We are using the powerful genetics of the nematode Caenorhabditis elegans to address to distinct questions: 1) What are the molecular mechanisms whereby general anesthetics disrupt nervous system function? 2) What are the genetic determinants of hypoxic cell death?

 C. elegans is a small non-parasitic worm with a well-characterized nervous system that directs a number of complex behaviors. Through both classical and quantitative genetic techniques, we have identified single gene mutations and quantitative trait loci that markedly alter the sensitivity of C. elegans to volatile anesthetics. In particular, mutations in a number of presynaptic proteins render C. elegans either resistant or hypersensitive to anesthetics. Through primarily genetic and biochemical approaches, we are defining how these gene products regulate anesthetic action. In the end, we should know what are the molecular targets of general anesthetics in C. elegans, and, perhaps, those operant in the vertebrate nervous system.

Our work on hypoxic death is motivated by the fact that stroke and heart attack, a result of hypoxic cell death, are together the number one cause of human death in the US. C. elegans genetics has made seminal contributions for apoptotic cell death yet is relatively untapped for hypoxic death. We have found several mutant strains that are hypoxia resistant. The mutant genes thusfar identified fall into two categories: those that diminish insulin/IGF receptor signaling and those that diminish Ca++-mediated necrotic cell death. The insulin receptor mutants produce profound protection from hypoxia through a PIP3-kinase cascade leading to phosphorylation of a forkhead-type transcription factor. The hypoxic protection provided by the Ca++-mediated necrotic cell death mutants suggest that the mechanisms of hypoxic cell death in vertebrates and C. elegans overlap since divalent cations have been implicated in hypoxic death of vertebrate neurons.

 

Selected Publications:

 van Swinderen B, Metz LB, Shebester LD, Crowder CM. A C. elegans pheromone antagonizes volatile anesthetic action through a Go-coupled pathway. Genetics, in press.

 van Swinderen B, Shebester LD, Mendel JE, Sternberg PW, Crowder CM. Gao regulates of volatile anesthetic action in Caenorhabditis. elegans. Genetics 158:643-55, 2001.

 Crowder CM, Westover EJ, Kumar AS, Ostlund RE, Covey DF. Enantiospecificity of cholesterol function in vivo. J Biol Chem 276:44369-72, 2001.

 van Swinderen B, Saifee O, Shebester L, Roberson R, Nonet ML, Crowder CM. A neomorphic syntaxin mutation blocks volatile-anesthetic action in Caenorhabditis elegans. Proc Natl Acad Sci USA 96: 2479-84, 1999.

 van Swinderen B, Ebert RH, Shook DR, Cherkasova VA, Johnson TE, Shmookler Reis RJ, and Crowder CM. Quantitative trait loci controlling halothane sensitivity in Caenorhabditis elegans, Proc Natl Acad Sci USA 94:8232-37, 1997.