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Am J Physiol Cell Physiol 281: C407-C417, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 2, C407-C417, August 2001

Functionally distinct pools of actin in secretory cells

David A. Ammar, Phuong N. B. Nguyen, and John G. Forte

Department of Molecular and Cell Biology, University of California, Berkeley, California 94720

Acid secretion by the gastric parietal cell is controlled through movement of vesicles containing the proton pump, the H+-K+-ATPase (HK). We have used latrunculin B (Lat B), which binds to monomeric actin, to investigate actin turnover in the stimulated parietal cell. In isolated gastric glands, relatively high concentrations of Lat B were required to inhibit acid accumulation (ED50 ~70 µM). Cultured parietal cells stimulated in the presence of low Lat B (0.1-1 µM) have reduced lamellipodia formation and some aberrant punctate phalloidin-stained structures, but translocation of HK and vacuolar swelling appeared unaffected. High Lat B (10-50 µM) resulted in gross changes in actin organization (punctate phalloidin-stained structures throughout the cell and nucleus) and reduced translocation of HK and vacuolar swelling. Resting parietal cells treated with high Lat B showed minor effects on morphology and F-actin staining. If resting cells treated with high Lat B were washed immediately before stimulation, they exhibited a normal stimulated morphology. These data suggest distinct pools of parietal cell actin: a pool highly susceptible to Lat B primarily involved in motile function of cultured cells; and a Lat B-resistant pool, most likely microvillar filaments, that is essential for secretion. Furthermore, the stimulation process appears to accentuate the effects of Lat B, most likely through Lat B binding to monomer actin liberated by the turnover of the motile actin filament pool.

parietal cell; latrunculin B; secretion; cytoskeleton


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