Identification of Quantitative Trait Loci and candidate genes influencing ethanol sensitivity in honey bees.

Invertebrate models have greatly furthered our understanding of ethanol sensitivity and alcohol addiction. The honey bee (Apis mellifera), a widely used behavioral model, is valuable for comparative studies. A quantitative trait locus (QTL) mapping experiment was designed to identify QTL and genes influencing ethanol vapor sensitivity. A backcross mating between ethanol-sensitive and resistant lines resulted in worker offspring that were tested for sensitivity to the sedative effects of alcohol. A linkage map was constructed with over 500 amplified fragment length polymorphism (AFLP) and sequence-tagged site (STS) markers. Four QTL were identified from three linkage groups with log of odds ratio (LOD) scores of 2.28, 2.26, 2.23, and 2.02. DNA from markers within and near QTL were cloned and sequenced, and this data was utilized to integrate our map with the physical honey bee genome. Many candidate genes were identified that influence synaptic transmission, neuronal growth, and detoxification. Others affect lipid synthesis, apoptosis, alcohol metabolism, cAMP signaling, and electron transport. These results are relevant because they present the first search for QTL that affect resistance to acute ethanol exposure in an invertebrate, could be useful for comparative genomic purposes, and lend credence to the use of honey bees as biomedical models of alcohol metabolism and sensitivity.

Characterization of honey bee sensitivity to ethanol vapor and its correlation with aggression.

Several candidate genes identified from quantitative trait loci (QTL) for defensive behavior in honey bees (Apis mellifera L.) are homologous to genes known to influence ethanol sensitivity in other organisms. To investigate this possible link between aggression/defense and ethanol sensitivity, assays were developed to evaluate ethanol vapor responses in worker bees from a low-defensive (gentle) colony and a high-defensive colony. Defensive workers exhibited characteristic signs of ethanol-induced sedation significantly faster than gentle workers upon exposure to ethanol vapor. Backcross workers displayed ethanol sensitivity intermediate to the parental defensive and gentle lines, suggesting a genetic basis for the trait. Workers were screened with sequence-tagged site markers linked to three defensive-behavior QTL and their genotypes were tested for associations with ethanol sensitivity. There were no significant associations, indicating that the defensive QTL were not having a pleiotropic effect on ethanol sensitivity. It is possible that gentle-source alleles at these QTL are dominant with respect to sensitivity, one or more of these QTL were not segregating in the backcross family, or unidentified QTL are influencing alcohol sensitivity.