Zervas M, Dobrenis K, Walkley S U
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Neuropathol Exp Neurol. 2001 Jan;60(1):49-64. doi: 10.1093/jnen/60.1.49.
Niemann-Pick disease type C (NPC) is a lethal neurologic storage disorder of children most often caused by a defect in the protein NPC1. To better understand the disease we thoroughly characterized the cellular and morphological alterations occurring in murine, feline, and human NPC. Using immunocytochemistry and filipin histochemistry we show that both gangliosides and unesterified cholesterol are differentially stored in neurons of the cerebral cortex, cerebellum, and hippocampus, as well as in liver. Double fluorescence labeling revealed that GM2 ganglioside and unesterified cholesterol were partially co-localized in vesicular structures, and triple fluorescence labeling utilizing a LAMP-1 antibody identified many of these organelles as part of the late endosomal/lysosomal pathway. These observations, coupled with the proposed role of NPC1 in intracellular cholesterol movement, suggest that GM3 and GM2 gangliosides as well as unesterified cholesterol may be retrogradely cleared from late endosomes/lysosomes by an NPC1-dependent mechanism. Cellular consequences of the NPC metabolic defect as shown by parvalbumin immunocytochemistry and rapid Golgi staining, respectively, revealed characteristic axonal spheroids on GABAergic neurons and ectopic dendritogenesis that followed a species-specific gradient of: mouse < feline < human. These studies suggest that the homeostatic regulation of gangliosides and cholesterol in neurons is mediated by NPC1 and that perturbations in this mechanism cause a complex neuronal storage disorder.
尼曼-匹克C型病(NPC)是一种致命的儿童神经贮积症,最常见的病因是NPC1蛋白缺陷。为了更好地理解这种疾病,我们全面描述了小鼠、猫和人类NPC中发生的细胞和形态学改变。利用免疫细胞化学和荧光素组织化学,我们发现神经节苷脂和未酯化胆固醇在大脑皮层、小脑和海马体的神经元以及肝脏中存在差异贮积。双重荧光标记显示GM2神经节苷脂和未酯化胆固醇部分共定位于囊泡结构中,利用LAMP-1抗体进行的三重荧光标记将其中许多细胞器鉴定为晚期内体/溶酶体途径的一部分。这些观察结果,再加上NPC1在细胞内胆固醇转运中的假定作用,表明GM3和GM2神经节苷脂以及未酯化胆固醇可能通过NPC1依赖机制从晚期内体/溶酶体逆行清除。分别通过小白蛋白免疫细胞化学和快速高尔基染色显示的NPC代谢缺陷的细胞后果,揭示了GABA能神经元上典型的轴突球状体和异位树突形成,其遵循小鼠<猫<人类的物种特异性梯度。这些研究表明,神经元中神经节苷脂和胆固醇的稳态调节由NPC1介导,这种机制的扰动会导致复杂的神经元贮积症。