Colorado PROFILES, The Colorado Clinical and Translational Sciences Institute (CCTSI)
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This submission is in response to Program Announcement 96-026 on the Molecular Pharmacology of Anesthetic Action. The goal of this proposal is to identify the genetic differences that underlie quantitative trait loci (QTLs) that are associated with anesthetic sensitivity (as measured by loss of righting reflex) in mice. The focus will be on sequencing (using automated techniques) a 250 kb region found to carry a gene affecting propofol sensitivity phenotype in certain transgenic lines. Once putative genes are identified that are related to propofol sensitivity, gene sequences are to be identified by computer search of known databases. Following identification of the genes related to propofol sensitivity in the 250 kb region, this region will be sequenced in selectively bred mice resistant (SS) and sensitive (LS) to ethanol to detect possible mutations since these lines are also resistant and sensitive to propofol.

QTLs are to be identified for other general anesthetics (isoflurane, enflurane, etomidate, halothane, and ketamine) using both LS x SS recombinant inbred strains and F2 mice. Identification of mouse cDNAs that lie in the implicated QTL region will involve several techniques with emphasis on the use of known and developing data bases for DNA sequences of the mouse and human ESTs (Expressed-Sequence-Tags). It is then proposed to amplify the protein coding regions from cDNA with the polymerase chain reaction to find DNA sequence differences between LS and SS mice that produce differences in primary amino acid sequence. Such differences will represent candidate mutations for anesthetic sensitivity/resistance, and would eventually allow for expression of protein products and characterization of the function of these proteins and interaction with anesthetics.
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