Phenylthiocarbamide produces conditioned taste aversions in mice

Previous work has demonstrated that SWR/J (SW) mice avoid phenylthiocarbamide (PTC) to a greater degree than C3HeB/FeJ mice in 48 h, two-bottle preference tests given in ascending series. The authors hypothesized, based also on previous work, that SW mice might form a conditioned taste aversion over time due to the toxic properties of PTC. We directly tested this hypothesis by attempting to condition a taste aversion to sucrose by injections of PTC. In experiment 1, PTC was nearly as effective as a strong dose of LiCl in reducing sucrose drinking. In experiment 2, the sucrose aversions were parametrically modified by both sucrose concentration and PTC dose, a hallmark of conditioned taste aversion. We conclude that PTC can cause a conditioned taste aversion and discuss the importance of considering toxic effects of aversive tastants when analyzing behavioral strain differences.     

Formation of taste aversions in the rat

The presence of correlation between the initial attitude of animals to a certain taste agent and the intensity of conditioned taste aversion (CTA) after LiCl poisoning was studied in nonlineal male white rats. 4 types of behavioural reactions were revealed: 1) the nonspecific inhibitory reaction to the situation in which CTA was elaborated; 2) specific associative-adaptive connection of CTA with the taste agent used (0,1% saccharine solution); 3) complex reaction consisting of the two first ones; 4) the same drinking behaviour as that before CTA elaboration. Moreover, a correlation was found between the initial attitude of animals to certain taste agent and the intensity of CTA after pairing of this agent with subsequent poisoning.     

Differences in the taste quality of maltose and sucrose in rats: issues involving the generalization of conditioned taste aversions

The present study employed a conditioned taste aversion generalizationparadigm to test the hypothesis that maltose produces tastesensations in the rat which are qualitatively distinguishablefrom sucrose. Since stimulus generalization can occur in boththe quality and intensity domains, an intrachemical (acrossconcentration) generalization gradient was established to aidin the interpretation of the interchemical (across molecules)generalization gradient. Moreover, since the commonly used intaketest is vulnerable to nontaste post-ingestional influences,the present study measured immediate responses to 100 µlstimulus samples, thus increasing our confidence that the behaviorwas under orosensory control. In Experiment 1, naive water deprivedrats were trained in a specially designed gustometer to maintaindrinking-spout contact for intermittent water reinforcement.Following this, rats in the experimental group were given threeexposures to 0.1 M sucrose on separate days, with the firsttwo exposures immediately preceding an injection of LiCl. Acontrol group was treated identically but received distilledwater instead of sucrose. Rats were then tested in the gustometerfor their avoidance of three equimolar concentrations of sucroseand maltose. Rats received ten trials of each stimulus quasi-randomlypresented in two sessions. Results indicated that all sucroseconcentrations were avoided (in experimental group only), butonly the 0.3 M concentration of maltose was avoided. The lowestsucrose concentration was significantly less avoided than thehigher concentrations. Intensity generalization gradients aresuch that intensities weaker than the conditioned stimulus (CS)produce just as much or less of a conditioned response (CR)and intensities stronger than the CS produce just as much ora greater CR than that elicited by the CS itself. Therefore,based on the results of Experimental, it was predicted thatif 0.1 M maltose served as the CS, the order of avoidance shouldbe: 0.3 M sucrose 0.1 M sucrose 0.03 M sucrose 0.3 M maltose 0.1 M maltose 0.03 M maltose, if it were true that maltoseand sucrose produce identical sensations that differ only inintensity. Experiment 2 explicitly tested this prediction usingthe same procedure as Experiment 1 except that 0.1 M maltoseserved as the CS. The observed order of avoidance was 0.3 Mmaltose > 0.1 M maltose > 0.03 M maltose = 0.3 M sucrose= 0.1 M sucrose = 0.03 M sucrose. In both experiments the intrachemicalgeneralization gradient broadened and the interchemical generalizationgradient steepened upon retesting. In conclusion, qualitativedifferences between maltose and sucrose explain the outcomesof these experiments better than differences in the relativeintensity of these sugars at isomolar concentrations.     

Generalization of specific taste aversions to alcohol in the rat

Two experiments were conducted to examine the taste qualitiesof alcohol in the rat. In Experiment 1, rats were trained toavoid sucrose, quinine or a sucrose + quinine mixture. In Experiment2, rats were trained to avoid sucrose, hydrochloric acid ora sucrose + hydrochloric acid mixture. In both experiments ratswere then tested for generalization to 3, 6 and 9% (v/v) alcoholsolutions. Following the alcohol tests rats were tested withthe taste solutions used during training. Results showed thatrats trained to avoid sucrose (Experiments 1 and 2), quinineor the sucrose + quinine mixture generalized that aversion tothe 6% alcohol solution. No generalization was found to the3% alcohol solution. No generalization was displayed by ratstrained to avoid either the hydrochloric acid solution or thesucrose + hydrochloric acid solution. Following the alcoholtests, all trained rats exhibited strong aversions to the solutionthey were trained to avoid. In addition, rats trained to avoida single solution generalized that aversion to the mixture andrats trained to avoid a mixture generalized to the single components.     

Generalization of conditioned taste aversions in hamsters: evidence for multiple bitter receptor sites

Multiple bitter receptor sites appear to exist within the hamstergustatory system supporting the data of other investigatorson humans, rats and frogs. The sodium salts of four anions,m-nitrobenzene sulfonate (NBSA), picrate, m-nitrobenzoate (NBA)and cholate, were tested in two-bottle preference tests andfor generalization to a variety of stimuli in a conditionedtaste aversion (CTA) paradigm. All four of these anions arebitter to humans. One, NBSA, generalized to sucrose suggestinga sweet taste, while the remaining three appear to be bitterwith varying degrees of saltiness. The bitterness of these threeanions to hamsters appears to be perceptually different froma quinine-type bitterness. Separate bitter receptor sites areindicated for quinine and urea, plus a third site acceptingNBA, picrate and cholate. More bitter sites are plausible. Separatesites for quinine and urea appear to occur across species. Itwas also concluded that quinine does not serve as a prototypicbitter stimulus for all bitters in the CTA test.     

Estradiol accelerates extinction of lithium chloride-induced conditioned taste aversions through its illness-associated properties

Estradiol accelerates extinction of LiCl-induced conditioned taste aversions when it is present during a period that starts 2-3 days after acquisition and extends throughout extinction (before and during extinction). It has been suggested that estradiol acts before, not during, extinction and that its effect on extinction is associated with its illness-inducing properties. This hypothesis is based on previous work which shows an attenuation of conditioned taste aversion learning when rats are exposed to illness-inducing agents during a period that starts 2 days after acquisition and ends 2 days before extinction trials are initiated. Four experiments were designed to test elements of this hypothesis. The first two experiments demonstrated that if an estradiol-filled Silastic capsule is implanted before extinction of a LiCl-induced aversion, when the conditioned taste is not present, it accelerates extinction, but if it is implanted during extinction, when the conditioned taste is present, it prolongs extinction. The third experiment showed that the same dose of estradiol that accelerates extinction of a LiCl-induced aversion was effective in producing a conditioned taste aversion when it was present for 18 h after consumption of a novel sucrose solution. The fourth experiment indicated that serum levels of estradiol were elevated during the 18 h. These results are consistent with the hypothesis that the acceleration of extinction by estradiol is associated with its illness-inducing properties. It is suggested that estradiol acts on neural areas that mediate illness information and that one of these areas, the area postrema is necessary for estradiol to accelerate extinction of a LiCl-induced aversion.     

Attenuation of a radiation-induced conditioned taste aversion after the development of ethanol tolerance

An attempt to reduce a radiation-induced conditioned taste aversion (CTA) was undertaken by rendering animals tolerant to ethanol. Ethanol tolerance, developed over 5 days, was sufficient to block a radiation-induced taste aversion, as well as an ethanol-induced CTA. Several intermittent doses of ethanol, which did not induce tolerance but removed the novelty of the conditioning stimulus, blocked an ethanol-induced CTA but not the radiation-induced CTA. A CTA induced by doses of radiation up to 500 rads was attenuated. These data suggest that radioprotection developing in association with ethanol tolerance is a result of a physiological response to the chronic presence of ethanol not to the ethanol itself.     

Characterization of orexin A immunoreactivity in the rat area postrema

The distribution of orexin A immunoreactivity and the synaptic relationships of orexin A-positive neurons in the rat area postrema were studied using both light and electron microscopy techniques. At the light microscope level, numerous orexin A-like immunoreactive fibers were found within the area postrema. Using electron microscopy, immunoreactivity within fibers was confined primarily to the axon terminals, most of which contained dense-cored vesicles. Both axo-somatic and axo-dendritic synapses made by orexin A-like immunoreactive axon terminals were found, with these synapses being both symmetric and asymmetric in form. Orexin A-like immunoreactive axon terminals could be found presynaptic to two different immunonegative profiles including the perikarya and dendrites. Occasionally, some orexin A-like immunoreactive profiles, most likely to be dendrites, could be seen receiving synaptic inputs from immunonegative or immunopositive axon terminals. The present results suggest that the physiological function of orexin A in the area postrema depends on synaptic relationships with other immunopositive and immunonegative neurons, with the action of orexin A mediated via a self-modulation feedback mechanism.     

Learned aversions and taste qualities in hamsters

Interralations among taste perceptions in gloden hamsters (Mesocricetusauratus) were examined using generalizaions of learned tasteaversions. If stimulus A is avoided given a taste aversion hasbeen established to stimulus B, and vice versa, A and B ‘cross-generalize’.Stimuli within five groups cross-generalized. The groups ofcompounds were (i) sweeteners (fructose, saccharin, sucrose);(ii) sodium salts (NaCl, NaNO₃, Na₂SO₄): (iii) non-sodium salts(KCl, MgSO₄ NH₄Cl) plus quinine HCl; (iv) acids (acetic, hydrochloric,citric); and (v) urea. Only two pairs of stimuli from differentgroups cross-generalized (HCl—NH₄Cl. quinine HCl—urea).Neural patterns of response recorded form chorda tympam nervefibers in hamsters suggest that taste receptors on the anteriortongue distinguish among three groups of taste stimuli: sweeteners,sodium salts, and a group including non-sodium salts, acids,quinine HCl and urea. Neurons innervating other taste fieldsare likely to provide the information that hamsters use to discriminateamong the tastes of non-sodium-salt and non-sweetener stimuli.     

Cholecystokinin-octapeptide like immunoreactivity in the area postrema of the rat and cat

The distribution of cholecystokinin-8 (CCK-8)-like immunoreactivity in the area postrema of the rat and cat was visualized using the peroxidase, antiperoxidase technique. In the rat the greatest amount of immunostaining occurred in peripheral regions of the area postrema at intermediate and rostral levels. Caudally, scattered immunoreactivity predominated. After colchicine treatment, numerous immunoreactive somata were observed throughout the area postrema. The cat area postrema had a different and more complex pattern of immunostaining than the rat. Moderate to dense accumulations of immunostaining occurred in the ventromedial region of the area postrema bordering the solitary tract and dorsal vagal nuclei. The central region of the area postrema possessed scattered amounts of immunoreactivity at rostral levels. Following colchicine treatment, no visible CCK-8-like immunoreactive cell bodies were observed in the cat area postrema. Results of the present investigation provide morphological evidence for the role of CCK-8 in cardiovascular regulation and satiety. The difference in the distribution of CCK-8 in the rat and cat suggest a possible role in the emetic reflex.     

Distribution of neurotensin-like immunoreactivity in rat and cat area postrema

The distribution of neurotensin-like immunoreactive (NT-LI) fibers and cell bodies in the area postrema (AP) of rat and cat, utilizing the peroxidase, antiperoxidase (PAP) technique, is described. In the rat, the greatest accumulation of NT-LI fibers were present along the borders of the AP, while there were very few NT-LI fibers in central regions. In the cat, scattered NT-LI fibers occupied the majority of the central AP, while moderate numbers of NT-LI fibers were present at the ventromedial border. In more rostral cat AP levels, the number of NT-LI fibers decreased. NT-LI somata were present in rat AP, but were lacking in cat AP. The localization of NT within the AP suggests that the cardiovascular and gastric effects attributed to NT may be mediated, in part, through the AP.     

Ultrastructural localization of monoamines and peptides in rat area postrema

An overall schema for the synaptic interactions of monoaminergic and peptidergic neurons and their relation to the ventricle and to blood vessels within the rat area postrema is presented. The specific markers include: 1) the immunocytochemical localization of the catecholamine-synthesizing enzyme tyrosine hydroxylase, and the neuropeptides enkephalin and substance P; and 2) the radioautographic localization of [3H]serotonin (5-hydroxytryptamine) and 3H-labeled amino acids anterogradely transported from the nodose ganglion.     

Excitatory effect of ATP on rat area postrema neurons

ATP-induced inward currents and increases in the cytosolic Ca²⁺ concentration ([Ca]_in) were investigated in neurons acutely dissociated from rat area postrema using whole-cell patch-clamp recordings and fura-2 microfluorometry, respectively. The ATP-induced current (I _ATP) and [Ca]_in increases were mimicked by 2-methylthio-ATP and ATP-γS, and were inhibited by P2X receptor (P2XR) antagonists. The current–voltage relationship of the I _ATP exhibited a strong inward rectification, and the amplitude of the I _ATP was concentration-dependent. The I _ATP was markedly reduced in the absence of external Na⁺, and the addition of Ca²⁺ to Na⁺-free saline increased the I _ATP. ATP did not increase [Ca]_in in the absence of external Ca²⁺, and Ca²⁺ channel antagonists partially inhibited the ATP-induced [Ca]_in increase, indicating that ATP increases [Ca]_in by Ca²⁺ influx through both P2XR channels and voltage-dependent Ca²⁺ channels. There was a negative interaction between P2XR- and nicotinic ACh receptor (nAChR)-channels, which depended on the amplitude and direction of current flow through either channel. Current occlusion was observed at V _hs between −70 and −10 mV when the I _ATP and ACh-induced current (I _ACh) were inward, but no occlusion was observed when these currents were outward at a V _h of +40 mV. The I _ATP was not inhibited by co-application of ACh when the I _ACh was markedly decreased either by removal of permeant cations, by setting V _h close to the equilibrium potential of I _ACh, or by the addition of d-tubocurarine or serotonin. These results suggest that the inhibitory interaction is attributable to inward current flow of cations through the activated P2XR- and nAChR-channels.     

Immunohistological studies of masked indoleamine cells in the area postrema of the rat

Masked indoleamine cells (MICS) in the area postrema and adjacent areas in the rat were immunohistochemically studied (the peroxidase-antiperoxidase method) using a serotonin antiserum. After pretreatment with nialamide (200-300 mg/kg), immunoreactive MICS could be observed. They were small cells (about 12 micron in diameter) with several processes and were distributed in nearly all parts of the area postrema and also in the nucleus tructus solitarii. Following a single intraventricular injection of 75 micrograms 5,6-dihydroxytryptamine, the immunoreactivity of these cells conspicuously decreased for several days. The submicroscopical structure of the cells was investigated using immunoelectron microscopy. Immunoreactive products were observed in the cytoplasm as particles with a diameter of 25-40 nm and high electron density, but these were not found in the nucleus or cell organelles.     

Fine structure of ependymal cysts in and around the area postrema of the rat

Summary Peculiar cells forming cysts were observed in the area postrema and sometimes also in the choroid plexus and the tela chorioidea near the area postrema, and were studied in detail by electron microscopy. The cytological features of the cyst cell and its junctional relationship to neighboring cells imply that cyst cells are derived from ependymal and choroid epithelial cells. The cyst cells usually contact directly the perivascular spaces of postremal, choroidal or pial capillaries, where the cytoplasm is often considerably attenuated. The cystic lumen is commonly filled with a flocculent material. The limiting membrane of the cystic lumen, which frequently bears cilia and microvilli, has the same thickness as the surface cell membrane. In many cases, the cyst is surrounded by the cytoplasm of a single cell. In some cases, however, two cells participate in the formation of the cyst, although one is only a slender process and joined by a zonula occludens with the main cyst cell. Horseradish peroxidase (HRP) injected into the cerebrospinal fluid (CSF) space failed to enter the cystic lumen. A possible significance of the cyst in relation to the CSF and blood circulation was considered.     

Electron microscopic observation of mu-opioid receptor in the rat area postrema.

A simple preembedding avidin-biotin-peroxidase complex technique was used to study the ultrastructural localization of mu-opioid receptor in the rat area postrema. By using low concentrations of the first antiserum for incubation with a short reaction time to 3,3'-diaminobenzidine, the immunostaining was faint at the light microscopic level. However, at the electron microscopic level, strong immunoreaction was observed. Mu-Opioid receptors were found to be localized on the postsynaptic membrane of dendrites, extrasynaptic plasma membrane, and the surface of the small, clear vesicles in axon terminals. Of the total 283 immunopositive profiles observed, 68.2% (193 of 283) were dendrites, 29.3% (83 of 283) were axon terminals, and 2.5% (7 of 283) were myelinated axons. No immunostained neuron bodies were found in the present study; 109 mu-opioid receptor immunoreactive dendrites received synapses (56.5%, 109 of 193) from nonimmunoreactive (84.4%, 92 of 109) or immunoreactive (15.6%, 17 of 109) axon terminals, whereas 84 dendrites (43.5%, 84 of 193) were found without receiving synapses. The present study shows that the mu-opioid receptor in the area postrema plays a role mainly at the synapses.     

Immunohistological studies of masked indoleamine cells in the area postrema of the rat

Summary Masked indoleamine cells (MICS) in the area postrema and adjacent areas in the rat were immunohistochemically studied (the peroxidase-antiperoxidase method) using a serotonin antiserum. After pretreatment with nialamide (200–300 mg/kg), immunoreactive MICS could be observed. They were small cells (about 12 m in diameter) with several processes and were distributed in nearly all parts of the area postrema and also in the nucleus tructus solitarii. Following a single intraventricular injection of 75 g 5,6-dihydroxytryptamine, the immunoreactivity of these cells conspicuously decreased for several days. The submicroscopical structure of the cells was investigated using immunoelectron microscopy. Immunoreactive products were observed in the cytoplasm as particles with a diameter of 25–40 nm and high electron density, but these were not found in the nucleus or cell organelles.This work was supported by a grant (No. 57214028) from the Ministry of Education, Science and Culture, Japan     

Enhanced consumption of an aversively conditioned taste following the presentation of a “medicine” taste

Rats given presentations of a citric acid solution while recovering from LiCl-induced illness (i.e., a “medicine effect” treatment) subsequently drank more of an aversively conditioned NaCl solution at test, when the NaCl presentation was immediately preceded by citric acid. That is, citric acid passed a summation test of conditioned inhibition. Such an effect was not observed in a group given explicitly unpaired presentations of LiCl and citric acid. It is proposed that enhanced consumption of an aversive taste due to the previous presentation of a “medicine” taste can provide an animal model of human maladaptive behavior in regards to food consumption.