躯体和鳃动神经元相比虽显著,但较弱。此外,脊髓前角的运动神经元也强烈标记有 TASK-3 免疫反应性。根据形态学标准,TASK-3 存在于运动神经元的体树突区室。使用甲基绿进行细胞染色和与抗囊泡乙酰胆碱转运蛋白(抗-vAChT)的免疫荧光双重标记表明,TASK-3 在运动神经元中普遍表达,但在其他脑区中,TASK-3 则广泛存在但并非普遍存在。使用 TASK-3 特异性核糖探针的原位杂交在 mRNA 水平上验证了 TASK-3 在运动神经元中的表达。

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