title: Inducible nitric oxide synthase in neutrophils and endothelium contributes to ischemic brain injury in mice.
abstract: NO produced by inducible NO synthase (iNOS) contributes to ischemic brain injury, but the cell types expressing iNOS and mediating tissue damage have not been elucidated. To examine the relative contribution of iNOS in resident brain cells and peripheral leukocytes infiltrating the ischemic brain, we used bone marrow (BM) chimeric mice in which the middle cerebral artery was occluded and infarct volume was determined 3 d later. iNOS(-/-) mice engrafted with iNOS(+/+) BM exhibited larger infarcts (44 ± 2 mm(3); n = 13; mean ± SE) compared with autologous transplanted iNOS(-/-) mice (24 ± 3 mm(3); n = 10; p < 0.01), implicating blood-borne leukocytes in the damage. Furthermore, iNOS(+/+) mice transplanted with iNOS(-/-) BM had large infarcts (39 ± 6 mm(3); n = 13), similar to those of autologous transplanted iNOS(+/+) mice (39 ± 4 mm(3); n = 14), indicating the resident brain cells also play a role. Flow cytometry and cell sorting revealed that iNOS is highly expressed in neutrophils and endothelium but not microglia. Surprisingly, postischemic iNOS expression was enhanced in the endothelium of iNOS(+/+) mice transplanted with iNOS(-/-) BM and in leukocytes of iNOS(-/-) mice with iNOS(+/+) BM, suggesting that endothelial iNOS suppresses iNOS expression in leukocytes and vice versa. To provide independent evidence that neutrophils mediate brain injury, neutrophils were isolated and transferred to mice 24 h after stroke. Consistent with the result in chimeric mice, transfer of iNOS(+/+), but not iNOS(-/-), neutrophils into iNOS(-/-) mice increased infarct volume. The findings establish that iNOS in both neutrophils and endothelium mediates tissue damage and identify these cell types as putative therapeutic targets for stroke injury.
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and.
authors.forename: Lidia
authors.initials: L
authors.lastname: Garcia-Bonilla
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and.
authors.forename: Jamie M
authors.initials: JM
authors.lastname: Moore
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Racchumi
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York, NY 10021.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Butler
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and joa2005@med.cornell.edu coi2001@med.cornell.edu.
authors.forename: Costantino
authors.initials: C
authors.lastname: Iadecola
authors.affiliation: Feil Family Brain and Mind Research Institute, Weill Cornell Medical College, New York, NY 10021; and joa2005@med.cornell.edu coi2001@med.cornell.edu.
authors.forename: Josef
authors.initials: J
authors.lastname: Anrather
mesh_terms: D000818:Animals; D020520:Brain Infarction / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D042783:Endothelial Cells / Q000276:immunology / Q000473:pathology; D004730:Endothelium, Vascular / Q000276:immunology* / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics / Q000276:immunology; D051379:Mice; D018345:Mice, Knockout; D009504:Neutrophils / Q000276:immunology* / Q000473:pathology; D009569:Nitric Oxide / Q000235:genetics / Q000276:immunology*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000276:immunology*; D020521:Stroke / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D013997:Time Factors
chemical_list: D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse