Identification and functional characterization of the pheromone biosynthesis activating neuropeptide receptor isoforms from Mamestra brassicae

József Fodor, J. Joe Hull, Gabriella Köblös, Emmanuelle Jacquin-Joly, Tamás Szlanka, Adrien Fónagy

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In most moth species, including Mamestra brassicae, pheromone biosynthesis activating neuropeptide (PBAN) regulates pheromone production. Generally, PBAN acts directly on the pheromone gland (PG) cells via its specific G protein-coupled receptor (i.e. PBANR) with Ca2+ as a second messenger. In this study, we identified cDNAs encoding three variants (A, B and C) of the M. brassicae PBANR (Mambr-PBANR). The full-length coding sequences were transiently expressed in cultured Trichoplusia ni cells and Sf9 cells for functional characterization. All three isoforms dose-dependently mobilized extracellular Ca2+ in response to PBAN analogs with Mambr-PBANR-C exhibiting the greatest sensitivity. Fluorescent confocal microscopy imaging studies demonstrated binding of a rhodamine red-labeled ligand (RR10CPBAN) to all three Mambr-PBANR isoforms. RR10CPBAN binding did not trigger ligand-induced internalization in cells expressing PBANR-A, but did in cells expressing the PBANR-B and -C isoforms. Furthermore, activation of the PBANR-B and -C isoforms with the 18 amino acid Mambr-pheromonotropin resulted in co-localization with a Drosophila melanogaster arrestin homolog (Kurtz), whereas stimulation with an unrelated peptide had no effect. PCR-based profiling of the three transcripts revealed a basal level of expression throughout development with a dramatic increase in PG transcripts from the day of adult emergence with PBANR-C being the most abundant.

Original languageEnglish
Pages (from-to)60-69
Number of pages10
JournalGeneral and Comparative Endocrinology
Volume258
DOIs
Publication statusPublished - Mar 1 2018

Keywords

  • Ca imaging
  • Confocal microscopy
  • Mamestra brassicae
  • PBAN
  • Pheromone biosynthesis activating neuropeptide receptor (PBANR)
  • Receptor internalization

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Endocrinology

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