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Biomedical subjects

K E Akerman

Publications and source records attributed to K E Akerman.

139 records · Page 8Linked to original sources
PubMed1979

Induction of exocytosis from glomus cells by incubation of the carotid body of the rat with calcium and ionophore A23187.

Carotid bodies from adult rats were electron microscopically studied after incubation in glucose-containing salt solutions containing calcium and/or ionophore A23187 or neither. In the absence of the ionophore, adding or omitting calcium had no effect on the fine structure of the glomus cells. Incubation in the medium containing both 1 mM calcium and the ionophore caused the appearance of exocytotic membrane profiles in several glomus cells. Exocytosis was not seen when only A23187 and endogenous calcium was present. For exocytosis to occur, calcium appeared to be essential and the event seemed to be due to a rise in the intracellular calcium concentration caused by the ionophore.

Animals↗
PubMed1978

Changes in membrane potential during calcium ion influx and efflux across the mitochondrial membrane.

1. A depolarisation of the membrane of rat liver mitochondria, as measured with the safranine method, is seen during Ca2+ uptake. The depolarisation is followed by a slow repolarisation, the rate of which can be increased by the addition of EGTA or phosphate. 2. Plots relating the initial rate of calcium ion (Ca2+) uptake and the decrease in membrane potential (delta psi) to the Ca2+ concentration show a half-maximal change at less than 10 micron Ca2+ and a saturation above 20 micron Ca2+. 3. Plots relating the initial rate of Ca2+ uptake to delta psi are linear. 4. Addition of Ca2+ chelators, nitriloacetate or EGTA, to deenergized mitochondria equilibrated with Ca2+ causes a polarisation of the mitochondrial membrane due to a diffusion potential created by electrogenic Ca2+ efflux. 5. If the extent of the response induced by different nitriloacetate concentrations is plotted against the expected membrane potential a linear plot is obtained up to 70 mV with a slope corresponding to two-times the extent of the response induced by valinomycin in the presence of different potassium ion gradients. This suggests that the Ca2+ ion is transferred across the membrane with one net positive charge in present conditions.

Acetates↗
PubMed1978

Measurements of membrane potentials using the dye safranine.

A metachromatic shift in the spectrum of the cationic dye safranine occurs upon induction of electrical potentials across the mitochondrial membrane by adding respiratory substrate, ATP or a cation conductor valinomycin (when a potassium gradient exists across the membrane) to a mitochondrial suspension. The extent of spectral change correlates linearily to the membrane potential. During the spectral change safranine is taken up by the mitochondria and most of the dye can be recovered in the pellet after centrifugation. By measuring the spectral changes a fairly good estimate of membrane potentials in mitochondria and bacteria is obtained. This method is compared to other optical methods of studying membrane potentials and is possible applications in different systems in the future is discussed.

Adenosine Triphosphate↗
PubMed1977

Effect of inhibitors on the sigmoidicity of the calcium ion transport kinetics in rat liver mitochondria.

The kinetic plot (initial rate of Ca2+ transport versus concentration) of mitochondrial Ca2+ transport is hyperbolic in a sucrose medium. The plot becomes sigmoidal in the presence of competitive inhibitors of Ca2+ binding to low affinity sites of the membrane surface such as Mg2+ and K+. The plot also becomes sigmoidal in the presence of Ba2+. Ba2+ is a competitive inhibitor of both Ca2+ transport and Ca2+ binding to the low affinity sites. The 5i for the inhibition of Ca2+ transport by Ba2+ increases in the presence of K+ and Mg2+, which suggests a competition for the low affinity sites between the cations. The plot is still hyperbolic in the presence of La3+, which inhibits Ca2+ transport competitively. Ruthenium red which is a pure non-competitive inhibitor of mitochondrial Ca2+ transport, does not affect the shape of the kinetic plot. These results indicate that the surface potential, which depends on the ions bound to the low affinity sites, determines whether the kinetics of Ca2+ uptake in mitochondria is sigmoidal or hyperbolic.

Animals↗
PubMed1977

Effect of Mg2+ and spermine on the kinetics of Ca2+ transport in rat-liver mitochondria.

Plots relating the initial rate of mitochondrial Ca2+ transport to the Ca2+ concentration (kinetic plots) have a hyperbolic shape in a Ca2+ concentration range of 2.5-100 muM as measured in sucrose or KCl media. In the presence of Mg2+ or a polyamine spermine, which both are competitive inhibitors of Ca2+ binding to low affinity sites at the membrane surface, the shape of the plots becomes sigmoidal. At higher concentrations of these agents linear kinetic plots are obtained as measured in a sucrose medium. In a KCl medium the sigmoidality of the kinetic plots is enhanced by an increase in the Mg2+ or spermine concentration. It is suggested that Mg2+ and spermine affect the kinetics of Ca2+ transport by interfering with Ca2+ binding to low affinity sites of the membrane surface and that the binding of Ca2+ to these sites is the first step of the mitochondrial Ca2+ transport.

Animals↗
PubMed1977

Effect of propranolol and related drugs on transmembraneous pH differences in liposomes.

Propranolol (1-isopropylamino-3-(1-naphtoloxy)-propan-2-ol) a beta-adrenergic receptor blocking agent was found to cause changes of transmembraneous pH in liposomes prepared from Soy-lecithin and cardiolipin. When the external pH was neutral and the internum of the liposomes acidic, the drug decreased the pH gradient. When the externum was acidic and the internum neutral, the gradient was increased by the drug. The effect of butacaine was similar to that of propranolol, while procaine, timolol and practolol were ineffective. It is suggested that the charged form of propranolol is bound to the membrane and dislocates protons from binding sites in the membrane and that the uncharged form of propranolol penetrates the membrane. After penetration it could associate with protons in the intraliposomal compartment and hence increase the pH of the interior. Depending on the direction of the pre-existing proton gradient propranolol would thus be able to increase or decrease the pH difference across the liposomal membrane.

4-Aminobenzoic Acid↗
PubMed1976

Stacking of safranine in liposomes during valinomycin-induced efflux of potassium ions.

Liposomes were prepared from phosphatidylcholine and cardiolipin in a KCl medium and suspended in a choline chloride medium with safranine. When efflux of K+ was induced by valinomycin, spectral shifts characteristic of stacking were observed. Ca2+ inhibited the rate of stacking in a competitive manner with a Ki of about 200 muM, while La3+ was about 10 times more potent. When liposomes were prepared from phospholipids with a higher ratio of cardiolipin to phosphatidylcholine the inhibition was more potent. No effect on the stacking phenomena was seen when CA2+ was added after the stacking was completed. When CA2+ or an organic cation with four charges, spermine was trapped in the intraliposomal compartment, no significant change in the rate of stacking was seen. However, the extent of stacking was decreased. It is suggested that safranine is driven by a diffusion potential to a site that is inaccessible to CA2+ in the medium, presumably to the inner boundaries of the liposomal membranes.

Binding Sites↗
PubMedDate not supplied

Kinetic evidence for different mechanisms of interaction of black mamba toxins MT alpha and MT beta with muscarinic receptors.

By studying the influence of two toxins from the black mamba Dendroaspis polylepis on the kinetics of [3H]-N-methylscopolamine binding to muscarinic acetylcholine receptors from rat cerebral cortex, it was revealed that these toxins, MT alpha and MT beta, interact with the receptors via kinetically distinct mechanisms. MT beta bound to receptors in a one-step, readily reversible process with the dissociation constant K(d)=5.3 microM. The binding mechanism of MTalpha was more complex, involving at least two consecutive steps. A fast receptor-toxin complex formation (K(T)=3.8 microM) was followed by a slow process of isomerisation of this complex (k(i)=1.8 x 10(-2) s(-1), half-time 39 s). A similar two-step interaction mechanism has been established for a related toxin, MT2 from the green mamba D. angusticeps (K(T)=1.4 microM, k(i)=8.3 x 10(-4) s(-1), half-time 840 s). The slow isomerisation process delays the effect of MT alpha and MT2, but increases their apparent potency compared to toxins unable to induce the isomerisation process.

Animals↗