Dataset Information

Regulation of mechanosensitive biliary epithelial transport by the epithelial Na(+) channel.

ABSTRACT: Intrahepatic biliary epithelial cells (BECs), also known as cholangiocytes, modulate the volume and composition of bile through the regulation of secretion and absorption. While mechanosensitive Cl(-) efflux has been identified as an important secretory pathway, the counterabsorptive pathways have not been identified. In other epithelial cells, the epithelial Na(+) channel (ENaC) has been identified as an important contributor to fluid absorption; however, its expression and function in BECs have not been previously studied. Our studies revealed the presence of ?, ?, and ? ENaC subunits in human BECs and ? and ? subunits in mouse BECs. In studies of confluent mouse BEC monolayers, the ENaC contributes to the volume of surface fluid at the apical membrane during constitutive conditions. Further, functional studies using whole-cell patch clamp of single BECs demonstrated small constitutive Na(+) currents, which increased significantly in response to fluid-flow or shear. The magnitude of Na(+) currents was proportional to the shear force, displayed inward rectification and a reversal potential of +40 mV (ENa+ ?=?+60 mV), and were abolished with removal of extracellular Na(+) (N-methyl-d-glucamine) or in the presence of amiloride. Transfection with ENaC? small interfering RNA significantly inhibited flow-stimulated Na(+) currents, while overexpression of the ? subunit significantly increased currents. ENaC-mediated currents were positively regulated by proteases and negatively regulated by extracellular adenosine triphosphate.These studies represent the initial characterization of mechanosensitive Na(+) currents activated by flow in biliary epithelium; understanding the role of mechanosensitive transport pathways may provide strategies to modulate the volume and composition of bile during cholestatic conditions. (Hepatology 2016;63:538-549).

SUBMITTER: Li Q

PROVIDER: S-EPMC4780683 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Regulation of mechanosensitive biliary epithelial transport by the epithelial Na(+) channel.

Li Qin Q Kresge Charles C Bugde Abhijit A Lamphere Michelle M Park Jason Y JY Feranchak Andrew P AP

Hepatology (Baltimore, Md.) 20151214 2

<h4>Unlabelled</h4>Intrahepatic biliary epithelial cells (BECs), also known as cholangiocytes, modulate the volume and composition of bile through the regulation of secretion and absorption. While mechanosensitive Cl(-) efflux has been identified as an important secretory pathway, the counterabsorptive pathways have not been identified. In other epithelial cells, the epithelial Na(+) channel (ENaC) has been identified as an important contributor to fluid absorption; however, its expression and f ...[more]

PMID: 26475057

Similar Datasets

Project description:Mutations that lead to muscular dystrophy often create deficiencies in cytoskeletal support of the muscle sarcolemma causing hyperactive mechanosensitive cation channel (MSC) activity and elevated intracellular Ca(2+). Caveolae are cholesterol-rich microdomains that form mechanically deformable invaginations of the sarcolemma. Mutations to caveolin-3, the main scaffolding protein of caveolae in muscle, cause Limbe-Girdle muscular dystrophy. Using genetic and acute chemical perturbations of developing myotubes we investigated whether caveolae are functionally linked to MSCs. MSC sensitivity was assayed using suction application to patches and probe-induced indentation during whole-cell recordings. Membrane mechanical stress in patches was monitored using patch capacitance/impedance. Cholesterol depletion disrupted caveolae and caused a large increase in MSC current. It also decreased the membrane mechanical relaxation time, likely reflecting cytoskeleton dissociation from the bilayer. Reduction of Cav3 expression with miRNA also increased MSC current and decreased patch relaxation time. In contrast Cav3 overexpression produced a small decrease in MSC currents. To acutely and specifically inhibit Cav3 interactions, we made a chimeric peptide containing the antennapedia membrane translocation domain and the Cav3 scaffolding domain (A-CSD3). A-CSD3 action was time dependent initially producing a mild Ca(2+) leak and increased MSC current, while longer exposures decreased MSC currents coinciding with increased patch stiffening. Images of GFP labeled Cav3 in patches showed that Cav3 doesn't enter the pipette, showing patch composition differed from the cell surface. However, disruption via cholesterol depletion caused Cav3 to become uniformly distributed over the sarcolemma and Cav3 appearance in the patch dome. The whole-cell indentation currents elicited under the different caveolae modifying conditions mirror the patch response supporting the role of caveolae in MSC function. These studies show that normal expression levels of Cav3 are mechanoprotective to the sarcolemma through multiple mechanisms, and Cav3 upregulation observed in some dystrophies may compensate for other mechanical deficiencies.

Dataset Information

Regulation of mechanosensitive biliary epithelial transport by the epithelial Na(+) channel.

Publications

Regulation of mechanosensitive biliary epithelial transport by the epithelial Na(+) channel.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets