TY - JOUR
T1 - Water Transporting Properties of Hepatocyte Basolateral and Canalicular Plasma Membrane Domains
AU - Marinelli, Raul A.
AU - Tietz, Pamela S.
AU - Caride, Ariel J.
AU - Huang, Bing Q.
AU - LaRusso, Nicholas F.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2003/10/31
Y1 - 2003/10/31
N2 - Previous work from our laboratory supports an important role for aquaporins (AQPs), a family of water channel proteins, in bile secretion by hepatocytes. To further define the pathways and molecular mechanisms for water movement across hepatocytes, we directly assessed osmotic water permeability (Pf) and activation energy (Ea) in highly purified, rat hepatocytes basolateral membrane vesicles (BLMV) and canalicular membrane (CMV) vesicles by measuring scattered light intensity using stopped-flow spectrophotometry. The time course of scattered light for BLMV and CMV fit well to a single-exponential function. In BLMV, Pf was 108 ± 4 μm·s-1 (25 °C) with an Ea of 7.7 kcal/mol; in CMV, Pf was 86 ± 5 μm·s-1 (25 °C) with an Ea of 8.0 kcal/mol. The AQP blocker, dimethyl sulfoxide, significantly inhibited the Pf of both basolateral (81 ± 4 μm·s-1; -25%) and canalicular (59 ± 4 μm·s-1 -30%) membrane vesicles. When CMV were isolated from hepatocytes treated with dibutyryl cAMP, a double-exponential fit was needed, implying two functionally different vesicle populations; one population had Pf and Ea values similar to those of CMV from untreated hepatocytes, but the other population had a very high Pf (655 ± 135 μm·s-1, 25 °C) and very low E a (2.8 kcal/mol). Dimethyl sulfoxide completely inhibited the high Pf value in this second vesicle population. In contrast, P f and Ea of BLMV were unaltered by cAMP treatment of hepatocytes. Our results are consistent with the presence of both lipid- and AQP-mediated pathways for basolateral and canalicular water movement across the hepatocyte plasma membrane barrier. Our data also suggest that the hepatocyte canalicular membrane domain is rate-limiting for transcellular water transport and that this domain becomes more permeable to water when hepatocytes are exposed to a choleretic agonist, presumably by insertion of AQP molecules. These data suggest a molecular mechanism for the efficient coupling of osmotically active solutes and water transport during canalicular bile formation.
AB - Previous work from our laboratory supports an important role for aquaporins (AQPs), a family of water channel proteins, in bile secretion by hepatocytes. To further define the pathways and molecular mechanisms for water movement across hepatocytes, we directly assessed osmotic water permeability (Pf) and activation energy (Ea) in highly purified, rat hepatocytes basolateral membrane vesicles (BLMV) and canalicular membrane (CMV) vesicles by measuring scattered light intensity using stopped-flow spectrophotometry. The time course of scattered light for BLMV and CMV fit well to a single-exponential function. In BLMV, Pf was 108 ± 4 μm·s-1 (25 °C) with an Ea of 7.7 kcal/mol; in CMV, Pf was 86 ± 5 μm·s-1 (25 °C) with an Ea of 8.0 kcal/mol. The AQP blocker, dimethyl sulfoxide, significantly inhibited the Pf of both basolateral (81 ± 4 μm·s-1; -25%) and canalicular (59 ± 4 μm·s-1 -30%) membrane vesicles. When CMV were isolated from hepatocytes treated with dibutyryl cAMP, a double-exponential fit was needed, implying two functionally different vesicle populations; one population had Pf and Ea values similar to those of CMV from untreated hepatocytes, but the other population had a very high Pf (655 ± 135 μm·s-1, 25 °C) and very low E a (2.8 kcal/mol). Dimethyl sulfoxide completely inhibited the high Pf value in this second vesicle population. In contrast, P f and Ea of BLMV were unaltered by cAMP treatment of hepatocytes. Our results are consistent with the presence of both lipid- and AQP-mediated pathways for basolateral and canalicular water movement across the hepatocyte plasma membrane barrier. Our data also suggest that the hepatocyte canalicular membrane domain is rate-limiting for transcellular water transport and that this domain becomes more permeable to water when hepatocytes are exposed to a choleretic agonist, presumably by insertion of AQP molecules. These data suggest a molecular mechanism for the efficient coupling of osmotically active solutes and water transport during canalicular bile formation.
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U2 - 10.1074/jbc.M305899200
DO - 10.1074/jbc.M305899200
M3 - Article
C2 - 12939275
AN - SCOPUS:0242384909
SN - 0021-9258
VL - 278
SP - 43157
EP - 43162
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 44
ER -