Biochemical and histological analysis of two Müller cell antibodies in developing and adult cat and rat central nervous system.
We investigated the binding characteristics of two monoclonal antibodies, 4F3 and 3F8, which in the retina specifically stain Müller cells, both with protein blots and immunohistochemically in sections of various regions of the central nervous system of neonatal and adult cats and rats. Clear differences emerged between the two antibodies. In addition, some species-specific as well as developmental differences within the staining pattern of each individual antibody were evident. The epitopes recognized by 4F3 lay mainly in the 57-63 kDa range. Histologically, 4F3 labelled mainly glia cells: oligodendrocytes and astrocytes in optic nerve, astrocytes in neocortex and cerebellum, Bergmann glia in the cerebellum and radial glia in neonatal animals. This was confirmed by double-immunofluorescence with the astrocyte marker GFAP. By contrast, 3F8 epitopes lay mainly in the 47-49 kDa range. Histologically, 3F8 labelled oligodendrocytes in the optic nerve, but only neurons in cerebellum and neocortex as confirmed by double-labelling with neuronal markers. Neither 4F3 nor 3F8 recognized GFAP or vimentin. These results clearly indicate (1) that the two antibodies identify new epitopes/molecules, (2) that the antigens are not retina-specific, and (3) that Müller cells share epitopes with other glial cells as well as with neurons outside the retina.
Distler
C
C
Allgemeine Zoologie und Neurobiologie, Ruhr-Universität Bochum, Postfach 102148, D-44780 Bochum, Germany. distler@neurobiologie.ruhr-uni-bochum.de
Bronzel
M
M
Paas
I
I
Wahle
P
P
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D001253:Astrocytes / Q000737:chemistry / Q000276:immunology; D015153:Blotting, Western; D001921:Brain / Q000254:growth & development*; D001923:Brain Chemistry; D002415:Cats; D002531:Cerebellum / Q000737:chemistry / Q000254:growth & development; D002540:Cerebral Cortex / Q000737:chemistry / Q000254:growth & development; D000939:Epitopes / Q000276:immunology; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009457:Neuroglia / Q000737:chemistry / Q000276:immunology*; D009474:Neurons / Q000737:chemistry / Q000276:immunology; D009900:Optic Nerve / Q000737:chemistry / Q000254:growth & development; D051381:Rats; D012160:Retina / Q000166:cytology*; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes