Intracellular free Ca2+ and vasoconstriction determined simultaneously in the perfused rat tail artery.
To measure, simultaneously, intracellular free Ca2+ ([Ca2+]i) and vasoconstriction in a perfused vessel, we used the fluorescent Ca2+ indicator fura 2 with a dual-wavelength excitation method. One-centimeter-long segments of the caudal artery were dissected from 12-mo-old male Wistar rats. The endothelium was removed by gentle rubbing. The artery was mounted in a specially constructed spectrofluorometer cuvette, perfused with oxygenated physiological saline solution at 37 degrees C, and loaded by perfusion with fura 2 acetoxymethyl ester (5 microM) over a 90-min period. This paper is a description of the technique and the experiments that validate it as a useful method for examining Ca(2+)-related vascular reactivity in an intact perfused vessel.
Capdeville-Atkinson
C
C
Laboratoire de Pharmacologie Cardio-Vasculaire, Unité de Formation et de Recherche des Sciences, Pharmaceutiques et Biologiques, Nancy, France.
Oster
L
L
Thorin-Trescases
N
N
Robert
A
A
Boutinet
S
S
Atkinson
J
J
D000818:Animals; D001158:Arteries / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D005456:Fluorescent Dyes; D016257:Fura-2 / Q000031:analogs & derivatives; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D011190:Potassium Cyanide / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D013623:Tail / Q000098:blood supply; D013997:Time Factors; D014661:Vasoconstriction / Q000502:physiology*
D005456:Fluorescent Dyes; C049925:fura-2-am; D011190:Potassium Cyanide; D002118:Calcium; D016257:Fura-2