Document Detail


Heterogeneous Accumulation of Fluorescent Bile Acids in Primary Rat Hepatocytes Does Not Correlate with Their Homogenous Expression of Ntcp.
MedLine Citation:
PMID:  21474652     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Sodium taurocholate co-transporting polypeptide (ntcp) is considered to be a major determinant of bile acid uptake into hepatocytes. However the regulation of ntcp and the degree that it participates in the accumulation of specific substrates is not well understood. We utilized fluorescent bile acid derivatives and direct quantitation of fluorescent microscopy images to examine the regulation of ntcp and its role in the cell-to-cell variability of fluorescent bile acid accumulation. Primary-cultured rat hepatocytes rapidly accumulated the fluorescent bile acids, chenodeoxycholylglycylamidofluorescein (CDCGamF), 7-beta-NBD 3-alpha hydroxy 5-beta cholan-24-oic acid (NBD-CA), and cholyl-glycylamido-fluorescein (CGamF). However in stably-transfected HeLa cells, ntcp preferred CDCGamF, while the organic anion transporter, oatp1a1, preferred NBD-CA, and neither ntcp nor oatp1a1 showed strong accumulation of CGamF by these methods. Ntcp-mediated transport of CDCGamF was inhibited by taurocholate, cyclosporin, actin depolymerization, and an inhibitor of atypical protein kinase C zeta. The latter two agents altered the cellular distribution of ntcp as visualized in ntcp-GFP transfected cells. Although fluorescent bile acid accumulation was reproducible by the imaging assays, individual cells showed variable accumulation that was not due to changes in membrane permeability or cell viability. In HeLa cells this was accounted for by variable levels of ntcp while in hepatocytes, ntcp expression was uniform and low accumulation was seen in a large portion of cells despite the presence of ntcp. These studies indicate that single-cell imaging can provide insight into previously unrecognized details of anion transport in the complex environment of polarized hepatocytes.
Authors:
John William Murray; Amar J Thosani; Pijun Wang; Allan W Wolkoff
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-4-7
Journal Detail:
Title:  American journal of physiology. Gastrointestinal and liver physiology     Volume:  -     ISSN:  1522-1547     ISO Abbreviation:  -     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-4-8     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100901227     Medline TA:  Am J Physiol Gastrointest Liver Physiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
1Albert Einstein College of Medicine.
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