Heterogeneity of subcellular localization and electrophoretic mobility of survival motor neuron (SMN) protein in mammalian neural cells and tissues
Heterogeneity of subcellular localization and electrophoretic mobility of survival motor neuron (SMN) protein in mammalian neural cells and tissues
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DOI：
10.1073/pnas.95.11.6492
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发表时间：
1998-05-26
影响因子：
11.1
通讯作者：
Brown, RH
中科院分区：
文献类型：
--
作者：
Francis, JW;Sandrock, AW;Brown, RH
脊髓性肌萎缩是由存活运动神经元（SMN）基因缺陷引起的。为了更好地了解神经元细胞和组织中SMN的表达模式，我们从SMN外显子2合成了一种寡肽，并构建了一种多克隆抗体（abSMN）。神经母细胞瘤细胞、小鼠和人中枢神经系统（CNS）的AbSMN免疫染色显示出强烈的核“宝石”标记，以及小鼠和人中枢神经系统组织中突出的核仁免疫反应性。在人类脊髓运动神经元的核周和近端树突中观察到强烈的细胞质标记，但在其轴突中没有。免疫印迹分析显示，在来自人类SY5Y神经母细胞瘤细胞、胚胎小鼠脊髓培养和人类中枢神经系统组织的不溶性蛋白组分中存在34 kda的物种。相比之下，在SY5Y细胞的细胞质部分检测到38-kDa种。我们得出结论，SMN蛋白在脑和脊髓的多种类型神经元的细胞质和细胞核中都有显著表达，这一发现与SMN作为神经元活力的决定因素的作用一致。
Spinal muscular atrophy is caused by defects in the survival motor neuron (SMN) gene. To better understand the patterns of expression of SMN in neuronal cells and tissues, we raised a polyclonal antibody (abSMN) against a synthetic oligopeptide from SMN exon 2. AbSMN immunostaining in neuroblastoma cells and mouse and human central nervous system (CNS) showed intense labeling of nuclear "gems," along with prominent nucleolar immunoreactivity in mouse and human CNS tissues. Strong cytoplasmic labeling was observed in the perikarya and proximal dendrites of human spinal motor neurons but not in their axons, Immunoblot analysis revealed a 34-kDa species in the insoluble protein fractions from human SY5Y neuroblastoma cells, embryonic mouse spinal cord cultures, and human CNS tissue. By contrast, a 38-kDa species was detected in the cytosolic fraction of SY5Y cells, We conclude that SMN protein is expressed prominently in both the cytoplasm and nucleus in multiple types of neurons in brain and spinal cord, a finding consistent with a role for SMN as a determinant of neuronal viability.