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.