Agonist-induced coordinated trafficking of functionally related transport proteins for water and ions in cholangiocytes.
We previously proposed that ductal bile formation is regulated by secretin-responsive relocation of aquaporin 1 (AQP1), a water-selective channel protein, from an intracellular vesicular compartment to the apical membrane of cholangiocytes. In this study, we immunoisolated AQP1-containing vesicles from cholangiocytes prepared from rat liver; quantitative immunoblotting revealed enrichment in these vesicles of not only AQP1 but also cystic fibrosis transmembrane regulator (CFTR) and AE2, a Cl- channel and a Cl-/HCO3- exchanger, respectively. Dual labeled immunogold electron microscopy of cultured polarized mouse cholangiocytes showed significant colocalization of AQP1, CFTR, and AE2 in an intracellular vesicular compartment; exposure of cholangiocytes to dibutyryl-cAMP (100 microm) resulted in co-redistribution of all three proteins to the apical cholangiocyte plasma membrane. After administration of secretin to rats in vivo, bile flow increased, and AQP1, CFTR, and AE2 co-redistributed to the apical cholangiocyte membrane; both events were blocked by pharmacologic disassembly of microtubules. Based on these in vitro and in vivo observations utilizing independent and complementary approaches, we propose that cholangiocytes contain an organelle that sequesters functionally related proteins that can account for ion-driven water transport, that this organelle moves to the apical cholangiocyte membrane in response to secretory agonists, and that these events account for ductal bile secretion at a molecular level.
Tietz
Pamela S
PS
Center for Basic Research in Digestive Diseases, Department of Internal Medicine, Mayo Medical School, Clinic and Foundation, Rochester, Minnesota 55905, USA.
Marinelli
Raul A
RA
Chen
Xian-Ming
XM
Huang
Bing
B
Cohn
Jonathan
J
Kole
Jolanta
J
McNiven
Mark A
MA
Alper
Seth
S
LaRusso
Nicholas F
NF
D000818:Animals; D027321:Anion Transport Proteins; D017920:Antiporters; D051398:Aquaporin 1; D020346:Aquaporins / Q000378:metabolism; D001653:Bile Ducts, Intrahepatic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D017136:Ion Transport; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D021381:Protein Transport; D051381:Rats; D011916:Rats, Inbred F344; D064606:SLC4A Proteins; D012633:Secretin / Q000494:pharmacology*; D014867:Water / Q000378:metabolism*
D027321:Anion Transport Proteins; D017920:Antiporters; C496022:Aqp1 protein, rat; D020346:Aquaporins; D008565:Membrane Proteins; D064606:SLC4A Proteins; D014867:Water; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012633:Secretin; D051398:Aquaporin 1