Ganglioside content of astroglia and neurons isolated from maturing rat brain: consideration of the source of astroglial gangliosides.
Previous biochemical and histochemical studies have failed to clarify the nature or quantity of gangliosides in CNS astrocytes. Using improved methodologies for bulk isolation of both neurons and astrocytes as well as for ganglioside purification, we find significantly higher ganglioside concentration in astrocytes and very similar thin-layer chromatography (TLC) patterns for the two cell types. However, in vivo labeling of glycoconjugates via intracerebral injection of [3H]glucosamine prior to cell isolation revealed a different picture: whereas glycoproteins were well-labeled in both cell types after labeling periods of 1-2 h, gangliosides were appreciably labeled only in neurons. With longer time periods (8-48 h) between injection and sacrifice, there was convergence of specific radioactivity of gangliosides from the two isolated cell preparations. These changes are compared to those observed in synaptosomes and microsomes that were isolated simultaneously. The results suggest limited ganglioside synthetic ability in astrocytes as compared to neurons, a conclusion supported by assay of UDP-galNAc:GM3 N-acetylgalactosaminyltransferase in the isolated cells. Nevertheless, the presence of ganglioside GM1 in a substantial portion of bulk-isolated astrocytes was demonstrated by indirect immunofluorescent detection of cholera toxin binding. Ideas on the reconciliation of these apparently contradictory phenomena, including the possibility of intercellular transfer and/or phagocytosis are discussed.
Byrne
M C
MC
Department of Neurology, Albert Einstein College of Medicine, Bronx, NY 10461.
Farooq
M
M
Sbaschnig-Agler
M
M
Norton
W T
WT
Ledeen
R W
RW
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001921:Brain / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005700:Galactosyltransferases / Q000378:metabolism; D005732:Gangliosides / Q000096:biosynthesis* / Q000302:isolation & purification; D005944:Glucosamine / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis; D066298:In Vitro Techniques; D007700:Kinetics; D017350:N-Acetylgalactosaminyltransferases; D009474:Neurons / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000378:metabolism; D000097763:Polypeptide N-acetylgalactosaminyltransferase
D005700:Galactosyltransferases; D005732:Gangliosides; D005944:Glucosamine; D006023:Glycoproteins; D017350:N-Acetylgalactosaminyltransferases; D011509:Proteoglycans; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase