cDNA-AFLP analysis in levamisole-resistant Haemonchus contortus reveals alternative splicing in a nicotinic acetylcholine receptor subunit

Mol Biochem Parasitol. 2010 Apr;170(2):105-7. doi: 10.1016/j.molbiopara.2009.11.007. Epub 2009 Nov 22.

Abstract

The cDNA-AFLP (cDNA-amplified fragment length polymorphism) method comparing transcripts from levamisole-resistant and susceptible Haemonchus contortus isolates has led to the successful identification of a number of potentially useful levamisole-resistance markers. In the present study, we report the characterization of the transcript-derived fragment (TDF) named HAX which was confirmed to be specifically expressed in three levamisole-resistant isolates by RT-PCR experiments. Cloning and sequencing of the full-length cDNA sequence of HAX revealed high similarity to the Caenorhabditis elegans acr-8 gene and its putative H. contortus orthologue encoding a nicotinic acetylcholine receptor subunit. This Hco-acr-8b short isoform corresponded to a spliced variant of Hco-acr-8 mRNA containing the two first exons and a part of intron 2. As nicotinic acetylcholine receptors constitute the pharmacological target of levamisole, Hco-acr-8b may potentially be involved in the molecular mechanisms leading to levamisole-resistance acquisition in H. contortus.

MeSH terms

  • Alternative Splicing*
  • Amplified Fragment Length Polymorphism Analysis
  • Animals
  • Antinematodal Agents / pharmacology*
  • Caenorhabditis elegans / genetics
  • DNA, Complementary / genetics
  • DNA, Helminth / chemistry
  • DNA, Helminth / genetics
  • Drug Resistance*
  • Haemonchus / drug effects*
  • Haemonchus / genetics*
  • Levamisole / pharmacology*
  • Molecular Sequence Data
  • Protein Subunits
  • Receptors, Nicotinic / biosynthesis*
  • Receptors, Nicotinic / genetics*
  • Sequence Analysis, DNA
  • Sequence Homology

Substances

  • Antinematodal Agents
  • DNA, Complementary
  • DNA, Helminth
  • Protein Subunits
  • Receptors, Nicotinic
  • Levamisole

Associated data

  • GENBANK/GU168769
  • GENBANK/GU168770