Measurement of the membrane potential component of the transmembrane proton electrochemical gradient in Crithidia fasciculata

1980 ◽  
Vol 8 (3) ◽  
pp. 307-308 ◽  
Author(s):  
MELVIN MIDGLEY ◽  
MARIS C. STEPHENSON
Keyword(s):  
Membrane Potential ◽  
Electrochemical Gradient ◽  
Crithidia Fasciculata ◽  
Potential Component

scholarly journals Membrane potential of mitochondria in intact lymphocytes during early mitogenic stimulation

1984 ◽  
Vol 217 (2) ◽  
pp. 453-459 ◽  
Author(s):  
M D Brand ◽  
S M Felber
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Concanavalin A ◽  
Mitochondrial Membrane ◽  
Ph Gradient ◽  
Electrochemical Gradient ◽  
Inner Membrane ◽  
Mitogenic Stimulation ◽  
Ion Movements

The mitochondrial membrane potential (delta psi m) in intact lymphocytes was calculated by measuring the distribution of radiolabelled methyltriphenylphosphonium cation. The value obtained was 120 mV. The pH gradient across the mitochondrial membrane in situ (delta pH m) was estimated to be 73 mV (1.2 pH units). Thus the electrochemical gradient of protons was about 190 mV. Addition of the mitogen concanavalin A did not alter delta psi m, showing that, if movement of Ca2+ across the inner membrane of lymphocyte mitochondria occurs when concanavalin A is added, it is accompanied by charge-compensating ion movements.

The generation of electric currents in cardiac fibers by Na/Ca exchange

1979 ◽  
Vol 236 (3) ◽  
pp. C103-C110 ◽  
Author(s):  
L. J. Mullins
Keyword(s):  
Membrane Potential ◽  
Duty Cycle ◽  
Action Potentials ◽  
Reversal Potential ◽  
Resting Membrane Potential ◽  
Electrochemical Gradient ◽  
Ca Current ◽  
Cardiac Action ◽  
Cardiac Fiber ◽  
Cardiac Fibers

The presence of a detectable Ca current during the excitation of a cardiac fiber implies that the Ca lost during the resting interval of the duty cycle must also be detectable. Ca outward movement appears to be effected by Na/Ca exchange when more Na enters than Ca leaves per cycle, thus making the mechanism electrogenic. Since Na/Ca exchange can move Ca either inward or outward depending on the direction of the electrochemical gradient for Na, a potential exists where there is no electric current generated by the Na/Ca exchange mechanism, i.e., a reversal potential ER. Cardiac fibers appear to have a reversal potential that is about midway between their resting membrane potential and their plateau. Carrier currents both inward and outward are therefore generated during cardiac action potentials. The implications of the conditions stated above are explored.

Total contents and net movements of magnesium in the rat uterus

1971 ◽  
Vol 220 (6) ◽  
pp. 2067-2067
Author(s):  
A. H. Moawad ◽  
E. E. Daniel
Keyword(s):  
Membrane Potential ◽  
Active Transport ◽  
Extracellular Space ◽  
Transport Process ◽  
Electrochemical Gradient ◽  
Rat Uterus ◽  
Active Transport Process

Page 75: A. H. Moawad and E. E. Daniel. "Total contents and net movements of magnesium in the rat uterus." Page 80, column 2, line 44, involving the calculation of Vm the answer to the equation, –0.067 V, should read, "–0.012 V." Page 80, column 2, lines 49–54 should read, "The calculated magnesium equilibrum potential is less than the observed membrane potential, which is about 0.050 V. Therefore, some of the tissue magnesium may be excluded by an active transport process against an electrochemical gradient or by loose binding in the extracellular space."

Acute reductions in PO2 depolarize pulmonary artery endothelial cells and decrease [Ca2+]i

1994 ◽  
Vol 266 (4) ◽  
pp. H1416-H1421 ◽  
Author(s):  
T. Stevens ◽  
D. N. Cornfield ◽  
I. F. McMurtry ◽  
D. M. Rodman
Keyword(s):  
Endothelial Cells ◽  
Membrane Potential ◽  
Pulmonary Artery ◽  
Driving Force ◽  
Primary Cultures ◽  
K Channel ◽  
Channel Blockers ◽  
Electrochemical Gradient ◽  
Similar Reduction ◽  
Fura 2

Whereas pulmonary artery endothelial cells (PAECs) are sensitive to oxygen, neither the effect of an acute reduction in PO2 on PAEC membrane potential nor its effect on intracellular free Ca2+ ([Ca2+]i) is known. We hypothesized that in confluent primary cultures of PAECs, an acute decrease in PO2 would depolarize the cell membrane, inhibit Ca2+ influx, and reduce [Ca2+]i. To test this hypothesis, the membrane-sensitive fluorophore bis (1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4, 1 microM) and [Ca2+]i-sensitive probe fura 2 (3 microM) were used. A decrease in PO2 from 125 to 35 mmHg caused membrane depolarization and a 60 +/- 8% (data are means +/- SE) reduction in Ca2+ influx, estimated by manganese quenching of fura 2 fluorescence. While basal [Ca2+]i was 79 +/- 5 nM in normoxic cells, it decreased to 31 +/- 2 nM after 15 min of hypoxia. Decreasing the electrochemical gradient for Ca2+ entry with either low extracellular Ca2+, the K+ channel blockers tetraethylammonium or charybdotoxin, or blockade of Ca2+ entry with lanthanum decreased [Ca2+]i by 54-71% of that observed during an acute reduction in PO2. These results demonstrate that an acute reduction in PO2 1) depolarizes PAECs, 2) reduces Ca2+ influx, and 3) decreases [Ca2+]i, and that a similar reduction in [Ca2+]i was observed with interventions designed to reduce the electrochemical driving force for Ca2+ entry.

Mechanisms by which mitochondria transport calcium

1990 ◽  
Vol 258 (5) ◽  
pp. C755-C786 ◽  
Author(s):  
T. E. Gunter ◽  
D. R. Pfeiffer
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Permeability Transition ◽  
Electrochemical Gradient ◽  
Matrix Space ◽  
Available Energy ◽  
Outward Transport ◽  
Wide Range ◽  
The Matrix ◽  
Efflux Mechanism

It has been firmly established that the rapid uptake of Ca2+ by mitochondria from a wide range of sources is mediated by a uniporter which permits transport of the ion down its electrochemical gradient. Several mechanisms of Ca2+ efflux from mitochondria have also been extensively discussed in the literature. Energized mitochondria must expend a significant amount of energy to transport Ca2+ against its electrochemical gradient from the matrix space to the external space. Two separate mechanisms have been found to mediate this outward transport: a Ca2+/nNa+ exchanger and a Na(+)-independent efflux mechanism. These efflux mechanisms are considered from the perspective of available energy. In addition, a reversible Ca2(+)-induced increase in inner membrane permeability can also occur. The induction of this permeability transition is characterized by swelling of the mitochondria, leakiness to small ions such as K+, Mg2+, and Ca2+, and loss of the mitochondrial membrane potential. It has been suggested that the permeability transition and its reversal may also function as a mitochondrial Ca2+ efflux mechanism under some conditions. The characteristics of each of these mechanisms are discussed, as well as their possible physiological functions.

scholarly journals The transport of α-aminoisobutyrate into Crithidia fasciculata

1978 ◽  
Vol 174 (1) ◽  
pp. 191-202 ◽  
Author(s):  
M Midgley
Keyword(s):  
Kinetic Data ◽  
Adenosine Triphosphatase ◽  
Cytoplasmic Membrane ◽  
Electrochemical Gradient ◽  
Anaerobic Conditions ◽  
Crithidia Fasciculata ◽  
Aerobic And Anaerobic ◽  
Marked Dependence ◽  
Aerobic And Anaerobic Conditions ◽  
Broad Specificity

The transport of alpha-aminoisobutyrate into Crithidia fasciculata was characterized under aerobic and anaerobic conditions. Kinetic data for alpha-aminoisobutyrate transport were consistent with the operation of a single system of broad specificity that showed no marked dependence on Na+. Under anaerobic conditions alpha-aminoisobutyrate transport was inhibited by uncouplers such as 2,4-dinitrophenol, lipophilic cations such as methyltriphenylphosphonium ion and adenosine triphosphatase inhibitors such as dicyclohexylcarbodi-imide and NaN3. A working model in which alpha-aminoisobutyrate enters this organism by an H+-symport mechanism, the electrochemical gradient of protons being maintained by an H+-translocating adenosine triphosphatase on the cytoplasmic membrane, is proposed.

scholarly journals The regulation of extramitochondrial free calcium ion concentration by rat liver mitochondria

1978 ◽  
Vol 176 (2) ◽  
pp. 463-474 ◽  
Author(s):  
David G. Nicholls
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Weak Acid ◽  
Ion Concentration ◽  
Rat Liver Mitochondria ◽  
Free Calcium ◽  
Electrochemical Gradient

The mechanism whereby rat liver mitochondria regulate the extramitochondrial concentration of free Ca2+ was investigated. At 30°C and pH7.0, mitochondria can maintain a steady-state pCa2+0 (the negative logarithm of the free extramitochondrial Ca2+ concentration) of 6.1 (0.8μm). This represents a true steady state, as slight displacements in pCa2+0 away from 6.1 result in net Ca2+ uptake or efflux in order to restore pCa2+0 to its original value. In the absence of added permeant weak acid, the steady-state pCa2+0 is virtually independent of the Ca2+ accumulated in the matrix until 60nmol of Ca2+/mg of protein has been taken up. The steady-state pCa2+0 is also independent of the membrane potential, as long as the latter parameter is above a critical value. When the membrane potential is below this value, pCa2+0 is variable and appears to be governed by thermodynamic equilibration of Ca2+ across a Ca2+ uniport. Permeant weak acids increase, and N-ethylmaleimide decreases, the capacity of mitochondria to buffer pCa2+0 in the region of 6 (1μm-free Ca2+) while accumulating Ca2+. Permeant acids delay the build-up of the transmembrane pH gradient as Ca2+ is accumulated, and consequently delay the fall in membrane potential to values insufficient to maintain a pCa2+0 of 6. The steady-state pCa2+0 is affected by temperature, incubation pH and Mg2+. The activity of the Ca2+ uniport, rather than that of the respiratory chain, is rate-limiting when pCa2+0 is greater than 5.3 (free Ca2+ less than 5μm). When the Ca2+ electrochemical gradient is in excess, the activity of the uniport decreases by 2-fold for every 0.12 increase in pCa2+0 (fall in free Ca2+). At pCa2+0 6.1, the activity of the Ca2+ uniport is kinetically limited to 5nmol of Ca2+/min per mg of protein, even when the Ca2+ electrochemical gradient is large. A steady-state cycling of Ca2+ through independent influx and efflux pathways provides a model which is kinetically and thermodynamically consistent with the present observations, and which predicts an extremely precise regulation of pCa2+0 by liver mitochondria in vivo.

Bioenergetics of membrane transport in Synechococcus R-2 (Anacystis nidulans, S.leopoliensis) PCC7942

1998 ◽  
Vol 76 (6) ◽  
pp. 1127-1145
Author(s):  
Raymond J Ritchie
Keyword(s):  
Membrane Potential ◽  
Intracellular Ph ◽  
Anacystis Nidulans ◽  
Transport Systems ◽  
Electrochemical Gradient ◽  
Accumulation Ratio ◽  
Electrogenic Transport ◽  
Wide Range ◽  
Transport Of Ions ◽  
Photosynthesis And Respiration

Specialized chemical probe techniques need to be used to measure the membrane potential (delta psii,o) or the intracellular pH (pHi) of the cyanobacterium Synechococcus R-2 (PCC7942). The pHi of Synechococcus is essentially a set point (7.3) over a wide range of extracellular pH (pHo) from 7 to 11. Maintenance of the pHo is strongly Na+-dependent and the cells cannot tolerate acid pHo. The 86Rb+-valinomycin method of measuring the delta psii,o has inherent limitations, the most obvious being that the valinomycin treatment itself might alter the membrane potential. 201Tl+ has been found in Synechococcus to distribute across the plasmalemma passively, and so the accumulation ratio of the ion ([Tl+]i/[Tl+]o or Tl+i,o) can be used to calculate the apparent delta psii,o. The two types of probe give comparable results in Synechococcus. Polarizations of the delta psii,o of cells, because of electrogenic transport of ions, can be detected from its effects upon the uptake rate of permeant cations using both the 86Rb+-valinomycin and 201Tl+ methods. HCO3- hyperpolarized delta psii,o, whereas NH4+, CH3NH3+, and K+ led to depolarization. Most active transport systems, including the HCO3- pump, in cyanophytes appear to be ATP binding cassette (ABC) type ATP pumps. Few cotransport (H+ or Na+) driven mechanisms have been identified. A substantial proportion of the power available from photosynthesis and respiration is used to maintain ionic gradients and the membrane potential and in the light a large part (10%) is used to import inorganic carbon.Key words: cyanobacteria, membrane potential, intracellular pH, electrochemical gradient, bioenergetics.

Extracellular acidification at the surface of depolarized voltage-clamped snail neurones detected with eccentric combination pH microelectrodes

1987 ◽  
Vol 65 (5) ◽  
pp. 1001-1005 ◽  
Author(s):  
R. C. Thomas
Keyword(s):  
Membrane Potential ◽  
Intracellular Ph ◽  
Extracellular Ph ◽  
Electrochemical Gradient ◽  
Extracellular Acidification ◽  
Surface Ph ◽  
Buffering Power ◽  
Snail Neurones ◽  
Series Of Experiments ◽  
Ph Microelectrodes

A new design of double micropipette was used to measure intracellular pH, membrane potential, and surface pH of superfused snail neurones. A third double micropipette was used to control the membrane potential via a CsCl-filled barrel and inject HCl iontophoretically. In one series of experiments the surface pH fell by up to one-third of a pH unit when the membrane potential was clamped to 20 mV, pHi was initially 6.7, and extracellular pH was about 7.4 in a medium buffered either with 2 mM HEPES or 2.7% CO2 and 20 mM bicarbonate. In a second series in which surface pH was observed during brief depolarizations to different potentials with different pHi, the potential at which the surface began to acidify varied with pHi with a slope of 32 mV per pH unit. The results confirm that H+ ions leave depolarized snail neurones if the electrochemical gradient is favourable and show that CO2–bicarbonate buffered solutions have a low effective extracellular buffering power for rapid additions of acid.

Sign in / Sign up

Export Citation Format

Share Document