Astrocyte proteome from the rat substantia nigra. Study in animal model of fluorocitrate-induced astrocyte dysfunction and microglia activation.

Dysfunction of astrocytic support for prolonged time can be harmful for nervous system. Inflammation and glia activation are among hallmarks of most neurodegenerative diseases. Therefore, astrocytes are considered as druggable targets for disease modifying strategies. Partial lesion of dopaminergic neurons is compensated functionally, but which mechanisms within each type of cells are responsible is unknown. The aim was to check how neuron vs astrocyte degeneration affected remaining astrocytes proteome and which mechanisms were regulated. In a rat model of 7-day infusion by osmotic minipumps of fluorocitrate (FC) into the substantia nigra (SN) (doi: 10.1007/s12035-017-0529-z; doi: 10.1111/jnc.14605; doi: 10.1016/j.mito.2018.12.002) astrocytes become dysfunctional and die inducing activation of remaining astrocytes and microglia. 6-OHDA injected into medial forebrain bundle caused selective degeneration of dopaminergic neurons in the SN. Controls were sham operated. Astrocytes were sorted out from dissociated SN tissue using FACS, and processed for mass spectrometry proteome analysis. Understanding the role of individual cell type in the diseased tissue cellular context is essential to understand disease pathomechanisms and identify better pharmacological targets.

长期星形胶质细胞（astrocyte）支持功能异常可对神经系统造成损害。炎症反应与胶质细胞活化是多数神经退行性疾病的典型特征之一，因此星形胶质细胞被视为疾病修饰治疗策略的可靶向靶点。多巴胺能神经元的部分损伤可通过功能代偿得以弥补，但目前尚不明确各类细胞内究竟是何种机制介导了这一过程。本研究旨在探究神经元与星形胶质细胞变性分别对残留星形胶质细胞蛋白质组的影响，以及受调控的相关分子机制。本研究采用两种大鼠模型：其一为通过渗透微型泵向黑质（substantia nigra, SN）内持续7天输注氟柠檬酸（fluorocitrate, FC）（相关文献DOI：10.1007/s12035-017-0529-z；10.1111/jnc.14605；10.1016/j.mito.2018.12.002），该模型可导致星形胶质细胞功能异常并死亡，进而诱发残留星形胶质细胞与小胶质细胞活化；其二为向内侧前脑束注射6-羟基多巴胺（6-OHDA），以造成黑质内多巴胺能神经元的选择性变性。对照组动物仅接受假手术处理。研究人员采用荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）从解离的黑质组织中分离星形胶质细胞，随后对其进行质谱蛋白质组学分析。明确疾病组织微环境中各类细胞的功能作用，是阐明疾病发病机制、发掘更优药理靶点的关键前提。