Institute of Neuroscience, College of Life and Health Sciences, Northeastern University, 110819, Shenyang, P. R. China.
Department of Neurology, Skåne University Hospital, Lund University, Lund, Sweden.
Acta Neuropathol Commun. 2020 Jun 3;8(1):77. doi: 10.1186/s40478-020-00949-y.
Prominent features of HD neuropathology are the intranuclear and cytoplasmic inclusions of huntingtin and striatal and cortical neuronal cell death. Recently, synaptic defects have been reported on HD-related studies, including impairment of neurotransmitter release and alterations of synaptic components. However, the definite characteristics of synapse dysfunction and the underlying mechanisms remain largely unknown. We studied the gene expression levels and patterns of a number of proteins forming the cytoskeletal matrix of the presynaptic active zones in HD transgenic mice (R6/1), in hippocampal neuronal cultures overexpressing mutant huntingtin and in postmortem brain tissues of HD patients. To investigate the interactions between huntingtin and active proteins, we performed confocal microscopic imaging and immunoprecipitation in mouse and HEK 293 cell line models. The mRNA and protein levels of Bassoon were reduced in mouse and cell culture models of HD and in brain tissues of patients with HD. Moreover, a striking re-distribution of a complex of proteins including Bassoon, Piccolo and Munc 13-1 from the cytoplasm and synapses into intranuclear huntingtin aggregates with loss of active zone proteins and dendritic spines. This re-localization was age-dependent and coincided with the formation of huntingtin aggregates. Using co-immunoprecipitation, we demonstrated that huntingtin interacts with Bassoon, and that this interaction is likely mediated by a third linking protein. Three structural proteins involved in neurotransmitter release in the presynaptic active zones of neurons are altered in expression and that the proteins are redistributed from their normal functional site into mutant huntingtin aggregates.
亨廷顿病神经病理学的突出特征是核内和细胞质中的亨廷顿蛋白包涵体以及纹状体和皮质神经元细胞死亡。最近,在与亨廷顿病相关的研究中报道了突触缺陷,包括神经递质释放受损和突触成分改变。然而,突触功能障碍的确切特征和潜在机制在很大程度上仍不清楚。我们研究了一些形成突触前活性区细胞骨架基质的蛋白质的基因表达水平和模式,这些蛋白质在亨廷顿病转基因小鼠(R6/1)、过度表达突变亨廷顿蛋白的海马神经元培养物和亨廷顿病患者的死后脑组织中。为了研究亨廷顿蛋白与活性蛋白之间的相互作用,我们在小鼠和 HEK 293 细胞系模型中进行了共聚焦显微镜成像和免疫沉淀。在亨廷顿病的小鼠和细胞培养模型以及亨廷顿病患者的脑组织中,Bassoon 的 mRNA 和蛋白水平降低。此外,包括 Bassoon、Piccolo 和 Munc 13-1 在内的蛋白质复合物从细胞质和突触重新分布到含有活性区蛋白和树突棘丢失的核内亨廷顿蛋白聚集体中。这种重新定位是年龄依赖性的,与亨廷顿蛋白聚集体的形成一致。通过共免疫沉淀,我们证明亨廷顿蛋白与 Bassoon 相互作用,并且这种相互作用可能由第三种连接蛋白介导。神经元突触前活性区中涉及神经递质释放的三种结构蛋白的表达发生改变,并且这些蛋白从其正常功能部位重新分布到突变亨廷顿蛋白聚集体中。