VESICLE TARGETING TO THE APICAL DOMAIN REGULATES BILE EXCRETORY FUNCTION IN ISOLATED RAT HEPATOCYTE COUPLETS

Background & Aims: Plasma membrane solute transport may be regulated in many epithelial cells by vesicle traffic to and from the site of residence of the transporter. The aim of this study was to determine if this phenomenon may also play a role in the regulation of canalicular transport of bile acids. Methods: Confocal microscopy and image analysis were performed to quantitatively assess changes in secretory capacity and vesicle targeting in isolated rat hepatocyte couplets that had been exposed to fluorescent bile acid after pretreatment with dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) and/or nocodazole. Results: DBcAMP stimulated bile acid secretion by 240% while significantly increasing canalicular circumference. Nocodazole decreased secretion by 410% and significantly decreased canalicular circumference. When DBcAMP was added to nocodazole-treated couplets, a slight but significant increase was found in both fluorescent bile acid secretion and canalicular circumference as compared with nocodazole alone. Finally, DBcAMP stimulated translocation of vesicles to the canalicular membrane as determined by immunocytochemical localization of a putative bile acid transporter, Ca2+, Mg2+-ecto-adenosine triphosphatase. Conclusions: The findings support the view that apical membrane transport activity in the rat hepatocyte is highly regulated by the insertion of vesicles into this domain and that this process involves both microtubule-dependent and -independent mechanisms.