INSERM, Research Centre U836, F-38042 Grenoble, France.
J Cell Sci. 2010 Sep 1;123(Pt 17):2943-54. doi: 10.1242/jcs.068817. Epub 2010 Aug 10.
The highly conserved ESCRT-III complex is responsible for deformation and cleavage of membranes during endosomal trafficking and other cellular activities. In humans, dominant mutations in the ESCRT-III subunit CHMP2B cause frontotemporal dementia (FTD). The decade-long process leading to this cortical degeneration is not well understood. One possibility is that, akin to other neurodegenerative diseases, the pathogenic protein affects the integrity of dendritic spines and synapses before any neuronal death. Using confocal microscopy and 3D reconstruction, we examined whether expressing the FTD-linked mutants CHMP2B(intron5) and CHMP2B(Delta10) in cultured hippocampal neurons modified the number or structure of spines. Both mutants induced a significant decrease in the proportion of large spines with mushroom morphology, without overt degeneration. Furthermore, CHMP2B(Delta10) induced a drop in frequency and amplitude of spontaneous excitatory postsynaptic currents, suggesting that the more potent synapses were lost. These effects seemed unrelated to changes in autophagy. Depletion of endogenous CHMP2B by RNAi resulted in morphological changes similar to those induced by mutant CHMP2B, consistent with dominant-negative activity of pathogenic mutants. Thus, CHMP2B is required for spine growth. Taken together, these results demonstrate that a mutant ESCRT-III subunit linked to a human neurodegenerative disease can disrupt the normal pattern of spine development.
高度保守的 ESCRT-III 复合物负责在胞内体运输和其他细胞活动期间膜的变形和分裂。在人类中,ESCRT-III 亚基 CHMP2B 的显性突变导致额颞叶痴呆(FTD)。导致这种皮质变性的长达十年的过程尚不清楚。一种可能性是,类似于其他神经退行性疾病,致病蛋白在任何神经元死亡之前影响树突棘和突触的完整性。使用共聚焦显微镜和 3D 重建,我们检查了在培养的海马神经元中表达 FTD 相关突变体 CHMP2B(intron5)和 CHMP2B(Delta10)是否改变了棘的数量或结构。这两种突变体都诱导蘑菇形态的大棘的比例显著降低,而没有明显的退化。此外,CHMP2B(Delta10)诱导自发兴奋性突触后电流的频率和幅度下降,表明更有效的突触丢失。这些影响似乎与自噬的变化无关。通过 RNAi 耗尽内源性 CHMP2B 导致与突变 CHMP2B 诱导的形态变化相似,与致病性突变体的显性负性活性一致。因此,CHMP2B 是棘突生长所必需的。总之,这些结果表明与人类神经退行性疾病相关的突变 ESCRT-III 亚基可以破坏正常的棘突发育模式。