Transmembrane 45Ca fluxes in isolated smooth muscle cells: basal Ca2+ fluxes

Abstract

Methods are described for monitoring unidirectional fluxes of 45Ca in smooth muscle cells in suspension. Compartmental analysis of 45Ca influx data indicate that 45Ca exchange consists of two kinetically distinguishable components in the toad stomach muscle cells: a relatively small (approximately 100-pmol X cm-2) component with a rapid rate of exchange (t1/2 approximately 1.4 min) and a larger (approximately 930-pmol X cm-2) component, which exchanges more slowly (t1/2 congruent to 87 min). The rate of exchange of the latter but not the former Ca2+ pool is increased on exposure of the cells to elevated K+ levels; thus the “slow” component of uptake appears to reflect transmembrane Ca2+ flux, whereas the “rapid” component may reflect exchange of surface-bound label. Consistent with this interpretation is the finding that under conditions in which surface 45Ca is rapidly and completely displaced, 45Ca efflux occurs as a simple monoexponential process with a slow rate of exchange (k = 7.80 X 10(-5) s-1). The apparent rate of transmembrane Ca2+ flux in smooth muscle cells at rest is approximately 0.1 pmol X cm-2 X s-1. Cellular processes that could give rise to an apparent transmembrane flux rate of this magnitude are discussed, and a model is presented which appears to describe cellular 45Ca exchange in the isolated smooth muscle cells.