Cloning, expression and functional analysis of an octopamine receptor from Periplaneta americana.
Octopamine regulates multiple physiological functions in invertebrates. The biological effects of octopamine and the pharmacology of octopamine receptors have been extensively studied in the American cockroach, Periplaneta americana. This paper reports the cloning of the first octopamine receptor from Periplaneta americana. A cDNA encoding a putative 7 transmembrane receptor was isolated from the head of Periplaneta americana. The encoded protein contains 628 amino acids and has sequence similarity to other biogenic amine receptors. This protein was expressed in COS-7 cells for radioligand binding studies using the antagonist 3H-yohimbine. Competitive binding comparing biogenic amines that could potentially function as endogenous ligands demonstrated this receptor had the highest affinity for octopamine (Ki = 13.3 microM) followed by tyramine, dopamine, serotonin and histamine. Octopamine increased both cAMP levels (EC50 = 1.62 microM) and intracellular concentrations of calcium through the receptor expressed in HEK-293 cells. Tyramine increased levels of both of these second messengers but only at significantly higher concentrations than octopamine. The cAMP increase by octopamine was independent of the increase in calcium. Competitive binding with antagonists revealed this receptor is similar to Lym oa1 from Lymnaea stagnalis. The data indicate that this cDNA is the first octopamine receptor cloned from Periplaneta americana and therefore has been named Pa oa1.
Bischof
Larry J
LJ
Department of Biochemistry, Vanderbilt University School of Medicine, 635A Light Hall, Nashville, TN 37232, USA.
Enan
Essam E
EE
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D001679:Biogenic Amines / Q000378:metabolism / Q000494:pharmacology; D019556:COS Cells; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D009655:Octopamine / Q000494:pharmacology; D010526:Periplaneta / Q000235:genetics / Q000378:metabolism*; D011869:Radioligand Assay; D017982:Receptors, Biogenic Amine / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016415:Sequence Alignment; D015016:Yohimbine / Q000378:metabolism / Q000494:pharmacology
D001679:Biogenic Amines; D002614:Chelating Agents; D017982:Receptors, Biogenic Amine; C027604:norsynephrine receptor; D009655:Octopamine; D015016:Yohimbine; D004533:Egtazic Acid; D000242:Cyclic AMP; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium