An UNC-49 GABA receptor subunit from the parasitic nematode Haemonchus contortus is associated with enhanced GABA sensitivity in nematode heteromeric channels

J Neurochem. 2010 Jun;113(5):1113-22. doi: 10.1111/j.1471-4159.2010.06651.x. Epub 2010 Feb 23.

Abstract

We have identified two genes from the parasitic nematode Haemonchus contortus, Hco-unc-49B and Hco-unc-49C that encode two GABA-gated chloride channel subunits. Electrophysiological analysis revealed that this channel has properties similar to those of the UNC-49 channel from the free-living nematode Caenorhabditis elegans. For example, the Hco-UNC-49B subunit forms a functional homomeric channel that responds to GABA and is highly sensitive to picrotoxin. Hco-UNC-49C alone does not respond to GABA but can assemble with Hco-UNC-49B to form a heteromeric channel with a lower sensitivity to picrotoxin. However, we did find that the Hco-UNC-49B/C heteromeric channel is significantly more responsive to agonists compared to the Hco-UNC-49B homomeric channel, which is the opposite trend to what has been found previously for the C. elegans channel. To investigate the subunit requirements for high agonist sensitivity, we generated cross-assembled channels by co-expressing the H. contortus subunits with UNC-49 subunits from C. elegans (Cel-UNC-49). Co-expressing Cel-UNC-49B with Hco-UNC-49C produced a heteromeric channel with a reduced sensitivity to GABA compared to that of the Cel-UNC-49B homomeric channel. In contrast, co-expressing Hco-UNC-49B with Cel-UNC-49C produced a heteromeric channel that, like the Hco-UNC-49B/C heteromeric channel, exhibits an increased sensitivity to GABA. These results suggest that the Hco-UNC-49B subunit is the key determinant for the high agonist sensitivity of heteromeric channels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chloride Channels / drug effects
  • Chloride Channels / genetics
  • Chloride Channels / metabolism*
  • Cloning, Molecular
  • Electrophysiology
  • GABA Agonists / pharmacology
  • Haemonchus / metabolism*
  • Helminth Proteins / drug effects
  • Helminth Proteins / genetics
  • Helminth Proteins / metabolism*
  • Ion Channel Gating / drug effects
  • Ion Channels / physiology*
  • Molecular Sequence Data
  • Muscimol / pharmacology
  • Oocytes / metabolism
  • Patch-Clamp Techniques
  • Receptors, GABA / drug effects
  • Receptors, GABA / genetics
  • Receptors, GABA / metabolism*
  • Synaptic Transmission / physiology
  • Xenopus laevis
  • gamma-Aminobutyric Acid / pharmacology*

Substances

  • Chloride Channels
  • GABA Agonists
  • Helminth Proteins
  • Ion Channels
  • Receptors, GABA
  • Muscimol
  • gamma-Aminobutyric Acid

Associated data

  • GENBANK/EU049602
  • GENBANK/EU939734