CD9 of mouse brain is implicated in neurite outgrowth and cell migration in vitro and is associated with the alpha 6/beta 1 integrin and the neural adhesion molecule L1.
We describe here a novel monoclonal antibody (mab H6) which recognizes CD9, an integral cell surface constituent previously described in cells of the hematopoietic lineage and involved in the aggregation of platelets. Mab H6 was raised against membranes of immature mouse astrocytes and reacted with a protein of 25-27 kD in detergent extracts of adult mouse brain membranes. Sequence analysis of the N-terminal amino acids revealed an identity of 96% with CD9 from mouse kidney. CD9 was localized in the central and peripheral mouse nervous systems: in the spinal cord of 11-day-old mouse embryos, CD9 was strongly expressed in the floor and roof plates. In the adult mouse sciatic nerve, myelin sheaths were highly CD9-immunoreactive. Mab H6 reacted with the cell surfaces of both glial cells and neurons in culture and inhibited migration of neuronal cell bodies, neurite fasciculation and outgrowth of astrocytic processes from cerebellar microexplants. Neurite outgrowth from isolated small cerebellar neurons was increased in the presence of mab H6 on substrate-coated laminin, but not on substrate-coated poly-L-lysine. Addition of mab H6 elicited an increase in intracellular Ca2+ concentration in these cells on substrate-coated laminin. Immunoprecipitates of CD9 from cultured mouse neuroblastoma N2A cells contained the alpha 6/beta 1 integrin. Moreover, preparations of CD9 immunoaffinity-purified from adult mouse brain using a mab H6 column contained the neural adhesion molecule L1, but not other neural adhesion molecules. CD9 bound to L1, but not to NCAM or MAG. Both the alpha 6/beta 1 integrin and L1 could be induced to coredistribute with CD9 on the surface of cultured neuroblastoma N2A cells. The combined observations suggest that CD9 can associate with L1 and alpha 6/beta 1 integrin to influence neural cell interactions in vitro.
Schmidt
C
C
Department of Neurobiology, Swiss Federal Institute of Technology, Zürich, Switzerland.
Künemund
V
V
Wintergerst
E S
ES
Schmitz
B
B
Schachner
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology / Q000502:physiology*; D000951:Antigens, Neoplasm / Q000032:analysis; D001923:Brain Chemistry; D002118:Calcium / Q000032:analysis; D002417:Cattle; D002450:Cell Communication; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D007152:Immunologic Capping; D007158:Immunologic Techniques; D039121:Integrin alpha6beta1; D016023:Integrins / Q000378:metabolism*; D007797:Laminin; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000502:physiology*; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism*; D016501:Neurites / Q000502:physiology*; D009447:Neuroblastoma / Q000473:pathology; D009457:Neuroglia / Q000737:chemistry; D009474:Neurons / Q000737:chemistry; D011107:Polylysine; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013116:Spinal Cord / Q000737:chemistry / Q000196:embryology; D060245:Tetraspanin 29; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000951:Antigens, Neoplasm; C554869:CD9 protein, human; C554870:Cd9 protein, mouse; D039121:Integrin alpha6beta1; D016023:Integrins; D007797:Laminin; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; D060245:Tetraspanin 29; D011107:Polylysine; D002118:Calcium