title: Astrocyte GRK2 as a novel regulator of glutamate transport and brain damage.
abstract: G protein-coupled receptor (GPCR) kinase 2 (GRK2) regulates cellular signaling via desensitization of GPCRs and by direct interaction with intracellular signaling molecules. We recently described that ischemic brain injury decreases cerebral GRK2 levels. Here we studied the effect of astrocyte GRK2-deficiency on neonatal brain damage in vivo. As astrocytes protect neurons by taking up glutamate via plasma-membrane transporters, we also studied the effect of GRK2 on the localization of the GLutamate ASpartate Transporter (GLAST). Brain damage induced by hypoxia-ischemia was significantly reduced in GFAP-GRK2(+/-) mice, which have a 60% reduction in astrocyte GRK2 compared to GFAP-WT littermates. In addition, GRK2-deficient astrocytes have higher plasma-membrane levels of GLAST and an increased capacity to take up glutamate in vitro. In search for the mechanism by which GRK2 regulates GLAST expression, we observed increased GFAP levels in GRK2-deficient astrocytes. GFAP and the cytoskeletal protein ezrin are known regulators of GLAST localization. In line with this evidence, GRK2-deficiency reduced phosphorylation of the GRK2 substrate ezrin and enforced plasma-membrane GLAST association after stimulation with the group I mGluR-agonist DHPG. When ezrin was silenced, the enhanced plasma-membrane GLAST association in DHPG-exposed GRK2-deficient astrocytes was prevented. In conclusion, we identified a novel role of astrocyte GRK2 in regulating plasma-membrane GLAST localization via an ezrin-dependent route. We demonstrate that the 60% reduction in astrocyte GRK2 protein level that is observed in GFAP-GRK2(+/-) mice is sufficient to significantly reduce neonatal ischemic brain damage. These findings underline the critical role of GRK2 regulation in astrocytes for dampening the extent of brain damage after ischemia.
authors: Laboratory of Neuroimmunology and Developmental Origins of Disease (NIDOD), University Medical Center Utrecht, KC03.068.0, Lundlaan 6, 3584 EA, Utrecht, The Netherlands. C.Nijboer@umcutrecht.nl
authors: Cora H
authors: CH
authors: Nijboer
authors: Cobi J
authors: CJ
authors: Heijnen
authors: Vincent
authors: V
authors: Degos
authors: Hanneke L D M
authors: HL
authors: Willemen
authors: Pierre
authors: P
authors: Gressens
authors: Annemieke
authors: A
authors: Kavelaars
mesh_terms: D027322:Amino Acid Transport System X-AG / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D005455:Fluorescent Antibody Technique; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism*; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism*; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D034741:RNA, Small Interfering; D000099149:Ezrin
chemical_list: D027322:Amino Acid Transport System X-AG; D003598:Cytoskeletal Proteins; D054769:G-Protein-Coupled Receptor Kinase 2; D018698:Glutamic Acid; D034741:RNA, Small Interfering; D000099149:Ezrin; C496568:GRK2 protein, mouse