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The Ca2+-gated channel TMEM16A amplifies capillary pericyte contraction and reduces cerebral blood flow after ischemia
Nils Korte, … , David Attwell, Paolo Tammaro
Nils Korte, … , David Attwell, Paolo Tammaro
Published March 22, 2022
Citation Information: J Clin Invest. 2022;132(9):e154118. https://doi.org/10.1172/JCI154118.
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Research Article Cell biology Vascular biology

The Ca2+-gated channel TMEM16A amplifies capillary pericyte contraction and reduces cerebral blood flow after ischemia

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Abstract

Pericyte-mediated capillary constriction decreases cerebral blood flow in stroke after an occluded artery is unblocked. The determinants of pericyte tone are poorly understood. We show that a small rise in cytoplasmic Ca2+ concentration ([Ca2+]i) in pericytes activated chloride efflux through the Ca2+-gated anion channel TMEM16A, thus depolarizing the cell and opening voltage-gated calcium channels. This mechanism strongly amplified the pericyte [Ca2+]i rise and capillary constriction evoked by contractile agonists and ischemia. In a rodent stroke model, TMEM16A inhibition slowed the ischemia-evoked pericyte [Ca2+]i rise, capillary constriction, and pericyte death; reduced neutrophil stalling; and improved cerebrovascular reperfusion. Genetic analysis implicated altered TMEM16A expression in poor patient recovery from ischemic stroke. Thus, pericyte TMEM16A is a crucial regulator of cerebral capillary function and a potential therapeutic target for stroke and possibly other disorders of impaired microvascular flow, such as Alzheimer’s disease and vascular dementia.

Authors

Nils Korte, Zeki Ilkan, Claire L. Pearson, Thomas Pfeiffer, Prabhav Singhal, Jason R. Rock, Huma Sethi, Dipender Gill, David Attwell, Paolo Tammaro

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Figure 2

Vasoconstricting GqPCR agonists raise pericyte [Ca2+]i and constrict capillaries at pericyte somata in acute cortical slices.

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Vasoconstricting GqPCR agonists raise pericyte [Ca2+]i and constrict cap...
(A) Representative bright-field images of a live rat cortical capillary pericyte before and after 15 minutes exposure to endothelin 1 (ET-1; 10 nM). Red arrowheads indicate the pericyte soma, and yellow lines indicate where the internal capillary diameter was measured. Scale bar: 10 μm. (B) Mean internal capillary diameter at pericyte somata during exposure to ET-1 (10 nM), normalized to the diameter measured in the absence of ET-1 (n = 10). The ET-1–evoked capillary constriction was not dependent on the sex of rats (Supplemental Figure 3A). (C) Representative bright-field images of a live capillary pericyte, as in A. The thromboxane A2 analog U46619 (200 nM) was applied. Scale bar: 10 μm. (D) Mean internal capillary diameter at pericyte somata during exposure to U46619 (200 nM), normalized to the diameter measured in the absence of U46619 (n = 8). (E) Two-photon microscopy images (maximum intensity projections) of SMCs on a PA and pericytes on first- to third-order capillary branches in acute cortical slices obtained from NG2-CreERT2-GCaMP5G mice. ET-1 raised the somatic [Ca2+]i of SMCs and pericytes (encircled with white dashed lines). Scale bar: 20 μm. The pericyte [Ca2+]i rise coincides with capillary constriction, as indicated by the white line across the vessel lumen in the higher magnification image (scale bar: 10 μm). (F) ET-1 significantly raises [Ca2+]i in first- to third-order pericyte somata and evoked the greatest [Ca2+]i rise in third-order pericytes. The mean GCaMP5G fluorescence in pericyte somata (points indicate individual pericytes from 5 mice; baseline, n = 22; first, n = 13; second, n = 4; third, n = 5) was normalized to the mean GCaMP5G fluorescence of the 17 minutes baseline (Fbaseline) with aCSF (1-way ANOVA with Tukey’s post hoc test).

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