PMID: 7373277Feb 1, 1980

Spectrophotometric measurements of transmembrane potential and pH gradients in chromaffin granules

The Journal of General Physiology
G SalamaA Scarpa

Abstract

The electrical potential (delta psi) and proton gradient (alpha pH) across the membranes of isolated bovine chromaffin granules and ghosts were simultaneously and quantitatively measured by using the membrane-permeable dyes 3,3'dipropyl-2,2'thiadicarbocyanine (diS-C3-(5)) to measure delta psi and 9-aminoacridine or atebrin to measure delta pH. Increases or decreases in the delta psi across the granular membrane could be monitored by fluorescence or transmittance changes of diS-C3-(5). Calibration of the delta psi was achieved by utilization of the endogenous K+ and H+ gradients, and valinomycin or carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), respectively, with the optical response of diS-C3-(5) varying linearly with the Nernst potential for H+ and K+ over the range -60 to +90 mV. The addition of chromaffin granules to a medium including 9-aminoacridine or atebrin resulted in a rapid quenching of the dye fluorescence, which could be reversed by agents known to cause collapse of pH gradients. From the magnitude of the quenching and the intragranular water space, it was possible to calculate the magnitude of the alpha pH across the chromaffin granule membrane. The time-course of the potential-dependent transmittance ...Continue Reading

References

Dec 1, 1976·The Journal of General Physiology·R G Johnson, A Scarpa
Jan 1, 1976·Neuroscience·C L BashfordG A Ritchie
May 1, 1975·Journal of Bioenergetics·H Rottenberg
Jun 30, 1976·The Journal of Membrane Biology·A S Waggoner
Dec 17, 1973·European Journal of Biochemistry·E Padan, H Rottenberg
Jul 1, 1971·The Review of Scientific Instruments·B Chance, N Graham
Sep 1, 1967·British Journal of Pharmacology and Chemotherapy·F H SchneiderH Winkler

Citations

Jul 1, 1988·Biulleten' eksperimental'noĭ biologii i meditsiny·I M Antonikov, R N Glebov
Mar 1, 1990·Trends in Neurosciences·P R MaycoxR Jahn
May 21, 1999·The European Journal of Neuroscience·C BoyerJ Lehouelleur
Nov 1, 1981·European Journal of Biochemistry·W C HorneE R Simons
May 1, 1980·Acta Physiologica Scandinavica·H Abrahamsson, E Gylfe
Nov 1, 1983·The Journal of Clinical Investigation·M S Klempner, B Styrt
Jan 1, 1982·Annals of the New York Academy of Sciences·M F BeersA Scarpa
Jan 1, 1985·Annals of the New York Academy of Sciences·R G JohnsonA Scarpa
Jan 1, 1987·Annals of the New York Academy of Sciences·D E Knight, P F Baker
Apr 1, 1984·Bioscience Reports·P F Baker, D E Knight
Apr 14, 2016·European Biophysics Journal : EBJ·Jose A Alvarez-Bustamante, Victor V Lemeshko
Mar 17, 2007·Journal of Neuroscience Methods·Andrew D Powell, Neil V Marrion
Feb 29, 1984·Biochemical and Biophysical Research Communications·D D FayJ L Costa
May 1, 1987·Bioscience Reports·D E Knight
Jan 1, 1983·The American Journal of Physiology·C S PaceJ S Smith

Related Concepts

Acridines
Adrenal Glands
Bos indicus
Chromaffin Granule
Chromaffin System
Red Cell Ghost
Fluorochromes
Hydrogen-Ion Concentration
Resting Potentials

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