Cultured granule cells and astrocytes from cerebellum differ in metabolizing sphingosine.
Sphingosine metabolism was studied in primary cultures of differentiated cerebellar granule cells and astrocytes. After a 2-h pulse with [C3-(3)H]sphingosine at different doses (0.1-200 nmol/mg of cell protein), both cell types efficiently incorporated the long chain base; the percentage of cellular [(3)H]sphingosine over total label incorporation was extremely low at sphingosine doses of <10 nmol/mg of cell protein and increased at higher doses. Most of the [(3)H]sphingosine taken up underwent metabolic processing by N-acylation, 1-phosphorylation, and degradation (assessed as (3)H(2)O released in the medium). The metabolic processing of exogenous sphingosine was extremely efficient in both cells, granule cells and astrocytes being able to metabolize, respectively, an amount of sphingosine up to 80- and 300-fold the cellular content of this long chain base in 2 h. At the different doses, the prevailing metabolic route of sphingosine was different. At lower doses and in a wide dose range, the major metabolic fate of sphingosine was N-acylation. With increasing doses, there was first increased sphingosine degradation and then increased levels of sphingosine-1-phosphate. The data demonstrate that, in neurons and astrocytes, the metabolic machinery devoted to sphingosine processing is different, astrocytes possessing an overall higher capacity to synthesize the bioactive compounds ceramide and sphingosine-1-phosphate.
Riboni
L
L
Department of Medical Chemistry and Biochemistry, Study Center for the Functional Biochemistry of Brain Lipids, University of Milan, Milan, Italy. laura.riboni@unimi.it
Viani
P
P
Bassi
R
R
Giussani
P
P
Tettamanti
G
G
D000215:Acylation; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism; D007700:Kinetics; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D051381:Rats; D012588:Scintillation Counting; D014316:Tritium; D000099157:Sphingosine Kinase
D005964:Glucosyltransferases; D009419:Nerve Tissue Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D014316:Tritium; D000099157:Sphingosine Kinase; C023246:ceramide glucosyltransferase