Anti‐ACSA‐2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes

• Published in: Glia 2017-06, Vol.65 (6), p.990-1004
• Main Authors: Kantzer, Christina G., Boutin, Camille, Herzig, Ina D., Wittwer, Carolina, Reiß, Sandy, Tiveron, Marie Catherine, Drewes, Jan, Rockel, Thomas D., Ohlig, Stefanie, Ninkovic, Jovica, Cremer, Harold, Pennartz, Sandra, Jungblut, Melanie, Bosio, Andreas
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Antibodies, Monoclonal - immunology; antibody; Antibody Specificity; Antigens, Surface - analysis; Antigens, Surface - immunology; Antigens, Surface - metabolism; astrocyte cell surface antigen‐2; Astrocytes - cytology; Astrocytes - immunology; Brain - cytology; Brain - growth & development; cell separation; cell surface marker; Cells, Cultured; Cellular Biology; Development Biology; Endothelial Cells - cytology; Endothelial Cells - immunology; Erythrocytes - cytology; Erythrocytes - metabolism; Excitatory Amino Acid Transporter 1 - analysis; Genes; GLAST; Immunomagnetic Separation - methods; Leukocytes - cytology; Leukocytes - immunology; Life Sciences; Medicin och hälsovetenskap; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Microglia - cytology; Microglia - immunology; Neural Stem Cells - immunology; Neurons - cytology; Neurons - metabolism; Oligodendroglia - cytology; Oligodendroglia - immunology; Rats, Wistar
• ISSN: 0894-1491; 1098-1136; 1098-1136
• DOI: 10.1002/glia.23140

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. Main Points Novel anti‐astrocyte cell surface antigen‐2 antibody (Anti‐ACSA‐2) labels specifically murine astrocytes, radial glia, neural stem and glial progenitor cells and allows their fast, prospective, acute and viable purification from neonatal and adult brain.