The role of Ca2+ in volume regulation induced by Na+-coupled alanine uptake in single proximal tubule cells isolated from frog kidney.

1. It has been suggested that epithelial cells maintain cell volume and function, in the face of changes in the rate of transepithelial transport, by activation of volume-regulatory pathways. 2. The aim of the following study was to examine directly the effect of an alteration in Na+-coupled alanine transport on cell length in single proximal tubule cells isolated from frog kidney. 3. An optical technique was used to examine the change in cell length induced by 5 mM L-alanine. 4. On addition of L-alanine to the bath there was an initial increase in cell length to a peak value. This was followed by two types of response. In eighteen out of thirty-one cells a typical volume-regulatory response was observed. The remaining cells showed no volume regulation. 5. Volume regulation was not affected by the removal of extracellular Ca2+. The mean degrees of recovery were 159 +/- 21 % (n = 18) and 144 +/- 18 % (n = 8) in the presence and absence of Ca2+, respectively. 6. Volume regulation was inhibited by depletion of intracellular Ca2+ stores, or in the presence of either Gd3+ or DIDS. The mean degrees of regulation were 55.4 +/- 9.2 % (n = 7), 68.2 +/- 18.8 % (n = 7) and 69.1 +/- 14.3 % (n = 7), respectively. 7. The alanine-induced increases in cell length were both stereospecific and Na+ dependent. 8. The evidence suggests that volume regulation induced by Na+-coupled alanine uptake may be dependent on the release of Ca2+ from intracellular stores. This is in contrast to volume regulation induced by hypotonic shock, which appears to require extracellular Ca2+. Results obtained using a hypotonic shock should, therefore, be viewed with caution.