An immunocytochemical technique with monoclonal antibodies to glycosphingolipids in rat primary cerebellar cultures: influence of detergent permeabilization.
Glycosphingolipids (GSLs) are highly expressed in the vertebrate central nervous system. GSLs have been implicated in a variety of phenomena involving cell-cell recognition, neurite outgrowth, synaptogenesis, transmembrane signalling and cell growth and differentiation. We recently determined the distribution of GSLs in rat brain tissues and in primary rat cerebellar cultures as well as using a number of MAbs to GSLs, which were generated and characterized in our laboratory. These results suggested that (i) the expression of GSLs was highly localized to a specific cell type and layer in the rat brain tissues and (ii) some GSLs may be useful markers for identifying cells in the primary cultures. In the present paper, we describe in detail an immunofluorescence technique for the detection of GSL expression in the primary cultures. We demonstrate that the localization of GSLs can be greatly influenced by detergent treatments.
Kawashima
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Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Honkomagome, Bunkyo-ku, Tokyo 113, Japan.
Tai
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T
D000818:Animals; D000911:Antibodies, Monoclonal; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002531:Cerebellum / Q000737:chemistry* / Q000166:cytology; D003902:Detergents / Q000493:pharmacokinetics*; D006028:Glycosphingolipids / Q000032:analysis* / Q000276:immunology; D007150:Immunohistochemistry; D009457:Neuroglia / Q000737:chemistry*; D009474:Neurons / Q000737:chemistry*; D017830:Octoxynol / Q000493:pharmacokinetics*; D051381:Rats; D017208:Rats, Wistar
D000911:Antibodies, Monoclonal; D003902:Detergents; D006028:Glycosphingolipids; D017830:Octoxynol