Discrimination of Isointense Bitter Stimuli in a Beer Model System

Prior work suggests humans can differentiate between bitter stimuli in water. Here, we describe three experiments that test whether beer consumers can discriminate between different bitterants in beer. In Experiment 1 (n = 51), stimuli were intensity matched; Experiments 2 and 3 were a difference from control (DFC)/check-all-that-apply (CATA) test (n = 62), and an affective test (n = 81). All used a commercial non-alcoholic beer spiked with Isolone (a hop extract), quinine sulfate dihydrate, and sucrose octaacetate (SOA). In Experiment 1, participants rated intensities on general labeled magnitude scales (gLMS), which were analyzed via ANOVA. In Experiment 2, participants rated how different samples were from a reference of Isolone on a 7-point DFC scale, and endorsed 13 attributes in a CATA task. DFC data were analyzed via ANOVA with Dunnett’s test to compare differences relative to a blind reference, and CATA data were analyzed via Cochran’s Q test. In Experiment 3, liking was assessed on labeled affective magnitude scales, and samples were also ranked. Liking was analyzed via ANOVA and rankings were analyzed with a Cochran–Mantel–Haenszel test. Experiment 1 confirmed that samples were isointense. In Experiment 2, despite being isointense, both quinine (p = 0.04) and SOA (p = 0.03) were different from Isolone, but no significant effects were found for CATA descriptors (all p values > 0.16). In Experiment 3, neither liking (p = 0.16) or ranking (p = 0.49) differed. Collectively, these data confirm that individuals can discriminate perceptually distinct bitter stimuli in beer, as shown previously in water, but these differences cannot be described semantically, and they do not seem to influence hedonic assessments.

Rats are unable to discriminate quinine from diverse bitter stimuli

Compounds described by humans as “bitter” are sensed by a family of type 2 taste receptors (T2Rs). Previous work suggested that diverse bitter stimuli activate distinct receptors, which might allow for perceptually distinct tastes. Alternatively, it has been shown that multiple T2Rs are expressed on the same taste cell, leading to the contrary suggestion that these stimuli produce a unitary perception. Behavioral work done to address this in rodent models is limited to Spector and Kopka (Spector AC, Kopka SL. J Neurosci 22: 1937–1941, 2002), who demonstrated that rats cannot discriminate quinine from denatonium. Supporting this finding, it has been shown that quinine and denatonium activate overlapping T2Rs and neurons in both the mouse and rat nucleus of the solitary tract (NTS). However, cycloheximide and 6-n-propylthiouracil (PROP) do not appear to overlap with quinine in the NTS, suggesting that these stimuli may be discriminable from quinine and the denatonium/quinine comparison is not generalizable. Using the same procedure as Spector and Kopka, we tasked animals with discriminating a range of stimuli (denatonium, cycloheximide, PROP, and sucrose octaacetate) from quinine. We replicated and expanded the findings of Spector and Kopka; rats could not discriminate quinine from denatonium, cycloheximide, or PROP. Rats showed a very weak ability to discriminate between quinine and sucrose octaacetate. All animals succeeded in discriminating quinine from KCl, demonstrating they were capable of the task. These data suggest that rats cannot discriminate this suite of stimuli, although they appear distinct by physiological measures.

Bitter-Induced Salivary Proteins Increase Detection Threshold of Quinine, But Not Sucrose

Exposures to dietary tannic acid (TA, 3%) and quinine (0.375%) upregulate partially overlapping sets of salivary proteins which are concurrent with changes in taste-driven behaviors, such as rate of feeding and brief access licking to quinine. In addition, the presence of salivary proteins reduces chorda tympani responding to quinine. Together these data suggest that salivary proteins play a role in bitter taste. We hypothesized that salivary proteins altered orosensory feedback to bitter by decreasing sensitivity to the stimulus. To that end, we used diet exposure to alter salivary proteins, then assessed an animal’s ability to detect quinine, using a 2-response operant task. Rats were asked to discriminate descending concentrations of quinine from water in a modified forced-choice paradigm, before and after exposure to diets that alter salivary protein expression in a similar way (0.375% quinine or 3% TA), or 1 of 2 control diets. Control animals received either a bitter diet that does not upregulate salivary proteins (4% sucrose octaacetate), or a nonbitter diet. The rats exposed to salivary protein-inducing diets significantly decreased their performance (had higher detection thresholds) after diet exposure, whereas rats in the control conditions did not alter performance after diet exposure. A fifth group of animals were trained to detect sucrose before and after they were maintained on the 3% TA diet. There was no significant difference in performance, suggesting that these shifts in threshold are stimulus specific rather than task specific. Taken together, these results suggest that salivary proteins reduce sensitivity to quinine.

Bivariate genome-wide association analysis strengthens the role of bitter receptor clusters on chromosomes 7 and 12 in human bitter taste

Human perception of bitter substances is partially genetically determined. Previously we discovered a single nucleotide polymorphism (SNP) within the bitter taste receptor gene TAS2R19 on chromosome 12 that accounts for 5.8% of the variance in the perceived intensity rating of quinine, and we strengthened the classic association between TAS2R38 genotype and the bitterness of propylthiouracil (PROP). Here we performed a genome-wide association study (GWAS) using a 40% larger sample (n = 1999) together with a bivariate approach to detect previously unidentified common variants with small effects on bitter perception. We identified two signals, both with small effects (< 2%), within the bitter taste receptor clusters on chromosomes 7 and 12, which influence the perceived bitterness of denatonium benzoate and sucrose octaacetate respectively. We also provided the first independent replication for an association of caffeine bitterness on chromosome 12. Furthermore, we provided evidence for pleiotropic effects on quinine, caffeine, sucrose octaacetate and denatonium benzoate for the three SNPs on chromosome 12 and the functional importance of the SNPs for denatonium benzoate bitterness. These findings provide new insights into the genetic architecture of bitter taste and offer a useful starting point for determining the biological pathways linking perception of bitter substances.