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.

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.

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.

scholarly journals The concentration of glycine by preparations of the yeast Saccharomyces Carlsbergensis depleted of adenosine triphosphate: Effects of proton gradients and uncoupling agents

1976 ◽  
Vol 154 (3) ◽  
pp. 669-676 ◽  
Author(s):  
A Seaston ◽  
G Carr ◽  
A. A Eddy
Keyword(s):  
Membrane Potential ◽  
Intracellular Ph ◽  
Adenosine Triphosphate ◽  
Proton Conductors ◽  
Equivalent Amount ◽  
Accumulation Ratio ◽  
Proton Gradients ◽  
Glycine Uptake ◽  
Proton Symport ◽  
Low Rate

1. At pH 4.5 and 30degreesC, yeast preparations depleted of ATP in the presence of antimycin and deoxyglucose spontaneously lost K+, gaining roughly an equivalent amount of H+. 2. Five proton conductors including azide and 2,4-dinitrophenol accelerated this process, as did [14C]glycine, which was absorbed with two extra equivalents of H+. 3. The rate of glycine uptake at pH 4.5 diminished fourfold when cellular K+ fell by 20%. 4. The distribution of [14C]propionate indicated that the intracellular pH fell from 6.2 to 5.7 when the cellular content of K+ fell by 30%. 5. Glycine uptake from a 5 muM solution was about 400 times faster at pH 4.5 than it was at pH 7.4 with 100mM-KCl present ostensibly to lower the membrane potential. 6. Yeast preparations containing 2mM-[14C]glycine absorbed a further amount from a 0.1 muM solution at pH 4.5. After about 10 min a net movement of [14C]glycine out of the yeast occurred. The ratio of the cellular [14Ia1glycine concentration to the concentration outside the yeast reached 4 × 104 in these assays, whereas at pH 7.4 in the presence of 100mM-KCl it did not exceed 15 in 3h. Dimitrophenol lowered the accumulation ratio at pH 4.5, apparently by causing proton conduction. 7. The observations are consistent with the notion that glycine uptake is driven by a proton symport mechanism. 8. Possible factors governing the strikingly low rate of glycine efflux as opposed to its optimum rate of influx are discussed.

scholarly journals Transmembrane electrical potential and transmembrane pH gradient in the acidophile Thiobacillus ferro-oxidans

1979 ◽  
Vol 178 (1) ◽  
pp. 195-200 ◽  
Author(s):  
J C Cox ◽  
D G Nicholls ◽  
W J Ingledew
Keyword(s):  
Membrane Potential ◽  
Intracellular Ph ◽  
Co2 Fixation ◽  
Electrical Potential ◽  
Sole Source ◽  
Ph Gradient ◽  
Electrochemical Gradient ◽  
Transmembrane Electrical Potential ◽  
External Ph

Thiobacillus ferro-oxidans is capable of using the oxidation of Fe2+ by O2 at pH 2.0 as the sole source of energy for growth and CO2 fixation. The bacterium maintains an intracellular pH of 6.5 over a range of external pH from 1.0 to 8.0, as measured by [14C]acetate and [3H]methylamine distribution. The membrane potential was estimated by the distribution of the lipid-soluble cation dibenzyldimethylammonium and the anion SCN-. At pH 2.0 (the pH of growth) during Fe2+ oxidation the transmembrane pH gradient is 4.5 units with an opposing membrane potential of -10mV, giving a proton electrochemical gradient of +256mV. This gradient is actively maintained.

scholarly journals Use of simulation model for measurement of MilkRun system performance

2019 ◽  
Vol 9 (1) ◽  
pp. 600-605 ◽  
Author(s):  
Gabriel Fedorko ◽  
Martin Vasil ◽  
Michaela Bartosova
Keyword(s):  
Simulation Model ◽  
Production Systems ◽  
Operational Performance ◽  
Transport Systems ◽  
Indirect Methods ◽  
Operational Characteristics ◽  
Wide Range ◽  
Effective Operation ◽  
And Performance ◽  
Plant Simulation

AbstractIntra-plant transport systems within their operation directly impact on the performance of production systems. For their effective operation, it is, therefore, necessary to realize evaluation of operational performance and effectivity. For the realization of this type of evaluation, in addition to a wide range of sensors that can be difficult for installation and operation, we can also use indirect methods that are equally able to provide reliable operational characteristics. Indirect analytical methods are presented above all by the approach which is based on the use of simulation methods. The method of computer simulation provides a wide range of options for the evaluation of efficiency and performance. The paper describes the use of a simulation model created in the program Tecnomatix Plant Simulation for analyzing the supply of production workplaces within the MilkRun system.

scholarly journals Cytoplasmic [Ca2+] and intracellular pH in lymphocytes. Role of membrane potential and volume-activated Na+/H+ exchange.

1987 ◽  
Vol 89 (2) ◽  
pp. 185-213 ◽  
Author(s):  
S Grinstein ◽  
S Cohen
Keyword(s):  
Protein Kinase C ◽  
Membrane Potential ◽  
Protein Kinase ◽  
Intracellular Ph ◽  
Enzyme Treatment ◽  
Membrane Hyperpolarization ◽  
Kinase C ◽  
Free Media ◽  
Stimulation Of

The effect of elevating cytoplasmic Ca2+ [( Ca2+]i) on the intracellular pH (pHi) of thymic lymphocytes was investigated. In Na+-containing media, treatment of the cells with ionomycin, a divalent cation ionophore, induced a moderate cytoplasmic alkalinization. In the presence of amiloride or in Na+-free media, an acidification was observed. This acidification is at least partly due to H+ (equivalent) uptake in response to membrane hyperpolarization since: it was enhanced by pretreatment with conductive protonophores, it could be mimicked by valinomycin, and it was decreased by depolarization with K+ or gramicidin. In addition, activation of metabolic H+ production also contributes to the acidification. The alkalinization is due to Na+/H+ exchange inasmuch as it is Na+ dependent, amiloride sensitive, and accompanied by H+ efflux and net Na+ gain. A shift in the pHi dependence underlies the activation of the antiport. The effect of [Ca2+]i on Na+/H+ exchange was not associated with redistribution of protein kinase C and was also observed in cells previously depleted of this enzyme. Treatment with ionomycin induced significant cell shrinking. Prevention of shrinking largely eliminated the activation of the antiport. Moreover, a comparable shrinking produced by hypertonic media also activated the antiport. It is concluded that stimulation of Na+/H+ exchange by elevation of [Ca2+]i is due, at least in part, to cell shrinking and does not require stimulation of protein kinase C.

Models of Respiratory Rhythm Generation in the Pre-Bötzinger Complex. I. Bursting Pacemaker Neurons

1999 ◽  
Vol 82 (1) ◽  
pp. 382-397 ◽  
Author(s):  
Robert J. Butera ◽  
John Rinzel ◽  
Jeffrey C. Smith
Keyword(s):  
Membrane Potential ◽  
Respiratory Rhythm ◽  
Rhythm Generation ◽  
Minimal Models ◽  
Dynamic Features ◽  
Bursting Neurons ◽  
Wide Range ◽  
Bötzinger Complex ◽  
Voltage Dependent ◽  
Respiratory Rhythm Generation

A network of oscillatory bursting neurons with excitatory coupling is hypothesized to define the primary kernel for respiratory rhythm generation in the pre-Bötzinger complex (pre-BötC) in mammals. Two minimal models of these neurons are proposed. In model 1, bursting arises via fast activation and slow inactivation of a persistent Na+ current I NaP-h. In model 2, bursting arises via a fast-activating persistent Na+ current INaP and slow activation of a K+ current IKS. In both models, action potentials are generated via fast Na+ and K+currents. The two models have few differences in parameters to facilitate a rigorous comparison of the two different burst-generating mechanisms. Both models are consistent with many of the dynamic features of electrophysiological recordings from pre-BötC oscillatory bursting neurons in vitro, including voltage-dependent activity modes (silence, bursting, and beating), a voltage-dependent burst frequency that can vary from 0.05 to >1 Hz, and a decaying spike frequency during bursting. These results are robust and persist across a wide range of parameter values for both models. However, the dynamics of model 1 are more consistent with experimental data in that the burst duration decreases as the baseline membrane potential is depolarized and the model has a relatively flat membrane potential trajectory during the interburst interval. We propose several experimental tests to demonstrate the validity of either model and to differentiate between the two mechanisms.

Ammonia movement and distribution after exercise across white muscle cell membranes in rainbow trout

1996 ◽  
Vol 271 (3) ◽  
pp. R738-R750 ◽  
Author(s):  
Y. Wang ◽  
G. J. Heigenhauser ◽  
C. M. Wood
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Intracellular Ph ◽  
Muscle Cell ◽  
Cell Membranes ◽  
White Muscle ◽  
Extracellular Ph ◽  
Posterior Portion ◽  
Arterial Inflow ◽  
Muscle Cell Membrane

Manipulations of pH and electrical gradients in a perfused preparation were used to analyze the factors controlling ammonia distribution and flux in trout white muscle after exercise. Trout were exercised to exhaustion, and then an isolated-perfused white muscle preparation with discrete arterial inflow and venous outflow was made from the posterior portion of the tail. The tail-trunks were perfused with low (7.4)-, medium (7.9)-, and high (8.4)-pH saline, achieved by varying HCO3- concentration ([HCO3-]) at constant Pco2. Intracellular and extracellular pH, ammonia, CO2, K+, Na+, and Cl- were measured. Muscle intracellular pH was not affected by changes in extracellular pH. Increasing extracellular pH caused a decrease in the transmembrane NH3 partial pressure (PNH3) gradient and a decrease in ammonia efflux. When extracellular K+ concentration was increased from 3.5 to 15 mM in the medium-pH group, a depolarization of the muscle cell membrane potential from -92 to -60 mV and a 0.1-unit depression in intracellular pH occurred. Ammonia efflux increased despite a marked reduction in the PNH3 gradient. Amiloride (10(-4) M) had no effect, indicating that Na+/H(+)-NH4+ exchange does not participate in ammonia transport in this system. A comparison of observed intracellular-to-extracellular ammonia distribution ratios with those modeled according to either pH or Nernst potential distributions supports a model in which ammonia distribution across white muscle cell membranes is affected by both pH and electrical gradients, indicating that the membranes are permeable to both NH3 and NH4+. Membrane potential, acting to retain high levels of NH4+ in the intracellular compartment, appears to have the dominant influence during the postexercise period. However, at rest, the pH gradient may be more important, resulting in much lower intracellular ammonia levels and distribution ratios. We speculate that the muscle cell membrane NH3-to-NH4+ permeability ratio in trout may change between the rest and postexercise condition.

scholarly journals Optical Estimation of Absolute Membrane Potential Using One- and Two-Photon Fluorescence Lifetime Imaging Microscopy

2021 ◽  
Author(s):  
Julia R. Lazzari-Dean ◽  
Evan W. Miller
Keyword(s):  
Membrane Potential ◽  
Fluorescence Lifetime ◽  
Single Photon ◽  
Photon Counting ◽  
Fluorescence Lifetime Imaging ◽  
Single Photon Counting ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Wide Range ◽  
Relative Measurements

AbstractBackgroundMembrane potential (Vmem) exerts physiological influence across a wide range of time and space scales. To study Vmem in these diverse contexts, it is essential to accurately record absolute values of Vmem, rather than solely relative measurements.Materials & MethodsWe use fluorescence lifetime imaging of a small molecule voltage sensitive dye (VF2.1.Cl) to estimate mV values of absolute membrane potential.ResultsWe test the consistency of VF2.1.Cl lifetime measurements performed on different single photon counting instruments and find that they are in striking agreement (differences of <0.5 ps/mV in the slope and <50 ps in the y-intercept). We also demonstrate that VF2.1.Cl lifetime reports absolute Vmem under two-photon (2P) illumination with better than 20 mV of Vmem resolution, a nearly 10-fold improvement over other lifetime-based methods.ConclusionsWe demonstrate that VF-FLIM is a robust and portable metric for Vmem across imaging platforms and under both one-photon and two-photon illumination. This work is a critical foundation for application of VF-FLIM to record absolute membrane potential signals in thick tissue.

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