Document Detail


alpha2-adrenergic agonist-induced inhibition of cyclic AMP formation in transfected cell lines using a microtiter-based scintillation proximity assay.
MedLine Citation:
PMID:  8903964     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human embryonic kidney cells (HEK-7) were transfected with the gene for the human alpha2C receptor (alpha2C4). Cells were grown in 96-well microtiter plates and cyclic AMP levels were measured by scintillation proximity assay, a modified radioimmunoassay technique. Radioactivity was quantified using a TopCount Scintillation detector. Cyclic AMP was increased in a dose-dependent manner by the addition of exogenous forskolin. The forskolin-induced enhancement of cyclic AMP was inhibited dose-dependently by the addition of alpha2-adrenergic agonists, and this inhibition was blocked by the addition of adrenergic antagonists. The extent of the inhibitory response caused by alpha2-adrenoceptor agonists was related to the receptor density in clonal cell lines derived from the transfected parental HEK-7 cells. By using cells grown in microtiter format, and employing the technological advantages of scintillation proximity assay and TopCount detection, it was possible to simultaneously evaluate the effects of multiple experimental permutations on cellular production of cyclic AMP with minimal disturbance of the cells and minimal and/or automated manipulation of the cyclic AMP formed. This combination of techniques should allow rapid testing of the actions of adrenergic agonists and antagonists on cells transfected with receptors linked to cyclic AMP formation.
Authors:
A A Hancock; A D Vodenlich; C Maldonado; R Janis
Related Documents :
7821704 - Functional expression of a stably transfected parathyroid hormone/parathyroid hormone r...
6205914 - On the mechanism of 'escape' from desensitization of the cyclic amp response to tsh in ...
6253064 - Expression of a cell surface glycoprotein (p180) related to cell-substratum adhesion du...
19184434 - Human mesenchymal stem cells form multicellular structures in response to applied cycli...
6225654 - Levels of filamentous and globular actin in chinese hamster ovary cells throughout the ...
10371414 - Leukocytes as modulators of stellate cell activation.
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of receptor and signal transduction research     Volume:  15     ISSN:  1079-9893     ISO Abbreviation:  J. Recept. Signal Transduct. Res.     Publication Date:    1995 Jan-Mar
Date Detail:
Created Date:  1997-03-12     Completed Date:  1997-03-12     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  9509432     Medline TA:  J Recept Signal Transduct Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  557-79     Citation Subset:  IM    
Affiliation:
Abbott Laboratories, Pharmaceutical Products Division, Abbott Park, IL 60064, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Cell Line
Clone Cells
Clonidine / pharmacology
Cyclic AMP / biosynthesis*
Forskolin / pharmacology
Humans
Norepinephrine / pharmacology
Phentolamine / pharmacology
Radioligand Assay
Receptors, Adrenergic, alpha-2 / agonists*,  genetics,  metabolism
Scintillation Counting / methods*
Transfection
Chemical
Reg. No./Substance:
0/Receptors, Adrenergic, alpha-2; 4205-90-7/Clonidine; 50-60-2/Phentolamine; 51-41-2/Norepinephrine; 60-92-4/Cyclic AMP; 66428-89-5/Forskolin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Characteristics of a membrane-associated steroid binding site in rat liver.
Next Document:  [123I/125I]-Ro 43-0463, a site specific tracer for MAO-B mapping with autoradiography as well as wit...