Microglial Cells Prevent Hemorrhage in Neonatal Focal Arterial Stroke

• Published in: The Journal of neuroscience Vol. 36; no. 10; pp. 2881 - 2893
• Main Authors: Fernández-López, David, Faustino, Joel, Klibanov, Alexander L, Derugin, Nikita, Blanchard, Elodie, Simon, Franziska, Leib, Stephen L, Vexler, Zinaida S
• Format: Journal Article
• Language: English
• Published: United States Society for Neuroscience 09.03.2016
• Subjects: Age Factors; Animals; Animals, Newborn; Benzamides - pharmacology; Bone Density Conservation Agents - pharmacology; Caspase 3 - metabolism; Clodronic Acid - toxicity; Dioxoles - pharmacology; Disease Models, Animal; Endothelial Cells - pathology; Female; Gene Expression Regulation, Developmental - drug effects; Gene Expression Regulation, Developmental - genetics; Infarction, Middle Cerebral Artery - complications; Intracranial Hemorrhages - etiology; Intracranial Hemorrhages - prevention & control; Male; Mice; Mice, Transgenic; Microglia - drug effects; Microglia - physiology; Rats; Signal Transduction - drug effects; Signal Transduction - physiology; Smad2 Protein - metabolism; Transforming Growth Factor beta1 - antagonists & inhibitors; Transforming Growth Factor beta1 - metabolism; postnatal; TGFβ1; blood–brain barrier; inflammation; extracellular matrix; middle cerebral artery occlusion
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/JNEUROSCI.0140-15.2016

Perinatal stroke leads to significant morbidity and long-term neurological and cognitive deficits. The pathophysiological mechanisms of brain damage depend on brain maturation at the time of stroke. To understand whether microglial cells limit injury after neonatal stroke by preserving neurovascular integrity, we subjected postnatal day 7 (P7) rats depleted of microglial cells, rats with inhibited microglial TGFbr2/ALK5 signaling, and corresponding controls, to transient middle cerebral artery occlusion (tMCAO). Microglial depletion by intracerebral injection of liposome-encapsulated clodronate at P5 significantly reduced vessel coverage and triggered hemorrhages in injured regions 24 h after tMCAO. Lack of microglia did not alter expression or intracellular redistribution of several tight junction proteins, did not affect degradation of collagen IV induced by the tMCAO, but altered cell types producing TGFβ1 and the phosphorylation and intracellular distribution of SMAD2/3. Selective inhibition of TGFbr2/ALK5 signaling in microglia via intracerebral liposome-encapsulated SB-431542 delivery triggered hemorrhages after tMCAO, demonstrating that TGFβ1/TGFbr2/ALK5 signaling in microglia protects from hemorrhages. Consistent with observations in neonatal rats, depletion of microglia before tMCAO in P9 Cx3cr1(GFP/+)/Ccr2(RFP/+) mice exacerbated injury and induced hemorrhages at 24 h. The effects were independent of infiltration of Ccr2(RFP/+) monocytes into injured regions. Cumulatively, in two species, we show that microglial cells protect neonatal brain from hemorrhage after acute ischemic stroke.