David W. Pittman

Detection of free fatty acids following a conditioned taste aversion in rats.

A gustatory transduction mechanism for free fatty acids (FFAs) has been described in isolated rat taste receptor cells; however, the ability of behaving rats to detect FFAs has not been characterized. Through conditioned taste aversion (CTA) methodology, this study defines the ability of rats to detect and avoid the two principal FFA components of corn oil, linoleic and oleic acid. Following taste aversion conditioning, rats avoided both linoleic and oleic acid at greater than or equal to 66 muM and failed to avoid either 44 muM linoleic or oleic acid. Rats demonstrated generalized avoidances between 88 muM linoleic and oleic acid irrespective of presenting the FFAs as either unesterified acids dissolved in 5 mM ethanol or aqueous sodium salts, sodium linoleate and sodium oleate. Following a CTA to linoleic acid, rats did not show generalized avoidance of NaCl or ethanol, two potentially concomitant tastants in the oral cavity. A CTA to linoleic or oleic acid did produce a generalized avoidance to the other FFA. These results support the ability of rats to detect linoleic and oleic acid (>44 muM) and suggest that the two FFAs share common orosensory properties. Furthermore, it is unlikely that the detection of the FFAs is due to an enhancement of other concomitant tastants such as saliva or the delivery solution.

Chorda tympani nerve transection impairs the gustatory detection of free fatty acids in male and female rats.

Sprague-Dawley rats with intact (SHAM) and bilaterally transected chorda tympani nerves (CTX) received conditioned taste aversions (CTAs) to the free fatty acids (FFAs), linoleic and oleic acid, at micromolar quantities. Two-bottle preference tests showed that CTX eliminated avoidance of 88 muM linoleic acid but did not affect CTA avoidance of corn oil or 250 mM sucrose. Short-duration stimulus tests following single-pairing CTAs revealed that 8-s stimulus durations resulted in higher detection thresholds for linoleic acid than 30-s trials. In these short-duration tests, CTX rats showed 2-fold elevations in threshold for linoleic acid compared to the SHAM rats. A single-pairing CTA did not produce avoidance of oleic acid during the short-duration tests; however, 3 consecutive days of CTA pairings did produce avoidance of oleic acid in both male and female rats. Finally, both male and female rats received SHAM or CTX surgery after demonstrating successful CTAs to either 100 microM linoleic or oleic acid. The ability to detect and avoid linoleic and oleic acid was eliminated by CTX for both sexes. Differences in the ability of rats to form CTAs to linoleic and oleic acid suggest that linoleic acid is a more salient stimulus than oleic acid. Our results suggest that FFAs stimulate afferent taste signals in the chorda tympani nerve of male and female rats and that these signals play an important role in the gustatory behavior of accepting or avoiding taste stimuli following a conditioned taste aversion.

Orosensory Detection of Fatty Acids by Obesity-Prone and Obesity-Resistant Rats: Strain and Sex Differences

A series of brief-access (15s) behavioral assays following the formation of a conditioned taste aversion (CTA) to linoleic acid were performed in order to follow up on observations showing differences in the chemosensory responses to dietary fat in obesity-prone (Osborne-Mendel [O-M]) and obesity-resistant (S5B/Pl) rat strains. Strong aversions to linoleic acid (conditioned stimulus 100 microM) were generated in both O-M and S5B/Pl rats to concentrations as low as 2.5 microM. Observed strain differences were in contrast to expectations based upon electrophysiological studies previously showing greater fatty acid-induced inhibition of delayed rectifying K+ channels in S5B/Pl rats. In the CTA assays, the O-M rats showed aversions at lower fatty acid concentrations with more resistance to extinction in brief-access orosensory tests, suggesting that the obesity-prone strain may be more sensitive in the detection and subsequent avoidance of linoleic acid than the obesity-resistant strain. The independent variable of sex produced even greater differences in the avoidance of linoleic acid following conditioning than the effects of strain. Female rats of both strains were significantly more sensitive to fatty acids, showed greater cross-generalization from linoleic to oleic acid, and showed greater avoidance of linoleic acid than male counterparts. These findings suggest genetic influences on yet to be identified mechanisms potentially within the gustatory system that affect the sensitivity to detect the fatty acid chemicals found in dietary fat during brief-access orosensory testing.

Magnetic Field Conditioned Taste Aversion In Rats

Conditioned taste aversion is a common classic conditioning procedure used to identify noxious stimuli. When a rat is given a taste solution, the conditioned stimulus (CS), followed by an unpleasant experience, the unconditioned stimulus (US), the rat will avoid consumption of the CS in future presentations. These experiments use the taste aversion procedure to examine the effect of exposure to a high magnetic field. A solution consisting of 3.0 g glucose and 1.25 g saccharin per 1 L of solution (G+S) was used as the CS and a 9.4-T magnet served as the US. In Experiment 1, all rats received a 10 min presentation of the G+S solution followed by either a 30 min exposure to the magnetic field (Magnet, n = 8), a 30-min exposure in a container with similar conditions but lacking the magnetic field (Sham, n = 8), or no exposure (Control, n = 8). The Magnet Group showed a taste aversion on the first day of preference testing (p < 0.05). Experiment 2 employed the same US-CS protocol for 3 consecutive days of conditioning. The Magnet Group demonstrated a taste aversion for the postexposure Days 1-8 (p < 0.01). There was no difference between the Sham and Control Groups in either experiment. The results of this study clearly demonstrate that the rats associated the G+S solution with the experience of being exposed to the high magnetic field and avoided the solution in subsequent presentations.

Rearing on basal or high dietary NaCl modifies chorda tympani nerve responses in rats

Two experiments were conducted to determine the influence of dietary NaCl level on the integrated responses of the chorda tympani (CT) nerve to salt stimulation alone and mixed with the sodium-channel blocker, amiloride hydrochloride. Five groups of adult male rats were reared on regular chow containing either basal 0.1%, intermediate 1.0%, or high 3.0% NaCl from conception to postnatal day (PD) 30 or from conception to adulthood. Adult rats reared from conception to adulthood on basal dietary NaCl demonstrated a reduction in the CT nerve response to NaCl due to a decrease in the amiloride sensitive transduction mechanism. However, the CT nerve responses of adult rats reared on basal dietary NaCl to PD30 and then switched to intermediate dietary NaCl were similar to those of rats reared for a lifetime on intermediate dietary NaCl. Similarly, the CT nerve responses to NaCl in animals reared on high dietary NaCl from conception to PD30 and then switched to an intermediate NaCl diet were comparable to animals reared on intermediate and basal dietary NaCl. However, we found that exposure to high dietary NaCl led to a greater amiloride inhibition of NaCl responses. Thus, there is critical association between dietary NaCl level over two different exposure periods and CT nerve responsiveness to NaCl specifically regarding the degree of amiloride inhibition.

Role for epithelial Na+channels and putative Na+/H+ exchangers in salt taste transduction in rats

The effects of the epithelial Na+ channel antagonists amiloride and benzamil and the Na+/H+ exchange antagonist 5-(N,N-dimethyl)-amiloride (DMA)-Cl on the integrated responses of the chorda tympani nerve to 30, 75, 150, 300, and 500 mM concentrations of NaCl, KCl, and NH4Cl were assessed in male Sprague-Dawley rats. Based on evidence from other systems, 1 and 25 microM amiloride and benzamil were chosen to selectively inhibit epithelial Na+ channels and 1 microM DMA was chosen to selectively inhibit Na+/H+ exchange. When added to stimulating salt solutions, amiloride, benzamil, and DMA were each effective in inhibiting responses to all three salts. The degree of inhibition varied with drug, salt, and salt concentration, but not drug dose. Amiloride suppressed NaCl responses to a greater degree than KCl and NH4Cl responses, whereas DMA suppressed NH4Cl responses to a greater degree than NaCl and KCl responses. In all but one case (25 microM amiloride added to KCl), drug suppression of taste nerve responses decreased with an increase in salt concentration. The present results suggest that 1) epithelial Na+ channels in rat taste receptor cells may play a role in KCl and NH4Cl taste transduction; 2) a Na+/H+ exchange protein may be present in taste receptor cells, representing a putative component, in addition to epithelial Na+ channels, in salt taste transduction; and 3) salt taste detection and transduction may depend on the utilization of a combination of common and distinct transcellular pathways.The effects of the epithelial Na+channel antagonists amiloride and benzamil and the Na+/H+exchange antagonist 5-( N, N-dimethyl)-amiloride (DMA)-Cl on the integrated responses of the chorda tympani nerve to 30, 75, 150, 300, and 500 mM concentrations of NaCl, KCl, and NH4Cl were assessed in male Sprague-Dawley rats. Based on evidence from other systems, 1 and 25 μM amiloride and benzamil were chosen to selectively inhibit epithelial Na+ channels and 1 μM DMA was chosen to selectively inhibit Na+/H+exchange. When added to stimulating salt solutions, amiloride, benzamil, and DMA were each effective in inhibiting responses to all three salts. The degree of inhibition varied with drug, salt, and salt concentration, but not drug dose. Amiloride suppressed NaCl responses to a greater degree than KCl and NH4Cl responses, whereas DMA suppressed NH4Cl responses to a greater degree than NaCl and KCl responses. In all but one case (25 μM amiloride added to KCl), drug suppression of taste nerve responses decreased with an increase in salt concentration. The present results suggest that 1) epithelial Na+ channels in rat taste receptor cells may play a role in KCl and NH4Cl taste transduction; 2) a Na+/H+exchange protein may be present in taste receptor cells, representing a putative component, in addition to epithelial Na+ channels, in salt taste transduction; and 3) salt taste detection and transduction may depend on the utilization of a combination of common and distinct transcellular pathways.

Cost-free and sustainable incentive increases healthy eating decisions during elementary school lunch

We aimed to develop a cost-free and sustainable program to influence healthier eating decisions during elementary school lunch. Baseline food and beverage choices were assessed for 9 days during lunch service at two racially and economically diverse elementary schools in Spartanburg County, SC, USA. After being informed that the labeled items on the daily lunch menu represented the healthiest choice, students were allowed to ring a call bell in the cafeteria for public recognition when they chose all of the identified healthiest food and beverage items during lunch service. Using menus matched to the baseline phase, food and beverage choices were measured during a 9-day intervention phase. After 30 days, food and beverage choices were reassessed during a 3-day follow-up phase. Healthiest food & beverage choices increased 49% with >60% of students choosing non-flavored milk over flavored milk during the intervention phase. There was no difference in the success of the program between the two schools. The program continued and healthy eating decisions were significantly sustained at a 30-day follow-up assessment. Public recognition through bell ringing appears to be an effective practice to sustain increases in healthy eating decisions during elementary school lunch and warrants expansion to larger scale, longitudinal trials.

Multiple Processes Underlie Benzodiazepine-Mediated Increases in the Consumption of Accepted and Avoided Stimuli

Hyperphagia is a reported side effect of anxiolytic benzodiazepines such as chlordiazepoxide (CDP). Prior research has focused primarily on the ingestive responses to sweet or solid foods. We examined CDP effects on licking for normally accepted and avoided taste solutions across a range of concentrations. The effect of CDP (10 mg/kg) versus saline on the licking patterns of water-restricted rats for water and 3 concentrations of sucrose, saccharin, NaCl, monosodium glutamate (MSG), citric acid, and quinine (Q-HCl) solutions was evaluated during 1 h tests. CDP increased meal size for all tastants except citric acid. Analysis of licking microstructure revealed 3 dissociable effects of CDP. CDP affected oromotor coordination as indicated by a uniform increase in the modal interlick interval for all stimuli. CDP increased meal size as indicated by shorter pauses during consumption of water, MSG, and weaker saccharin concentrations, and by fewer long interlick intervals (250-2000 ms) for normally avoided tastants. CDP also increased meal size by increasing burst size, burst duration, and the initial rate of licking for most solutions, suggesting increased hedonic taste evaluation. CDP did not affect variables associated with postingestive feedback such as meal duration or number of bursts, and the results also suggest that CDP did not enhance the perceived taste intensity. We hypothesize that the reduction of pause duration is consistent with an increased motivation to sample the stimulus that synergizes with changes in taste-mediated responsiveness to some but not all stimuli to yield increases in the consumption of both normally accepted and avoided taste stimuli.

High-Fat Diet Alters the Orosensory Sensitivity to Fatty Acids in Obesity- Resistant but not Obesity-Prone Rats

Gene-environment interactions play a role in the development of obesity but specific effects of diet on the orosensory detection of fatty acids have yet to be clarified. The objective of this study is to characterize the effect of prolonged (5-week) exposure to a high-fat (60%) diet on the behavioral sensitivity to the fatty acid linoleate following a conditioned taste aversion in obesity-prone and obesity-resistant rats. Exposure to the high-fat diet significantly enhanced the sensitivity of obesity-resistant (S5B/Pl) rats to linoleate while producing no effect on the fatty acid sensitivity for obesity-prone rats. Specifically, high-fat diet fed S5B/Pl rats showed stronger initial avoidance of linoleate and slower extinction rates than their normal diet cohorts. Our study suggests that prolonged dietary fat consumption may alter the behavioral sensitivity to fatty acids particularly in obesity-resistant animals.

The Palatability of Lopinavir and Ritonavir Delivered by an Innovative Freeze-Dried Fast-Dissolving Tablet Formulation

Negative hedonic sensory qualities of HIV antiretroviral drugs often reduce patient adherence particularly in pediatric populations requiring oral consumption. This study examines the palatability of an innovative delivery mechanism utilizing a freeze-drying-in-blister approach to create fast-dissolving tablets (FDTs) containing a fixed-dose combination of lopinavir and ritonavir (LPV/r). Consumption patterns of solutions during brief-access and long-term testing and baby foodstuff consumption were analyzed to evaluate the orosensory detection and avoidance of placebo FDTs containing no LPV/r (FDT−) and FDTs containing LPV/r (FDT+). Rats showed no change in consumption patterns for the placebo FDT− compared with control solutions. Rats can detect but do not avoid FDT+ at body-weight-adjusted dosages in both brief-access (30-s) and long-term (23 h) consumption tests. There is an aversive response to concentrated doses of FDT+ during brief-access tests that cannot be masked by 25% sucrose. However, the strongest FDT+ concentration was not rejected when mixed with 50 g of applesauce, banana sauce, or rice cereal baby foodstuffs. The averseness of the FDT+ was associated with the presence of LPV/r and not the FDT− formulation itself. The novel FDT formulation appears to be a palatable delivery mechanism for oral antiretroviral pharmaceuticals especially when mixed with baby foodstuffs.