Use of bioluminescent aequorin for the pharmacological characterization of 5HT receptors.
A convenient functional assay for 5HT2a and 5HT2c receptors is reported utilizing the bioluminescent aequorin to detect intracellular calcium changes. Using this assay, the pharmacological properties of many 5HT ligands can be determined in a 96-well format. The data indicate that the aequorin detection method is superior to the inositol phosphate assay with regard to speed and scope. This system is also appropriate for kinetic studies of receptor desensitization. We showed that the human 5HT2c receptor desensitizes in a biphasic manner, with a fast desensitization of approximately 90% of the total response occurring within 15 minutes while the remaining 10% response remains for at least 3 hours.
Schaeffer
M T
MT
Merck Research Laboratories, Rahway, NJ 07065, USA.
Cully
D
D
Chou
M
M
Liu
J
J
Van der Ploeg
L H
LH
Fong
T M
TM
D000331:Aequorin; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D008024:Ligands; D008163:Luminescent Measurements; D011985:Receptors, Serotonin / Q000032:analysis* / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000378:metabolism / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects
D008024:Ligands; D011985:Receptors, Serotonin; D017366:Serotonin Receptor Agonists; D012701:Serotonin; D000331:Aequorin; D002118:Calcium