Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, 21941-902, Brazil.
Mol Neurobiol. 2018 Jan;55(1):751-762. doi: 10.1007/s12035-016-0343-z. Epub 2017 Jan 3.
Astrocytes, the most abundant glial cells in the central nervous system (CNS), comprise a heterogeneous population of cells. However, how this heterogeneity impacts their function within brain homeostasis and response to injury and disease is still largely unknown. Recently, astrocytes have been recognized as important regulators of synapse formation and maturation. Here, we analyzed the synaptogenic property of astrocytes from different regions of the CNS. The effect of conditioned medium derived from astrocytes (astrocyte-conditioned medium (ACM)) from cerebral cortex, hippocampus, midbrain and cerebellum, in synapse formation, was evaluated. Synapse formation was analyzed by quantification of pre- and postsynaptic proteins, synaptophysin, and postsynaptic density protein 95 (PSD-95). ACM from the four regions increased significantly the number of synaptophysin/PSD-95 puncta on neurons from the same and different brain regions. Differences on astrocytic synaptogenic potential between the regions were observed according to ACM protein concentration. Thus, cerebellar astrocytes have higher synaptogenic effect when ACM is less concentrated. Also, heterotypical co-culture assays revealed that neurons from cerebral cortex and midbrain equally respond to ACM, indicating that differences in synapse effect are unlike to be neuron-autonomous. The expression profile of the synaptogenic molecules secreted by astrocytes from distinct brain regions was analyzed by qPCR. Gene expression of glypicans 4 and 6, hevin, and secreted protein-acidic and rich in cysteine (SPARC) greatly varies between astrocytes from different brain regions. Furthermore, in vivo analysis of hevin protein confirmed that variance. These findings highlight the heterogeneity of astrocytes and suggest that their synaptogenic potential may be different in each brain region, mainly due to distinct gene expression profiles.
星形胶质细胞是中枢神经系统 (CNS) 中最丰富的神经胶质细胞,由异质性细胞组成。然而,这种异质性如何影响它们在大脑内稳态中的功能以及对损伤和疾病的反应,在很大程度上仍然未知。最近,星形胶质细胞已被认为是突触形成和成熟的重要调节因子。在这里,我们分析了来自中枢神经系统不同区域的星形胶质细胞的突触发生特性。评估了来自大脑皮层、海马体、中脑和小脑的星形胶质细胞条件培养基 (astrocyte-conditioned medium (ACM)) 对突触形成的影响。通过定量分析突触前和突触后蛋白、突触小泡蛋白和突触后密度蛋白 95 (PSD-95) 来分析突触形成。来自四个区域的 ACM 显著增加了来自相同和不同脑区神经元的突触小泡蛋白/PSD-95 斑点的数量。根据 ACM 蛋白浓度观察到不同区域的星形胶质细胞突触发生潜能存在差异。因此,当 ACM 浓度较低时,小脑星形胶质细胞具有更高的突触发生作用。此外,异质共培养测定表明,来自大脑皮层和中脑的神经元对 ACM 的反应相同,表明突触效应的差异不太可能是神经元自主的。通过 qPCR 分析了来自不同脑区的星形胶质细胞分泌的突触发生分子的表达谱。糖蛋白 4 和 6、hevin 和富含半胱氨酸的酸性分泌蛋白 (SPARC) 的基因表达在来自不同脑区的星形胶质细胞之间存在很大差异。此外,体内分析 hevin 蛋白证实了这种差异。这些发现突出了星形胶质细胞的异质性,并表明它们的突触发生潜力可能在每个脑区都不同,主要是由于不同的基因表达谱。