Voltage- and time-dependent block of I(f) by Sr2+ in rabbit sino-atrial node cells

The hyperpolarization-activated cation current (I(f)) was recorded in single myocytes dissociated from the rabbit sino-atrial node and the Sr(2+)-mediated block of I(f) examined. In the presence of 0.1-10 mM Sr2+, the activation phase of I(f) was followed by a slower decay during hyperpolarization. In the steady state I/V diagram, the Sr2+ block progressed with increasing hyperpolarization. Ba2+ also blocked I(f), but no time dependency could be observed. The blocking effect of Ca2+ was weak, and Mg2+ had little effect on I(f). The relationship between extracellular Sr2+ concentration [Sr2+]o and the block was described by a Hill coefficient of 1. The half-saturating [Sr2+]o were 2.0, 1.6, 1.1 and 0.65 mM at -90, -100, -110 and -120 mV, respectively. The rapid application of Sr2+ during the full activation of I(f) using the jet-stream method induced an exponential decline of I(f). The reciprocal time constants were linearly correlated to [Sr2+]o, suggesting 1:1 binding stoichiometry. The fractional electrical distance for the binding site was approximately 0.5 from the external side of the channel. Based on the multiple closed and open states for the I(f) channel, a mathematical model for the Sr2+ block was constructed in which the time course of I(f) in the presence of Sr2+ was described by the sum of three exponential functions. Fitting the model to the original traces revealed blocking and unblocking rates similar to those obtained from the jet-stream method. At -110 mV, the blocking rate was 410 M-1 s-1 while the unblocking rate was 0.16 s-1.(ABSTRACT TRUNCATED AT 250 WORDS)