title: Activation of Fas receptor is required for the increased formation of the disialoganglioside GD3 in cultured cerebellar granule cells committed to apoptotic death.
abstract: Apoptosis was induced in cultured cerebellar granule cells by lowering extracellular K+ concentrations (usually from 25 to 10 mM). The apoptotic phenotype was preceded by an early and transient increase in the intracellular levels of the disialoganglioside, GD3, which behaves as a putative pro-apoptotic factor. We examined whether activation of Fas receptor mediates the increase in GD3 formation in granule cells committed to die. Degenerating granule cells showed increased expression of both Fas receptor and its ligand (Fas-L), at times that coincided with the increase in GD3 levels and the induction of GD3 synthase mRNA. Addition of neutralizing anti-Fas-L antibodies reduced the extent of 'low-K+'-induced apoptosis and abolished the increase in GD3 levels and GD3 synthase mRNA. Similar reductions were observed in cultures prepared from gld or lpr mice, which harbor loss-of-function mutations of Fas-L and Fas receptor, respectively. In addition, exogenous application of soluble Fas-L further enhanced both the increase in GD3 formation and cell death in cultured granule cells switched from 25 into 10 mM K+. We conclude that activation of Fas receptor is entirely responsible for the increase in GD3 levels and contributes to the development of apoptosis by trophic deprivation in cultured cerebellar granule cells.
authors.affiliation: Department of Human Physiology and Pharmacology, University of Rome La Sapienza, Piazzale Aldo Moro, 5, 00185, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Castiglione
authors.forename: P
authors.initials: P
authors.lastname: Spinsanti
authors.forename: L
authors.initials: L
authors.lastname: Iacovelli
authors.forename: L
authors.initials: L
authors.lastname: Lenti
authors.forename: F
authors.initials: F
authors.lastname: Martini
authors.forename: R
authors.initials: R
authors.lastname: Gradini
authors.forename: V
authors.initials: V
authors.lastname: Di Giorgi Gerevini
authors.forename: A
authors.initials: A
authors.lastname: Caricasole
authors.forename: A
authors.initials: A
authors.lastname: Caruso
authors.forename: R
authors.initials: R
authors.lastname: De Maria
authors.forename: F
authors.initials: F
authors.lastname: Nicoletti
authors.forename: D
authors.initials: D
authors.lastname: Melchiorri
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western / Q000379:methods; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D002518:Ceramides / Q000032:analysis; D002531:Cerebellum / Q000166:cytology*; D002843:Chromatin / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D053222:Fas Ligand Protein; D005455:Fluorescent Antibody Technique / Q000379:methods; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000276:immunology; D051379:Mice; D008817:Mice, Mutant Strains; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism*
chemical_list: D000906:Antibodies; D002518:Ceramides; D002843:Chromatin; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; C505662:Faslg protein, rat; D005732:Gangliosides; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; C026226:ganglioside, GD3; D005700:Galactosyltransferases; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D011188:Potassium