Localization of the Slack potassium channel in the rat central nervous system
Localization of the Slack potassium channel in the rat central nervous system
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DOI：
10.1002/cne.10439
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发表时间：
2002-12-16
影响因子：
2.5
通讯作者：
Kaczmarek, LK
中科院分区：
文献类型：
--
作者：
Bhattacharjee, A;Gan, L;Kaczmarek, LK
Slack基因编码一个电压依赖性钾离子通道，其单位电导约为60 pS。来自异源表达研究的证据表明，Slack通道亚基也可以与Slo亚基联合收割机组合以产生具有较大电导的Ca 2+激活的K+通道。尽管如此，Slack在大脑中的功能仍有待确定。我们现在已经产生了一个亲和纯化的抗体对N-末端的大鼠松弛，生化和免疫组化研究。该抗体通过免疫细胞化学和Western印迹分析识别瞬时转染的CHO细胞中的Slack。该抗体还在大鼠脑细胞膜中检测到单一条带。然后使用抗体确定Slack在大鼠脑切片中的定位。最突出的Slack免疫反应性发生在脑干，特别是三叉神经系统和网状结构，其中在相关核的细胞体和轴突纤维中发现非常强烈的染色。前庭核和眼神经核的标记也很强。在听觉系统内，小斜方内侧核有一个强大的信号与巨大的突触前末梢的染色一致。强Slack免疫反应存在于嗅球，红核，小脑深核。在丘脑、黑质和杏仁核中也有标记。唯一检测到Slack免疫反应的皮质区域是额叶皮质。Slack的亚细胞和区域分布与先前报道的Slo通道亚基不同，表明Slack在调节神经元的放电特性方面也可能具有自主作用。
The Slack gene encodes a voltage-dependent K+ channel that has a unitary conductance of similar to60 pS. Evidence from heterologous expression studies suggests that Slack channel subunits can also combine with the Slo subunit to generate Ca2+-activated K+ channels of larger conductances. Nonetheless, the function of Slack in the brain remains to be identified. We have now generated an affinity-purified antibody against the N-terminal of rat Slack, for biochemical and immunohistochemical studies. The antibody recognized Slack in transiently transfected CHO cells both by immunocytochemistry and by Western blot analysis. The antibody also detected a single band in rat brain membranes. The localization of Slack in rat brain slices was then determined using the antibody. Most prominent Slack immunoreactivity occurs in the brainstem, in particular the trigeminal system and reticular formation, where very intense staining was found in both cell bodies and axonal fibers of associated nuclei. Labeling was also very strong in the vestibular and oculomotor nuclei. Within the auditory system, the medial nucleus of the trapezoid had a robust signal consistent with staining of the giant presynaptic terminals. Strong Slack immunoreactivity was present in the olfactory bulb, red nucleus, and deep cerebellar nuclei. There was labeling also in the thalamus, substantia nigra, and amygdala. The only cortical region in which Slack immunoreactivity was detected was the frontal cortex. The subcellular and regional distribution of Slack differs from that previously reported for the Slo channel subunit and suggests that Slack may also have an autonomous role in regulating the firing properties of neurons.