Multi-point measurement of intracellular chloride concentrations.

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1. Cl^--sensitive probe: A Cl^--sensitive fluorescent dye,N-(6-methoxyquinolyl) acetoethyl ester (MQAE, purity > 99%) was synthesized by the method of Verkman.2. Multi-channel window photometer:The fluorescence intensity of a MQAE-loaded cell was continuously recorded on an inverted microscope equipped with a night vision camera-based multi-channel window photometer, which was devised to measure the photo intensity at desired multiple windows in a microscopic view. Using this equipment, changes in the intracellular Cl^- concentration ([Cl^-]_i) were monitored to elucidate the mechanisms for its regulation.3. Corneal endothelial cell:Stationary [Cl^-]_i of rabbit corneal endothelial cells was 30.1*1.6 mM. Efflux of Cl^- with alpha_2^-adrenoceptor stimulation,and an increase in Cl^- permeability of plasma membranes with beta-adrenoceptor stimulation were suggested.4. Retinal pigment epithelial cell (RPE): Stationary [Cl^-]_i of rabbit RPE was approx. 90 mM. Atrial natriuretic peptide was demonstrated to accumulate through activation of Na^+/K^+/Cl^- co-transporter regulated via cGMP-dependent protein kinase.5. Hippocampal pyramidal cell:Stationary [Cl^-]_i of rat hippocampal pyramidal cells was 6.63*1.56 mM. An ATP-driven Cl^- pump extrudes Cl^- and Na^+/K^+/Cl^- co-transporter accumulates the same anion.