Death and survival of heterozygous Lurcher Purkinje cells in vitro.
The differentiation and survival of heterozygous Lurcher (+/Lc) Purkinje cells in vitro was examined as a model system for studying how chronic ionic stress affects neuronal differentiation and survival. The Lurcher mutation in the delta2 glutamate receptor (GluRdelta2) converts an orphan receptor into a membrane channel that constitutively passes an inward cation current. In the GluRdelta2(+/Lc) mutant, Purkinje cell dendritic differentiation is disrupted and the cells degenerate following the first week of postnatal development. To determine if the GluRdelta2(+/Lc) Purkinje cell phenotype is recapitulated in vitro, +/+, and +/Lc Purkinje cells from postnatal Day 0 pups were grown in either isolated cell or cerebellar slice cultures. GluRdelta2(+/+) and GluRdelta2(+/Lc) Purkinje cells appeared to develop normally through the first 7 days in vitro (DIV), but by 11 DIV GluRdelta2(+/Lc) Purkinje cells exhibited a significantly higher cation leak current. By 14 DIV, GluRdelta2(+/Lc) Purkinje cell dendrites were stunted and the number of surviving GluRdelta2(+/Lc) Purkinje cells was reduced by 75% compared to controls. However, treatment of +/Lc cerebellar cultures with 1-naphthyl acetyl spermine increased +/Lc Purkinje cell survival to wild type levels. These results support the conclusion that the Lurcher mutation in GluRdelta2 induces cell autonomous defects in differentiation and survival. The establishment of a tissue culture system for studying cell injury and death mechanisms in a relatively simple system like GluRdelta2(+/Lc) Purkinje cells will provide a valuable model for studying how the induction of a chronic inward cation current in a single cell type affects neuronal differentiation and survival.
Zanjani
Hadi S
HS
Equipe Développement et Vieillissement du Système Nerveux, 75005, Paris, France.
McFarland
Rebecca
R
Cavelier
Pauline
P
Blokhin
Andrei
A
Gautheron
Vanessa
V
Levenes
Carole
C
Bambrick
Linda L
LL
Mariani
Jean
J
Vogel
Michael W
MW
D000818:Animals; D002412:Cations / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D002525:Cerebellar Cortex / Q000254:growth & development / Q000378:metabolism* / Q000503:physiopathology; D004195:Disease Models, Animal; D005260:Female; D015640:Ion Channel Gating / Q000235:genetics; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000235:genetics; D051379:Mice; D008818:Mice, Neurologic Mutants; D009154:Mutation / Q000235:genetics; D009410:Nerve Degeneration / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D009924:Organ Culture Techniques; D010641:Phenotype; D011689:Purkinje Cells / Q000378:metabolism* / Q000473:pathology; D017470:Receptors, Glutamate / Q000235:genetics* / Q000378:metabolism*; D013096:Spermine / Q000031:analogs & derivatives / Q000494:pharmacology
D002412:Cations; D007473:Ion Channels; D017470:Receptors, Glutamate; C107913:glutamate receptor delta 2; C067506:1-naphthylacetylspermine; D013096:Spermine