The characteristics of Ca -activated Cl- channels of the salt-tolerant charophyte Lamprothamnium

Abstract

The dependence of the Ca++-activated Cl- channels on potential difference (PD) was extracted from current-voltage (I/V) profiles recorded at the time of hypotonic regulation while the large conductance (G) K+ channels were blocked by tetraethylammonium (TEA). The total clamp current (I) was dominated by the Cl- I, i(Cl), with small contribution from the background I (i(background)). The i(Cl) was fitted by the Goldman-Hodgkin-Katz (GHK) model with enhanced PD dependence simulated by Boltzmann probability distributions. The i(background) was modelled by an empirical equation. The i(Cl) responded to PD changes within tens of milliseconds. The G maxima were located between -20 and -150 mV. The Cl- channel number and channel permeability parameter, N(Cl)P(Cl), decreased as a function of time in a hypotonic medium (from 0.45 x 10(-7) to 0.17 x 10(-7) ms(-1) in 19 min), with the positive half activation PD, V50+, shifting from +35 to -65 mV, and the negative half activation PD, V50-, shifting from -134 to -310 mV. The fitted Cl- concentration [Cl-]cyt at the time of hypotonic regulation indicated rapid equalization of vacuolar and cytoplasmic concentrations. Excellent data obtained under similar experimental conditions in a previous study enabled us to infer [Ca++]cyt influences on the Cl- channel characteristics. Thick sulphated polysaccharide mucilage, found on Lamprothamnium cells acclimated to more saline media, eliminated the activation of the i(Cl) at the time of the hypotonic regulation. This effect was reversed by the application of the enzyme heparinase. The characteristics of the i(Cl) were found to be consistent with a component of the excitation Is at the time of the action potential (AP). The short duration of the excitation transients was contrasted with that of the hypotonic regulation. The mechanisms for Cl- channel activation (and hence the Ca++ channel activation) were considered.