A technique for simultaneous measurement of Ca2+, FRET fluorescence and force in intact mouse small arteries.
FRET (Forster resonance energy transfer)-based biosensor molecules are powerful tools to reveal specific molecular interactions in cells. Typically however, they are used in cultured cells that (inevitably) express different genes than their counterparts in intact organisms. In such cells it may be impossible to administer physiological stimuli and measure physiological outputs. Here, through the use of transgenic mice that express a FRET-based myosin light chain kinase (MLCK) biosensor molecule, we report a technique for dynamically observing activation and regulation of MLCK within the smooth muscle cells of intact, functioning small arteries, together with measurement of arterial force production and intracellular [Ca(2+)].
Wier
W Gil
WG
Department of Physiology, School of Medicine, University of Maryland Baltimore, 655 West Baltimore St, Baltimore, MD 21201, USA. gwier001@umaryland.edu
Rizzo
Mark A
MA
Raina
Hema
H
Zacharia
Joseph
J
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D066298:In Vitro Techniques; D008638:Mesenteric Arteries / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009219:Myosin-Light-Chain Kinase / Q000032:analysis / Q000096:biosynthesis; D014661:Vasoconstriction / Q000502:physiology
D009219:Myosin-Light-Chain Kinase; D002118:Calcium