title: Glial dysfunction in parkin null mice: effects of aging.
abstract: Parkin mutations in humans produce parkinsonism whose pathogenesis is related to impaired protein degradation, increased free radicals, and abnormal neurotransmitter release. The role of glia in parkin deficiency is little known. We cultured midbrain glia from wild-type (WT) and parkin knock-out (PK-KO) mice. After 18-20 d in vitro, PK-KO glial cultures had less astrocytes, more microglia, reduced proliferation, and increased proapoptotic protein expression. PK-KO glia had greater levels of intracellular glutathione (GSH), increased mRNA expression of the GSH-synthesizing enzyme gamma-glutamylcysteine synthetase, and greater glutathione S-transferase and lower glutathione peroxidase activities than WT. The reverse happened in glia cultured in serum-free defined medium (EF12) or in old cultures. PK-KO glia was more susceptible than WT to transference to EF12 or neurotoxins (1-methyl-4-phenylpyridinium, blockers of GSH synthesis or catalase, inhibitors of extracellular signal-regulated kinase 1/2 and phosphatidylinositol 3 kinases), aging of the culture, or combination of these insults. PK-KO glia was less susceptible than WT to Fe2+ plus H2O2 and less responsive to protection by deferoxamine. Old WT glia increased the expression of heat shock protein 70, but PK-KO did not. Glia conditioned medium (GCM) from PK-KO was less neuroprotective and had lower levels of GSH than WT. GCM from WT increased the levels of dopamine markers in midbrain neuronal cultures transferred to EF12 more efficiently than GCM from PK-KO, and the difference was corrected by supplementation with GSH. PK-KO-GCM was a less powerful suppressor of apoptosis and microglia in neuronal cultures. Our data prove that abnormal glial function is critical in parkin mutations, and its role increases with aging.
authors.affiliation: Department of Neurobiology and Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas, Hospital Ramón y Cajal, 28034 Madrid, Spain.
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Solano
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Casarejos
authors.forename: Jamie
authors.initials: J
authors.lastname: Menéndez-Cuervo
authors.forename: Jose A
authors.initials: JA
authors.lastname: Rodriguez-Navarro
authors.forename: Justo
authors.initials: J
authors.lastname: García de Yébenes
authors.forename: Maria A
authors.initials: MA
authors.lastname: Mena
mesh_terms: D000375:Aging / Q000502:physiology*; D000704:Analysis of Variance; D000818:Animals; D018845:CD11 Antigens / Q000378:metabolism; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008636:Mesencephalon / Q000166:cytology; D051379:Mice; D018345:Mice, Knockout; D009457:Neuroglia / Q000737:chemistry / Q000145:classification / Q000473:pathology* / Q000502:physiology*; D009474:Neurons / Q000145:classification / Q000187:drug effects / Q000502:physiology; D018384:Oxidative Stress / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics*; D051020:bcl-X Protein / Q000378:metabolism
chemical_list: C494812:Bcl2l1 protein, mouse; D018845:CD11 Antigens; D017077:Culture Media, Conditioned; D005904:Glial Fibrillary Acidic Protein; D051020:bcl-X Protein; D007770:L-Lactate Dehydrogenase; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D005978:Glutathione