In vivo assessment of artery smooth muscle [Ca2+]i and MLCK activation in FRET-based biosensor mice.

American Journal of Physiology. Heart and Circulatory Physiology
Jin ZhangW Gil Wier

Abstract

The cellular mechanisms that control arterial diameter in vivo, particularly in hypertension, are uncertain. Here, we report a method that permits arterial intracellular Ca(2+) concentration ([Ca(2+)](i)), myosin light-chain kinase (MLCK) activation, and artery external diameter to be recorded simultaneously with arterial blood pressure (BP) in living mice under 1.5% isofluorane anesthesia. The method also enables an assessment of local receptor activity on [Ca(2+)](i), MLCK activity, and diameter in arteries, uncomplicated by systemic effects. Transgenic mice that express, in smooth muscle, a Ca(2+)/calmodulin-activated, Förster resonance energy transfer (FRET)-based "ratiometric", exogenous MLCK biosensor were used. Vasoactive substances were administered either intravenously or locally to segments of exposed femoral or cremaster arteries. In the basal state, mean BP was approximately 90 mmHg, femoral arteries were constricted to 65% of their passive diameter, MLCK fractional activation was 0.14, and [Ca(2+)](i) was 131 nM. Phenylephrine (300 ng/g wt iv) elevated mean BP transiently to approximately 110 mmHg, decreased heart rate, increased femoral artery [Ca(2+)](i) to 244 nM and fractional MLCK activation to 0.24, and decre...Continue Reading

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Citations

Nov 8, 2011·American Journal of Physiology. Heart and Circulatory Physiology·Mordecai P BlausteinW Gil Wier
Jan 19, 2011·Journal of Cell Science·Evelyne BeerlingJacco van Rheenen
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Mar 23, 2011·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Pooneh BagherSteven S Segal
Nov 22, 2018·American Journal of Physiology. Heart and Circulatory Physiology·Jin ZhangMordecai P Blaustein
May 27, 2014·American Journal of Physiology. Heart and Circulatory Physiology·Joseph R H MaubanWithrow Gil Wier
Apr 19, 2015·American Journal of Physiology. Heart and Circulatory Physiology·Joseph R H MaubanWithrow Gil Wier
Nov 5, 2013·The Journal of Physiology·Michael A Hill, Gerald A Meininger
Oct 31, 2020·American Journal of Physiology. Heart and Circulatory Physiology·Youhua WangMordecai P Blaustein
Oct 19, 2020·The American Journal of Pathology·Ayako ImanishiKenta Terai

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