title: A cell cycle alteration precedes apoptosis of granule cell precursors in the weaver mouse cerebellum.
abstract: A missense mutation in the gene coding for the G-protein-activated inwardly rectifying potassium (GIRK) channel, GIRK2, is responsible for apoptosis in the external germinal layer (EGL) of the cerebellum and a nonapoptotic death of midbrain dopaminergic neurons in the weaver (wv) mouse. Failure of axonogenesis and migration are considered to be the primary consequences of GIRK2 channel malfunction in the cerebellum. We investigated whether a disruption of the cell cycle precedes the failure of migration and axonogenesis and leads to massive apoptosis. To this end, immunohistochemistry and immunoblotting for PCNA, Cdk4, cyclin D, cyclin A, and the Cdk inhibitor p27/kip1, as well as in situ end-labeling for apoptotic DNA fragmentation, were applied to cerebella of P7-P21+/+, wv/+, and wv/wv mice. In +/+ and wv/+ mice, the expression of cell cycle proteins was limited to the outer, premigratory zone of the EGL. Antibodies to p27, a marker of cell differentiation, gave a reverse staining pattern. Due to migration delay, patches of p27-positive cells persisted in the outer EGL in P21 wv/+ mice. On the contrary, marked cell cycle up-regulation and absence of p27 occurred throughout the EGL at all ages in wv/wv mice, indicating an inability to switch off the cell cycle. Mitotic index evaluation showed that cell cycle activation was unrelated to proliferative events. Cell cycle proteins were not expressed in the substantia nigra, suggesting that nonapoptotic death of mature dopaminergic neurons is not preceded by abortive cell cycle re-entry. Our data show that abnormalities of the cell cycle in wv/wv cerebellum represent a major and early consequence of GIRK2 channel malfunction and may strongly influence the susceptibility of EGL cells to apoptosis. These observations may help in understanding the pathogenesis of human neurological channelopathies.
authors.affiliation: Department of Neuroscience, Laboratory of Neuropathology, University of Turin, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Migheli
authors.forename: R
authors.initials: R
authors.lastname: Piva
authors.forename: S
authors.initials: S
authors.lastname: Casolino
authors.forename: C
authors.initials: C
authors.lastname: Atzori
authors.forename: S R
authors.initials: SR
authors.lastname: Dlouhy
authors.forename: B
authors.initials: B
authors.lastname: Ghetti
mesh_terms: D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002531:Cerebellum / Q000378:metabolism* / Q000502:physiology*; D019925:Cyclin A / Q000032:analysis / Q000378:metabolism*; D056741:Cyclin D; D051358:Cyclin-Dependent Kinase 4; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016213:Cyclins / Q000032:analysis / Q000378:metabolism*; D004249:DNA Damage; D053938:DNA Fragmentation; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007473:Ion Channels / Q000502:physiology; D051379:Mice; D008818:Mice, Neurologic Mutants; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000378:metabolism*; D008938:Mitosis; D008940:Mitotic Index; D015221:Potassium Channels / Q000378:metabolism; D024661:Potassium Channels, Inwardly Rectifying; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis* / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D013378:Substantia Nigra / Q000033:anatomy & histology / Q000502:physiology; D025521:Tumor Suppressor Proteins
chemical_list: C493961:Cdkn1b protein, mouse; D018797:Cell Cycle Proteins; D019925:Cyclin A; D056741:Cyclin D; D016213:Cyclins; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D007473:Ion Channels; D008869:Microtubule-Associated Proteins; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D018809:Proliferating Cell Nuclear Antigen; D011518:Proto-Oncogene Proteins; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; C495900:CDK4 protein, human; C495902:Cdk4 protein, mouse; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases