Subcellular localization and putative role of VPS13A/chorein in dopaminergic neuronal cells
Subcellular localization and putative role of VPS13A/chorein in dopaminergic neuronal cells
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DOI：
10.1016/j.bbrc.2012.02.047
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发表时间：
2012-03-16
影响因子：
3.1
通讯作者：
Kishida, Shosei
中科院分区：
文献类型：
--
作者：
Hayashi, Takehiro;Kishida, Michiko;Kishida, Shosei
棘细胞舞蹈症是一种罕见的遗传性神经退行性疾病，由编码舞蹈素的空泡蛋白13同源A(VPS13A)基因功能缺失突变引起。尽管Chorein功能缺陷会导致CHAC模型小鼠纹状体神经元的凋亡，但其详细的亚细胞定位和生理作用尚不清楚。在这项研究中，我们制备了两种抗舞蹈素的多克隆抗体，并检测了内源性舞蹈素在神经细胞中的定位。免疫细胞化学显示，在分化的PC12细胞中，Chorein位于突起终末，并与突触素I部分共存。蔗糖密度梯度分离的亚细胞定位分析表明，Chorein和Synaptopagmin I定位于含有多巴胺的致密核小泡(DCV)。此外，稳定表达舞蹈素羧基末端片段的PC12细胞增加了K+诱导的多巴胺释放。综上所述，这些结果表明舞蹈素参与了DCV的胞吐作用。(C)2012 Elsevier Inc.保留所有权利。
Chorea-acanthocytosis (ChAc) is a rare hereditary neurodegenerative disorder caused by loss of function mutations in the vacuolar protein sorting 13 homolog A (VPS13A)gene encoding chorein. Although a deficiency in chorein function leads to apoptosis of striatal neurons in ChAc model mouse, its detailed subcellular localization and physiological role remain unclear. In this study, we produced two anti-chorein polyclonal antibodies and examined the intracellular localization of endogenous chorein in neuronal cells. Immunocytochemically, chorein was observed in the termini of extended neurites and partially colocalized with synaptotagmin I in differentiated PC12 cells. Subcellular localization analysis by sucrose density gradient fractionation showed that chorein and synaptotagmin I were located in dense-core vesicles (DCVs), which contain dopamine. In addition, PC12 cells stably expressing carboxyterminal fragment of chorein increased K+-induced dopamine release. Taken together, these results suggest that chorein is involved in exocytosis of DCV. (C) 2012 Elsevier Inc. All rights reserved.