Isolation and functional characterization of an allatotropin receptor from Manduca sexta

• Published in: Insect biochemistry and molecular biology Vol. 41; no. 10; pp. 804 - 814
• Main Authors: Horodyski, Frank M, Verlinden, Heleen, Filkin, Nanda, Vandersmissen, Hans Peter, Fleury, Christophe, Reynolds, Stuart E, Kai, Zhen-peng, Broeck, Jozef Vanden
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.10.2011
• Subjects: adults; Allatotropin; alternative splicing; Amino Acid Sequence; Animals; Base Sequence; biosynthesis; Bombyx mori; Calcium - metabolism; Calcium Signaling; CHO Cells; complementary DNA; corpora allata; Cricetinae; Cricetulus; Cyclic AMP - metabolism; DNA, Complementary - isolation & purification; Female; G protein-coupled receptors; genes; hindgut; Insect Hormones - metabolism; Insect Proteins - isolation & purification; Insect Proteins - metabolism; insects; invertebrates; ion transport; Juvenile hormone; juvenile hormones; larvae; Lepidoptera; Male; Malpighian tubules; mammals; Manduca - chemistry; Manduca - metabolism; Manduca sexta; messenger RNA; midgut; Molecular Sequence Data; Neuropeptides; Neuropeptides - metabolism; receptors; Receptors, Neuropeptide - isolation & purification; Receptors, Neuropeptide - metabolism; testes; Manduca sexta; Neuropeptides; G protein-coupled receptors; Allatotropin; Juvenile hormone
• ISSN: 0965-1748; 1879-0240
• DOI: 10.1016/j.ibmb.2011.06.002

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 Ca2+ 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.