Rab11a redistributes to apical secretory canaliculus during stimulation of gastric parietal cells

Previous investigations in several systems have demonstratedthat Rab3 family members redistribute to soluble fractions on fusion ofsecretory granules with target plasma membranes. Rab proteins are thenrecycled back onto mature secretory vesicles after reinternalization ofthe membrane. Although this cycle is well established for Rab3, farless is known about redistribution of other Rab proteins during vesiclefusion and recycling. In the gastric parietal cell, Rab11a isassociated with H-K-ATPase-containing tubulovesicles, which fuse withthe apical plasma membrane (secretory canaliculus) in response toagonists such as histamine. We have analyzed distribution of Rab11a andother tubulovesicle proteins in resting and histamine-stimulated rabbitparietal cells. Stimulation of isolated gastric glands in the presenceof 100 µM histamine and 100 µM 3-isobutyl-1-methylxanthine did notcause a significant increase in soluble Rab11a. H-K-ATPase, Rab11a,Rab25, syntaxin 3, and SCAMPs increased immunoreactivity instimulus-associated vesicles prepared from rabbits treated withhistamine compared with those from ranitidine-treated animals. Thelarge GTPase dynamin was found in both vesicle preparations, but therewas no change in amount of immunoreactivity. Immunofluorescencestaining of resting and histamine-stimulated primary cultures ofparietal cells demonstrated redistribution of H-K-ATPase and Rab11a to F-actin-rich canalicular membranes. Dynamin was present on canalicular membranes in resting and stimulated cells. These results indicate thatRab11a does not cycle off the membrane during the process oftubulovesicle fusion with the secretory canaliculus. Thus Rab11a mayremain associated with recycling apical membrane vesicle populations.