Search PubMed⌕ Search
Home/Archive/PubMed

PubMed · 7977728

Convolution: a method for data analysis in perifusion systems.

R M Brand·A R Midgley·W J Williams
View original source ↗Export metadata ↓

Abstract

Considerable interest has developed in defining how imposed stimuli effect dynamic changes in cellular metabolism. We have developed a miniature perifusion system that can reveal alterations in extracellular protons within seconds after application of metabolic perturbants. This perifusion system contains two pH sensors: one before the cells records changes between medium and test solutions, and one, located just past the cells, records these alterations plus cellular modifications. Because distortion occurs as chemicals pass through perifusion systems, the shape of pH changes induced by switching from medium to test solutions is different at each electrode. This study describes and validates convolution to correct this distortion. Data from HeLa cells exposed to the metabolic uncoupler of oxidative phosphorylation carbonyl cyanide m-chlorophenyl hydrazone have been analyzed with convolution. Cellular response to oxidative phosphorylation removal is comprised of multiple components, is consistent with a rapid uncoupling, and is followed by cellular adaptation. Therefore convolutional analysis can provide an important adjunct to the analysis of data acquired by perifusion and can provide new insights into cellular responsiveness and metabolism.

Explore related subjects

Carbonyl Cyanide m-Chlorophenyl HydrazoneElectrodesEquipment DesignHeLa CellsHumansMathematicsModels, BiologicalPerfusionStatistics as TopicJournal ArticleResearch Support, U.S. Gov't, P.H.S.

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

R M Brand, A R Midgley, W J Williams. 1994. Convolution: a method for data analysis in perifusion systems.. https://doi.org/10.1152/ajpendo.1994.267.5.e759

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed1995

Isolation of the polar and lateral flagellum-defective mutants in Vibrio alginolyticus and identification of their flagellar driving energy sources.

Vibrio alginolyticus has two types of flagella (polar and lateral) in one cell. We isolated mutants with only a polar flagellum (Pof+ Laf-) or only lateral flagella (Pof- Laf+). Using these mutants, we demonstrated that the energy sources of the lateral and polar flagellar motors in V. alginolyticus are H+ and Na+ motive forces, respectively, as in the related species V. parahaemolyticus.

Carbonyl Cyanide m-Chlorophenyl Hydrazone↗
PubMed1994

The effect of pH on the growth and motility of Rhodobacter sphaeroides WS8 and the nature of the driving force of the flagellar motor.

Rhodobacter sphaeroides WS8 grew, and swam vigorously, over the pH range 6 to 9. Sustained motility was, however, observed in populations of cells resuspended at pH values between 4.9 and 10.4, although the mean run speed was reduced at the extremes of pH. The ability of R. sphaeroides to swim in strong alkaline conditions prompted the question of whether motility at alkaline pH was powered by a sodium motive force, as has been found in the facultative alkalophilic Bacillus and Vibrio species, particularly as motility was found to be sensitive to the sodium channel inhibitor amiloride. The nature of the driving force of the flagellar motor was therefore investigated. It was found that R. sphaeroides was motile over the same pH range in the absence and presence of sodium ions. The protonophore CCCP was found to inhibit motility under all conditions, whereas monensin, an inhibitor of sodium pumps, had no effect upon motility in the presence or absence of sodium. It was concluded that the delta p is the driving force for the flagellar motor in R. sphaeroides at all values of pH. Amiloride, a specific inhibitor of the sodium-driven flagellar motor in alkalophilic Bacillus and Vibrio was shown to act non-specifically on the proton driven motor of R. sphaeroides, reducing the swimming speed of this organism in media with and without sodium to the same extent and over the complete pH range. Measurement of the delta p by using the electrochromic absorbance change of the carotenoid pigments to measure delta psi and 31P-NMR to measure delta pH showed that the maximum delta p was about -215 mV. At pH 10 the cells swam more slowly and the delta p was about -90 mV. These data suggest that the flagellar motor of R. sphaeroides is proton-driven under all conditions with a threshold for motor rotation below -90 mV and saturation at above -90 mV and below -215 mV.

Carbonyl Cyanide m-Chlorophenyl Hydrazone↗
PubMed1994

Combination of the electrogenic ionophores, valinomycin and CCCP, can lead to non-electrogenic K+/H+ exchange on bilayer lipid membranes.

The method of pH shift measuring by means of a pH microelectrode was applied to measure hydrogen ion fluxes across a planar bilayer lipid membrane (BLM) in the presence of the potassium ion ionophore, valinomycin, and a protonophore, carbonylcyanide m-chlorophenylhydrazone (CCCP), under conditions of the voltage clamp. The voltage dependence of the flux was determined to be in the range of +/- 150 mV under the conditions of both symmetrical KCl as well as a KCl gradient across the BLM. Surprisingly, at a clamped zero voltage on BLM a significant hydrogen ion flux was observed in the presence of a KCL gradient and both valinomycin and CCCP. This finding was interpreted as a result of induction of non-electrogenic K+/H+ exchange in the presence of valinomycin and CCCP, presumably through the formation of electrically neutral complexes of these two ionophores and K+ (H+) ions: valinomycin-K(+)-CCCP- and/or possibly valinomycin-CCCP(-)-H+.

Carbonyl Cyanide m-Chlorophenyl Hydrazone↗