+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Intravital Multiphoton Microscopy with Fluorescent Bile Salts in Rats as an In Vivo Biomarker for Hepatobiliary Transport Inhibition



Intravital Multiphoton Microscopy with Fluorescent Bile Salts in Rats as an In Vivo Biomarker for Hepatobiliary Transport Inhibition



Drug Metabolism and Disposition: the Biological Fate of Chemicals 46(5): 704-718



The bile salt export pump (BSEP) is expressed at the canalicular domain of hepatocytes, where it mediates the elimination of monovalent bile salts into the bile. Inhibition of BSEP is considered a susceptibility factor for drug-induced liver injury that often goes undetected during nonclinical testing. Although in vitro assays exist for screening BSEP inhibition, a reliable and specific method for confirming Bsep inhibition in vivo would be a valuable follow up to a BSEP screening strategy, helping to put a translatable context around in vitro inhibition data, incorporating processes such as metabolism, protein binding, and other exposure properties that are lacking in most in vitro BSEP models. Here, we describe studies in which methods of quantitative intravital microscopy were used to identify dose-dependent effects of two known BSEP/Bsep inhibitors, 2-[4-[4-(butylcarbamoyl)-2-[(2,4-dichlorophenyl)sulfonylamino]phenoxy]-3-methoxyphenyl]acetic acid (AMG-009) and bosentan, on hepatocellular transport of the fluorescent bile salts cholylglycyl amidofluorescein and cholyl-lysyl-fluorescein in rats. Results of these studies demonstrate that the intravital microscopy approach is capable of detecting Bsep inhibition at drug doses well below those found to increase serum bile acid levels, and also indicate that basolateral efflux transporters play a significant role in preventing cytosolic accumulation of bile acids under conditions of Bsep inhibition in rats. Studies of this kind can both improve our understanding of exposures needed to inhibit Bsep in vivo and provide unique insights into drug effects in ways that can improve our ability interpret animal studies for the prediction of human drug hepatotoxicity.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 065254765

Download citation: RISBibTeXText

PMID: 29467212

DOI: 10.1124/dmd.117.079277


Related references

Using quantitative intravital multiphoton microscopy to dissect hepatic transport in rats. Methods 128: 40-51, 2017

Visualization of hepatobiliary excretory function by intravital multiphoton microscopy. Journal of Biomedical Optics 12(1): 014014, 2007

Hepatic transport of fluorescent molecules: in vivo studies using intravital TV microscopy. Hepatology 6(3): 444-449, 1986

Fluorescent liposomes for intravital staining of Kupffer cells to aid in vivo microscopy in rats. Methods and Findings in Experimental and Clinical Pharmacology 29(5): 321-327, 2007

Visualizing the Behavior of HIV-Infected T Cells In Vivo Using Multiphoton Intravital Microscopy. Methods in Molecular Biology 1354: 189-201, 2016

A Quantitative In-Vivo Multiphoton Microscopy Assay of Hepatic Organic Anion Transport in Rats. Gastroenterology 138(5): S-794-S-795, 2010

Multiphoton microscopy applied for real-time intravital imaging of bacterial infections in vivo. Methods in Enzymology 506: 35-61, 2012

Hepatobiliary transport of monoanionic and dianionic bile salts. Biological Chemistry Hoppe-Seyler 376(SPEC Suppl. ): S66, 1995

Fluorescent derivatives of bile salts. II. Suitability of NBD-amino derivatives of bile salts for the study of biological transport. Journal of Lipid Research 32(11): 1769-1779, 1991

Visualization of the transport of primary and secondary bile acids across liver tissue in rats: In vivo study with fluorescent bile acids. Journal of Hepatology 34(1): 4-10, 2001

The transport of antibiotics, lipids and bile salts by BCRP suggests a role in hepatobiliary secretion. FASEB Journal 18(4-5): Abst 793 5, 2004

New model for in vivo investigation after microvascular breakdown in burns: use of intravital fluorescent microscopy. Burns 31(2): 168-174, 2005

Evaluating Hepatobiliary Transport with 18 F-Labeled Bile Acids: The Effect of Radiolabel Position and Bile Acid Structure on Radiosynthesis and In Vitro and In Vivo Performance. Contrast Media and Molecular Imaging 2018: 6345412, 2018

Cell-based and in vivo spectral analysis of fluorescent proteins for multiphoton microscopy. Journal of Biomedical Optics 17(9): 96001, 2012

Intracellular distribution of fluorescent bile salts Studies with confocal microscopy. Hepatology 18(4 Part 2): 136A, 1993