Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (OnlineFirst PDF)
Right arrow All Versions of this Article:
1087057105274896v1
10/5/476    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Ghosh, R. N.
Right arrow Articles by Oakley, R. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ghosh, R. N.
Right arrow Articles by Oakley, R. H.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Article

Quantitative Cell-Based High-Content Screening for Vasopressin Receptor Agonists Using Transfluor® Technology

Richik N. Ghosh1*, Richard Debiasio1, Christine C. Hudson2, Everett R. Ramer1, Conrad L. Cowan2, Robert H. Oakley2

1 Cellomics, Inc., Pittsburgh, PA.
2 Xsira Pharmaceuticals, Inc. (formerly Norak Biosciences), Morrisville, NC.

* To whom correspondence should be addressed. E-mail: rghosh{at}cellomics.com.


  Abstract

The authors demonstrate the use of a simple, universal G-protein-coupled receptor (GPCR) assay to screen for agonists for a specific GPCR. Cells stably expressing a green fluorescent protein (GFP)--labeled {beta}-arrestin fusion protein and the vasopressin V2 receptor (V2R) were used in a high-content screening (HCS) assay to screen a small peptide library for V2R agonists. Cells were treated with the peptides at a final concentration of 500 nM for 30 min. Agonist stimulation causes V2R internalization into endosomes. GFP-{beta}-arrestin remains associated with the V2R in endosomes, resulting in a fluorescent pattern of intracellular spots. Assay plates were automatically imaged and quantitatively analyzed using an HCS imaging platform and a fast turnkey image analysis application optimized for detection of receptor activation and intracellular spots. Hits were further evaluated to determine their potency. The combination of unique biology, automated high-content analysis, and a powerful means of validating hits results in better leads.

Key Words: high-content screening, cell-based assays, vasopressin, {beta}-arrestin, G-protein-coupled receptors, Transfluor®

First published on August 29, 2005, doi:10.1177/1087057105274896

Journal of Biomolecular Screening 2005;10:476.

A more recent version of this article appeared on August 1, 2005

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:


Home page
 J Biomol ScreenHome page
M. Vrecl, P. K. Norregaard, D. L.C. Almholt, L. Elster, A. Pogacnik, and A. Heding
{beta}-Arrestin-Based Bret2 Screening Assay for the "Non"-{beta}-Arrestin Binding CB1 Receptor
J Biomol Screen, April 1, 2009; 14(4): 371 - 380.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
C. Doucette, K. Vedvik, E. Koepnick, A. Bergsma, B. Thomson, and T. C. Turek-Etienne
Kappa Opioid Receptor Screen with the TangoTM {beta}-Arrestin Recruitment Technology and Characterization of Hits with Second-Messenger Assays
J Biomol Screen, April 1, 2009; 14(4): 381 - 394.
[Abstract] [PDF]