Tl. Messier et al., HIGH-THROUGHPUT ASSAYS OF CLONED ADRENERGIC, MUSCARINIC, NEUROKININ, AND NEUROTROPHIN RECEPTORS IN LIVING MAMMALIAN-CELLS, Pharmacology & toxicology, 76(5), 1995, pp. 308-311
Many receptors stimulate proliferation of NIH 3T3 cells in a ligand de
pendent fashion. Based on this observation, we developed a high throug
hput assay of cloned receptor pharmacology. In this assay, receptors a
re transiently co-expressed with the marker enzyme beta-galactosidase.
Receptors that induce cellular proliferation select and amplify the c
ells that also express the marker, thus the ability of ligands to alte
r receptor activity are reported as changes in enzyme activity In the
present study, we used this assay to evaluate the ability of agonist l
igands to stimulate four cloned receptors. The agonists phenylephrine,
carbachol, substance P and nerve growth Factor selectively stimulated
cells transfected with the alpha-1b adrenergic, m4 muscarinic, NK1 ne
urokinin and trkA neurotrophin receptors, respectively. These data dem
onstrate that a high throughput colorimetric assay performed in 96 wel
l plates can be used to evaluate the pharmacology of ligands for clone
d receptors belonging to a wide range of functional and pharmacologica
l classes.