Selective knockout of astrocytic Na+/H+ exchanger isoform 1 reduces astrogliosis, BBB damage, infarction, and improves neurological function after ischemic stroke

• Published in: Glia Vol. 66; no. 1; pp. 126 - 144
• Main Authors: Begum, Gulnaz, Song, Shanshan, Wang, Shaoxia, Zhao, Hanshu, Bhuiyan, Mohammad Iqbal H., Li, Eric, Nepomuceno, Rachel, Ye, Qing, Sun, Ming, Calderon, Michael Joseph, Stolz, Donna B., St. Croix, Claudette, Watkins, Simon C., Chen, Yinhuai, He, Pingnian, Shull, Gary E., Sun, Dandan
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-01-2018
• Subjects: Animals; astrocyte end‐feet; Astrocytes; Astrocytes - metabolism; Astrocytes - ultrastructure; Blood flow; Blood-brain barrier; Blood-Brain Barrier - pathology; Blood-Brain Barrier - ultrastructure; Brain damage; Brain Infarction - diagnosis; Brain Infarction - etiology; Brain Infarction - genetics; Brain injury; Cerebral blood flow; cerebral edema; Cerebral infarction; Cerebrovascular Circulation - genetics; Cerebrovascular Circulation - physiology; Clonal deletion; Corn oil; Disease Models, Animal; Edema; Endothelial cells; Flox; Gelatinase B; Gene deletion; Gene Expression Regulation - genetics; Glial Fibrillary Acidic Protein - genetics; Glial Fibrillary Acidic Protein - metabolism; Gliosis; Gliosis - etiology; Gliosis - genetics; Gliosis - metabolism; Gliosis - pathology; Hydrogen; Hypertrophy; Infarction; Infarction, Middle Cerebral Artery - complications; Infarction, Middle Cerebral Artery - pathology; Inflammation; Injury prevention; Ischemia; Male; Matrix Metalloproteinase 9 - metabolism; Matrix metalloproteinases; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Electron, Transmission; MMP; Motor Activity - genetics; Na+/H+-exchanging ATPase; Neurologic Examination; Neurological diseases; neurovascular unit; Nhe1 gene; Occlusion; Recombinase; Reperfusion; Rodents; Sodium-Hydrogen Exchanger 1 - deficiency; Sodium-Hydrogen Exchanger 1 - genetics; Stroke; Tamoxifen; Tight junctions; blood-brain barrier; MMP; astrocyte end-feet; gliosis; cerebral edema; neurovascular unit
• ISSN: 0894-1491; 1098-1136
• DOI: 10.1002/glia.23232

Stimulation of Na+/H+ exchanger isoform 1 (NHE1) in astrocytes causes ionic dysregulation under ischemic conditions. In this study, we created a Nhe1flox/flox (Nhe1f/f) mouse line with exon 5 of Nhe1 flanked with two loxP sites and selective ablation of Nhe1 in astrocytes was achieved by crossing Nhe1f/f mice with Gfap‐CreERT2 Cre‐recombinase mice. Gfap‐CreERT2+/−;Nhe1f/f mice at postnatal day 60–90 were treated with either corn oil or tamoxifen (Tam, 75 mg/kg/day, i.p.) for 5 days. After 30 days post‐injection, mice underwent transient middle cerebral artery occlusion (tMCAO) to induce ischemic stroke. Compared with the oil‐vehicle group (control), Tam‐treated Gfap‐CreERT2+/−;Nhe1f/f (Nhe1 KO) mice developed significantly smaller ischemic infarction, less edema, and less neurological function deficits at 1–5 days after tMCAO. Immunocytochemical analysis revealed less astrocytic proliferation, less cellular hypertrophy, and less peri‐lesion gliosis in Nhe1 KO mouse brains. Selective deletion of Nhe1 in astrocytes also reduced cerebral microvessel damage and blood–brain barrier (BBB) injury in ischemic brains. The BBB microvessels of the control brains show swollen endothelial cells, opened tight junctions, increased expression of proinflammatory protease MMP‐9, and significant loss of tight junction protein occludin. In contrast, the Nhe1 KO mice exhibited reduced BBB breakdown and normal tight junction structure, with increased expression of occludin and reduced MMP‐9. Most importantly, deletion of astrocytic Nhe1 gene significantly increased regional cerebral blood flow in the ischemic hemisphere at 24 hr post‐MCAO. Taken together, our study provides the first line of evidence for a causative role of astrocytic NHE1 protein in reactive astrogliosis and ischemic neurovascular damage.
Main Points
Stimulation of astrocytic NHE1 protein at the BBB is detrimental in ischemic brains.
Deletion of astrocytic Nhe1 gene inhibits astrogliosis, reduces stroke volume, prevents BBB damage, and improves rCBF and neurological function after stroke.