ABSTRACT
Sulfatide is enriched in the myelin sheath and accounts for 5% of the total lipids in this membrane. In the present work we describe the production and characterization of mouse monoclonal antibodies against sulfatide. The antibodies were detected and characterized in a previously described ELISA test system. The clone AIC3IA2 produced antibodies of the IgG3 class with high specificity for sulfatide. These antibodies showed almost no cross-reactivity with galactocerebroside or with any of the other lipids we tested. When used with the peroxidase antiperoxidase technique the antibodies stained a cell population either in fixed or unfixed brain cell cultures, indicating a surface localization of sulfatide in the respective cell population. In double-staining experiments the stained cell population was identified as myelin basic protein-positive oligodendrocytes.
Subject(s)
Antibodies, Monoclonal/biosynthesis , Membrane Lipids/immunology , Myelin Sheath/immunology , Sulfoglycosphingolipids/immunology , Animals , Antibody Specificity , Clone Cells , Fluorescent Antibody Technique , Immunoenzyme Techniques , Mice , Mice, Inbred BALB C , Oligodendroglia/metabolism , Sulfoglycosphingolipids/metabolismABSTRACT
We investigated whether in cultures of mechanically dissociated brain cells from newborn mice the reduction of the number of oligodendrocytes influences their proliferation rate. 14-day-old cultures were subjected to complement-dependent anti-galactocerebroside (GC) antibody-mediated cytotoxicity. The cytotoxic treatment completely destroyed oligodendrocytes. Thereafter, GC+ oligodendrocytes progressively reappeared. Their number was 20 and 66% compared to controls, 3 and 7 days after cytotoxicity, respectively. Proliferating oligodendrocytes were detected 3 and 7 days after cytotoxicity by combining the immunostaining for GC with 3H-thymidine autoradiography. The proliferation rate of oligodendrocytes in treated cultures was increased by 100 and 76% compared to controls, 3 and 7 days after cytotoxicity, respectively. These data suggest that the proliferation rate of oligodendrocytes can be influenced by extrinsic factors.
Subject(s)
Neuroglia/cytology , Oligodendroglia/cytology , Animals , Brain/cytology , Cell Division , Cells, Cultured , Cytotoxicity, Immunologic , MiceABSTRACT
The expression of myelin basic protein (MBP) and galactocerebroside (GC), two antigenic markers for oligodendrocytes, was checked on 7-, 14-, 21- and 28-day-old dissociated mouse brain cell cultures (BCC) by using the indirect immunofluorescence method with double staining. The number of GC positive cells increased between the 7th and the 14th day of culture before a steady state was reached. In contrast to this, the MBP-positive cells appeared only on the 14th day of culture, and their number increased with the age of the culture. In double staining, the serum produced against isolated oligodendrocytes shows the same picture as the anti-GC serum, while only a part of GC-positive cells showed also the presence of MBP. Our data suggest that the GC appears very early on the membrane of the oligodendrocytes during development while cells exhibiting both GC and MBP probably represent a more differentiated oligodendrocyte population.
