This application both requests continued support for the research objectives of AA 011034 and responds to PA-03-162 -""""""""FINDING GENES FOR ALCOHOL-RELATED BEHAVIORS AND RISK FOR ALCOHOLISM"""""""" - in that we propose to implement a high throughput strategy for the detection of quantitative trait genes (QTGs). The new strategy will integrate several different experimental approaches, all of which are """"""""on-line"""""""" in the participating laboratories. These include multiple cross mapping [MCM] (Hitzemann et al. 2002, 2003a), fine mapping in heterogenous stock [HS] animals (Demarest et al. 2001; Hitzemann et al. 2002, 2003a), the integration of QTL analysis and functional genomics (Belknap et al. 2001; Hitzemann et al. 2003a,b), the use of short term selective breeding to confirm QTLs and follow gene segregation (Belknap et al. 1997; Metten et al. 1998) and QTL confirmation in BAC transgenic mice (Buck et al. - Section D).
The specific aims may be summarized: l. To fine map in heterogeneous stock (HS) animals (to a resolution of 1 cM or less) known QTLs for ethanol-induced activation, ethanol preference and acute ethanol withdrawal. 2. To integrate short term selective breeding and gene expression analysis. 3. To determine which genes within the QTL interval (aim 1) show the genotype x phenotype appropriate expression pattern. 4. To determine which genes within the QTL interval are cis-regulated. 5. To use all available strategies to provide additional proof of QTL --> QTG (see Belknap et al. 2001).
Aims 1 -4 build from our previous arguments (Belknap et al. 2001; Phillips et al. 2002; Hitzemann et al. 2002, 2003a) and describe a strategy which provides the requisite interlocking levels of proof to move in a timely fashion from QTL --> QTG. A candidate QTG must lie in the 1 cM interval, exhibit the appropriate pattern of segregation with short term selective breeding, show the QTL appropriate difference in expression among the inbred strains and be cis-regulated. We argue that such multiple sources of evidence will exclude the possibility of a false positive result.
Aim 5 will test this hypothesis ? ?
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