Molecular cloning and heterologous expression of an alpha-adrenergic-like octopamine receptor from the silkworm Bombyx mori.
A cDNA encoding an octopamine (OA) receptor (BmOAR1) was isolated from the nerve tissue of silkworm (Bombyx mori) larvae. Comparison of amino acid sequences showed that BmOAR1 is highly identical to OA receptors isolated from Periplaneta americana (Pa oa(1)), Apis mellifera (AmOA1), and Drosophila melanogaster (OAMB or DmOA1A). BmOAR1 was stably expressed in HEK-293 cells. OA above 1 microM led to an increase in intracellular cyclic AMP concentration ([cAMP](i)). The synthetic OA-receptor agonist demethylchlordimeform also elevated [cAMP](i) to the same maximal level (approximately 5-fold over the basal level) as that induced by OA. However, other biogenic amines, tyramine and dopamine, and chlordimeform were without effects. The [cAMP](i) level raised by OA was lowered by antagonists; the rank order of antagonist activity was chlorpromazine > mianserin = yohimbine. Cyproheptadine and metoclopramide had little effect. OA above 100 nM induced a transient or sustained increase in intracellular Ca(2+) concentration ([Ca(2+)](i)), depending on the concentration of OA. Sequence homology and functional analysis data indicate that BmOAR1 is an alpha-adrenergic-like OA receptor of B. mori.
Ohtani
A
A
Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Shimane, Japan.
Arai
Y
Y
Ozoe
F
F
Ohta
H
H
Narusuye
K
K
Huang
J
J
Enomoto
K
K
Kataoka
H
H
Hirota
A
A
Ozoe
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000737:chemistry / Q000235:genetics*; D020013:Calcium Signaling / Q000187:drug effects; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000096:biosynthesis; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D049750:Genome, Insect / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009655:Octopamine / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011941:Receptors, Adrenergic / Q000737:chemistry*; D017982:Receptors, Biogenic Amine / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014316:Tritium; D015016:Yohimbine / Q000378:metabolism / Q000494:pharmacology
D018076:DNA, Complementary; D012333:RNA, Messenger; D011941:Receptors, Adrenergic; D017982:Receptors, Biogenic Amine; C027604:norsynephrine receptor; D014316:Tritium; D009655:Octopamine; D015016:Yohimbine; D000242:Cyclic AMP