AQP4 Transfected into Mouse Cholangiocytes Promotes Water Transport in Biliary Epithelia

Patrick L. Splinter, Anatoliy I. Masyuk, Raul A. Marinelli, Nicholas F La Russo

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Rodent cholangiocytes express 6 of the 11 known channel proteins called aquaporins (AQPs) that are involved in transcellular water transport in mammals. However, clarifying the role of AQPs in mediating water transport in biliary epithelia has been limited in part because of the absence of physiologically relevant experimental models. In this study, we established a novel AQP4-transfected polarized mouse cholangiocyte cell line suitable for functional studies of transepithelial water transport, and, using this model, we define the importance of this AQP in water transport across biliary epithelia. Polarized normal mouse cholangiocytes (NMCs) lacking endogenous AQP4 were transfected stably with functional AQP4 or cotransfected with functional AQP4 and a transport-deficient AQP4 dominant negative mutant using a retroviral delivery system. In transfected NMCs, AQP4 is expressed on both the mRNA and protein levels and is localized at both the apical and basolateral membranes. In nontransfected NMCs, the transcellular water flow, Pf, value was relatively high (i.e., 16.4 ± 3.2 μm/sec) and likely was a reflection of endogenous expression of AQP1 and AQP8. In NMCs transfected with AQP4, P f increased to 75.7 ± 1.4 μm/sec, that is, by 4.6-fold, indicating the contribution of AQP4 in channel-mediated water transport across MNCs monolayer. In cotransfected NMCs, AQP4 dominant negative reduced P f twofold; no changes in Pf were observed in NMCs transfected with the empty vector. In conclusion, we developed a novel polarized mouse cholangiocyte monolayer model, allowing direct study of AQP4-mediated water transport by biliary epithelia and generated data providing additional support for the importance of AQP4 in cholangiocyte water transport.

Original languageEnglish (US)
Pages (from-to)109-116
Number of pages8
JournalHepatology
Volume39
Issue number1
DOIs
StatePublished - Jan 2004

Fingerprint

Epithelium
Aquaporins
Water
Transcytosis
Mammals
Rodentia
Proteins
Theoretical Models
Cell Line
Messenger RNA
Membranes

ASJC Scopus subject areas

  • Hepatology

Cite this

AQP4 Transfected into Mouse Cholangiocytes Promotes Water Transport in Biliary Epithelia. / Splinter, Patrick L.; Masyuk, Anatoliy I.; Marinelli, Raul A.; La Russo, Nicholas F.

In: Hepatology, Vol. 39, No. 1, 01.2004, p. 109-116.

Research output: Contribution to journalArticle

Splinter, Patrick L. ; Masyuk, Anatoliy I. ; Marinelli, Raul A. ; La Russo, Nicholas F. / AQP4 Transfected into Mouse Cholangiocytes Promotes Water Transport in Biliary Epithelia. In: Hepatology. 2004 ; Vol. 39, No. 1. pp. 109-116.
@article{715053bc8dfd41df9f1b1bff8b78911a,
title = "AQP4 Transfected into Mouse Cholangiocytes Promotes Water Transport in Biliary Epithelia",
abstract = "Rodent cholangiocytes express 6 of the 11 known channel proteins called aquaporins (AQPs) that are involved in transcellular water transport in mammals. However, clarifying the role of AQPs in mediating water transport in biliary epithelia has been limited in part because of the absence of physiologically relevant experimental models. In this study, we established a novel AQP4-transfected polarized mouse cholangiocyte cell line suitable for functional studies of transepithelial water transport, and, using this model, we define the importance of this AQP in water transport across biliary epithelia. Polarized normal mouse cholangiocytes (NMCs) lacking endogenous AQP4 were transfected stably with functional AQP4 or cotransfected with functional AQP4 and a transport-deficient AQP4 dominant negative mutant using a retroviral delivery system. In transfected NMCs, AQP4 is expressed on both the mRNA and protein levels and is localized at both the apical and basolateral membranes. In nontransfected NMCs, the transcellular water flow, Pf, value was relatively high (i.e., 16.4 ± 3.2 μm/sec) and likely was a reflection of endogenous expression of AQP1 and AQP8. In NMCs transfected with AQP4, P f increased to 75.7 ± 1.4 μm/sec, that is, by 4.6-fold, indicating the contribution of AQP4 in channel-mediated water transport across MNCs monolayer. In cotransfected NMCs, AQP4 dominant negative reduced P f twofold; no changes in Pf were observed in NMCs transfected with the empty vector. In conclusion, we developed a novel polarized mouse cholangiocyte monolayer model, allowing direct study of AQP4-mediated water transport by biliary epithelia and generated data providing additional support for the importance of AQP4 in cholangiocyte water transport.",
author = "Splinter, {Patrick L.} and Masyuk, {Anatoliy I.} and Marinelli, {Raul A.} and {La Russo}, {Nicholas F}",
year = "2004",
month = "1",
doi = "10.1002/hep.20033",
language = "English (US)",
volume = "39",
pages = "109--116",
journal = "Hepatology",
issn = "0270-9139",
publisher = "John Wiley and Sons Ltd",
number = "1",

}

TY - JOUR

T1 - AQP4 Transfected into Mouse Cholangiocytes Promotes Water Transport in Biliary Epithelia

AU - Splinter, Patrick L.

AU - Masyuk, Anatoliy I.

AU - Marinelli, Raul A.

AU - La Russo, Nicholas F

PY - 2004/1

Y1 - 2004/1

N2 - Rodent cholangiocytes express 6 of the 11 known channel proteins called aquaporins (AQPs) that are involved in transcellular water transport in mammals. However, clarifying the role of AQPs in mediating water transport in biliary epithelia has been limited in part because of the absence of physiologically relevant experimental models. In this study, we established a novel AQP4-transfected polarized mouse cholangiocyte cell line suitable for functional studies of transepithelial water transport, and, using this model, we define the importance of this AQP in water transport across biliary epithelia. Polarized normal mouse cholangiocytes (NMCs) lacking endogenous AQP4 were transfected stably with functional AQP4 or cotransfected with functional AQP4 and a transport-deficient AQP4 dominant negative mutant using a retroviral delivery system. In transfected NMCs, AQP4 is expressed on both the mRNA and protein levels and is localized at both the apical and basolateral membranes. In nontransfected NMCs, the transcellular water flow, Pf, value was relatively high (i.e., 16.4 ± 3.2 μm/sec) and likely was a reflection of endogenous expression of AQP1 and AQP8. In NMCs transfected with AQP4, P f increased to 75.7 ± 1.4 μm/sec, that is, by 4.6-fold, indicating the contribution of AQP4 in channel-mediated water transport across MNCs monolayer. In cotransfected NMCs, AQP4 dominant negative reduced P f twofold; no changes in Pf were observed in NMCs transfected with the empty vector. In conclusion, we developed a novel polarized mouse cholangiocyte monolayer model, allowing direct study of AQP4-mediated water transport by biliary epithelia and generated data providing additional support for the importance of AQP4 in cholangiocyte water transport.

AB - Rodent cholangiocytes express 6 of the 11 known channel proteins called aquaporins (AQPs) that are involved in transcellular water transport in mammals. However, clarifying the role of AQPs in mediating water transport in biliary epithelia has been limited in part because of the absence of physiologically relevant experimental models. In this study, we established a novel AQP4-transfected polarized mouse cholangiocyte cell line suitable for functional studies of transepithelial water transport, and, using this model, we define the importance of this AQP in water transport across biliary epithelia. Polarized normal mouse cholangiocytes (NMCs) lacking endogenous AQP4 were transfected stably with functional AQP4 or cotransfected with functional AQP4 and a transport-deficient AQP4 dominant negative mutant using a retroviral delivery system. In transfected NMCs, AQP4 is expressed on both the mRNA and protein levels and is localized at both the apical and basolateral membranes. In nontransfected NMCs, the transcellular water flow, Pf, value was relatively high (i.e., 16.4 ± 3.2 μm/sec) and likely was a reflection of endogenous expression of AQP1 and AQP8. In NMCs transfected with AQP4, P f increased to 75.7 ± 1.4 μm/sec, that is, by 4.6-fold, indicating the contribution of AQP4 in channel-mediated water transport across MNCs monolayer. In cotransfected NMCs, AQP4 dominant negative reduced P f twofold; no changes in Pf were observed in NMCs transfected with the empty vector. In conclusion, we developed a novel polarized mouse cholangiocyte monolayer model, allowing direct study of AQP4-mediated water transport by biliary epithelia and generated data providing additional support for the importance of AQP4 in cholangiocyte water transport.

UR - http://www.scopus.com/inward/record.url?scp=1442331010&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1442331010&partnerID=8YFLogxK

U2 - 10.1002/hep.20033

DO - 10.1002/hep.20033

M3 - Article

C2 - 14752829

AN - SCOPUS:1442331010

VL - 39

SP - 109

EP - 116

JO - Hepatology

JF - Hepatology

SN - 0270-9139

IS - 1

ER -