Title
Time-resolved quantitative analysis of CCK1 receptor-induced intracellular calcium increase Time-resolved quantitative analysis of CCK1 receptor-induced intracellular calcium increase
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Publication type
article
Publication
Fayetteville, N.Y. ,
Subject
Chemistry
Biology
Pharmacology. Therapy
Source (journal)
Peptides. - Fayetteville, N.Y.
Volume/pages
34(2012) :1 , p. 219-225
ISSN
0196-9781
ISI
000302851500029
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Abstract
Cholecystokinin (CCK) is a gastrointestinal hormone, which regulates many physiological functions such as satiety by binding to the CCK receptor (CCKR). Molecules, which recognize this receptor can mimic or block CCK signaling and thereby influence CCKR-mediated processes. We have set up a quantitative heterologous assay with CHO cells over-expressing the rat CCK1 receptor to screen for such candidate molecules. Receptor activation, induced by agonist binding, is followed by an intracellular calcium increase, which was monitored using a fluorescent sensor dye. For quantification of the calcium increase, a population average technique using a fluorescence plate reader was optimized and subsequently compared with a single-cell approach using confocal microscopy. With both strategies, dose-response curves were generated for the natural agonist CCK-8S, the partial agonist JMV-180 as well as the antagonist lorglumide. Significant differences were found between the ligands and a strong correspondence was observed between both methods in terms of maximum response and median effect concentrations. Both highly sensitive methods proved complementary: whereas the plate reader assay allowed faster, high throughput screening, the confocal microscopy identified single-cell variations and revealed factors that reduce specificity and sensitivity. (C) 2011 Elsevier Inc. All rights reserved.
E-info
https://repository.uantwerpen.be/docman/iruaauth/9ba93d/1e45958.pdf
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000302851500029&DestLinkType=RelatedRecords&DestApp=ALL_WOS&UsrCustomerID=ef845e08c439e550330acc77c7d2d848
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000302851500029&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=ef845e08c439e550330acc77c7d2d848
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000302851500029&DestLinkType=CitingArticles&DestApp=ALL_WOS&UsrCustomerID=ef845e08c439e550330acc77c7d2d848