Glutamate-induced neuronal death in primary cultures of cerebellar granule cells: protection by synthetic derivatives of endogenous sphingolipids.
The delayed neuronal death induced by a brief (15 min) application of glutamate to primary cultures of cerebellar granule cells can be prevented by pretreating the cultures with the natural ganglioside monosialoglycosylceramide (GM1), the semisynthetic GM1 with N-acetyl sphingosine (LIGA4), GM1 with N-dichloroacetyl sphingosine (LIGA20) and d-eritro 1,3-dihydroxy-2-dichloroacetylamide-4-trans-octadecene (PKS3). The semisynthetic lipids LIGA4, LIGA20 and PKS3 are more potent than the parent natural compounds. The rank order of potency for the protection against glutamate-induced neuronal death is: LIGA20 greater than or equal to LIGA4 greater than PKS3 greater than GM1; the corresponding EC50 values are 4.5 microM for LIGA20, 5 microM for LIGA4, 30 microM for PKS3 and 55 microM for GM1. The effect of the semisynthetic lipids is faster (maximal protection after a 5-min preincubation) and lasts longer (up to 24 hr) than that of the natural compound GM1. The protection from glutamate-induced neuronal death by the semisynthetic sphingolipids persists after thorough washout of free sphingolipid from the incubation medium. Therefore, LIGA4, LIGA20 and PKS3 are potent and efficacious antagonists of glutamate-induced neuronal death with a good separation between the doses needed for pharmacological action and the intrinsic neurotoxic activity. The natural (GM1) and semisynthetic (LIGA4, LIGA20 and PKS3) sphingolipids block neuronal death without affecting the function of glutamate-operated cationic channels. The protective action of sphingolipids appears to be associated with their insertion into membranes where they inhibit specific second messenger-mediated responses triggered by persistent stimulation of glutamate receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
FIDIA-Georgetown Institute for the Neurosciences, Georgetown University, Washington, District of Columbia.
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H
Manev
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Favaron
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Vicini
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Guidotti
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Costa
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects*; D005677:G(M1) Ganglioside / Q000494:pharmacology; D005971:Glutamates / Q000633:toxicity*; D018698:Glutamic Acid; D007473:Ion Channels / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter / Q000187:drug effects; D013107:Sphingolipids / Q000494:pharmacology*; D013110:Sphingosine / Q000494:pharmacology; D013329:Structure-Activity Relationship
D005971:Glutamates; D007473:Ion Channels; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter; D013107:Sphingolipids; D005677:G(M1) Ganglioside; D018698:Glutamic Acid; D011493:Protein Kinase C; D013110:Sphingosine