Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes
Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes
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DOI：
10.1002/glia.23140
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发表时间：
2017-06-01
期刊：
影响因子：
6.2
通讯作者：
Bosio, Andreas
中科院分区：
文献类型：
--
作者：
Kantzer, Christina G.;Boutin, Camille;Bosio, Andreas
星形胶质细胞是中枢神经系统中最丰富的细胞类型，涵盖广泛的功能。我们在此报告了一种新型单克隆抗体的产生，即抗星形胶质细胞表面抗原 2 (Anti-ACSA-2)。流式细胞术、免疫组织化学和免疫细胞化学显示，Anti-ACSA-2 与所有发育阶段的鼠星形胶质细胞的尚未鉴定的糖基化表面分子发生特异性反应。它没有显示任何非星形胶质细胞的标记，例如神经元、少突胶质细胞、NG2(+)细胞、小胶质细胞、内皮细胞、白细胞或红细胞。 GLAST 和 ACSA-2 的联合标记研究显示很大程度上重叠的表达。然而，中枢神经系统某些区域（例如小脑）的蛋白质表达水平和单阳性细胞亚群频率也存在显着差异，尤其是在出生后早期。在神经源性生态位、海马齿状回和室下区（SVZ）中，再次观察到表达水平存在轻微差异的总体重叠。与 GLAST 不同，ACSA-2 对基于木瓜蛋白酶的组织解离不敏感，并且基于使用直接与超顺磁性微珠偶联的抗 ACSA-2 的新快速分选程序，可以对活星形胶质细胞进行高效、急性、特异性和前瞻性纯化。总之，ACSA-2 似乎是星形胶质细胞、放射状胶质细胞、神经干细胞和双能神经胶质祖细胞的新表面标记，这为进一步剖析星形胶质细胞亚群和谱系的特征开辟了可能性。
Astrocytes are the most abundant cell type of the central nervous system and cover a broad range of functionalities. We report here the generation of a novel monoclonal antibody, anti-astrocyte cell surface antigen-2 (Anti-ACSA-2). Flow cytometry, immunohistochemistry and immunocytochemistry revealed that Anti-ACSA-2 reacted specifically with a not yet identified glycosylated surface molecule of murine astrocytes at all developmental stages. It did not show any labeling of non-astroglial cells such as neurons, oligodendrocytes, NG2(+) cells, microglia, endothelial cells, leukocytes, or erythrocytes. Co-labeling studies of GLAST and ACSA-2 showed largely overlapping expression. However, there were also notable differences in protein expression levels and frequencies of single-positive subpopulations of cells in some regions of the CNS such as cerebellum, most prominently at early postnatal stages. In the neurogenic niches, the dentate gyrus of the hippocampus and the subventricular zone (SVZ), again a general overlap with slight differences in expression levels were observed. ACSA-2 was unlike GLAST not sensitive to papain-based tissue dissociation and allowed for a highly effective, acute, specific, and prospective purification of viable astrocytes based on a new rapid sorting procedure using Anti-ACSA-2 directly coupled to superparamagnetic MicroBeads. In conclusion, ACSA-2 appears to be a new surface marker for astrocytes, radial glia, neural stem cells and bipotent glial progenitor cells which opens up the possibility of further dissecting the characteristics of astroglial subpopulations and lineages.