Simultaneous Registration of Contraction and Cytosolic Calcium ([Ca2+]i) of Smooth Muscle Strips Using Front-Surface Fluorimetry

Abstract

Using front-surface fluorimetry with fura-2-loaded smooth muscle strips, simultaneous registration of the cytosolic calcium concentration ([Ca²⁺]_i) changes and tension development was done under the action of 40 mM KCl and the myotropic peptide 10⁻⁶ M angiotensin II. The strips were mounted vertically, connected to a force transducer that keeps a basal isometric tension of 0.5 g, and maintained in a bathing solution oxygenated at 37°C. The fiber-optic platform FluoroMax-2 accessory 1950F was used to do the remote sensing for the samples. Light from the excitation spectrometer (FluoroMax-2), alternating between 340 and 380 nm, was focused onto the fiber-optic bundle and directed to the smooth muscle strip. The fluorescence (505 nm) was collected and redirected to the emission port of the fluorimeter FluoroMax-2. The ratiometric method (R340/380) was used as an index of [Ca²⁺]_i change during smooth muscle contraction. All data, R340/380 and tension, were recorded using a computerized data acquisition system: Soft & Solution and GRAMS/386 of Galatic Industries Corporation.