title: Neurons and astrocytes in an infantile neuroaxonal dystrophy (INAD) mouse model show characteristic alterations in glutamate-induced Ca2+signaling.
abstract: INAD (infantile neuroaxonal dystrophy, OMIM#256600), an autosomal recessive inherited degenerative disease, is associated with PLA2G6 mutations. PLA2G6 encodes Ca2+-independent phospholipase A2(VIA iPLA2). However, it is unclear how the PLA2G6-mutations lead to disease. Non-canonical functions, which were suggested for VIA iPLA2, such as regulation of cellular and mitochondrial Ca2+are promising candidates. Therefore, we investigate glutamate (Glu)-evoked Ca2+signals in neurons and astrocytes in co-culture obtained from three INAD mouse model strains with Pla2g6 mutations, (i) hypomorphic Pla2g6 allele with reduced transcript levels, (ii) knocked-out Pla2g6, and (iii) (G373R)-point mutation with inactive VIA iPLA2enzyme. Homozygous offspring from these strains develop pathology similar to that observed in INAD patients. We found that in mouse neurons the Pla2g6 mutation disrupted the dependency of Glu-induced extracellular Ca2+influx on mitochondrial Ca2+uptake. Thus, in neurons with Pla2g6 mutation we did not detect the characteristic reduction in Glu-induced Ca2+influx upon treatment with Ru360, a blocker of mitochondrial Ca2+uniporter, or with rotenone. In contrast to neurons, in astrocytes, both with Pla2g6 mutation or wild-type cells, the treatment with Ru360 or rotenone reduced the rate of Glu-induced Ca2+influx ∼2-fold. This Ca2+influx in astrocytes represents capacitative Ca2+entry. In astrocytes with Pla2g6 mutation, the Glu-induced Ca2+influx was ∼2-fold lower than in wild-type controls. We suggest that this is the mechanism for strongly decreased durations of Glu-induced Ca2+responses in astrocytes with Pla2g6 mutation. We could mimic the mutation by pharmacological inhibition of iPLA2using S-BEL. Thus, lack of VIA iPLA2activity caused effects in astrocytes. In summary, three INAD mouse models show comparable changes in Glu-induced Ca2+signaling, but specific for neurons or astrocytes. This finding helps to identify pathways altered during INAD and highlights non-canonical VIA iPLA2functions, like regulation of cellular Ca2+fluxes by mitochondria or capacitative Ca2+-entry.
authors.affiliation: Institut für Neurobiochemie (Institut für Inflammation und Neurodegeneration), Medizinische Fakultät, Otto-von-Guericke-Universität Magdeburg, Magdeburg, Germany.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Strokin
authors.affiliation: Institut für Neurobiochemie (Institut für Inflammation und Neurodegeneration), Medizinische Fakultät, Otto-von-Guericke-Universität Magdeburg, Magdeburg, Germany. Electronic address: georg.reiser@med.ovgu.de.
authors.forename: Georg
authors.initials: G
authors.lastname: Reiser
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D018920:Coculture Techniques; D004195:Disease Models, Animal; D005260:Female; D018698:Glutamic Acid / Q000633:toxicity*; D054522:Group VI Phospholipases A2 / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D019150:Neuroaxonal Dystrophies / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*
chemical_list: D018698:Glutamic Acid; D054522:Group VI Phospholipases A2; C516476:Pla2g6 protein, mouse
keywords: Astrocyte; Ca(2+) signaling; Glutamate; Infantile neuroaxonal dystrophy (INAD); Mouse models of INAD; Neuron; Phospholipase A(2)