Simultaneous Registration of Contraction and Cytosolic Calcium ([Ca2+]i) of Smooth Muscle Strips Using Front-Surface Fluorimetry |
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Authors: | Alice T. Ferreira Rogerio Neri Maria E. M. Oshiro Hideo Kanaide |
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Affiliation: | (1) Department of Biophysics, Universidade Federal de São Paulo (UNIFESP)/Escola Paulista de Medicina, São Paulo, Brazil;(2) Department of Biophysics, Universidade Federal de São Paulo (UNIFESP)/Escola Paulista de Medicina, São Paulo, Brazil;(3) Division of Molecular Cardiology, Institute of Angiocardiology, Faculty of Medicine, Kyushu University, Fukuoka, Japan |
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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 10–6M 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. |
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Keywords: | Fura-2 front surface smooth muscle cytosolic calcium |
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