Use of a semi-automated, robotic radioimmunoassay to measure cAMP generated by activation of DP-, EP2-, and IP-prostaglandin receptors in human ocular and other cell types

The aim of these studies was to compare the effects of several prostaglandin agonists on adenylyl cyclase activity in embryonic bovine tracheal (EBTr) cells, transformed human nonpigmented ciliary epithelial (NPE) cells and National Cancer Bank (NCB-20) cells. These cell types have been shown to exp...

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Bibliographic Details
Published in:Prostaglandins, leukotrienes and essential fatty acids leukotrienes and essential fatty acids, 1998-07, Vol.59 (1), p.77-82
Main Authors: CRIDER, J. Y, GRIFFIN, B. W, XU, S. X, SHARIF, N. A
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cattle
Cell receptors
Cell structures and functions
Cells, Cultured
Cyclic AMP - analysis
Epoprostenol - metabolism
Fundamental and applied biological sciences. Psychology
Ganglia, Sympathetic - cytology
Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors
Humans
Hybridomas - pathology
Iodine Radioisotopes
Mice
Molecular and cellular biology
Neuroblastoma - pathology
Prostaglandin D2 - metabolism
Radioimmunoassay
Receptors, Epoprostenol
Receptors, Immunologic
Receptors, Prostaglandin - metabolism
Receptors, Prostaglandin E - metabolism
Robotics - methods
Trachea - cytology
Trachea - embryology
Tumor Cells, Cultured
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Summary:The aim of these studies was to compare the effects of several prostaglandin agonists on adenylyl cyclase activity in embryonic bovine tracheal (EBTr) cells, transformed human nonpigmented ciliary epithelial (NPE) cells and National Cancer Bank (NCB-20) cells. These cell types have been shown to express DP, EP2 and IP prostaglandin (PG) receptors, respectively. Cyclic AMP (cAMP) generation was measured by manual and semi-automated radioimmunoassay (RIA) techniques. ZK118182 (EC50 = 10-27 nM), PGE2 (EC50 = 21-27 nM) and PGI2 (EC50 = 3.5-4 nM) had the highest potency at the DP, EP2 and IP receptors, respectively. A plot of potency (EC50) values generated with both techniques showed a high degree of correlation for all three receptors. These studies provide further characterization of prostanoid receptor functional responses in three cell types and demonstrate the advantages of a semi-automated RIA method for the analysis of the second messenger cAMP.
ISSN:0952-3278
1532-2823
DOI:10.1016/S0952-3278(98)90055-2