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Background and Purpose: Many medications taste intensely bitter. The innate aversion to bitterness affects medical compliance, especially in children. There is a clear need to develop bitter blockers to suppress the bitterness of vital medications. Bitter taste is mediated by TAS2R receptors. Because different pharmaceutical compounds activate distinct sets of TAS2Rs, targeting specific receptors may only suppress bitterness for certain, but not all, bitter-tasting compounds. Alternative strategies are needed to identify universal bitter blockers that will improve the acceptance of every medication. Taste cells in the mouth transmit signals to afferent gustatory nerve fibres through the release of ATP, which activates the gustatory nerve-expressed purine receptors P2X2/P2X3. We hypothesized that blocking gustatory nerve transmission with P2X2/P2X3 inhibitors (e.g. 5-(5-iodo-4-methoxy-2-propan-2-ylphenoxy)pyrimidine-2,4-diamine [AF-353]) would reduce bitterness for all medications and bitter compounds., Experimental Approach: Human sensory taste testing and mouse behavioural analyses were performed to determine if oral application of AF-353 blocks perception of bitter taste and other taste qualities but not non-gustatory oral sensations (e.g. tingle)., Key Results: Rinsing the mouth with AF-353 in humans or oral swabbing it in mice suppressed the bitter taste and avoidance behaviours of all compounds tested. We further showed that AF-353 suppressed other taste qualities (i.e. salt, sweet, sour and savoury) but had no effects on other oral or nasal sensations (e.g, astringency and oral tingle)., Conclusion and Implications: This is the first time a universal, reversible taste blocker in humans has been reported. Topical application of P2X2/P2X3 inhibitor to suppress bitterness may improve medical compliance., (© 2024 The Authors. British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Objective: Food processing greatly contributed to increased food safety, diversity, and accessibility. However, the prevalence of highly palatable and highly processed food in our modern diet has exacerbated obesity rates and contributed to a global health crisis. While accumulating evidence suggests that chronic consumption of such foods is detrimental to sensory and neural physiology, it is unclear whether its short-term intake has adverse effects. Here, we assessed how short-term consumption (<2 months) of three diets varying in composition and macronutrient content influence olfaction and brain metabolism in mice., Methods: The diets tested included a grain-based standard chow diet (CHOW; 54% carbohydrate, 32% protein, 14% fat; #8604 Teklad Rodent diet , Envigo Inc.), a highly processed control diet (hpCTR; 70% carbohydrate, 20% protein, 10% fat; #D12450B, Research Diets Inc.), and a highly processed high-fat diet (hpHFD; 20% carbohydrate, 20% protein, 60% fat; #D12492, Research Diets Inc.). We performed behavioral and metabolic phenotyping, electro-olfactogram (EOG) recordings, brain glucose metabolism imaging, and mitochondrial respirometry in different brain regions. We also performed RNA-sequencing (RNA-seq) in the nose and across several brain regions, and conducted differential expression analysis, gene ontology, and network analysis., Results: We show that short-term consumption of the two highly processed diets, but not the grain-based diet, regardless of macronutrient content, adversely affects odor-guided behaviors, physiological responses to odorants, transcriptional profiles in the olfactory mucosa and brain regions, and brain glucose metabolism and mitochondrial respiration., Conclusions: Even short periods of highly processed food consumption are sufficient to cause early olfactory and brain abnormalities, which has the potential to alter food choices and influence the risk of developing metabolic disease., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

The detection of nutrients in the gut influences ongoing and future feeding behavior as well as the development of food preferences. In addition to nutrient sensing in the intestine, the hepatic portal vein plays a considerable role in detecting ingested nutrients and conveying this information to brain nuclei involved in metabolism, learning, and reward. Here, we review mechanisms underlying hepatic portal vein sensing of nutrients, particularly glucose, and how this is relayed to the brain to influence feeding behavior and reward. We additionally highlight several gaps where future research can provide new insights into the effects of portal nutrients on neural activity in the brain and feeding behavior., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

To investigate the contributions of carbohydrate and fat to obesity we measured the body weight, body composition and food intake of adult C57BL/6J mice fed ad libitum with various combinations of two semisynthetic diets that differed in carbohydrate and fat but not in protein, micronutrient or energy content. In Experiment 1, involving male mice, body weights were similar in groups fed diets comprised of (by energy) 20% protein, 75% carbohydrate and 5% fat (C75-F5) or 20% protein, 5% carbohydrate and 75% fat (C5-F75). However, mice fed a 50:50 composite mixture of the C75-F5 and C5-F75 diets (i.e., a C40-F40 diet) became substantially more obese. Mice that could choose between the C75-F5 and C5-F75 diets ate equal amounts of each diet and gained almost as much weight as did the group fed C40-F40 diet. Mice switched every day between the C75-F5 and C5-F75 diets gained no more weight than did those fed either diet exclusively. In Experiment 2, male and female mice were fed chow or one of 8 isocaloric diets that differed parametrically in carbohydrate and fat content. Groups fed diets in the middle of the range (i.e., C35-F45 or C45-F35) weighed significantly more and were significantly fatter than were those fed diets with more extreme proportions of carbohydrate and fat (e.g., C75-F5, C5-F75), an effect that was more pronounced in males than females. In Experiment 3 and 4, male mice fed versions of the C40-F40 formulation gained more weight than did those fed the C75-F5 or C5-F75 formulations irrespective of whether the carbohydrate was predominantly sucrose or predominantly starch, or whether the fat was vegetable shortening, corn oil, palm oil or canola oil; the type of carbohydrate or fat had little or no impact on body weight. In all four experiments, energy intakes differed among the diet groups but could not account for the differences in body weight. These results demonstrate that the proportion of carbohydrate and fat in the diet influences body weight independently of energy content, and that the type of carbohydrate or fat has little impact on body weight. Consuming carbohydrate and fat simultaneously or in close temporal proximity exacerbates obesity., (Copyright © 2021 Elsevier Inc. All rights reserved.)

A major conceptual gap in taste biology is the lack of a general framework for understanding the evolution of different taste modalities among animal species. We turn to two complementary nutritional frameworks, biological stoichiometry theory and nutritional geometry, to develop hypotheses for the evolution of different taste modalities in animals. We describe how the attractive tastes of Na-, Ca-, P-, N-, and C-containing compounds are consistent with principles of both frameworks based on their shared focus on nutritional imbalances and consumer homeostasis. Specifically, we suggest that the evolution of multiple nutritive taste modalities can be predicted by identifying individual elements that are typically more concentrated in the tissues of animals than plants. Additionally, we discuss how consumer homeostasis can inform our understanding of why some taste compounds (i.e., Na, Ca, and P salts) can be either attractive or aversive depending on concentration. We also discuss how these complementary frameworks can help to explain the evolutionary history of different taste modalities and improve our understanding of the mechanisms that lead to loss of taste capabilities in some animal lineages. The ideas presented here will stimulate research that bridges the fields of evolutionary biology, sensory biology, and ecology., Competing Interests: The authors of this manuscript declare no competing interests between the listed authors, institutions, or any third parties concerning this manuscript., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

We have previously used crosses between C57BL/6ByJ (B6) and 129P3/J (129) inbred strains to map a quantitative trait locus (QTL) on mouse chromosome (Chr) 4 that affects behavioral and neural responses to sucrose. We have named it the sucrose consumption QTL 2 (Scon2), and shown that it corresponds to the Tas1r3 gene, which encodes a sweet taste receptor subunit TAS1R3. To discover other sucrose consumption QTLs, we have intercrossed B6 inbred and 129.B6-Tas1r3 congenic mice to produce F 2 hybrids, in which Scon2 (Tas1r3) does not segregate, and hence does not contribute to phenotypical variation. Chromosome mapping using this F 2 intercross identified two main-effect QTLs, Scon3 (Chr9) and Scon10 (Chr14), and an epistatically interacting QTL pair Scon3 (Chr9)-Scon4 (Chr1). Using serial backcrosses, congenic and consomic strains, we conducted high-resolution mapping of Scon3 and Scon4 and analyzed their epistatic interactions. We used mice with different Scon3 or Scon4 genotypes to understand whether these two QTLs influence sucrose intake via gustatory or postoral mechanisms. These studies found no evidence for involvement of the taste mechanisms, but suggested involvement of energy metabolism. Mice with the B6 Scon4 genotype drank less sucrose in two-bottle tests, and also had a higher respiratory exchange ratio and lower energy expenditure under basal conditions (when they had only chow and water available). Our results provide evidence that Scon3 and Scon4 influence mouse-to-mouse variation in sucrose intake and that both likely act through a common postoral mechanism.

Mice of the C57BL/6ByJ (B6) strain have higher consumption of sucrose, and stronger peripheral neural responses to it, than do mice of the 129P3/J (129) strain. To identify quantitative trait loci (QTLs) responsible for this strain difference and to evaluate the contribution of peripheral taste responsiveness to individual differences in sucrose intake, we produced an intercross (F 2 ) of 627 mice, measured their sucrose consumption in two-bottle choice tests, recorded the electrophysiological activity of the chorda tympani nerve elicited by sucrose in a subset of F 2 mice, and genotyped the mice with DNA markers distributed in every mouse chromosome. We confirmed a sucrose consumption QTL (Scon2, or Sac) on mouse chromosome (Chr) 4, harboring the Tas1r3 gene, which encodes the sweet taste receptor subunit TAS1R3 and affects both behavioral and neural responses to sucrose. For sucrose consumption, we also detected five new main-effect QTLs, Scon6 (Chr2), Scon7 (Chr5), Scon8 (Chr8), Scon3 (Chr9), and Scon9 (Chr15), and an epistatically interacting QTL pair Scon4 (Chr1) and Scon3 (Chr9). No additional QTLs for the taste nerve responses to sucrose were detected besides Scon2 (Tas1r3) on Chr4. Identification of the causal genes and variants for these sucrose consumption QTLs may point to novel mechanisms beyond peripheral taste sensitivity that could be harnessed to control obesity and diabetes.

Binding of sweet, umami, and bitter tastants to G protein-coupled receptors (GPCRs) in apical membranes of type II taste bud cells (TBCs) triggers action potentials that activate a voltage-gated nonselective ion channel to release ATP to gustatory nerves mediating taste perception. Although calcium homeostasis modulator 1 (CALHM1) is necessary for ATP release, the molecular identification of the channel complex that provides the conductive ATP-release mechanism suitable for action potential-dependent neurotransmission remains to be determined. Here we show that CALHM3 interacts with CALHM1 as a pore-forming subunit in a CALHM1/CALHM3 hexameric channel, endowing it with fast voltage-activated gating identical to that of the ATP-release channel in vivo. Calhm3 is co-expressed with Calhm1 exclusively in type II TBCs, and its genetic deletion abolishes taste-evoked ATP release from taste buds and GPCR-mediated taste perception. Thus, CALHM3, together with CALHM1, is essential to form the fast voltage-gated ATP-release channel in type II TBCs required for GPCR-mediated tastes., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Mechanical stimulation of airway epithelial cells causes apical release of ATP, which increases ciliary beat frequency (CBF) and speeds up mucociliary clearance. The mechanisms responsible for this ATP release are poorly understood. CALHM1, a transmembrane protein with shared structural features to connexins and pannexins, has been implicated in ATP release from taste buds, but it has not been evaluated for a functional role in the airway. In the present study, Calhm1 knockout, Panx1 knockout, and wild-type mouse nasal septal epithelial cells were grown at an air-liquid interface (ALI) and subjected to light mechanical stimulation from an air puff. Apical ATP release was attenuated in Calhm1 knockout cultures following mechanical stimulation at a pressure of 55 mmHg for 50 milliseconds (p < 0.05). Addition of carbenoxolone, a PANX1 channel blocker, completely abolished ATP release in Calhm1 knockout cultures but not in wild type or Panx1 knockout cultures. An increase in CBF was observed in wild-type ALIs following mechanical stimulation, and this increase was significantly lower (p < 0.01) in Calhm1 knockout cultures. These results demonstrate that CALHM1 plays a newly defined role, complementary to PANX1, in ATP release and downstream CBF modulation following a mechanical stimulus in airway epithelial cells.

Rodents consume solutions of phosphates and pyrophosphates in preference to water. Recently, we found that the preference for trisodium pyrophosphate (Na3HP2O7) was greater in T1R3 knockout (KO) mice than wild-type (WT) controls, suggesting that T1R3 is a pyrophosphate detector. We now show that this heightened Na3HP2O7 preference of T1R3 KO mice extends to disodium phosphate (Na2HPO4), disodium and tetrasodium pyrophosphate (Na2H2PO4 and Na4H2PO4), a tripolyphosphate (Na5P3O10), a non-sodium phosphate [(NH4)2HPO4], and a non-sodium pyrophosphate (K4P2O7) but not to non-P salts with large anions (sodium gluconate, acetate, or propionate). Licking rates for Na3HP2O7 are higher in T1R2 KO mice than WT controls; Na3HP2O7 preference scores are increased even more in T1R2 KO mice and T1R2+T1R3 double KO mice than in T1R3 KO mice; preference scores for Na3HP2O7 are normal in T1R1 KO mice. These results implicate each subunit of the T1R2+T1R3 dimer in the behavioral response to P-containing taste compounds., (© The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Many people avidly consume foods and drinks containing caffeine, despite its bitter taste. Here, we review what is known about caffeine as a bitter taste stimulus. Topics include caffeine's action on the canonical bitter taste receptor pathway and caffeine's action on noncanonical receptor-dependent and -independent pathways in taste cells. Two conclusions are that (1) caffeine is a poor prototypical bitter taste stimulus because it acts on bitter taste receptor-independent pathways, and (2) caffeinated products most likely stimulate "taste" receptors in nongustatory cells. This review is relevant for taste researchers, manufacturers of caffeinated products, and caffeine consumers., Competing Interests: No competing financial interests exist.

Does eating good-tasting food influence body weight? To investigate, we first established some concentrations of sucralose and mineral oil in chow that mice strongly preferred. Then, in Experiment 1, we compared groups of 16 mice fed plain chow (i.e., chow with no additives) to groups fed chow with added (a) sucralose, (b) mineral oil, (c) sucralose and mineral oil, or (d) sucralose on odd days and mineral oil on even days. During a 6-week test, the body weights and body compositions of the five groups never differed. In Experiment 2, we compared groups of 18 mice fed plain chow or plain high-fat diet to groups fed these diets with added sucralose. During a 9-week test, the high-fat diet caused weight gain, but the body weights of mice fed the sucralose-sweetened diets did not differ from those fed the corresponding plain versions. Two-cup choice tests conducted at the end of each experiment showed persisting strong preferences for the diets with added sucralose and/or mineral oil. In concert with earlier work, our results challenge the hypothesis that the orosensory properties of a food influence body weight gain. A good taste can stimulate food intake acutely, and guide selection toward nutrient-dense foods that cause weight gain, but it does not determine how much is eaten chronically., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Taste intensity and quality affect the liking of foods, and determine food choice and consumption. We aimed to 1) classify commonly consumed foods based on recalled taste intensity for bitter, sweet, salty, sour, and fatty taste, and 2) examine the associations among recalled taste intensity, liking, and habitual consumption of foods. In Stage 1, 62 Canadian adults recalled the taste intensity of 120 common foods. Their responses were used to identify sets of 20-25 foods classified as strongly bitter, sweet, salty, sour or fatty-tasting. In Stage 2, 287 U.S. adults validated these selections, and let us reduce them to sets of 11-13 foods. Ratings of recalled taste intensity were consistent across age, sex and overweight status, with the exceptions that sweet, bitter and fatty-tasting foods were rated as more intense by women than by men. The recalled intensity ratings of the most bitter, salty and fatty foods (but not sour or sweet foods) were inversely correlated with liking and intake. The negative correlation between fatty taste intensity and fatty food liking was stronger among normal weight than among overweight participants. Our results suggest that the recalled taste intensity of foods is associated with food liking and habitual consumption, but the strength of these relationships varies by taste. The food lists based on taste intensity ratings provide a resource to efficiently calculate indices of exposure to the different tastes in future studies., Competing Interests: None., (Copyright © 2016. Published by Elsevier Ltd.)

Obesity is a major public health problem. An in-depth knowledge of the molecular mechanisms of oro-sensory detection of dietary lipids may help fight it. Humans and rodents can detect fatty acids via lipido-receptors, such as CD36 and GPR120. We studied the implication of the MAPK pathways, in particular, ERK1/2, in the gustatory detection of fatty acids. Linoleic acid, a dietary fatty acid, induced via CD36 the phosphorylation of MEK1/2-ERK1/2-ETS-like transcription factor-1 cascade, which requires Fyn-Src kinase and lipid rafts in human taste bud cells (TBCs). ERK1/2 cascade was activated by Ca 2+ signaling via opening of the calcium-homeostasis modulator-1 (CALHM1) channel. Furthermore, fatty acid-evoked Ca 2+ signaling and ERK1/2 phosphorylation were decreased in both human TBCs after small interfering RNA knockdown of CALHM1 channel and in TBCs from Calhm1 -/- mice. Targeted knockdown of ERK1/2 by small interfering RNA or PD0325901 (MEK1/2 inhibitor) in the tongue and genetic ablation of Erk1 or Calhm1 genes impaired preference for dietary fat in mice. Lingual inhibition of ERK1/2 in healthy volunteers also decreased orogustatory sensitivity for linoleic acid. Our data demonstrate that ERK1/2-MAPK cascade is regulated by the opening of CALHM1 Ca 2+ channel in TBCs to modulate orogustatory detection of dietary lipids in mice and humans.-Subramaniam, S., Ozdener, M. H., Abdoul-Azize, S., Saito, K., Malik, B., Maquart, G., Hashimoto, T., Marambaud, P., Aribi, M., Tordoff, M. G., Besnard, P., Khan, N. A. ERK1/2 activation in human taste bud cells regulates fatty acid signaling and gustatory perception of fat in mice and humans., (© FASEB.)

The consumption of amino acids by animals is controlled by both oral and postoral mechanisms. We used a genetic approach to investigate these mechanisms. Our studies have shown that inbred mouse strains differ in voluntary amino acid consumption, and these differences depend on sensory and nutritive properties of amino acids. Like humans, mice perceive some amino acids as having a sweet (sucrose-like) taste and others as having an umami (glutamate-like) taste. Mouse strain differences in the consumption of some sweet-tasting amino acids (d-phenylalanine, d-tryptophan, and l-proline) are associated with polymorphisms of a taste receptor, type 1, member 3 gene (Tas1r3), and involve differential peripheral taste responsiveness. Strain differences in the consumption of some other sweet-tasting amino acids (glycine, l-alanine, l-glutamine, and l-threonine) do not depend on Tas1r3 polymorphisms and so must be due to allelic variation in other, as yet unknown, genes involved in sweet taste. Strain differences in the consumption of l-glutamate may depend on postingestive rather than taste mechanisms. Thus, genes and physiologic mechanisms responsible for strain differences in the consumption of each amino acid depend on the nature of its taste and postingestive properties. Overall, mouse strain differences in amino acid taste and appetite have a complex genetic architecture. In addition to the Tas1r3 gene, these differences depend on other genes likely involved in determining the taste and postingestive effects of amino acids. The identification of these genes may lead to the discovery of novel mechanisms that regulate amino acid taste and appetite., (© 2016 American Society for Nutrition.)

Rodents are strongly attracted to the taste(s) of maltodextrins. A first step toward discovery of the underlying genes involves identifying phenotypic differences among inbred strains of mice. To do this, we used 5-s brief-access tests and 48-h 2-bottle choice tests to survey the avidity for the maltodextrin, Maltrin M040, of mice from 8 inbred strains (129S1/SvImJ, A/J, CAST/EiJ, C57BL/6J, NOD/ShiLTJ, NZO/HlLtJ, PWK/PhJ, and WSB/EiJ). In brief-access tests, the CAST and PWK strains licked significantly less maltodextrin than equivalent concentrations of sucrose, whereas the other strains generally licked the 2 carbohydrates equally. Similarly, in 2-bottle choice tests, the CAST and PWK strains drank less 4% maltodextrin than 4% sucrose, whereas the other strains had similar intakes of these 2 solutions; the CAST and PWK strains did not differ from the C57, NOD, or NZO strains in 4% sucrose intake. In sum, we have identified strain variation in maltodextrin perception that is distinct from variation in sucrose perception. The phenotypic variation characterized here will aid in identifying genes responsible for maltodextrin acceptance. Our results identify C57 × PWK mice or NZO × CAST mice as informative crosses to produce segregating hybrids that will expose quantitative trait loci underlying maltodextrin acceptance and preference., (© The Author 2015. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Consuming a fructose-rich diet leads to hyperinsulinemia, impaired glucose tolerance, and insulin resistance. In humans, the consumption of high levels of refined sugars often coincides with a diet containing suboptimal levels of calcium. Calcium and carbohydrate metabolism interact, so there is potential for fructose to have different health outcomes depending on whether the diet is calcium-rich or calcium-poor., Methods: We evaluated the metabolic effects of feeding fructose to rats that were maintained on either a calcium-replete diet or a low-calcium diet. Growing male Sprague Dawley rats were fed diets based on the AIN-93G formulation, with the main source of carbohydrate derived either from a mixture of cornstarch and sucrose or from fructose. Half the rats given each carbohydrate source were fed calcium at recommended levels (125 mmol/kg Ca(2+)); the others were fed a diet low in calcium (25 mmol/kg Ca(2+)). At various times, glucose and insulin tolerance tests were conducted to assess glucose metabolism., Results: Rats fed low-calcium diet had lower fasting insulin levels irrespective of the carbohydrate source they ate. They had a normal glycemic response to a glucose load and did not develop hyperinsulinemia under conditions of fructose feeding. The drop in blood glucose levels in response to insulin injection was larger in rats fed low-calcium diet than in those fed calcium-replete diet., Conclusions: Low-calcium diet prevented fructose-induced hyperinsulinemia and improved glucose handling under conditions of fructose feeding. Potential mechanisms underlying these effects of the low-calcium diet remain to be determined, but possibilities include impairment of insulin release from the pancreas and improved peripheral insulin sensitivity.

Taste compounds detected by G protein-coupled receptors on the apical surface of Type 2 taste cells initiate an intracellular molecular cascade culminating in the release of ATP. It has been suggested that this ATP release is accomplished by pannexin 1 (PANX1). However, we report here that PANX1 knockout mice do not differ from wild-type controls in response to representative taste solutions, measured using 5-s brief-access tests or 48-h two-bottle choice tests. This implies that PANX1 is unnecessary for taste detection and consequently that ATP release from Type 2 taste cells does not require PANX1., (© The Author 2015. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Neurons that fire in bursts have been well-characterized in vision and other neural systems, but not in taste systems. We therefore examined whether brain stem gustatory neurons fire in bursts during spontaneous activity and, if so, whether such cells differ from nonbursting cells in other characteristics. We looked at neurons in the nucleus of the solitary tract (NST) of C57BL/6ByJ (B6) and 129P3/J (129) mice, and in the NST and parabrachial nucleus (PBN) of Sprague-Dawley rats. Many NST cells fired frequently with short intervals characteristic of bursting, and such neurons differed from others in their responsiveness to taste compounds. In B6 mice and rats, there was a significant positive correlation between the prevalence of short-interval firing and the net spikes evoked by application of NaCl. In contrast, in 129 mice the prevalence of short intervals was positively correlated with the size of sucrose responses. We also compared breadth-of-tuning measures based on counting either all spikes or only those following short intervals, and we found narrower tuning for the latter in the NST of B6 mice and rats. There was little evidence of spontaneous bursting in the rat PBN, and firing patterns in this nucleus were not related to the size of taste-evoked responses. We suggest that bursting may be a strategy employed by the NST to amplify the postsynaptic impact of particular taste stimuli, depending on an animal's needs. Another function may be to sharpen breadth-of-tuning and thus enhance the contrast between stimuli of different taste qualities., (Copyright © 2015 the American Physiological Society.)

Laboratory rats and mice prefer some concentrations of tri- and tetrasodium pyrophosphate (Na3HP2O7 and Na4P2O7) to water, but how they detect pyrophosphates is unknown. Here, we assessed whether T1R3 is involved. We found that relative to wild-type littermate controls, Tas1r3 knockout mice had stronger preferences for 5.6-56mM Na3HP2O7 in 2-bottle choice tests, and they licked more 17.8-56mM Na3HP2O7 in brief-access tests. We hypothesize that pyrophosphate taste in the intact mouse involves 2 receptors: T1R3 to produce a hedonically negative signal and an unknown G protein-coupled receptor to produce a hedonically positive signal; in Tas1r3 knockout mice, the hedonically negative signal produced by T1R3 is absent, leading to a heightened avidity for pyrophosphate., (© The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Many animals thrive when given a choice of separate sources of macronutrients. How they do this is unknown. Here, we report some studies comparing the spontaneous choices between carbohydrate- and fat-containing food sources of seven inbred mouse strains (B6, BTBR, CBA, JF1, NZW, PWD and PWK) and three mouse models with genetic ablation of taste transduction components (T1R3, ITPR3 and CALHM1). For 8days, each mouse could choose between sources of carbohydrate (CHO-P; sucrose-cornstarch) and fat (Fat-P; vegetable shortening) with each source also containing protein (casein). We found that the B6 and PWK strains markedly preferred the CHO-P diet to the Fat-P diet, the BTBR and JF1 strains markedly preferred the Fat-P diet to the CHO-P diet, and the CBA, NZW and PWD strains showed equal intakes of the two diets (by weight). Relative to their WT littermates, ITPR3 and CALHM1 KO mice had elevated Fat-P preferences but T1R3 KO mice did not. There were differences among strains in adaption to the diet choice and there were differences in response between males and females on some days. These results demonstrate the diverse responses to macronutrients of inbred mice and they point to the involvement of chemosensory detectors (but not sweetness) as contributors to macronutrient selection., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Genetic ablation of calcium homeostasis modulator 1 (CALHM1), which releases adenosine triphosphate from Type 2 taste cells, severely compromises the behavioral and electrophysiological responses to tastes detected by G protein-coupled receptors, such as sweet and bitter. However, the contribution of CALHM1 to salty taste perception is less clear. Here, we evaluated several salty taste-related phenotypes of CALHM1 knockout (KO) mice and their wild-type (WT) controls: 1) In a conditioned aversion test, CALHM1 WT and KO mice had similar NaCl avoidance thresholds. 2) In two-bottle choice tests, CALHM1 WT mice showed the classic inverted U-shaped NaCl concentration-preference function but CALHM1 KO mice had a blunted peak response. 3) In brief-access tests, CALHM1 KO mice showed less avoidance than did WT mice of high concentrations of NaCl, KCl, NH(4)Cl, and sodium lactate (NaLac). Amiloride further ameliorated the NaCl avoidance of CALHM1 KO mice, so that lick rates to a mixture of 1000 mM NaCl + 10 µM amiloride were statistically indistinguishable from those to water. 4) Relative to WT mice, CALHM1 KO mice had reduced chorda tympani nerve activity elicited by oral application of NaCl, NaLac, and sucrose but normal responses to HCl and NH(4)Cl. Chorda tympani responses to NaCl and NaLac were amiloride sensitive in WT but not KO mice. These results reinforce others demonstrating that multiple transduction pathways make complex, concentration-dependent contributions to salty taste perception. One of these pathways depends on CALHM1 to detect hypertonic NaCl in the mouth and signal the aversive taste of concentrated salt., (© The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We investigated how preferred and nonpreferred tastes influence the disposition of fat. Adult male Sprague Dawley rats were infused with 5 ml of 20% intralipid through an intragastric catheter and with 0.3 ml of a taste solution through an intraoral catheter. At 120 min postinfusion, plasma concentrations of fat fuels (triglycerides and non-esterified fatty acids) were either unchanged or slightly higher after rats tasted a preferred sweet taste solution (0.125% saccharin +3% glucose) than after they tasted water. They were markedly lower after rats tasted a non-preferred solution-either a bitter solution (0.15% quinine hydrochloride) or a sweet solution that had previously been the conditioned stimulus for lithium-induced taste aversion. The distribution of 14C-triolein mixed with the gastric load was determined at 4 h postinfusion. Rats that received a non-preferred bitter taste had significantly more 14C remaining in the stomach than did those that received a preferred sweet taste. These results suggest that taste hedonics--either unconditioned or conditioned aversive tastes--influence fat disposition by altering gastric emptying.

We investigated whether maternal influences during the suckling period alter the avidity for calcium, using as models mice from the calcium-preferring PWK/PhJ strain and the calcium-avoiding C57BL/6J strain. We found that milk collected from PWK/PhJ dams had higher calcium concentrations than did milk collected from C57BL/6J dams. Despite this, cross-strain fostering had no effect on adult calcium preferences relative to mice of the same strain that were within-strain fostered or not fostered. Our results indicate that calcium avoidance by C57BL/6J mice and acceptance by PWK/PhJ mice are unaffected by maternal environment during the suckling period., (© 2013.)

The BTBR T+ tf/J (BTBR) mouse strain is indifferent to exemplars of sweet, Polycose, umami, bitter, and calcium tastes, which share in common transduction by G protein-coupled receptors (GPCRs). To investigate the genetic basis for this taste dysfunction, we screened 610 BTBR×NZW/LacJ F2 hybrids, identified a potent QTL on chromosome 17, and isolated this in a congenic strain. Mice carrying the BTBR/BTBR haplotype in the 0.8-Mb (21-gene) congenic region were indifferent to sweet, Polycose, umami, bitter, and calcium tastes. To assess the contribution of a likely causative culprit, Itpr3, the inositol triphosphate receptor 3 gene, we produced and tested Itpr3 knockout mice. These were also indifferent to GPCR-mediated taste compounds. Sequencing the BTBR form of Itpr3 revealed a unique 12 bp deletion in Exon 23 (Chr 17: 27238069; Build 37). We conclude that a spontaneous mutation of Itpr3 in a progenitor of the BTBR strain produced a heretofore unrecognized dysfunction of GPCR-mediated taste transduction.

Obesity is a heritable trait caused by complex interactions between genes and environment, including diet. Gene-by-diet interactions are difficult to study in humans because the human diet is hard to control. Here, we used mice to study dietary obesity genes, by four methods. First, we bred 213 F2 mice from strains that are susceptible [C57BL/6ByJ (B6)] or resistant [129P3/J (129)] to dietary obesity. Percent body fat was assessed after mice ate low-energy diet and again after the same mice ate high-energy diet for 8 weeks. Linkage analyses identified QTLs associated with dietary obesity. Three methods were used to filter candidate genes within the QTL regions: (a) association mapping was conducted using >40 strains; (b) differential gene expression and (c) comparison of genomic DNA sequence, using two strains closely related to the progenitor strains from Experiment 1. The QTL effects depended on whether the mice were male or female or which diet they were recently fed. After feeding a low-energy diet, percent body fat was linked to chr 7 (LOD=3.42). After feeding a high-energy diet, percent body fat was linked to chr 9 (Obq5; LOD=3.88), chr 12 (Obq34; LOD=3.88), and chr 17 (LOD=4.56). The Chr 7 and 12 QTLs were sex dependent and all QTL were diet-dependent. The combination of filtering methods highlighted seven candidate genes within the QTL locus boundaries: Crx, Dmpk, Ahr, Mrpl28, Glo1, Tubb5, and Mut. However, these filtering methods have limitations so gene identification will require alternative strategies, such as the construction of congenics with very small donor regions.

The food, water and sodium intake of laboratory rats fluctuates over the circadian and estrous cycles. Blood calcium and calcium-regulating hormones also wax and wane in response to these cycles, raising the possibility that the same might be true of calcium intake. To investigate this, we monitored the fluid intakes of female Long-Evans rats given a choice between water and 10mM CaCl2 solution for two consecutive estrous cycles. We found that calcium solution intake changed over the circadian cycle in a similar manner to water intake; the preference scores for CaCl2 solution remained stable. We did not detect any changes in calcium solution intake or preference scores during the estrous cycle despite a decrease in fluid intake at estrus. Thus, fluctuations in intake of calcium solution during the circadian cycle appear to be nonspecific and probably the result of changes in fluid balance. Estrous changes either do not influence calcium intake or their effects are masked by other factors, resulting in stable levels of calcium intake., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Recognition of sweet, bitter and umami tastes requires the non-vesicular release from taste bud cells of ATP, which acts as a neurotransmitter to activate afferent neural gustatory pathways. However, how ATP is released to fulfil this function is not fully understood. Here we show that calcium homeostasis modulator 1 (CALHM1), a voltage-gated ion channel, is indispensable for taste-stimuli-evoked ATP release from sweet-, bitter- and umami-sensing taste bud cells. Calhm1 knockout mice have severely impaired perceptions of sweet, bitter and umami compounds, whereas their recognition of sour and salty tastes remains mostly normal. Calhm1 deficiency affects taste perception without interfering with taste cell development or integrity. CALHM1 is expressed specifically in sweet/bitter/umami-sensing type II taste bud cells. Its heterologous expression induces a novel ATP permeability that releases ATP from cells in response to manipulations that activate the CALHM1 ion channel. Knockout of Calhm1 strongly reduces voltage-gated currents in type II cells and taste-evoked ATP release from taste buds without affecting the excitability of taste cells by taste stimuli. Thus, CALHM1 is a voltage-gated ATP-release channel required for sweet, bitter and umami taste perception.

The tf (tufted) locus is responsible for a classic phenotype of hair loss and regrowth in mice. It is a characteristic of the BTBR strain. Here, we use a combination of positional cloning methods and complementation mapping to identify Itpr3, the inositol triphosphate receptor type 3, as the gene responsible for the tf locus.

We explored genetic influences on the perception of taste and smell stimuli. Adult twins rated the chemosensory aspects of water, sucrose, sodium chloride, citric acid, ethanol, quinine hydrochloride, phenylthiocarbamide (PTC), potassium chloride, calcium chloride, cinnamon, androstenone, Galaxolide™, cilantro, and basil. For most traits, individual differences were stable over time and some traits were heritable (h(2) from 0.41 to 0.71). Subjects were genotyped for 44 single nucleotide polymorphisms within and near genes related to taste and smell. The results of these association analyses confirmed previous genotype-phenotype results for PTC, quinine, and androstenone. New associations were detected for ratings of basil and a bitter taste receptor gene, TAS2R60, and between cilantro and variants in three genes (TRPA1, GNAT3, and TAS2R50). The flavor of ethanol was related to variation within an olfactory receptor gene (OR7D4) and a gene encoding a subunit of the epithelial sodium channel (SCNN1D). Our study demonstrates that person-to-person differences in the taste and smell perception of simple foods and drinks are partially accounted for by genetic variation within chemosensory pathways.

Monosodium glutamate (MSG) is pervasively consumed as a flavor enhancer so there are important implications to understanding its physiological actions, particularly its effects on body weight. Previous studies suggest that MSG increases, decreases, or has no effect on the body weight of rodents. However, most of these studies involved administration of MSG to immature rodents and consequently may not be relevant for understanding human obesity. We report here five experiments in which we measured the body weights of a total of 32 groups of 10-12 adult rats or mice given various diets to eat and MSG to eat or drink. We found no evidence that MSG influenced body weight, energy intake, or body composition. To the extent that experiments in rodents illuminate mechanisms involved in human obesity and body weight control, our results suggest that MSG is unlikely to be a useful anti-obesity supplement but neither is it responsible for exacerbating obesity., (Copyright © 2012 Elsevier Inc. All rights reserved.)

There has been scant work to investigate the mechanisms influencing macronutrient selection by mice. Here, we measured the consumption and choice of carbohydrate- and fat-containing diets by NZW/LacJ (NZW) and BTBR/T+ tf/J (BTBR) strains. We found that NZW mice voluntarily ate more carbohydrate and less fat than did BTBR mice. Mice with a BTBR background and a heterozygous (BTBR/NZW) congenic region on chromosome 17 between 25.7 and 27.5 Mb (N10 generation) or 26.7 and 27.5 Mb (N12 generation) also ate more carbohydrate and less fat than did homozygous (BTBR/BTBR) littermate controls. Of the 21 known and predicted genes in the congenic interval between 26.7 and 27.5 Mb, we raise for consideration as a causative candidate Itpr3, the inositol triphosphate receptor type 3 gene, which is a component of the GPCR-mediated taste transduction cascade. We speculate that a mutation in Itpr3 influences food choice by impairing the detection of nutrients in the macronutrient-containing diets., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Calcium intake depends on orosensory factors, implying the presence of a mechanism for calcium detection in the mouth. To better understand how information about oral calcium is conveyed to the brain, we examined the effects of chorda tympani nerve transection on calcium chloride (CaCl(2)) taste preferences and thresholds in male Wistar rats. The rats were given bilateral transections of the chorda tympani nerve (CTX) or control surgery. After recovery, they received 48-h two-bottle tests with an ascending concentration series of CaCl(2). Whereas control rats avoided CaCl(2) at concentrations of 0.1mM and higher, rats with CTX were indifferent to CaCl(2) concentrations up to 10mM. Rats with CTX had significantly higher preference scores for 0.316 and 3.16 mM CaCl(2) than did control rats. The results imply that the chorda tympani nerve is required for the normal avoidance of CaCl(2) solution., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Many animals can detect the taste of calcium but it is unclear how or whether humans have this ability. We show here that calcium activates hTAS1R3-transfected HEK293 cells and that this response is attenuated by lactisole, an inhibitor of hT1R3. Moreover, trained volunteers report that lactisole reduces the calcium intensity of calcium lactate. Thus, humans can detect calcium by taste, T1R3 is a receptor responsible for this, and lactisole can reduce the taste perception of calcium by acting on T1R3.

The goal of this study was to determine the adiposity of a range of rat strains, including a panel of consomics, to estimate heritability. To that end, we assessed the body fat distribution and organ weights of groups of adult male rats from 3 outbred strains, 11 inbred strains and 22 consomic strains. We measured the weights of the gonadal, retroperitoneal, mesenteric, femoral, subscapular and pericardial white fat depots, the subscapular brown fat depot, the kidneys, liver, heart, spleen, and brain. Strains were compared for the measured weight of each of these adipose depots and organs, and also for these weights adjusted statistically for body size. All individual adipose depot and organ weights were highly heritable, in most cases h(2)>0.50. The fourteen inbred and outbred rat strains were not very different in body length but there was a three-fold difference in body weight, and up to a twenty-fold difference in the weight of some adipose depots. Comparison of the FHH-Chr n(BN) consomic strains with the FHH host strain revealed 98 quantitative trait loci (QTLs) for body composition and organ weight, with the introgressed chromosome reducing weight or adiposity in most cases. These results can be used to guide the choice of appropriate rat strains for future studies of the genetic architecture of obesity and body size., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We used the C57BL/6J (B6) and PWD/PhJ (PWD) mouse strains to investigate the controls of calcium intake. Relative to the B6 strain, the PWD strain had higher preferences in two-bottle choice tests for CaCl(2), calcium lactate (CaLa), MgCl(2), citric acid and quinine hydrochloride, but not for sucrose, KCl or NaCl. We also measured taste-evoked chorda tympani (CT) nerve activity in response to oral application of these compounds. Electrophysiological results paralleled the preference test results, with larger responses in PWD than in B6 mice for those compounds that were more highly preferred for the former strain. The strain differences were especially large for tonic, rather than phasic, chorda tympani activity. These data establish the PWD strain as a "calcium-preferring" strain and suggest that differences between B6 and PWD mice in taste transduction or a related peripheral event contributes to the differences between the strains in preferences for calcium solutions., (Copyright © 2011 Elsevier B.V. All rights reserved.)

There has been extensive work to elucidate the behavioral and physiological mechanisms responsible for taste preferences of the rat but little attempt to delineate the underlying genetic architecture. Here, we exploit the FHH-Chr n(BN)/Mcwi consomic rat strain set to identify chromosomes carrying genes responsible for taste preferences. We screened the parental Fawn Hooded Hypertensive (FHH) and Brown Norway (BN) strains and 22 FHH-Chr n(BN) consomic strains, with 96-h 2-bottle tests, involving a choice between water and each of the following 16 solutions: 10 mM NaCl, 237 mM NaCl, 32 mM CaCl(2), 1 mM saccharin, 100 mM NH(4)Cl, 32 mM sucrose, 100 mM KCl, 4% ethanol, 1 mM HCl, 10 mM monosodium glutamate, 1 mM citric acid, 32 microM quinine hydrochloride, 1% corn oil, 32 microM denatonium, 1% Polycose, and 1 microM capsaicin. Depending on the taste solution involved, between 1 and 16 chromosomes were implicated in the response. Few of these chromosomes carried genes believed to mediate taste transduction in the mouse, and many chromosomes with no candidate taste genes were revealed. The genetic architecture of taste preferences is considerably more complex than has heretofore been acknowledged.

This article provides an overview of our studies of variation in voluntary glutamate consumption in mice. In 2-bottle preference tests, mice from the C57BL/6ByJ (B6) strain consume more monosodium l-glutamate (MSG) than do mice from the 129P3/J (129) strain. We used these mice to study physiologic and genetic mechanisms that underlie the strain differences in glutamate intake. Our genetic analyses showed that differences between B6 mice and 129 mice in MSG consumption are unrelated to strain variation in consumption of sodium or sweeteners and therefore are attributed to mechanisms specific for glutamate. These strain differences could be due to variation in responses to either taste or postingestive effects of glutamate. To examine the role of taste responsiveness, we measured MSG-evoked activity in gustatory nerves and showed that it is similar in B6 and 129 mice. On the other hand, strain-specific postingestive effects of glutamate were evident from our finding that exposure to MSG increases its consumption in B6 mice and decreases its consumption in 129 mice. We therefore examined whether B6 mice and 129 mice differ in postingestive metabolism of glutamate. We showed that, after intragastric administration of MSG, the MSG is preferentially metabolized through gluconeogenesis in B6 mice, whereas thermogenesis is the predominant process for 129 mice. We hypothesize that a process related to gluconeogenesis of the ingested glutamate generates the rewarding stimulus, which probably occurs in the liver before glucose enters the general circulation, and that the glutamate-induced postingestive thermogenesis generates an aversive stimulus. Our animal model studies raise the question of whether humans also vary in glutamate metabolism in a manner that influences their glutamate preference, consumption, and postingestive processing.

Clay consumption can occur during illness but there has been little work to understand why. To investigate whether consuming clay confers an advantage to the sick animal, we compared the recovery from illness of adult male rats with or without access to kaolin. Illness was induced by injection of 6 mg/kg, ip, cisplatin, a toxic chemotherapy agent, and recovery was assessed by changes in daily food intake, water intake, and body weight. Relative to saline-injected controls, cisplatin-injected rats reduced food and water intake and lost weight. However, those with access to kaolin ate more food and lost less body weight than did those without access to kaolin. Thus, clay consumption appeared beneficial in that it either protected the rats from illness or enhanced recovery and might prove useful as an adjunct therapy for other animals, including humans, experiencing visceral malaise.

In the U.S. and Europe, most people do not consume the recommended amounts of either calcium or vegetables. We investigated whether there might be a connection; specifically, whether the taste of calcium in vegetables contributes to their bitterness and thus acceptability. We found a strong correlation between the calcium content of 24 vegetables, based on USDA Nutrient Database values, and bitterness, based on the average ratings of 35 people (r = 0.93). Correlations between the content of other nutrients and bitterness were lower and most were not statistically significant. To assess whether it is feasible that humans can detect calcium in vegetables we tested two animal models known to display a calcium appetite. Previous work indicates that calcium solutions are preferentially ingested by PWK/PhJ mice relative to C57BL/6J mice, and by rats deprived of dietary calcium relative to replete controls. In choice tests between collard greens, a high-calcium vegetable, and cabbage, a low-calcium vegetable, the calcium-favoring animals had higher preferences for collard greens than did controls. These observations raise the possibility that the taste of calcium contributes to the bitterness and thus acceptability of vegetables.

Two-bottle choice tests were used to assess the taste preferences of 8 male and 8 female rats from 3 outbred strains (SD, LE, WI) and 11 inbred strains (BN, BUF, COP, DA, Dahl-S, F344, FHH, LEW, Noble, PVG, SHR). Each rat received a series of 109 48-h tests with a choice between water and a "taste solution". Four to eight concentrations of the following compounds were tested: NaCl, CaCl2, NH4Cl, KCl, MgCl2, saccharin, sucrose, ethanol, HCl, citric acid, quinine hydrochloride (QHCl), caffeine, denatonium, monosodium glutamate (MSG), Polycose, corn oil, and capsaicin. Strain differences (p<0.001) were observed in preferences for at least one concentration of all compounds tested except denatonium (p=0.0015). There were also strain differences in the following ancillary measures: fungiform papillae number, water intake, food intake, and body weight. There were sex differences in food intake and body weight but no concerted sex differences in any of the other measures, including preferences for any taste solution. This comprehensive source of information can be used to guide the choice of appropriate rat strains and taste solution concentrations for future genetic studies.

Calcium and magnesium are essential for survival but it is unknown how animals detect and consume enough of these minerals to meet their needs. To investigate this, we exploited the PWK/PhJ (PWK) strain of mice, which, in contrast to the C57BL/6J (B6) and other inbred strains, displays strong preferences for calcium solutions. We found that the PWK strain also has strong preferences for MgCl2 and saccharin solutions but not representative salty, sour, bitter, or umami taste compounds. A genome scan of B6 x PWK F2 mice linked a component of the strain difference in calcium and magnesium preference to distal chromosome 4. The taste receptor gene, Tas1r3, was implicated by studies with 129.B6ByJ-Tas1r3 congenic and Tas1r3 knockout mice. Most notably, calcium and magnesium solutions that were avoided by wild-type B6 mice were preferred (relative to water) by B6 mice null for the Tas1r3 gene. Oral calcium elicited less electrophysiological activity in the chorda tympani nerve of Tas1r3 knockout than wild-type mice. Comparison of the sequence of Tas1r3 with calcium and saccharin preferences in inbred mouse strains found 1) an inverse correlation between calcium and saccharin preference scores across primarily domesticus strains, which was associated with an I60T substitution in T1R3, and 2) a V689A substitution in T1R3 that was unique to the PWK strain and thus may be responsible for its strong calcium and magnesium preference. Our results imply that, in addition to its established roles in the detection of sweet and umami compounds, T1R3 functions as a gustatory calcium-magnesium receptor.

This review makes the case that gene discovery is a worthwhile approach to the study of ingestive behavior in general and to calcium appetite in particular. A description of the methods used to discover genes is provided for non-geneticists. Areas covered include the characterization of an appropriate phenotype, the choice of suitable mouse strains, the generation of a hybrid cross, interval mapping, congenic strain production, and candidate gene analysis. The approach is illustrated with an example involving mice of the C57BL/6J and PWK/PhJ strains, which differ in avidity for calcium solutions. The variation between the strains can be attributed to at least seven quantitative trait loci (QTLs). One of these QTLs is most likely accounted for by Tas1r3, which is a gene involved in the detection of sweet and umami tastes. The discovery of a novel function for a gene with no previously known role in calcium consumption illustrates the power of gene discovery methods to uncover novel mechanisms.

To characterize the genetic basis of voluntary calcium consumption, we tested C57BL/6J mice (B6; with low avidity for calcium), PWK/PhJ mice (PWK; with high avidity for calcium) and their F(1) and F(2) hybrids. All mice received a series of 96-h two-bottle preference tests with a choice between water and the following: 50 mm CaCl(2), 50 mm calcium lactate, 50 mm MgCl(2), 100 mm KCl, 100 mm NH(4)Cl, 100 mm NaCl, 5 mm citric acid, 30 microm quinine hydrochloride and 2 mm saccharin. Most frequency distributions of the parental and F(1) but not F(2) groups were normally distributed, and there were few sex differences. Reciprocal cross analysis showed that B6 x PWK F(1) mice had a non-specific elevation of fluid intake relative to PWK x B6 F(1) mice. In the F(2) mice, trait correlations were clustered among the divalent salts and the monovalent chlorides. A genome screen involving 116 markers showed 30 quantitative trait loci (QTLs), of which six involved consumption of calcium chloride or lactate. The results show pleiotropic controls of calcium and magnesium consumption that are distinct from those controlling consumption of monovalent chlorides or exemplars of the primary taste qualities.

There has been little work on the specificity and mechanisms underlying the appetite of potassium (K(+)) deprived rats, and there are conflicting results. To investigate the contribution of oral factors to changes in intake induced by K(+) deficiency, we conducted two experiments using 20-s "brief access" tests. In Experiment 1, K(+)-deprived rats licked less for water than did replete rats. After adjusting for this difference, K(+)-deprived rats exhibited increased licking for 100 mM CaCl(2), 100 mM MgCl(2), and 100 mM FeCl(2) compared with K(+)-replete rats. In Experiment 2, which used larger rats, the K(+)-deprived and replete groups licked equally for water, 500 mM Na.Gluconate, 350 mM KCl, 500 mM KHCO(3), and 1 mM quinine.HCl, but the K(+)-deprived rats licked more for 500 mM KCl, 500 mM CsCl, and 500 mM NaCl than did the replete rats. Licking was unaffected by addition to NaCl of 200 muM amiloride, an epithelial Na(+) channel (ENaC) blocker, or 100 muM ruthenium red, a vanilloid receptor 1 (VR-1) antagonist, or by addition to KCl of 50 muM 4-aminopyridine, a K(+) channel blocker. These findings suggest that K(+)-deprivation produces a non-specific appetite that is guided by oral factors. We found no evidence that this response was mediated by ENaC, VR-1, or K(+) channels in taste receptor cells.

Background: During a search for obesity candidate genes in a small region of the mouse genome, we noticed that many genes when knocked out influence body weight. To determine whether this was a general feature of gene knockout or a chance occurrence, we surveyed the Jackson Laboratory Mouse Genome Database for knockout mouse strains and their phenotypes. Body weights were not available for all strains so we also obtained body weight information by contacting a random sample of investigators responsible for a knockout strain., Results: We classified each knockout mouse strain as (1) lighter and smaller, (2) larger and heavier, or (3) the same weight, relative to control mice. We excluded knockout strains that died early in life, even though this type of lethality is often associated with a small embryo or reduced body size. Based on a dataset of 1,977 knockout strains, we found that that 31% of viable knockout mouse strains weighed less and an additional 3% weighed more than did controls., Conclusion: Body weight is potentially a latent variable in about a third of experiments that use knockout mice and should be considered in interpreting experimental outcomes, e.g., in studies of hypertension, drug and hormone metabolism, organ development, cell proliferation and apoptosis, digestion, heart rate, or atherosclerosis. If we assume that the knockout genes we surveyed are representative then upward of 6,000 genes are predicted to influence the size of a mouse. Body weight is highly heritable, and numerous quantitative trait loci have been mapped in mice, but "multigenic" is an insufficient term for the thousands of loci that could contribute to this complex trait.

To determine the genetic variation that contributes to body composition in the mouse, we interbred a wild-derived strain (PWK/PhJ; PWK) with a common laboratory strain (C57BL/6J; B6). The parental, F(1), and F(2) mice were phenotyped at 18 weeks old for body weight and composition using dual-energy X-ray absorptiometry (DEXA). A total of 479 (244 male and 235 female) F(2) mice were genotyped for 117 polymorphic markers spanning the autosomes. Twenty-eight suggestive or significant linkages for four traits (body weight, adjusted lean and fat weight, and percent fat) were detected. Of these, three QTLs were novel: one on the proximal portion of Chr 5 for body weight (Bwq8; LOD = 4.7), one on Chr 3 for lean weight (Bwtq13; LOD = 3.6), and one on Chr 11 for percent fat (Adip19; LOD = 5.8). The remaining QTLs overlapped previously identified linkages, e.g., Adip5 on Chr 9. One QTL was sex-specific (present in males only) and seven were sex-biased (more prominent in one sex than the other). Most alleles that increased body weight were contributed by the B6 strain, and most alleles that increased percent fat were contributed by the PWK strain. Eight pairs of interacting loci were identified, none of which exactly overlapped the main-effect QTLs. Many of the QTLs found in the B6 x PWK cross map to the location of previously reported linkages, suggesting that some QTLs are common to many strains (consensus QTLs), but three new QTLs appear to be particular to the PWK strain. The location and type of QTLs detected in this new cross will assist in future efforts to identify the genetic variation that determines the ratio of lean to fat weight as well as body size in mice.

To examine whether age influences taste solution preferences, we measured taste preferences of C57BL/6J and 129X1/SvJ mice given a series of 48-h 2-bottle tests with a choice between water and one of the following taste solutions: 2 mM saccharin, 5 mM citric acid, 30 microM quinine hydrochloride, 75 mM sodium chloride (NaCl), 10 mM inosine monophosphate (IMP), 50 mM calcium chloride (CaCl(2)), and 10% ethanol. We tested separate groups of male mice fed Teklad 8604 chow at ages 4, 6, 9, 12, 15, 20, 25, 30, 40, and 50 weeks and retested some of these mice at 54, 75, and 100 weeks and again at 125 weeks. Female mice fed chow were tested at ages 4, 12, 25, and 50 weeks and retested at 54, 75, 100, and 125 weeks. Male mice fed AIN-93G semisynthetic diet were tested at ages 4, 12, 25, and 50 weeks and retested at 54, 75, and 100 weeks. Concentration-response functions for each taste solution were collected from male and female mice fed chow aged 8 or 125 weeks. In general, the results showed that age had little effect on taste preferences. Exceptions included 1) a small increase in quinine hydrochloride preference between 54 and 125 weeks in mice of both strains and sexes, 2) a marked increase in NaCl preference between 4 and 12 weeks in female B6 mice, 3) a gradual decrease in IMP preference between 4 and 125 weeks in male and female 129 mice, 4) a marked decrease in CaCl(2) preference between 54 and 125 weeks in male and female 129 mice, and 5) a marked reduction in ethanol preference between 4 and 12 weeks in male B6 mice fed AIN-93G diet but not chow. These results show that over a wide range and with the exceptions noted, age contributes little to the variation in taste preferences observed in C57BL/6J and 129X1/SvJ mice.