Arachidonic acid release in cell lines transfected with muscarinic receptors: a simple functional assay to determine response of agonists.
Muscarinic agonists stimulated arachidonic acid release from 10- to 32-fold in Chinese hamster ovary (CHO) cells transfected with muscarinic M1, M3 and M5 receptor subtypes. Muscarinic agonists liberated arachidonic acid from the cAMP-coupled M2 and M4 cells only in the presence of ATP. Partial agonists were less efficacious at liberating arachidonic acid than full agonists. The ability of muscarinic agonists to liberate arachidonic acid and stimulate phosphoinositide hydrolysis in the same CHO M1, M3 and M5 cells was well correlated; however, partial agonists were more efficacious at stimulating phosphoinositide hydrolysis than arachidonic acid release. The efficacy and potency of 13 muscarinic agonists to liberate arachidonic acid was characterised. Influx of external calcium was required for arachidonic acid release even after initiation of agonist-induced release. It is concluded that arachidonic acid release is a simple assay suitable for evaluation of muscarinic agonists, antagonists and the flux of external calcium into cells.
Bymaster
F P
FP
Lilly Neuroscience Research, Lilly Corporate Center, Indianapolis, IN 46285, USA. f.bymaster@lilly.com
Calligaro
D O
DO
Falcone
J F
JF
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D016466:CHO Cells; D002118:Calcium / Q000502:physiology; D002614:Chelating Agents / Q000494:pharmacology; D006224:Cricetinae; D004533:Egtazic Acid / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D018721:Muscarinic Agonists / Q000378:metabolism* / Q000494:pharmacology; D018727:Muscarinic Antagonists / Q000494:pharmacology; D010095:Oxotremorine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D010890:Pirenzepine / Q000494:pharmacology; D011976:Receptors, Muscarinic / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D002614:Chelating Agents; D018721:Muscarinic Agonists; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D016718:Arachidonic Acid; D010890:Pirenzepine; D004533:Egtazic Acid; D010095:Oxotremorine; C042743:oxotremorine M; D002118:Calcium