Immunofluorescent Patterns of Dissociated Rat Embryo Cerebral Cells during Development in Surface Culture: Distinctive Reactions with Neurite and Perikaryon Cell Membranes

Abstract

The presence of antigenic determinants on surface cultures of dissociated cells from rat embryo cerebral tissue has been investigated with the immunofluorescence technique. Antisera raised in rabbits against mature mouse brain reacted with the embryonic cells and enabled the identification of two distinct populations of immunofluorescent cells. One population clearly distinguishable by its ring fluorescence was involved in early cellular reaggregation and neurite formation. A second population which displayed dispersed fluorescence over the whole cell, did not establish intercellular contacts for prolonged periods of time in culture. Indirect evidence suggests that after several days in culture a great portion of the latter population was found in close proximity to the early aggregate, to form an immunofluorescent-heterotypic aggregate. A second antiserum, raised against embryonic rat cerebral cells, displayed immunofluorescent labeling patterns similar to those observed with the mature mouse brain antiserum, when reacted with freshly dissociated cerebral cells. As the cells differentiated in surface culture, distinctive patterns of immunofluorescent reactions developed. Large neuronal cells were labeled on their neuritic cell membranes by the antiserum to adult brain but not by the antiserum to embryonic cells. The results suggest that the anti-mature brain serum possess a class of antibodies which is able to react with antigenic determinants localized specifically on the neuritic plasma membrane.

Deregulated Sphingolipid Metabolism and Membrane Organization in Neurodegenerative Disorders

“…These increases were accompanied by a dramatic shift from simple gangliosides (GM3 and GD3) to more complex species (GM1, GD1a, GD1b, GT1b). A similar increase in the quantity and in the complexity of gangliosides has also been observed during differentiation in cultured neurons of different origin and in mouse neural precursor cells [89,90,[92][93][94][95][96][97]. In humans, the phase of rapid ganglioside increase started from the sixth month of gestation and reached the maximum value at about 5 years of age [91].…”

Nervous System‐Specific Proteins in Developing Rat Cerebral Cells in Culture

“…The procedure employed for D2 staining of viable cells has been described previously by Yavin and Yavin (1978). In brief, cells grown on polylysine-precoated coverslips were reacted with the antiserum raised against excised D2 precipitates obtained by immunoelectrophoresis.…”

Secondary Alterations of Sphingolipid Metabolism in Lysosomal Storage Diseases

“…For example, the total ganglioside content increased several fold from the embryonic stages to the postnatal life in vertebrate brain [2][3][4], as well as during in vitro differentiation of cultured neurons of different origin and in mouse neural precursor cells [2,3,[5][6][7][8][9], with a concomitant shift from simple gangliosides (GM3 and GD3) to more complex species (GM1, GD1a, GD1b, GT1b). Sphingomyelin (SM) content also dramatically increased during in vitro differentiation of rat cerebellar neurons [10].…”