Unit

UNIT 15.17 Analysis of Polarized Membrane Traffic in Hepatocytes and Hepatic Cell Lines

  1. Julie G. In1,
  2. Gudrun Ihrke2,
  3. Pamela L. Tuma1

Published Online: 1 MAR 2012

DOI: 10.1002/0471143030.cb1517s54

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

In, J. G., Ihrke, G. and Tuma, P. L. 2012. Analysis of Polarized Membrane Traffic in Hepatocytes and Hepatic Cell Lines. Current Protocols in Cell Biology. 54:15.17:15.17.1–15.17.19.

Author Information

  1. 1

    Department of Biology, The Catholic University of America, Washington, DC

  2. 2

    Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAR 2012
  2. Published Print: MAR 2012

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. Curr. Protoc. Cell Biol. 54:15.17.1-15.17.19. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • hepatocytes;
  • apical;
  • basolateral;
  • transcytosis;
  • membrane;
  • traffic