Mitochondrial complex I function affects halothane sensitivity in Caenorhabditis elegans.
BACKGROUND
: The gene gas-1 encodes a subunit of complex I of the mitochondrial electron transport chain in Caenorhabditis elegans. A mutation in gas-1 profoundly increases sensitivity of C. elegans to volatile anesthetics. It is unclear which aspects of mitochondrial function account for the hypersensitivity of the mutant.


METHODS
: Oxidative phosphorylation was determined by measuring mitochondrial oxygen consumption using electron donors specific for either complex I or complex II. Adenosine triphosphate concentrations were determined by measuring luciferase activity. Oxidative damage to mitochondrial proteins was identified using specific antibodies.


RESULTS
: Halothane inhibited oxidative phosphorylation in isolated wild-type mitochondria within a concentration range that immobilizes intact worms. At equal halothane concentrations, complex I activity but not complex II activity was lower in mitochondria from mutant (gas-1) animals than from wild-type (N2) animals. The halothane concentrations needed to immobilize 50% of N2 or gas-1 animals, respectively, did not reduce oxidative phosphorylation to identical rates in the two strains. In air, adenosine triphosphate concentrations were similar for N2 and gas-1 but were decreased in the presence of halothane only in gas-1 animals. Oxygen tension changed the sensitivity of both strains to halothane. When nematodes were raised in room air, oxidative damage to mitochondrial proteins was increased in the mutant animal compared with the wild type.


CONCLUSIONS
: Rates of oxidative phosphorylation and changes in adenosine triphosphate concentrations by themselves do not control anesthetic-induced immobility of wild-type C. elegans. However, they may contribute to the increased sensitivity to volatile anesthetics of the gas-1 mutant. Oxidative damage to proteins may be an important contributor to sensitivity to volatile anesthetics in C. elegans.
Kayser
Ernst-Bernhard
EB
Department of Anesthesiology, University Hospitals, Case Western Reserve University, Cleveland, Ohio, USA.
Morgan
Phil G
PG
Sedensky
Margaret M
MM
D000244:Adenosine Diphosphate / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000502:physiology*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D042967:Electron Transport Complex I / Q000187:drug effects* / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects; D006221:Halothane / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D008293:Malates / Q000378:metabolism; D016877:Oxidants / Q000633:toxicity; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation / Q000187:drug effects; D013386:Succinates / Q000378:metabolism
D018685:Anesthetics, Inhalation; D029742:Caenorhabditis elegans Proteins; D008293:Malates; D016877:Oxidants; D013386:Succinates; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D042967:Electron Transport Complex I; D006221:Halothane