We undertook functional studies of four candidate loci, COL6A3 and LOC339975 based on the GWS signals in discovery or replication analyses and KLF12 and RYR3 based on discovery q<0.1 and prior evidence of implicating these loci in alcohol-related phenotypes (reviewed below). We used established behavioral paradigms to test orthologs of COL6A3, KLF12 and RYR3 for effects on behavioral responses to ethanol in C. elegans and Drosophila where orthologous genes were present in the models and genetic reagents and information were available. We analyzed correlation between brain gene expression and alcohol-related phenotypes in BXD mouse data for all three genes. All these studies are summarized in Table 2 and we report here the results of all studies performed. We additionally tested the effect of the ryanodine receptor antagonist dantrolene on ethanol self-administration in rats. The fourth gene, LOC339975, is primate-specific and was taken forward for study in human post-mortem brain.
