title: ATP depletion in rat cholangiocytes leads to marked internalization of membrane proteins.
abstract: Intrahepatic bile ducts (BD) are a critical target of injury in the postischemic liver. Decreased vascular perfusion causes characteristic changes in the morphology of the ductular epithelia including a loss of secondary membrane structures and a decrease in plasma membrane surface area. Using adenosine triphosphate (ATP) depletion of cultured normal rat cholangiocytes (NRC) to model ischemic ducts, the present studies examined the fate of apical membrane proteins to determine whether membrane recycling might contribute to rapid functional recovery. Apical proteins, including gamma-glutamyl transpeptidase (GGT), Na(+)-glucose cotransporter (SGLT1), and apically biotinylated proteins, were not shed into the luminal space during ATP depletion. Instead, labeling of surface proteins after ATP depletion showed a significant decrease in GGT and SGLT1, consistent with membrane internalization. Similarly, z-axis confocal microscopy of biotinylated apical proteins also showed protein internalization. During ATP recovery, SGLT1 transport activity remained profoundly depressed even after 24 hours of recovery, indicating that the function of the internalized apical proteins is not rapidly recovered. These studies suggest that the membrane internalization in ATP-depleted cholangiocytes is a unidirectional process that contributes to prolonged functional deficits after restoration of normal cellular ATP levels. This sustained decrease in transport capacity may contribute to the development of ductular injury in postischemic livers.
authors.affiliation: University of Colorado Health Sciences Center, Denver, CO 80262, USA. brian.doctor@uchs.edu
authors.forename: R B
authors.initials: RB
authors.lastname: Doctor
authors.forename: R H
authors.initials: RH
authors.lastname: Dahl
authors.forename: K D
authors.initials: KD
authors.lastname: Salter
authors.forename: L
authors.initials: L
authors.lastname: Fouassier
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: J G
authors.initials: JG
authors.lastname: Fitz
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001653:Bile Ducts, Intrahepatic / Q000166:cytology / Q000378:metabolism*; D007511:Ischemia / Q000378:metabolism; D008099:Liver / Q000098:blood supply; D016031:Liver Transplantation; D008562:Membrane Glycoproteins / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D051269:Sodium-Glucose Transporter 1; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
chemical_list: D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; C495676:Slc5a1 protein, rat; D051269:Sodium-Glucose Transporter 1; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase