A membrane potential-sensitive dye for vascular smooth muscle cells assays

Changes in membrane potential of rat aorta smooth muscle cells were investigated using the bis-oxonol sensitive probe DIBAC 2(3). We compared the changes in membrane potential induced by a high external KCl concentration in aorta smooth muscle cells from normotensive 2 kidney (2K) and from renal hyp...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2003-01, Vol.301 (1), p.113-118
Main Authors: Sguilla, Fabiana S, Tedesco, Antonio C, Bendhack, Lusiane M
Format: Article
Language:English
Subjects:
Animals
Aorta - anatomy & histology
Bis-oxonol
Blood Pressure - physiology
Calibration
Fluorescence intensity
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
Hypertension - metabolism
Male
Membrane Potentials - physiology
Microscopy, Confocal
Myocytes, Smooth Muscle - cytology
Myocytes, Smooth Muscle - metabolism
Potentiometric optical probe
Rats
Rats, Inbred Strains
Rats, Wistar
Renal hypertension
Thiobarbiturates - chemistry
Thiobarbiturates - metabolism
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Summary:Changes in membrane potential of rat aorta smooth muscle cells were investigated using the bis-oxonol sensitive probe DIBAC 2(3). We compared the changes in membrane potential induced by a high external KCl concentration in aorta smooth muscle cells from normotensive 2 kidney (2K) and from renal hypertensive 2 kidney-1 clip (2K-1C) rats. The spectral properties of the membrane potential were first characterized in aqueous buffers and in cultured smooth muscle cells from 2K and 2K-1C rat aortas. Fluorescence emission and the images were recorded using a laser scanning confocal microscope. The relationship between fluorescence intensity (FI) and membrane potential ( ψ m) as a function of the increasing extracellular KCl concentration was linear in the 5–40 mmol/L KCl range in both 2K and 2K-1C rat aorta cells. Cell membranes from 2K-1C rat aorta cells were more depolarized (−55 mV) than 2K rat aorta cells (−65 mV). The results show that in 2K-1C aorta cells only 10 mmol/L KCl was needed to induce complete membrane depolarization while in 2K cells 40 mmol/L KCl was needed to induce a similar effect. This study clearly shows that the method is suitable to measure the membrane potential in cultured smooth muscle cells.
ISSN:0006-291X
1090-2104
DOI:10.1016/S0006-291X(02)02973-X