Na+ currents through low-voltage-activated Ca2+ channels of chick sensory neurones: block by external Ca2+ and Mg2+

1. Whole-cell currents through low-voltage-activated (LVA) Ca2+ channels carried by monovalent cations were studied in chick dorsal root ganglion (DRG) cells. 2. With 120 mM [Na+] on both sides of the membrane and [Ca2+]o less than or equal to 100 microM, the currents reversed at 0 mV. Their half-times of activation and inactivation were strictly voltage-dependent and decreased to near-constant values of 0.6-0.85 and 40 ms, respectively, at positive membrane potentials. The longer activation times were observed with [Ca2+]o greater than or equal to 50 microM. 3. The selectivity of the Ca2+ channel for monovalent ions with reference to internal Na+ was evaluated from the reversal potential. The Li+ and Na+ permeabilities were similar. The permeability ratios of K+ and Rb+ were 0.45, and 0.33 for Cs+. 4. Micromolar increases in [Ca2+]o produced small voltage shifts of half-times of activation (less than or equal to +3 mV at 10 microM and +10 mV at 500 microM), but strongly depressed the Na+ current. The Ca2(+)-induced block of Na+ current satisfied a 1:1 stoichiometry with an apparent KD of 1.8 microM at -20 mV. The block was, however, relieved with more positive and negative potentials, with KDs of 55 and 8.5 microM at +90 and -110 mV, respectively. 5. Relaxation time constants of block and unblock of Na+ currents through the LVA Ca2+ channel were measured on step changes to and from membrane potentials at which pronounced Ca2(+)-induced block occurred. 6. At -20 mV, the time constants of block decreased with micromolar increase in [Ca2+]o in line with a blocking rate coefficient of 1.9 x 10(8) M-1 s-1, but settled to values of 0.18 ms at [Ca2+]o beyond 50 microM. The Na+ currents were unblocked with time constant (tau u) of around 0.25 ms at strongly positive and negative membrane potentials at 22 degrees C. 7. Tau u failed to show any obvious dependence on [Ca2+]o up to the millimolar range. This finding contradicts suggestions that removal of the block occurs in a [Ca2+]o-dependent manner as a result of an increased probability of Ca2+ ion mobilization by repulsive forces with increased Ca2+ occupation of the channel. 8. The time course of unblock of Na+ currents was strongly temperature-dependent showing a Q10 of 2.5 for tau u. 9. The voltage dependence of the Na+ current block by Ca2+ ions is best accounted for by a single, centrally located Ca2+ binding site. The kinetic features of block and unblock suggest ion-specific but concentration-independent transitions between channel conformations with different ionic selectivities. A state model based on these assumptions was able to reproduce the observed voltage- and concentrationdependent behaviour of the channel.