Monoclonal antibody markers for amphibian oligodendrocytes and neurons.
Few immunocytochemical probes have been developed for cold-blooded vertebrates, thus hampering analyses of cellular processes in these species. Those developed from mammalian and avian tissue often fail either to react or to show similar specificities in poikilotherms. Therefore, we have begun raising monoclonal antibodies (mabs) in mice against frog and tadpole brain tissue. The following analyses of two of these mabs suggest that these antibodies represent specific probes for frog axons and oligodendrocytes. Mab Olig recognizes all the myelinated axon tracts of the mature frog brain and spinal cord, as well as the tracts of the developing tadpole CNS once they have become myelinated. Axons cut in cross section show characteristic o-shaped staining around individual axons when processed with this antibody. Particularly easy to visualize in the tadpole are immunoreactive cell bodies and processes, seen in continuity with the myelin sheath. Occasionally, in this developing tissue, cells with highly branched processes characteristic of immature oligodendrocytes are observed. No other cells or processes within the brain or spinal cord react with this antibody. Mab Linc stains numerous filaments in all axonal projections. Occasionally, a thin rim of filamentous staining is observed in cell somata, but many regions rich in neuronal somata or dendrites are unreactive to this antibody. This in vivo staining pattern suggests that the Linc antigen is differentially distributed within neurons and exhibits a high concentration in axons. Linc immunoreactivity is robust in the processes of a subpopulation of dissociated tectal cells in culture. These Linc-positive cells are characterized as neurons on morphological criteria. Also, intense Linc immunoreactivity is present in the bundles of retinal axons that extend from retinal explants. Olig immunoreactivity, however, has not been detected in tectal cultures or retinal explants. Improved staining following Triton X-100 treatment of tissue sections suggests that neither of the mabs recognizes lipid antigens and that both are probably localized within the cell cytoplasm. Only the Linc mab reacts on Western blots of denatured brain protein. Linc consistently recognizes two Triton X-100-insoluble proteins with apparent molecular weights of 56 and 58 kD. The epitopes recognized by the Olig and Linc mabs have been surveyed in terms of their resistance to optic nerve crush and their consequent value in studies requiring such procedures. Possible homologies to known cell-type-specific molecules are discussed.
Steen
P
P
Department of Biology, Yale University, New Haven, Connecticut 06511.
Kalghatgi
L
L
Constantine-Paton
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology / Q000276:immunology*; D007150:Immunohistochemistry; D007814:Larva; D009409:Nerve Crush; D009413:Nerve Fibers, Myelinated / Q000276:immunology; D009474:Neurons / Q000276:immunology*; D009836:Oligodendroglia / Q000276:immunology*; D011894:Rana pipiens / Q000033:anatomy & histology* / Q000254:growth & development; D012160:Retina / Q000276:immunology*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000276:immunology*; D014795:Visual Pathways / Q000166:cytology
D000911:Antibodies, Monoclonal