title: The temporo-spatial localization of polymorphonuclear cells related to the neurovascular unit after transient focal cerebral ischemia.
abstract: Inflammatory responses after cerebral ischemia are important for the development of final infarct size but the role of polymorphonuclear cells (PMN) is still a matter of debate, since previously used antibodies were recently declared as non-specific. In the present study, we investigated the temporo-spatial localization of PMN related to the neurovascular unit using specific antibodies, 7/4 and Ly6G, and application of G-CSF to induce proliferation and mobilization of PMN precursors after transient focal cerebral ischemia in mice. Infarct volumes, sensorimotor function, neurological outcome and immunohistochemical analysis of PMN were performed after G-CSF administration or placebo treatment. G-CSF-treated mice showed reduced infarct size (51.15±15.68 mm(2) vs. 39.31±16.13 mm(2) at day 1; 50.11±16.68 mm(2) vs. 33.16±4.86 mm(2) at day 4; p<0.05). They showed improved motor-function recovery and had a significantly better outcome compared to placebo-treated animals. Comparison of the two PMN detecting antibodies showed no difference in saturation plots or cell quantification. Studying the basement membrane-associated localization revealed ca. 60% extravascular PMN, independent of G-CSF administration. Extravascular PMNs were without any connection to laminin, but all near to the vessels. We conclude that 7/4 is a suitable marker to investigate PMN compared to Ly6G, which confirms results from former studies using the 7/4-antibody. Furthermore we report the observation that PMN were detected outside the laminin barrier but almost exclusively in close vicinity to the neurovascular unit.
authors.affiliation: Department of Neurology, University Hospital Münster, Albert-Schweitzer-Campus 1, D-48149 Münster, Germany.
authors.forename: Nora
authors.initials: N
authors.lastname: Ullrich
authors.affiliation: Department of Neurology, University Hospital Münster, Albert-Schweitzer-Campus 1, D-48149 Münster, Germany.
authors.forename: Jan-Kolja
authors.initials: JK
authors.lastname: Strecker
authors.affiliation: Department of Neurology, University Hospital Münster, Albert-Schweitzer-Campus 1, D-48149 Münster, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Minnerup
authors.affiliation: Department of Neurology, University Hospital Münster, Albert-Schweitzer-Campus 1, D-48149 Münster, Germany. Electronic address: matthias.schilling@ukmuenster.de.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schilling
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D001929:Brain Edema / Q000209:etiology; D020520:Brain Infarction / Q000188:drug therapy / Q000209:etiology; D004195:Disease Models, Animal; D007839:Functional Laterality; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology / Q000627:therapeutic use; D002546:Ischemic Attack, Transient / Q000150:complications / Q000188:drug therapy / Q000473:pathology*; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009043:Motor Activity / Q000187:drug effects; D009460:Neurologic Examination; D018696:Neuroprotective Agents / Q000494:pharmacology / Q000627:therapeutic use; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010799:Phycoerythrin / Q000378:metabolism; D013997:Time Factors
chemical_list: D000950:Antigens, Ly; C480309:Ly6G antigen, mouse; D018696:Neuroprotective Agents; D010799:Phycoerythrin; D016179:Granulocyte Colony-Stimulating Factor
keywords: Experimental stroke; Granulocyte colony stimulating factor; Neurovascular unit; Polymorphonuclear cell