Polygenic determination of quinine aversion among mice

There are substantial differences among inbred mouse strainsin avoidance of quinine solutions in two-bottle preference tests.x A Mendelian cross-breeding experiment was conducted to testthe hypothesis that a single locus Qui has a major influenceon quinine aversion. Inbred strains C57B1/6J (B6, avoider) andC3HeB/FeJ (C3, indifferent) were progenitors of two segregatinggenerations. Phenotypic ratios for 100 µM and 30 µMquinine sulfate (QSO4) in these generations were not consistentwith ratios expected for a single gene. Frequency distributionsfor individual preference ratios were more characteristic ofa polygenic trait. An outbred strain (CFW/Crl) which displayssegregation for the Soa locus was tested for both QSO4 and sucroseocta-acetate (SOA) avoidance. Correlated avoidance patternsfor the two bitter compounds were found in these mice. A Soaeffect might not have been seen in the C3.B6 cross because bothstrains are relatively poor SOA avoiders. A second Mendeliancross was made between strains C3 and SWR/J (SW, SOA and QSO4avoider). One segregating generation was tested with both compounds.In these mice, as in the CFW population, QSO4 aversion was correlatedwith SOA aversion. These results suggest that quinine aversionis polygenic, that there is a relationship between SOA sensitivityand quinine sensitivity, and that this association may be theresult of variation at the Soa locus.     

A brief-access test for bitter taste in mice

Inbred mouse strains vary in their response to bitter-tasting compounds as assessed by 48 h preference tests. These differences are generally assumed to result from altered gustatory function, although such long-term tests could easily reflect additional factors. We developed a brief-access taste test and tested the responses of two inbred strains, as well as C3. SW congenic mice, to the bitter stimulus sucrose octaacetate (SOA). Water-deprived trained mice were tested with five concentrations of SOA (0.00018-0.18 mM) and distilled water in a Davis MS- 160 apparatus. Trials were 5 s in duration and stimuli were presented randomly within blocks; each stimulus trial was preceded by a water rinse trial. Each concentration was presented twice in a session and mice were repeatedly tested across consecutive days. SOA-taster mice, including the SWR/J (SW) inbred and C3. SW congenic taster (T) mice, avoided licking SOA at concentrations >0.003 mM. In comparison, C3HeB/FeJ (C3) and C3. SW demitaster mice (D) licked all concentrations at the same rate as water. Concentration-response functions were similar across strains for both the brief-access test and a parallel 48 h preference test run on separate groups of mice. Furthermore, concentration-response functions were similar whether or not the brief-access test was preceded by a 4 day, single concentration pretest with SOA. The brief-access test is a suitable assay for bitter taste function in mice because it minimizes possible post-ingestive influences on taste.     

SW.B6-Soab Nontaster Congenic Strains Completed and a Sucrose Octaacetate Congenic Quartet Tested with other Bitters

Ten SW.B6 SOA nontaster strains congenic with the SWR/J SOAtaster inbred strain were bred via repeated backcross-intercrosscycles, with selection for nontasting in each cycle. Preferenceratio distributions and phenotypic proportions across cyclesat 0.1 mM SOA were consistent with monogenic predictions. TheSW.B6 mice completed a congenic quartet with the SWR/J, B6.SWSOA taster and C57BL/6J SOA nontaster strains. The Soa locuscontrolled avoidance differences within the quartet for SOA,raffinose undecaacetate, glucose pentaacetate and brucine. Backgroundgenes not linked to Soa controlled avoidance differences forL-phenylalanine and ethanol. Avoidance of bitter picric acidwas influenced by the Soa locus, but avoidance of acetic acidwas not. The quartet pattern for quinine HCl was unclear, withindications of both Soa and background effects. Two forms ofribose tetraacetate yielded different patterns. Avoidance differencescontrolled by the Soa locus were found for the pyranose form;however, all four strains avoided the furanose form. The pleiotropiceffects of Soa allele substitution within the quartet were limitedto a subset of bitter compounds. Chem. Senses 21: 507–517,1996.     

A common polygenic basis for quinine and PROP avoidance in mice

Inbred strains of mice (Mus musculus) differ greatly in ability to taste various bitter compounds. For some compounds, the differences result from allelic variation at a single locus. However, segregation patterns incompatible with monogenic inheritance have been found for quinine avoidance. The Soa bitter sensitivity locus exerts some influence on this phenotype, but an unknown number of other loci also contribute. Relative avoidance patterns for quinine sulfate in panels of naive inbred strains resembled avoidance patterns for 6-n-propyl-2- thiouracil (PROP), suggesting a common genetic basis. In particular, C57BL/6J mice strongly avoided both 0.1 mM quinine sulfate and 1 mM PROP in two-bottle preference tests, whereas C3H/HeJ mice were indifferent to both. Therefore, 12 BXH/Ty recombinant inbred strains, derived from these strains, were tested with both solutions to begin identification of the unknown bitter loci. Naive mice were tested for four consecutive days with each compound (order counterbalanced). Some BXH/Ty strain means resembled those of the parent strains, but others were intermediate. This indicated recombination among loci affecting avoidance, and therefore polygenic inheritance. The strain means were highly correlated across compounds (r = 0.98), suggesting that the same polygenes controlled both phenotypes. The BXH/Ty means for both compounds were then compared with the strain genotypes at 212 chromosome position markers distributed throughout the genome. Eight markers on five chromosomes (3, 6, 7, 8 and 9) yielded significant correlations. Six of the markers were correlated with both phenotypes, again suggesting common polygenic inheritance. The marker with the highest correlation was Prp, tightly linked to Soa on chromosome 6. The correlated marker regions likely contain quantitative trait loci affecting bitter avoidance. The phenotypic similarity of PROP to quinine, rather than to phenylthiourea, apparently stemming from a common polygenic basis, indicates a difference between mice and humans in gustatory organization related to bitters.      

Taste sensitivities to PROP and PTC vary independently in mice

Mammals use common mechanisms to detect, transduce and process taste stimulus information. For example, they share families of receptors that respond to amino acids, and sweet- and bitter-tasting stimuli. Nonetheless, it also clear that different species exhibit unique taste sensitivities that may reflect specific genetic variations. In humans, sensitivities to the chemically similar, bitter-tasting compounds 6-n-propylthiouracil (PROP) and phenylthiocarbamide (PTC) are heritable and strongly correlated, suggesting a common genetic basis. However, it is unknown whether PROP and PTC taste sensitivities are similarly correlated in mice. Here we report that PROP and PTC taste sensitivities vary independently between two inbred strains of mice. In brief-access taste tests C3HeB/FeJ (C3) and SWR/J (SW) mice possess similar taste sensitivity to PTC, while SW mice are significantly more sensitive to PROP than are C3 mice. In two-bottle preference tests, however, SW mice display greater aversion to both compounds. This discrepancy may be explained by the observation that SW mice consumed taste solutions at a greater rate during the intake test than did C3 mice. Therefore, PTC avoidance is correlated with the amount of PTC consumed in the intake tests rather than the concentration of PTC tested. These findings suggest that post-ingestive factors play a significant role in PTC avoidance during intake tests and highlight an important advantage of brief-access tests over intake tests in resolving the gustatory and post-ingestive contributions to taste-related behaviors. Most strikingly, these results demonstrate that in mice, unlike in humans, PTC and PROP taste sensitivities vary independently, thereby suggesting a subtle functional diversity of bitter-taste mechanisms across mammalian species.     

Intermediate sucrose octa-acetate sensitivity suggests a third allele at mouse bitter taste locus Soa and Soa-Rua identity

Mice have been characterized as either tasters or non-tastersof the bitter compound sucrose octa-acetate(SOA). However, 11of 17 supposedly non-taster inbred strains were found to avoid1 mM SOA. All 17 strains were indifferent to 0.1 mM SOA. Tasterstrains avoided both concentrations. The intermediate phenotypewas dubbed demitaster. A consistent phenotypic dominance orderwas found in crosses among both inbred and outbred strains (taster> non-taster > demitaster). Demitasters were found (withtasters) in an outbred strain showing monogenic segregationfor SOA avoidance. This, plus monogenic segregation in a back-crossof taster to demitaster inbred strains, suggested a third alleleat the Soa locus (Soa^c). Demitaster allelism was supported bythe strong associations found in 15 strains between the threeSOA phenotypes and HindIII restriction fragment patterns forthe closely linked Prp (proline rich protein) loci. SOA demitasterstrains were also intermediate in raffinose undeca-acetate (RUA)avoidance. Furthermore, B6.SW-Soa² congenic mice avoided notonly SOA, but RUA and eight other acetylated sugars. A previouslyproposed separate RUA-sensitivity gene (Rua) thus appeared tobe redundant.     

High-resolution genetic mapping of the sucrose octaacetate taste aversion (Soa) locus on mouse Chromosome 6

An acetylated sugar, sucrose octaacetate (SOA), tastes bitter to humans and has an aversive taste to at least some mice and other animals. In mice, taste aversion to SOA depends on allelic variation of a single locus, Soa. Three Soa alleles determine `taster' (Soa <sup>a</sup> ), `nontaster' (Soa ^b ), and `demitaster' (Soa ^c ) phenotypes of taste sensitivity to SOA. Although Soa has been mapped to distal Chromosome (Chr) 6, the limits of the Soa region have not been defined. In this study, mice from congenic strains SW.B6-Soa ^b , B6.SW-Soa ^a , and C3.SW-Soa ^a/c and from an outbred CFW strain were genotyped with polymorphic markers on Chr 6. In the congenic strains, the limits of introgressed donor fragments were determined. In the outbred mice, linkage disequilibrium and haplotype analyses were conducted. Positions of the markers were further resolved by using radiation hybrid mapping. The results show that the Soa locus is contained in a ∼1-cM (3.3–4.9 Mb) region including the Prp locus. Received: 5 February, 2001 / Accepted: 1 May, 2001     

Effect of chorda tympani nerve transection on salt taste perception in mice

Effects of gustatory nerve transection on salt taste have been studied extensively in rats and hamsters but have not been well explored in the mouse. We examined the effects of chorda tympani (CT) nerve transection on NaCl taste preferences and thresholds in outbred CD-1 mice using a high-throughput phenotyping method developed in our laboratory. To measure taste thresholds, mice were conditioned by oral self-administration of LiCl or NaCl and then presented with NaCl concentration series in 2-bottle preference tests. LiCl-conditioned and control NaCl-exposed mice were given bilateral transections of the CT nerve (LiCl-CTX, NaCl-CTX) or were left intact as controls (LiCl-CNT, NaCl-CNT). After recovery from surgery, mice received a concentration series of NaCl (0-300 mM) in 48-h 2-bottle tests. CT transection increased NaCl taste thresholds in LiCl-conditioned mice and eliminated avoidance of concentrated NaCl in control NaCl-exposed mice. This demonstrates that in mice, the CT nerve is important for detection and recognition of NaCl taste and is necessary for the normal avoidance of high concentrations of NaCl. The results of this experiment also show that the method of high-throughput phenotyping of salt taste thresholds is suitable for detecting changes in the taste periphery in mouse genetic studies.     

Nerve regeneration-induced recovery of quinine avoidance after complete gustatory deafferentation of the tongue

The concentration-dependent decrease in quinine licking by rats is substantially attenuated by combined bilateral transection of the chorda tympani (CT) and glossopharyngeal (GL) nerves, but transection of either nerve alone produces marginal impairments at most. Here we tested whether regeneration of one or both of these nerves after combined transection would result in recovery of taste avoidance. Water-restricted rats were presented with a series of brief-access (5 s) taste trials (water and 0.003-3.0 mM quinine-HCl) in a 5-day test block of 40-min sessions both before nerve transection and starting 75-77 days after transection. Licking avoidance returned to presurgical levels when both nerves were allowed to regenerate. When only the GL was allowed to regenerate, performance did not differ from that of sham-transected animals. This suggests that even after considerable gustatory deafferentation, regeneration has the capacity to restore normal taste-guided behavior. Surprisingly, when only the CT was allowed to regenerate, avoidance behavior was severely impaired and was not different from that of rats in which regeneration of both nerves was prevented. Taking into account prior findings, it appears that the absence of the GL in the presence of an intact CT is fundamentally different from the absence of the GL in the presence of a regenerated CT with respect to some taste functions. This represents the first reported instance to our knowledge in which the capacity of a regenerated nerve to maintain taste-guided behavior was distinctly different from that of an intact nerve in a rodent model.     

NaCl taste thresholds in 13 inbred mouse strains

Molecular mechanisms of salty taste in mammals are not completely understood. We use genetic approaches to study these mechanisms. Previously, we developed a high-throughput procedure to measure NaCl taste thresholds, which involves conditioning mice to avoid LiCl and then examining avoidance of NaCl solutions presented in 48-h 2-bottle preference tests. Using this procedure, we measured NaCl taste thresholds of mice from 13 genealogically divergent inbred stains: 129P3/J, A/J, BALB/cByJ, C3H/HeJ, C57BL/6ByJ, C57BL/6J, CBA/J, CE/J, DBA/2J, FVB/NJ, NZB/BlNJ, PWK/PhJ, and SJL/J. We found substantial strain variation in NaCl taste thresholds: mice from the A/J and 129P3/J strains had high thresholds (were less sensitive), whereas mice from the BALB/cByJ, C57BL/6J, C57BL/6ByJ, CE/J, DBA/2J, NZB/BINJ, and SJL/J had low thresholds (were more sensitive). NaCl taste thresholds measured in this study did not significantly correlate with NaCl preferences or amiloride sensitivity of chorda tympani nerve responses to NaCl determined in the same strains in other studies. To examine whether strain differences in NaCl taste thresholds could have been affected by variation in learning ability or sensitivity to toxic effects of LiCl, we used the same method to measure citric acid taste thresholds in 4 inbred strains with large differences in NaCl taste thresholds but similar acid sensitivity in preference tests (129P3/J, A/J, C57BL/6J, and DBA/2J). Citric acid taste thresholds were similar in these 4 strains. This suggests that our technique measures taste quality-specific thresholds that are likely to represent differences in peripheral taste responsiveness. The strain differences in NaCl taste sensitivity found in this study provide a basis for genetic analysis of this phenotype.     

Oral amiloride treatment decreases taste sensitivity to sodium salts in C57BL/6J and DBA/2J mice

Sodium taste transduction is thought to occur via an amiloride-sensitive, sodium-selective pathway and an amiloride-insensitive, cation nonselective, anion-dependent pathway(s). It has been shown by others that amiloride, an epithelial sodium channel (ENaC) blocker, significantly reduces the chorda tympani nerve response to lingually applied NaCl in C57BL/6 (B6) mice but not in DBA/2 (D2) mice, suggesting that the latter strain might not possess functional ENaCs in taste receptor cells. We psychophysically measured and compared taste detection thresholds of NaCl and sodium gluconate (NaGlu) prepared with and without 100 microM amiloride in these two strains (eight/strain). Mice were trained and tested in a two-response operant signal detection procedure conducted in a gustometer. Surprisingly, no strain effect was found for the detection thresholds of both salts (approximately 0.05-0.06 M). Moreover, these thresholds were increased by almost an order of magnitude by amiloride adulteration of the solutions. This marked effect of amiloride on sodium detection thresholds suggests that ENaCs are necessary for normal sensitivity to sodium salts in both strains. In addition, because NaGlu is thought to stimulate primarily the amiloride-sensitive pathway, especially at low concentrations, the similarity of NaCl and NaGlu thresholds (r > 0.81 both strains) suggests that ENaCs are also sufficient to support the detection of sodium in weak solutions by B6 and D2 mice.     

Residual chemoresponsiveness to acids in the superior laryngeal nerve in "taste-blind" (P2X2/P2X3 double-KO) mice

Mice lacking both the P2X2 and the P2X3 purinergic receptors (P2X-dblKO) exhibit loss of responses to all taste qualities in the taste nerves innervating the tongue. Similarly, these mice exhibit a near total loss of taste-related behaviors in brief access tests except for a near-normal avoidance of acidic stimuli. This persistent avoidance of acids despite the loss of gustatory neural responses to sour was postulated to be due to continued responsiveness of the superior laryngeal (SL) nerve. However, chemoresponses of the larynx are attributable both to taste buds and to free nerve endings. In order to test whether the SL nerve of P2X-dblKO mice remains responsive to acids but not to other tastants, we recorded responses from the SL nerve in wild-type (WT) and P2X-dblKO mice. WT mice showed substantial SL responses to monosodium glutamate, sucrose, urea, and denatonium-all of which were essentially absent in P2X-dblKO animals. In contrast, the SL nerve of P2X-dblKO mice exhibited near-normal responses to citric acid (50 mM) although responsiveness of both the chorda tympani and the glossopharyngeal nerves to this stimulus were absent or greatly reduced. These results are consistent with the hypothesis that the residual avoidance of acidic solutions by P2X-dblKO mice may be attributable to the direct chemosensitivity of nerve fibers innervating the laryngeal epithelium and not to taste.     

Detection of Mls-antigens in various mouse strains using a new method

Mice of strains CBA and BALB/c, when injected with lymphocytes from theH-2-compatible Mls-antigen-incompatible strains C3H and DBA/2, respectively, develop a reduced lymphocyte reactivity against cells of the injected strains as measured in the mixed lymphocyte culture (MLC). The mechanism of the development of a depression of the MLC response against Mlsantigens is unknown. In this investigation we have tested the MLC response of lymphocytes from CBA mice preinjected with C3H lymphocytes against cells from 12 different strains. It was observed that the response decreased against cells from strains C3H, AKR, and A/Sn. Infusion of CBA mice with AKR lymphocytes decreased their MLC response against the same three strains. In contrast, infusion of CBA mice with A/Sn lymphocytes reduced their MLC responses against strains C3H, DBA/2, and the congenic strains A/Sn, A.SW, A.CA, and A.BY. BALB/c mice which were infused with DBA/2 lymphocytes developed reduced responses against DBA/2, C3H, and AKR. On the basis of these results we propose that mice of our strains C3H and AKR possess a common Mls-antigen which is strongly stimulatory, and that DBA/2 mice possess a second Mls-antigen which is also strongly stimulatory. The congenic strains A/Sn, A.SW, A.CA, and A.BY, which have differentH-2 complexes, possess a third Mls-antigen which is less stimulatory. The Mls-antigens of the strains listed above seem to exhibit extensive immunological crossreactivity.     

Training and extinction of passive avoidance in mice lacking monoamine oxidase A

The passive avoidance learning and memory trace retention in mice lacking monoamine oxidase A (MAO A) and control C3H strain were analyzed. It is shown that mice of both strains were well the passive avoidance learners. A delay of the memory trace extinction was found in lacking MAO A mice as compared with control C3H strain. Mice with a genetic MAO A knockout showed decreased amounts of transitions, rearings, looking to a dark compartment and appearing from it. These findings seem to reflect more expressed fear reaction to a dangerous compartment and increased anxiety promoting longer retention of memory trace on a high level of retrieval.     

Hydrolysis of sucrose octa-acetate: qualitative differences in taster and demistaster avoidance phenotypes

Calcium hydroxide and sodium hydroxide were used to hydrolysesucrose octa-acetate (SOA) as a means of evaluating the taster(Soa^a) and demitaster (Soa^c) allelic phenotypes of the geneticlocus Soa. The SWR/J (taster) inbred strain and the B6.SW Soa^a(taster) congenic strain were demonstrated to cease avoidingupon nearly complete hydrolysis of 10^–5 M SOA with calciumhydroxide Or sodium hydroxide and of 10^–4 M SOA with calciumhydroxide. The BALB/cByJ, C3HeB/FeJ and DBA/2J (demitaster)inbred strains were demonstrated to cease avoiding after onlya partial hydrolysis of 10^–3 M SOA using calcium hydroxide.It is suggested that specificity for the number or placementof the acetates of SOA underlies the difference between thetaster and demitaster phenotypes.     

Absence of anxiolytic response to chlordiazepoxide in two common background strains exposed to the elevated plus-maze: importance and implications of behavioural baseline

Although genetic background is acknowledged as a potentially important determinant of mutant phenotypes, publications on genetically modified mice far outnumber those on progenitor strains. We have recently reported major differences in basal anxiety levels (elevated plus-maze & light/dark exploration) among three strains (C57BL/6JOlaHsd, 129/SvEv and 129S2/SvHsd) employed as progenitor stock in European laboratories (Rodgers et al. in press). Furthermore, the phenotypes of these inbred strains differed significantly from that of an outbred strain (Swiss-Webster) commonly used in behavioural pharmacology. In view of these findings, the present study assessed possible differences in the anxiolytic efficacy of chlordiazepoxide (0, 7.5 & 15.0 mg/kg, IP) in three of these strains (Swiss-Webster (SW), C57BL/6JOIaHsd (C57) & 129S2/SvHsd (129)). Experimentally naive mice were exposed to the elevated plus-maze, sessions were videotaped and behaviour analysed using ethological software. The performance of control subjects confirmed significant strain differences in basal levels of activity (SW > C57 > 129) and anxiety-related behaviours (129 = SW > C57), with hypolocomotion dominating the 129 profile. SW mice displayed an anxioselective response to both doses of chlordiazepoxide (CDP), with significant reductions in open arm avoidance and risk assessment observed in the absence of any change in general activity. In direct contrast, the lower dose of CDP (7.5 mg/kg) was without effect in either inbred strain, whereas treatment with 15.0 mg/kg induced a profile indicative of muscle relaxation/mild sedation in C57 mice and virtually abolished all behavioural activity in 129 mice. Although the absence of an anxiolytic response to CDP in C57 mice may be attributed to their low basal anxiety levels, the profile of 129 mice strongly suggests an abnormality in benzodiazepine/GABAA receptor function. The implications of these findings for research on mutant mice are discussed.     

Bitter taste stimuli induce differential neural codes in mouse brain

A growing literature suggests taste stimuli commonly classified as "bitter" induce heterogeneous neural and perceptual responses. Here, the central processing of bitter stimuli was studied in mice with genetically controlled bitter taste profiles. Using these mice removed genetic heterogeneity as a factor influencing gustatory neural codes for bitter stimuli. Electrophysiological activity (spikes) was recorded from single neurons in the nucleus tractus solitarius during oral delivery of taste solutions (26 total), including concentration series of the bitter tastants quinine, denatonium benzoate, cycloheximide, and sucrose octaacetate (SOA), presented to the whole mouth for 5 s. Seventy-nine neurons were sampled; in many cases multiple cells (2 to 5) were recorded from a mouse. Results showed bitter stimuli induced variable gustatory activity. For example, although some neurons responded robustly to quinine and cycloheximide, others displayed concentration-dependent activity (p<0.05) to quinine but not cycloheximide. Differential activity to bitter stimuli was observed across multiple neurons recorded from one animal in several mice. Across all cells, quinine and denatonium induced correlated spatial responses that differed (p<0.05) from those to cycloheximide and SOA. Modeling spatiotemporal neural ensemble activity revealed responses to quinine/denatonium and cycloheximide/SOA diverged during only an early, at least 1 s wide period of the taste response. Our findings highlight how temporal features of sensory processing contribute differences among bitter taste codes and build on data suggesting heterogeneity among "bitter" stimuli, data that challenge a strict monoguesia model for the bitter quality.     

A novel mouse Chr5 locus <Emphasis Type="Italic">Diht</Emphasis> controls dopamine-induced hypothermia

Mice of the strain C3H.PRI-Flv^r, carrying genetically determined resistance to flaviviruses, have been shown to be more sensitive to the hypothermic effect of dopamine than congenic flavivirus-susceptible C3H/HeJARC mice. In the current study, the greater sensitivity to dopamine-induced hypothermia observed in flavivirus-resistant mice was shown to be dose-dependent, with strain differences being the most prominent at a moderate dose of apomorphine (1 mg/kg). In addition, hypothermic responses to apomorphine were shown to be under developmental regulation; aging increased the potency of apomorphine-induced hypothermia and abrogated strain and sex differences observed in young mice. Linkage analysis of mouse strain-dependent co-inheritance between flavivirus resistance and greater sensitivity to the hypothermic effect of dopamine was performed using two genetically unrelated flavivirus-susceptible and two highly congenic flavivirus-resistant mouse strains in parallel with C3H.PRI-Flv^r-and C3H/HeJARC reference strains. This study has revealed a clear segregation between flavivirus resistance conferred by the Flv locus and sensitivity to dopamine-controlled hypothermia conferred by a novel locus, Diht. Parallel studies in F1 and F2 heterozygote mice showed that the high sensitivity to hypothermic effect of dopamine (Diht^high) is inherited as the Chr5-linked dominant trait. The novel locus, Diht, has been mapped proximal to the Flv locus on a distal part of mouse Chr5 between microsatellite markers D5Mit41 and D5Mit158.     

Effects of isoproterenol treatment on gustatory neural responses in three inbred strains of mice

1. Treatment of a beta-agonist, isoproterenol, for 5 days reduced chorda tympani responses to sucrose by about 40% of the control without affecting responses to other taste stimuli, such as NaCl, HCl and quinine HCl, in balb CrSlc mice whereas such reduction of sucrose responses was not observed in C57BL/6-CrSlc and C3H/HeSlc mice, although in the latter two strains long-lasting off-responses to quinine HCl appeared after the treatment. 2. In BALB mice, the magnitude of reduction of sucrose responses by isoproterenol increased with prolonging the treatment from 1 to 5 days, although it reached almost its maximum level by the 3 days treatment. 3. BALB mice with the removal of the submandibular glands showed slightly greater control responses of the chorda tympani nerve to sucrose than BALB mice with the sham-operation or the removal of the sublingual glands, and showed no significant reduction of sucrose responses by isoproterenol treatment. 4. These results suggest that isoproterenol probably did not act directly on sweetener receptors of taste cell membranes but affect them through the submandibular salivary system.     

Immunodominance in the T cell response to multiple non-H-2 histocompatibility antigens. IV. Partial tissue distribution and mapping of immunodominant antigens

The immunization of C57BL/6 responder mice with spleen cells from H-2-matched BALB.B donors, which differ by multiple non-H-2 histocompatibility (H) antigens, results in the generation of cytotoxic T lymphocytes (CTL) that are specific for only a limited number of immunodominant antigens. Previous analysis of the genes encoding these dominant antigens has not mapped these genes to any of the non-H-2 H loci defined by congenic strains. It would have been expected that the histogenetic techniques employed for congenic strain selection would have preferentially identified the "strongest" H antigens. Therefore, we have investigated the possibility that immunodominant antigens do not belong to the class of non-H-2 H antigens encoded by genes mapping to H loci defined and mapped by congenic strains. The first experiments were aimed at identifying antigens that were expressed by independently derived inbred strains and were cross-reactive with the immunodominant cytotoxic T cell target (CTT-1) antigen of BALB.B. Strong cross-reaction with the C3H.SW (H-2b) strain was observed; the C3H gene encoding this antigen was mapped with BXH recombinant inbred strains. Contrary to the mapping of the CTT-1 gene to chromosome 1 in BALB.B, the C3H gene was shown to map to either chromosome 4 or chromosome 7. This result indicates that identical, or at least extensively cross-reactive, non-H-2 antigens may be encoded by genes mapping to independently segregating loci in different inbred strains. The tissue distribution of immunodominant antigens was approached by determining the reactivity of CTL specific for these antigens with either lymphoid-derived or fibroblast-derived targets. These CTL effectively lysed lymphoblast and lymphoid tumor targets but did not lyse an SV40-transformed fibroblast line that was shown to be efficiently lysed by CTL specific for non-H-2 H antigens defined by congenic strains. Therefore, it was concluded that immunodominant antigens detected by B6 anti-BALB.B CTL have a restricted tissue distribution in comparison to non-H-2 H antigens defined by congenic strains. The implications of these results for our understanding of the origin and heterogeneity of non-H-2 cell-surface antigen recognized by effector T cells are discussed.