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Neutrophil recruitment to the brain in mouse and human ischemic stroke

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Abstract

Neutrophils are rapidly recruited in response to local tissue infection or inflammation. Stroke triggers a strong inflammatory reaction but the relevance of neutrophils in the ischemic brain is not fully understood, particularly in the absence of reperfusion. We investigated brain neutrophil recruitment in two murine models of permanent ischemia induced by either cauterization of the distal portion of the middle cerebral artery (c-MCAo) or intraluminal MCA occlusion (il-MCAo), and three fatal cases of human ischemic stroke. Flow cytometry analyses revealed progressive neutrophil recruitment after c-MCAo, lesser neutrophil recruitment following il-MCAo, and absence of neutrophils after sham operation. Confocal microscopy identified neutrophils in the leptomeninges from 6 h after the occlusion, in the cortical basal lamina and cortical Virchow–Robin spaces from 15 h, and also in the cortical brain parenchyma at 24 h. Neutrophils showed signs of activation including histone-3 citrullination, chromatin decondensation, and extracellular projection of DNA and histones suggestive of extracellular trap formation. Perivascular neutrophils were identified within the entire cortical infarction following c-MCAo. After il-MCAo, neutrophils prevailed in the margins but not the center of the cortical infarct, and were intraluminal and less abundant in the striatum. The lack of collaterals to the striatum and a collapsed pial anastomotic network due to brain edema in large hemispheric infarctions could impair neutrophil trafficking in this model. Neutrophil extravasation at the leptomeninges was also detected in the human tissue. We concluded that neutrophils extravasate from the leptomeningeal vessels and can eventually reach the brain in experimental animal models and humans with prolonged arterial occlusion.

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Acknowledgments

Supported by the Spanish Ministries of Economy (SAF2011-30492) and Health (FIS PI12/01437), and FP7 EU projects ERANET-NEURON (PRI-PIMNEU-2011-1342) and InMIND (grant agreement no 278,850). We acknowledge Alba Hernández, Unai Perpiñá and Ana Guerrero for technical assistance. We thank the Cytometry and Image Platforms of IDIBAPS, and Dr. Maria Calvo and Anna Bosch of the Confocal Microscopy Unit of the Serveis Cientifico-Tècnics of the University of Barcelona for technical advice and support. We are indebted to the Neurological Tissue Bank of the Biobank-Hospital Clinic-IDIBAPS for sample and data procurement, and to patient’s relatives for giving consent to this study. We thank Dr. Vanessa Brait for helpful comments and editing.

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Correspondence to Anna M. Planas.

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I. Perez-de-Puig and F. Miró-Mur equally contributed to this work.

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Perez-de-Puig, I., Miró-Mur, F., Ferrer-Ferrer, M. et al. Neutrophil recruitment to the brain in mouse and human ischemic stroke. Acta Neuropathol 129, 239–257 (2015). https://doi.org/10.1007/s00401-014-1381-0

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  • DOI: https://doi.org/10.1007/s00401-014-1381-0

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