mV Of Hyperpolarization Research Articles

Biophysical properties of microvascular endothelium: Requirements for initiating and conducting electrical signals

Electrical signaling along the endothelium underlies spreading vasodilation and blood flow control. We use mathematical modeling to determine the electrical properties of the endothelium and gain insight into the biophysical determinants of electrical conduction. Electrical conduction data along endothelial tubes (40μm wide, 2.5mm long) isolated from mouse skeletal muscle resistance arteries were analyzed using cable equations and a multicellular computational model. Responses to intracellular current injection attenuate with an axial length constant (λ) of 1.2-1.4mm. Data were fitted to estimate the axial (ra ; 10.7MΩ/mm) and membrane (rm ; 14.5MΩ∙mm) resistivities, EC membrane resistance (Rm ; 12GΩ), and EC-EC coupling resistance (Rgj ; 4.5MΩ) and predict that stimulation of ≥30 neighboring ECs is required to elicit 1 mV of hyperpolarization at distance = 2.5 mm. Opening Ca2+ -activated K+ channels (KCa ) along the endothelium reduced λ by up to 55%. High Rm makes the endothelium sensitive to electrical stimuli and able to conduct these signals effectively. Whereas the activation of a group of ECs is required to initiate physiologically relevant hyperpolarization, this requirement is increased by myoendothelial coupling and KCa activation along the endothelium inhibits conduction by dissipating electrical signals.

Single channel properties of synaptic acetylcholine receptors in dystrophic fibers

Neuromuscular transmission in dystrophic mice has been extensively studied through analysis of nerve-evoked endplate potentials. In the present study, single channel ACh-induced activity recorded from endplates of fast twitch muscle fibers from mdx and dy dystrophic mice, was compared with activity recorded from wild-type fiber endplates to ascertain whether expression of these phenotypes leads to changes in ACh receptor properties and synaptic transmission. An 89 pS class of ACh-induced events predominated in recordings from wild-type and both strains of dystrophic mice. The mean open times for this class of events was well described by two exponential components, one with a voltage-independent time constant of approximately 0.3 ms and the other with a time constant of 2-5 ms which increased e-fold with approximately 120 mV of hyperpolarization. The expression of the mdx or the dy phenotype was not associated with significant differences in the conductance, distribution of open durations, or the voltage-dependence of the mean open time for this class of ACh-induced events.

Conus geographus toxins that discriminate between neuronal and muscle sodium channels.

We describe the properties of a family of 22-amino acid peptides, the mu-conotoxins, which are useful probes for investigating voltage-dependent sodium channels of excitable tissues. The mu-conotoxins are present in the venom of the piscivorous marine snail, Conus geographus L. We have purified seven homologs of the mu-conotoxin set and determined their amino acid sequences, as follows, where Hyp = trans-4-hydroxyproline. GIIIA R.D.C.C.T.Hyp.Hyp.K.K.C.K.D.R.Q.C.K.Hyp.Q.R.C.C.A-NH2 [Pro6]GIIIA R.D.C.C. T.P.Hyp.K.K.C.K.D.R.Q.C.K.Hyp.Q.R.C.C.A-NH2 [Pro7]GIIIA R.D.C.C.T.Hyp.P.K.K.C.K.D.R.Q.C.R.Hyp.Q.R.C.C.A-NH2 GIIIB R.D.C.C.T.Hyp.Hyp.R.K.C.K.D.R.R.C.K.Hyp.M.K.C.C.A-NH2 [Pro6]GIIIB R.D.C.C.T.P.Hyp.R.K.C.K.D.R.R. C.K.Hyp.M.K.C.C.A-NH2 [Pro7]GIIIB R.D.C.C.T.Hyp.P.R.K.C.K.D.R.R.C.K.Hyp.M.K.C.C.A-NH2 GIIIC R.D.C.C.T.Hyp.Hyp.K.K.C.K.D.R.R.C.K.Hyp.L.K.C.C.A-NH2. Using the major peptide (GIIIA) in electrophysiological studies on nerve-muscle preparations and in single channel studies using planar lipid bilayers, we have established that the toxin blocks muscle sodium channels, while having no discernible effect on nerve or brain sodium channels. In bilayers the blocking kinetics of GIIIA were derived by statistical analysis of discrete transitions between blocked and unblocked states of batrachotoxin-activated sodium channels from rat muscle. The kinetics conform to a single-site, reversible binding equilibrium with a voltage-dependent binding constant. The measured value of the equilibrium KD for GIIIA is 100 nM at OmV, decreasing e-fold/34 mV of hyperpolarization. This voltage dependence of blocking is similar to that of tetrodotoxin and saxitoxin as measured by the same technique. The tissue specificity and kinetic characteristics suggest that the mu-conotoxins may serve as useful ligands to distinguish sodium channel subtypes in different tissues.

The interaction of potassium with the activation of anomalous rectification in frog muscle membrane.

1. Inward rectification of frog muscle membrane was analysed with the Vaseline gap method. 2. Hyperpolarization, under voltage clamp, produced inward potassium currents, which had a component that activated with a time constant, tau K. 3. The activation time constant tau K of the inward potassium currents was voltage dependent. For a given external potassium concentration, the time constant was maximal for potentials near the potassium equilibrium potential, EK. 4. The potassium chord conductance gK, had a sigmoidal voltage dependency, increasing initially e-fold per 11.6 mV of hyperpolarization. 5. When the internal potassium concentration was fixed, raising external potassium induced a shift of the tau K-V and the gK-V relations in the positive direction along the voltage axis. That shift was comparable to the change in EK. 6. No shift of the tau K-V and the gK-V relations was observed when the internal potassium was reduced from 150 to 50 mM. 7. Changes of internal sodium concentration between 5 and 100 mM did not significantly effect the magnitude of inward rectification.