Cloning and characterization of a GABA receptor from Plutella xylostella (Lepidoptera: Plutellidae)

J Econ Entomol. 2008 Dec;101(6):1888-96. doi: 10.1603/0022-0493-101.6.1888.

Abstract

A full-length cDNA, with an open reading frame (ORF) of 1,449 bp, encoding a subunit of the gamma-aminobutyric acid (GABA)-activated chloride channel was isolated from Plutella xylostella (L.) (Lepidoptera: Plutellidae) (GenBank accession no. EF156251). The subunit gene encoded a 483-amino acid polypeptide that showed 84% sequence identity with DmRdl subunit (U02042) (Drosophila melanogaster resistant to dieldrin). When expressed in Xenopus laevis oocytes, the subunit assembled as a functional homomeric complex activated by GABA and abamectin in a dose-dependent manner. The EC50 value of GABA was 0.49 mM (0.41-0.58) (n = 5). However, the responses to abamectin were very robust, with an EC50 of 4.85 microM (4.02-5.89) (n = 6), indicating that abamectin was > 100-fold more potent in activating chloride currents than GABA. The results suggest that this subunit is vital to the formation of a functional channel and contains the binding site of abamectin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • Electric Conductivity
  • Insect Proteins / chemistry
  • Insect Proteins / drug effects*
  • Insect Proteins / genetics
  • Insecticides / pharmacology*
  • Ivermectin / analogs & derivatives*
  • Ivermectin / pharmacology
  • Molecular Sequence Data
  • Moths / genetics*
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Patch-Clamp Techniques
  • Receptors, GABA / chemistry
  • Receptors, GABA / drug effects*
  • Receptors, GABA / genetics
  • Sequence Alignment
  • Xenopus
  • gamma-Aminobutyric Acid / pharmacology

Substances

  • Insect Proteins
  • Insecticides
  • Receptors, GABA
  • gamma-Aminobutyric Acid
  • abamectin
  • Ivermectin