Microglial Hv1 proton channels promote white matter injuries after chronic hypoperfusion in mice

Microglia are critical in damage/repair processes during ischemic white matter injury (WMI). Voltage‐gated proton channel (Hv1) is expressed in microglia and contributes to nicotinamide adenine dinucleotide phosphate oxidase complex‐dependent production of reactive oxygen species (ROS). Recent findings have shown that Hv1 is involved in regulating luminal pH of M1‐polarized microglial phagosomes and inhibits endocytosis in microglia. We previously reported that Hv1 facilitated production of ROS and pro‐inflammatory cytokines in microglia and enhanced damage to oligodendrocyte progenitor cells from oxygen and glucose deprivation. To investigate the role of Hv1 in hypoperfusion‐induced WMI, we employed mice that were genetically devoid of Hv1 (Hv1‐/‐), as well as a model of subcortical vascular dementia via bilateral common carotid artery stenosis. Integrity of myelin was assessed using immunofluorescent staining and transmission electron microscopy, while cognitive impairment was assessed using an eight‐arm radial maze test. Hv1 deficiency was found to attenuate bilateral common carotid artery stenosis‐induced disruption of white matter integrity and impairment of working memory. Immunofluorescent staining and western blotting were used to assay changes in oligodendrocytes, OPCs, and microglial polarization. Compared with that in wild‐type (WT) mice, Hv1‐/‐ mice exhibited reduced ROS generation, decreased pro‐inflammatory cytokines production, and an M2‐dominant rather than M1‐dominant microglial polarization. Furthermore, Hv1‐/‐ mice exhibited enhanced OPC proliferation and differentiation into oligodendrocytes. Results of mouse‐derived microglia‐OPC co‐cultures suggested that PI3K/Akt signaling was involved in Hv1‐deficiency‐induced M2‐type microglial polarization and concomitant OPC differentiation. These results suggest that microglial Hv1 is a promising therapeutic target for reducing ischemic WMI and cognitive impairment. Voltage‐gated proton channel (Hv1) is expressed in microglia and contributes to nicotinamide adenine dinucleotide phosphate oxidase complex (NOX)‐dependent production of reactive oxygen species (ROS). Recent findings have shown that Hv1 is involved in regulating luminal pH of M1‐polarized microglial phagosomes and inhibits endocytosis in microglia. Some studies reported that Hv1 enhances brain damage from ischemic stroke. We proposed the following mechanism for microglia mediated white matter injuries induced by chronic hypoperfusion