Analysis of Polarized Membrane Traffic in Hepatocytes and Hepatic Cell Lines

Overview

Journal Curr Protoc Cell Biol

Specialty Cell Biology

Date 2012 Mar 17

PMID 22422475

Citations 5

Authors

Julie G In

Gudrun Ihrke

Pamela L Tuma

Affiliations

Soon will be listed here.

Abstract

The protocols described in this unit were developed to monitor membrane traffic in cultured cell monolayers that display hepatic polarity. In general, the assays are designed to visualize and/or quantitate membrane trafficking by monitoring the fates of antibodies bound to specific membrane proteins. We first describe how to infect cells with recombinant adenovirus, the preferred method for introducing exogenous genes into hepatic cells. We next provide a morphological assay to monitor basolateral to apical transcytosis. In a supporting protocol, we describe how to visualize apical recycling and/or retention. In an additional supporting protocol, we provide a semi-quantitative method to measure the relative extents of apical delivery. Finally, we describe quantitative assays to measure basolateral internalization and recycling. The methods presented in this unit provide a relatively simple, yet powerful approach to examining hepatic membrane traffic.

Citing Articles

Low Rho activity in hepatocytes prevents apical from basolateral cargo separation during trans-Golgi network to surface transport.

Lazaro-Dieguez F, Musch A Traffic. 2020; 21(5):364-374.

PMID: 32124512 PMC: 7959587. DOI: 10.1111/tra.12725.

Rab17 regulates apical delivery of hepatic transcytotic vesicles.

Striz A, Stephan A, Lopez-Coral A, Tuma P Mol Biol Cell. 2018; 29(23):2887-2897.

PMID: 30256711 PMC: 6249867. DOI: 10.1091/mbc.E18-07-0433.

A Serine/Threonine Kinase 16-Based Phospho-Proteomics Screen Identifies WD Repeat Protein-1 As A Regulator Of Constitutive Secretion.

Lopez-Coral A, Striz A, Tuma P Sci Rep. 2018; 8(1):13049.

PMID: 30158666 PMC: 6115458. DOI: 10.1038/s41598-018-31426-1.

Serine/threonine kinase 16 and MAL2 regulate constitutive secretion of soluble cargo in hepatic cells.

In J, Striz A, Bernad A, Tuma P Biochem J. 2014; 463(2):201-13.

PMID: 25084525 PMC: 4736628. DOI: 10.1042/BJ20140468.

Hepatocyte polarity.

Treyer A, Musch A Compr Physiol. 2013; 3(1):243-87.

PMID: 23720287 PMC: 3697931. DOI: 10.1002/cphy.c120009.

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