A rapid quantitative bioassay for evaluating the effects of ligands upon receptors that modulate cAMP levels in a melanophore cell line.
A new method for rapidly evaluating the effects of drugs on receptors that regulate intracellular cAMP in a cell line derived from Xenopus laevis melanophores has been developed. Melanophores were plated into sterile 96 well microtiter plates, and 3 days later the cells were treated with melatonin for 30 min to induce melanosome aggregation. Subsequent exposure to MSH or adrenergic agonists caused dose dependent pigment dispersion that peaked within 30 min. The cumulative pigment displacement from cells could be quantitated by using a microplate reader to measure changes in transmittance of light through the wells. The acquired data enabled detailed and reproducible dose response curves and time course analyses to be generated. In addition, the assay followed for the rapid characterization of the effects of antagonists upon the beta adrenergic receptor (beta AR). The assay has the potential to test the effects of ligands upon any receptor capable of mediating pigment translocation in the melanophore cell line.
Potenza
M N
MN
Department of Cell Biology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Lerner
M R
MR
D000818:Animals; D001681:Biological Assay; D000242:Cyclic AMP / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D009074:Melanocyte-Stimulating Hormones / Q000494:pharmacology; D008547:Melanophores / Q000187:drug effects / Q000378:metabolism*; D008550:Melatonin / Q000494:pharmacology; D009638:Norepinephrine / Q000494:pharmacology; D010860:Pigments, Biological / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000187:drug effects* / Q000502:physiology; D011979:Receptors, Pituitary Hormone / Q000187:drug effects* / Q000502:physiology; D014982:Xenopus laevis
D010860:Pigments, Biological; D011943:Receptors, Adrenergic, beta; D011979:Receptors, Pituitary Hormone; C029446:MSH receptor; D009074:Melanocyte-Stimulating Hormones; D000242:Cyclic AMP; D008550:Melatonin; D009638:Norepinephrine