title: c-Src function is necessary and sufficient for triggering microglial cell activation.
abstract: Microglial cells are the resident macrophages of the central nervous system. Their function is essential for neuronal tissue homeostasis. After inflammatory stimuli, microglial cells become activated changing from a resting and highly ramified cell shape to an amoeboid-like morphology. These morphological changes are associated with the release of proinflammatory cytokines and glutamate, as well as with high phagocytic activity. The acquisition of such phenotype has been associated with activation of cytoplasmic tyrosine kinases, including those of the Src family (SFKs). In this study, using both in vivo and in vitro inflammation models coupled to FRET-based time-lapse microscopy, lentiviruses-mediated shRNA delivery and genetic gain-of-function experiments, we demonstrate that among SFKs c-Src function is necessary and sufficient for triggering microglia proinflammatory signature, glutamate release, microglia-induced neuronal loss, and phagocytosis. c-Src inhibition in retinal neuroinflammation experimental paradigms consisting of intravitreal injection of LPS or ischemia-reperfusion injury significantly reduced microglia activation changing their morphology to a more resting phenotype and prevented neuronal apoptosis. Our data demonstrate an essential role for c-Src in microglial cell activation.
authors.affiliation: Instituto de Biologia Molecular e Celular (IBMC), Universidade do Porto, Porto, Portugal; Program of Neurosciences, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil; Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil; Faculty of Medicine, Centre of Ophthalmology and Vision Sciences, Institute for Biomedical Imaging and Life Sciences (IBILI), University of Coimbra, Coimbra, Portugal.
authors.forename: Renato
authors.initials: R
authors.lastname: Socodato
authors.forename: Camila C
authors.initials: CC
authors.lastname: Portugal
authors.forename: Ivan
authors.initials: I
authors.lastname: Domith
authors.forename: Nádia A
authors.initials: NA
authors.lastname: Oliveira
authors.forename: Vivian S M
authors.initials: VS
authors.lastname: Coreixas
authors.forename: Erick C
authors.initials: EC
authors.lastname: Loiola
authors.forename: Tânia
authors.initials: T
authors.lastname: Martins
authors.forename: Ana Raquel
authors.initials: AR
authors.lastname: Santiago
authors.forename: Roberto
authors.initials: R
authors.lastname: Paes-de-Carvalho
authors.forename: António F
authors.initials: AF
authors.lastname: Ambrósio
authors.forename: João B
authors.initials: JB
authors.lastname: Relvas
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D000081247:CSK Tyrosine-Protein Kinase; D002460:Cell Line; D002478:Cells, Cultured; D002645:Chickens; D005911:Gliosis / Q000201:enzymology / Q000473:pathology; D018698:Glutamic Acid / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007249:Inflammation / Q000201:enzymology / Q000473:pathology; D007511:Ischemia / Q000201:enzymology / Q000473:pathology; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D017628:Microglia / Q000201:enzymology* / Q000473:pathology; D009474:Neurons / Q000502:physiology; D010587:Phagocytosis / Q000502:physiology; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism*; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000201:enzymology / Q000473:pathology; D055351:Retinal Neurons / Q000473:pathology / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D018698:Glutamic Acid; D000081247:CSK Tyrosine-Protein Kinase; D016392:Proto-Oncogene Proteins pp60(c-src); D019061:src-Family Kinases; C000631346:CSK protein, human
keywords: FRET; glutamate; inflammation; retinal neurons; tyrosine kinase