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Two-photon Imaging of Intracellular Ca2+ Handling and Nitric Oxide Production in Endothelial and Smooth Muscle Cells of an Isolated Rat Aorta
Published on: June 10, 2015
A membrane potential-sensitive dye for vascular smooth muscle cells assays
Fabiana S Sguilla1, Antonio C Tedesco, Lusiane M Bendhack
1Laboratório de Farmacologia, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/no. 14040-903 Ribeirão Preto, SP, Brazil.
Biochemical and Biophysical Research Communications
|January 22, 2003
Summary
Hypertensive rat aorta smooth muscle cells exhibit altered membrane potential compared to normotensive cells. This study demonstrates a sensitive method for measuring membrane potential changes in these cells.
Area of Science:
- Physiology
- Cardiovascular Research
- Cell Biology
Background:
- Membrane potential is crucial for smooth muscle cell function.
- Renal hypertension alters vascular smooth muscle cell properties.
- Accurate measurement of membrane potential is essential for understanding cellular changes.
Purpose of the Study:
- To investigate changes in rat aorta smooth muscle cell membrane potential.
- To compare membrane potential in normotensive (2 kidney) and hypertensive (2 kidney-1 clip) rats.
- To validate the use of DIBAC2(3) probe for membrane potential measurements.
Main Methods:
- Utilized the bis-oxonol sensitive probe DIBAC2(3) to assess membrane potential.
- Employed laser scanning confocal microscopy to record fluorescence emission and images.
- Characterized spectral properties in aqueous buffers and cultured smooth muscle cells.
- Applied varying concentrations of extracellular KCl to induce membrane potential changes.
Main Results:
- A linear relationship was observed between fluorescence intensity and membrane potential across a KCl range of 5-40 mmol/L.
- Aorta smooth muscle cells from hypertensive rats (2K-1C) showed greater depolarization (-55 mV) than normotensive rats (2K) (-65 mV).
- Complete membrane depolarization occurred at 10 mmol/L KCl in 2K-1C cells, versus 40 mmol/L in 2K cells.
Conclusions:
- The DIBAC2(3) probe is suitable for measuring membrane potential in cultured rat aorta smooth muscle cells.
- Renal hypertension significantly alters the membrane potential of aorta smooth muscle cells.
- These findings highlight the utility of the DIBAC2(3) probe in cardiovascular research.
