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Comparison of FcRn- and pIgR-Mediated Transport in MDCK Cells by Fluorescence Confocal Microscopy

Jerdeva, Galina V. and Tesar, Devin B. and Huey-Tubman, Kathryn E. and Ladinsky, Mark S. and Fraser, Scott E. and Bjorkman, Pamela J. (2010) Comparison of FcRn- and pIgR-Mediated Transport in MDCK Cells by Fluorescence Confocal Microscopy. Traffic, 11 (9). pp. 1205-1220. ISSN 1398-9219. PMCID PMC2975666. http://resolver.caltech.edu/CaltechAUTHORS:20100830-141021105

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Abstract

Protein delivery across polarized epithelia is controlled by receptor-mediated transcytosis. Many studies have examined basolateral-to-apical trafficking of polymeric IgA (pIgA) by the polymeric immunoglobulin receptor (pIgR). Less is known about apical-to-basolateral transcytosis, the direction the neonatal Fc receptor (FcRn) transports maternal IgGs across intestinal epithelia. To compare apical-to-basolateral and basolateral-to-apical transcytosis, we co-expressed FcRn and pIgR in Madin-Darby canine kidney (MDCK) cells and used pulse-chase experiments with confocal microscopy to examine transport of apically applied IgG Fcγ and basolaterally applied pIgA. Fcγ and pIgA trafficking routes were initially separate but intermixed at later chase times. Fcγ was first localized near the apical surface, but became more equally distributed across the cell, consistent with concomitant transcytosis and recycling. By contrast, pIgA transport was strongly unidirectional: pIgA shifted from near the basolateral surface to an apical location with increasing time. Some Fcγ and pIgA fluorescence colocalized in early (EEA1-positive), recycling (Rab11a-positive), and transferrin (Tf)-positive common/basolateral recycling endosomes. Fcγ became more enriched in Tf-positive endosomes with time, whereas pIgA was sorted from these compartments. Live-cell imaging revealed that vesicles containing Fcγ or pIgA shared similar mobility characteristics and were equivalently affected by depolymerizing microtubules, indicating that both trafficking routes depended to roughly the same extent on intact microtubules.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1111/j.1600-0854.2010.01083.xDOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2975666/PubMed CentralArticle
ORCID:
AuthorORCID
Fraser, Scott E.0000-0002-5739-4026
Bjorkman, Pamela J.0000-0002-2277-3990
Additional Information:© 2010 John Wiley & Sons. Received 10 February 2010, revised and accepted for publication 24 May 2010, uncorrected manuscript published online 26 May 2010, published online 29 June 2010. We thank Marta Murphy for the help in making figures, Enrique Rodriguez-Boulan for the AC17 antibody, Keith Mostov for the MDCK II cells, Adrian Rice for unpublished ferroportin data, and Adrian Rice and members of the Bjorkman lab for critical reading of the manuscript. This work was supported by the National Institutes of Health (2 R37 AI041239-06A1 to P.J.B.).
Funders:
Funding AgencyGrant Number
NIH2 R37 AI041239-06A1
Subject Keywords:apical; basolateral; FcRn; live-cell imaging; Madin-Darby canine kidney (MDCK) cells; pIgR; pulse-chase; transcytosis
PubMed Central ID:PMC2975666
Record Number:CaltechAUTHORS:20100830-141021105
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20100830-141021105
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:19718
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Sep 2010 19:57
Last Modified:01 Jan 2016 02:40

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