We then test the physiological responses of all 31 labellar taste hairs to 16 diverse bitter tastants. The responses of different sensilla show extensive diversity both in magnitude and in response dynamics. We define four functional
classes of bitter neurons and the results provide a functional map of the organ. We then examine the expression of all 68 members of the Gr family of taste receptors. Based on receptor expression, the bitter neurons fall into four classes that coincide closely with the four classes based on Tyrosine Kinase Inhibitor Library physiological responses. The results provide a receptor-to-neuron-to-tastant map of the organ. Misexpression of a receptor confers bitter responses that agree with predictions of the map. Together, the results reveal a degree of complexity that greatly expands the capacity of the system to encode bitter taste; it allows for combinatorial coding and may enable discrimination or adaptive responses to selected bitter stimuli. We selected 14 compounds that have previously been described as bitter by virtue of their behavioral effects on various insect species (Koul, 2005 and Schoonhoven
et al., 2005). The 14 selected tastants include naturally occurring alkaloids, terpenoids, and phenolic compounds, as well as three synthetic compounds. Many of these compounds are toxic and many are perceived as bitter by humans. Some have been tested in Drosophila previously ( Hiroi et al., DAPT solubility dmso 2004, Lee et al., 2010, Marella et al., 2006, Meunier et al., 2003, Thorne et al., 2004 and Wang et al., 2004). We used a modification of a two-choice behavioral paradigm (Tanimura et al., 1982) in which a population of flies is allowed to feed on a microtiter plate containing alternating wells of 1 mM sucrose alone and 5 mM sucrose mixed with a bitter tastant (Figure 2A). Each of the two
solutions contains either red or blue dye, and Bay 11-7085 upon conclusion of the experiment a P.I. is calculated. The P.I. is based on the number of flies with red, blue, and purple abdomens, indicating ingestion of the solution with red dye, the solution with blue dye, or both solutions, respectively (P.I. = [Nblue + 0.5Npurple]/[Nred + Npurple + Nblue]). In our experiments, a P.I. of 1.0 indicates a complete preference for the 5 mM sucrose solution; a P.I. of 0 indicates a complete preference for the 1 mM sucrose solution. We found that in control experiments, flies given a choice between 1 mM sucrose and 5 mM sucrose alone, with no added bitter compounds, showed a P.I. of 0.71, indicating a preference for the 5 mM concentration. We tested a range of concentrations of the 14 tastants. Low concentrations of each tastant had little or no effect on the strong preference for 5 mM sucrose (Figure 2B and Figure S1, available online). However, with addition of increasing concentrations of each bitter tastant to the 5 mM solution, flies increasingly avoided the 5 mM sucrose-bitter mixture.