title: Enhanced survival of wild-type and Lurcher Purkinje cells in vitro following inhibition of conventional PKCs or stress-activated MAP kinase pathways.
abstract: Recent studies using both dissociated and organotypic cell cultures have shown that heterozygous Lurcher (Lc/+) Purkinje cells (PCs) grown in vitro share many of the same survival and morphological characteristics as Lc/+ PCs in vivo. We have used this established tissue culture system as a valuable model for studying cell death mechanisms in a relatively simple system where neurodegeneration is induced by a constitutive cation leak mediated by the Lurcher mutation in the δ2 glutamate receptor (GluRδ2). In this study, Ca(++) imaging and immunocytochemistry studies indicate that intracellular levels of Ca(++) are chronically increased in Lc/+ PCs and the concentration and/or distribution of the conventional PKCγ isoform is altered in degenerating Lc/+ PCs. To begin to characterize the molecular mechanisms that regulate Lc/+ PC death, the contributions of conventional PKC pathways and of two MAP kinase family members, JNK and p38, were examined in slice cultures from wild-type and Lc/+ mutant mouse cerebellum. Cerebellar slice cultures from P0 pups were treated with either a conventional PKC inhibitor, a JNK inhibitor, or a p38 inhibitor either from 0 to 14 or 7 to 14 DIV. Treatment with either of the three inhibitors from 0 DIV significantly increased wild type and Lc/+ PC survival through 14 DIV, but only Lc/+ PC survival was significantly increased following treatments from 7 to 14 DIV. The results suggest that multiple PC death pathways are induced by the physical trauma of making organotypic slice cultures, naturally-occurring postnatal cell death, and the GluRδ2 (Lc) mutation.
authors.affiliation: UMR 7102, CNRSet Université Pierre et Marie Curie, 9 Quai St. Bernard, Paris, France. hadi.zanjani@snv.jussieu.fr
authors.forename: Hadi S
authors.initials: HS
authors.lastname: Zanjani
authors.forename: Ann M
authors.initials: AM
authors.lastname: Lohof
authors.forename: Rebecca
authors.initials: R
authors.lastname: McFarland
authors.forename: Michael W
authors.initials: MW
authors.lastname: Vogel
authors.forename: Jean
authors.initials: J
authors.lastname: Mariani
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D064026:Calbindins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002531:Cerebellum / Q000166:cytology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009924:Organ Culture Techniques; D011493:Protein Kinase C / Q000378:metabolism*; D011689:Purkinje Cells / Q000187:drug effects*; D017470:Receptors, Glutamate / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics
chemical_list: D064026:Calbindins; D004791:Enzyme Inhibitors; D017470:Receptors, Glutamate; C107913:glutamate receptor delta 2; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D053148:Caspase 3; D002118:Calcium