Cellular defibrillation: interaction of micro-scale electric fields with voltage-gated ion channels.

We study the effect of micro-scale electric fields on voltage-gated ion channels in mammalian cell membranes. Such micro- and nano-scale electric fields mimic the effects of multiferroic nanoparticles that were recently proposed [1] as a novel way of controlling the function of voltage-sensing biomolecules such as ion channels. This article describes experimental procedures and initial results that reveal the effect of the electric field, in close proximity of cells, on the ion transport through voltage-gated ion channels. We present two configurations of the whole-cell patch-clamping apparatus that were used to detect the effect of external stimulation on ionic currents and discuss preliminary results that indicate modulation of the ionic currents consistent with the applied stimulus.

Wavelet-based protocols for ion channel electrophysiology.

BACKGROUND: Fluctuation-induced phenomena caused by both random and deterministic stimuli have been previously studied in a variety of contexts. They are based on the interplay between the spectro-temporal patterns of the signal and the kinetics of the system it is applied to. The aim of this study was to develop a method for designing fluctuating inputs into nonlinear system which would elicit the most desired system output and to implement the method to studies of ion channels. RESULTS: We describe an algorithm based on constructing the input as a superposition of wavelets and optimizing it according to a selected cost functional. The algorithm is applied to ion channel electrophysiology where the input is the fluctuating voltage delivered through a patch-clamp experimental apparatus and the output is the whole-cell ionic current. The algorithm is optimized to aid selection of Markov models of the gating kinetics of the voltage-gated Shaker K+ channel and tested by comparison of numerically obtained ionic currents predicted by different models with experimental data obtained from the Shaker K+ channels. Other applications and optimization criteria are also suggested. CONCLUSION: The method described in this paper can be useful in development and testing of models of ion channel gating kinetics, developing voltage inputs that optimize certain nonequilibrium phenomena in ion channels, such as the kinetic focusing, and potentially has applications to other fields.

Test of the nonequilibrium kinetic focusing of voltage-gated ion channels.

It has been shown numerically (Millonas and Chialvo 1996 Phys. Rev. Lett. 76 550) that voltage-gated ion channels can be focused into specific conformational states by the application of fluctuating voltages, such as dichotomous noise. We present results of an experimental test performed on Shaker K+ channels. We applied dichotomous noise with properties mimicking the numerical simulations as closely as physiologically feasible. We observed that the probability for intermediate states can be increased as much as 50% above the maximal value for any static voltages.

A study of porous structure of cellular membranes in human erythrocytes.

Properties of cell membrane of human erythrocytes are studied using the mechanistic formalism of membrane transport developed earlier. We estimate that an erythrocyte with a membrane surface of 176 x 10(6)nm2 has about 1900 water-permeable pores with cross-section areas ranging from 0.07 to 0.2 nm2.

Application of the ensemble nonequilibrium response spectroscopy to shaker potassium channel gating.

Standard electrophysiology techniques study relaxation transients in voltage-gated ion channels generated by discrete voltage steps. The nonequilibrium response spectroscopy involves analyzing responses to fluctuating potentials. We apply the ensemble NRS method to gating kinetics of Shaker potassium ion channels. We evaluate various proposed Markov models of channel gating from the nonequilibrium response viewpoint. These new NRS protocols can be used to test otherwise indistinguishable models or improve estimates for parameters of channel kinetics models.

Applications of nonequilibrium response spectroscopy to the study of channel gating. Experimental design and optimization.

A novel experimental technique known as non-equilibrium response spectroscopy (NRS) based on ion channel responses to rapidly fluctuating voltage waveforms was recently described (Millonas & Hanck, 1998a). It was demonstrated that such responses can be affected by subtle details of the kinetics that are otherwise invisible when conventional stepped pulses are applied. As a consequence, the kinetics can be probed in a much more sensitive way by supplementing conventional techniques with measurements of the responses to more complex voltage waveforms. In this paper we provide an analysis of the problem of the design and optimization of such waveforms. We introduce some methods for determination of the parametric uncertainty of a class of kinetic models for a particular data set. The parametric uncertainty allows for a characterization of the amount of kinetic information acquired through a set of experiments which can in turn be used to design new experiments that increase this information. We revisit the application of dichotomous noise (Millonas & Hanck, 1998a, b), and further consider applications of a more general class of continuous wavelet -based waveforms. A controlled illustration of these methods is provided by making use of a simplified "toy" model for the potassium channel kinetics.