Analysis of polarized membrane traffic in hepatocytes and hepatic cell lines

Julie G. In, Gudrun Ihrke, Pamela L. Tuma

Research output: Contribution to journalArticlepeer-review

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 preferredmethod 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.

Original languageEnglish (US)
Article number15.17
JournalCurrent Protocols in Cell Biology
Issue numberSUPPL.54
DOIs
StatePublished - Mar 2012

Keywords

  • Apical
  • Basolateral
  • Hepatocytes
  • Membrane
  • Traffic
  • Transcytosis

ASJC Scopus subject areas

  • Cell Biology

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