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dc.contributor.authorTsukamoto, Kosuke
dc.contributor.authorKocher, Olivier
dc.contributor.authorKrieger, Monty
dc.contributor.authorBuck, Lorenna Dianne
dc.contributor.authorGriffith, Linda G.
dc.contributor.authorInman, Samuel Walker
dc.date.accessioned2013-09-30T16:27:03Z
dc.date.available2013-09-30T16:27:03Z
dc.date.issued2013-07
dc.date.submitted2013-01
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/1721.1/81242
dc.description.abstractBackground: PDZK1 is a four PDZ-domain containing cytoplasmic protein that binds to a variety of membrane proteins via their C-termini and can influence the abundance, localization and/or function of its target proteins. One of these targets in hepatocytes in vivo is the HDL receptor SR-BI. Normal hepatic expression of SR-BI protein requires PDZK1 - <5% of normal hepatic SR-BI is seen in the livers of PDZK1 knockout mice. Progress has been made in identifying features of PDZK1 required to control hepatic SR-BI in vivo using hepatic expression of wild-type and mutant forms of PDZK1 in wild-type and PDZK1 KO transgenic mice. Such in vivo studies are time consuming and expensive, and cannot readily be used to explore many features of the underlying molecular and cellular mechanisms. Methodology/Principal Findings: Here we have explored the potential to use either primary rodent hepatocytes in culture using 2D collagen gels with newly developed optimized conditions or PDZK1/SR-BI co-transfected cultured cell lines (COS, HEK293) for such studies. SR-BI and PDZK1 protein and mRNA expression levels fell rapidly in primary hepatocyte cultures, indicating this system does not adequately mimic hepatocytes in vivo for analysis of the PDZK1 dependence of SR-BI. Although PDZK1 did alter SR-BI protein expression in the cell lines, its influence was independent of SR-BI’s C-terminus, and thus is not likely to occur via the same mechanism as that which occurs in hepatocytes in vivo. Conclusions/Significance: Caution must be exercised in using primary hepatocytes or cultured cell lines when studying the mechanism underlying the regulation of hepatic SR-BI by PDZK1. It may be possible to use SR-BI and PDZK1 expression as sensitive markers for the in vivo-like state of hepatocytes to further improve primary hepatocyte cell culture conditions.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant HL052212)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant HL066105)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant ES015241)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant GM068762)en_US
dc.language.isoen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofhttp://dx.doi.org/10.1371/journal.pone.0069725en_US
dc.rightsCreative Commons Attributionen_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.5/en_US
dc.sourcePLoSen_US
dc.titleChallenges in Using Cultured Primary Rodent Hepatocytes or Cell Lines to Study Hepatic HDL Receptor SR-BI Regulation by Its Cytoplasmic Adaptor PDZK1en_US
dc.typeArticleen_US
dc.identifier.citationTsukamoto, Kosuke, Lorenna Buck, Walker Inman, Linda Griffith, Olivier Kocher, and Monty Krieger. “Challenges in Using Cultured Primary Rodent Hepatocytes or Cell Lines to Study Hepatic HDL Receptor SR-BI Regulation by Its Cytoplasmic Adaptor PDZK1.” Edited by Matias A. Avila. PLoS ONE 8, no. 7 (July 23, 2013): e69725.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biologyen_US
dc.contributor.mitauthorTsukamoto, Kosukeen_US
dc.contributor.mitauthorKrieger, Montyen_US
dc.contributor.mitauthorBuck, Lorenna Dianneen_US
dc.contributor.mitauthorGriffith, Linda G.en_US
dc.contributor.mitauthorInman, Samuel Walkeren_US
dc.relation.journalPLoS ONEen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsTsukamoto, Kosuke; Buck, Lorenna; Inman, Walker; Griffith, Linda; Kocher, Olivier; Krieger, Montyen_US
dc.identifier.orcidhttps://orcid.org/0000-0003-4541-5181
dc.identifier.orcidhttps://orcid.org/0000-0002-1801-5548
mit.licensePUBLISHER_CCen_US


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