title: Isolation and functional characterization of an allatotropin receptor from Manduca sexta.
abstract: Manduca sexta allatotropin (Manse-AT) is a multifunctional neuropeptide whose actions include the stimulation of juvenile hormone biosynthesis, myotropic stimulation, cardioacceleratory functions, and inhibition of active ion transport. Manse-AT is a member of a structurally related peptide family that is widely found in insects and also in other invertebrates. Its precise role depends on the insect species and developmental stage. In some lepidopteran insects including M. sexta, structurally-related AT-like (ATL) peptides can be derived from alternatively spliced mRNAs transcribed from the AT gene. We have isolated a cDNA for an AT receptor (ATR) from M. sexta by a PCR-based approach using the sequence of the ATR from Bombyx mori. The sequence of the M. sexta ATR is similar to several G protein-coupled receptors from other insect species and to the mammalian orexin receptor. We demonstrate that the M. sexta ATR expressed in vertebrate cell lines is activated in a dose-responsive manner by Manse-AT and each Manse-ATL peptide in the rank order ATL-I > ATL-II > ATL-III > AT, and functional analysis in multiple cell lines suggest that the receptor is coupled through elevated levels of Ca(2+) and cAMP. In feeding larvae, Manse-ATR mRNA is present at highest levels in the Malpighian tubules, followed by the midgut, hindgut, testes, and corpora allata, consistent with its action on multiple target tissues. In the adult corpora cardiaca--corpora allata complex, Manse-ATR mRNA is present at relatively low levels in both sexes.
authors.affiliation: Department of Biomedical Sciences, College of Osteopathic Medicine, Ohio University, Athens, OH, USA. horodysk@ohio.edu
authors.forename: Frank M
authors.initials: FM
authors.lastname: Horodyski
authors.forename: Heleen
authors.initials: H
authors.lastname: Verlinden
authors.forename: Nanda
authors.initials: N
authors.lastname: Filkin
authors.forename: Hans Peter
authors.initials: HP
authors.lastname: Vandersmissen
authors.forename: Christophe
authors.initials: C
authors.lastname: Fleury
authors.forename: Stuart E
authors.initials: SE
authors.lastname: Reynolds
authors.forename: Zhen-peng
authors.initials: ZP
authors.lastname: Kai
authors.forename: Jozef Vanden
authors.initials: JV
authors.lastname: Broeck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000378:metabolism; D018076:DNA, Complementary / Q000302:isolation & purification; D005260:Female; D007301:Insect Hormones / Q000378:metabolism*; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D018402:Manduca / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism*; D018013:Receptors, Neuropeptide / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D007301:Insect Hormones; D019476:Insect Proteins; D009479:Neuropeptides; D018013:Receptors, Neuropeptide; C050669:allatotropin; D000242:Cyclic AMP; D002118:Calcium