Conditioned taste aversions.

When one becomes ill after consuming a meal, there is a propensity to target a particular taste as the cause of the illness. The qualities of the taste most likely targeted include more novel, less preferred, and higher protein content. This association between a particular taste and illness is a form of learning that is termed conditioned taste aversion (CTA). A consequence of the learned association is that the taste will become aversive. When experiencing the taste again, individuals will show aversive reactions such as expressions of loathing, will experience mimicked illness sensations such as nausea, and subsequently, will avoid further exposure to the taste. The ability to acquire CTA occurs across species and across ages within a species. In the rat animal model, however, age differences exist in the capability of acquiring CTAs when increasingly longer intervals are imposed between consumption of a novel sweet solution and onset of illness. Pups have a decreased ability compared to young adults while aged rats have an increased ability. Evidence suggests that the failure of pups to acquire CTA at longer intervals is due to an immature retrieval mechanism and the facilitated ability of aged rats is due to a compromised clock mechanism that tracks the passage of time. Learned taste-illness association serves the critical function of informing individuals of the toxic nature of certain foods, thus preventing further illness and potentially death. Additionally, it contributes to the hypophagia observed during cancer chemotherapy and may contribute to the hypophagia found while suffering from bacterial infection, chronic medical conditions such as cancer, and restrictive food intake disorders such as anorexia nervosa.

Dissociating the conditioning and the anorectic effects of estradiol in female rats.

The present series of experiments challenges the ability of the hormone estradiol to act as an unconditioned stimulus in the conditioned taste avoidance (CTA) learning paradigm. We hypothesize that reductions in sucrose consumption observed after pairing it with estradiol are not indicative of associative learning, but due to the unconditioned expression of estradiol's anorectic effects during the time of CTA assessment. Three experiments in which a sucrose solution was paired with estradiol were conducted to test this hypothesis. Experiment 1 demonstrated that female rats expressed a reduction in post-pairing sucrose consumption even though the anorectic effects of estradiol had subsided. Experiment 2 showed that although a low dose of estradiol produced anorexia, it did not elicit post-pairing reductions in sucrose consumption. Experiment 3 revealed that contingent pairing was a requirement for post-pairing reduction in sucrose consumption even when testing was done at a time when anorexia is expressed. These findings demonstrate the dissociability of the conditioning and anorectic effects of estradiol, providing evidence against the hypothesis. The results are discussed in terms of independent neural mechanisms underlying the disparate behaviors.

Intraoral cheek fistulae: a refined technique.

Taste reactivity testing (TRT), which entails infusing a solution into the oral cavity of subjects, is used across a wide range of studies. For laboratories inexperienced in the conventional technique of implanting cheek fistulae, the surgery can be problematic for both the subjects and the experimenter. We have proposed a refined method for fistulae implantation that is less invasive, thereby reducing the pain and distress of the animals. Using this refined technique, we were able to replicate the findings of previous TRT studies, namely that a high dose of lithium chloride produces an increase in aversive and a decrease in ingestive orofacial and somatic responses. Using indices of health, we demonstrate that unlike animals with the conventional method of fistulae implantation, subjects that receive the refined technique regain their pre-surgery body weights rapidly and show no physical signs of discomfort. Additional advantages of the refined technique are discussed.

Cortical cooling induces conditioned consumption reduction in male rats.

Previous studies have shown that male rats acquire a conditioned reduction in consumption of a sucrose solution when consumption of that taste solution is followed by cooling the caudate putamen. Because the shaft of the cold probe was not insulated, this cooling also included the cortex and meninges overlying the caudate putamen. When cooling the meninges was eliminated as a factor, the conditioned consumption reduction was weaker but it was not abolished. This suggests that meninges cooling contribute to the conditioned consumption reduction induced when all three structures are cooled, but it is not sufficient. Five experiments were designed to determine whether cooling the cortex also contributes. In the first experiment, the temperature of the cortex and meninges overlying the caudate putamen was measured during cooling. In the following three experiments the ability of male rats to acquire a conditioned consumption reduction was determined after pairing a sucrose solution with cooling the cortex and meninges overlying the caudate putamen, cooling the cortex with and without cooling the dura meninges membrane, and cooling the cortex with and without cooling the entire meninges. When the cortex was cooled without cooling the caudate putamen, dura, or entire meninges, a conditioned consumption reduction was acquired. The last experiment demonstrated that contingent pairing of sucrose and cortical cooling was required to obtain consumption reduction. These results clearly indicate that cortical cooling contributes to the acquisition of conditioned consumption reduction induced when the caudate putamen and overlying cortex and meninges are cooled. Two hypotheses are suggested to account for the ability of neural cooling to act as an unconditioned stimulus in the conditioned consumption reduction paradigm: (1) neuronal inactivation produces physiological changes that can serve as unconditioned stimuli and (2) cooling itself produces physiological changes that can serve as unconditioned stimuli.

High doses of vasopressin delay the onset of extinction and strengthen acquisition of LiCl-induced conditioned taste avoidance.

When low doses of vasopressin are given 50 min after pairing sucrose consumption with a high dose of LiCl, extinction of the LiCl-induced conditioned taste avoidance is accelerated. These low doses of vasopressin do not themselves induce conditioned taste avoidance when paired with sucrose consumption. Predicated on previous studies administering two avoidance-inducing agents after sucrose consumption, studies were designed to determine whether high doses of vasopressin capable of inducing conditioned taste avoidance would (1) delay rather than accelerate extinction of a conditioned taste avoidance induced by a high dose of LiCl and (2) strengthen acquisition of a conditioned taste avoidance induced by a low dose of LiCl. The results of three studies showed that doses of 9 and 18 microg/kg of vasopressin induced a conditioned taste avoidance when injected 50 min after sucrose consumption, delayed the onset of extinction when injected 50 min after pairing sucrose consumption with a high dose of LiCl, and strengthened acquisition of a conditioned taste avoidance when injected 50 min after pairing sucrose consumption with a low dose of LiCl. Taken together, these data suggest that the delay in onset of extinction is due to a strengthening of acquisition. It has been suggested that vasopressin is a mnemonic neuropeptide that delays extinction of learned tasks. However, for conditioned taste avoidance, the evidence for the effects of low doses of vasopressin on extinction do not support this hypothesis and the evidence for high doses of vasopressin can be accounted for by the avoidance-inducing properties of vasopressin.

The role of histamine in estradiol-induced conditioned consumption reductions.

Conditioned consumption reductions (CCRs) develop toward novel taste stimuli as a consequence of associating those tastes with certain physiological changes. Few studies have focused on the neurochemical basis of this learned behavior. The purpose of these experiments was to reexamine the role of histamine in CCRs elicited by estradiol. Previous studies have suggested that histamine mediates CCRs induced by radiation, centrifugal rotation, and estradiol. However, because the animals were trained in a drug state, but tested in a nondrug state, it is possible that state-dependent learning confounded the results of these studies. The following series of experiments was performed to test this possibility for estradiol-induced CCRs. Implementing our own methodologies in Experiment 1, we demonstrated that an estradiol-induced CCR was blocked by treatment with the histamine 1 receptor blocker, chlorpheniramine maleate, before sucrose consumption during acquisition. In Experiment 2, identical states were maintained during acquisition and extinction by administering chlorpheniramine prior to sucrose exposure during both phases. The results indicated that chlorpheniramine blocked the estradiol-induced CCR. However, circumventing state-dependency in Experiment 3 by administering chlorpheniramine following exposure to sucrose during acquisition augmented the estradiol CCR. Taken together, the results of these experiments suggest that the ability of chlorpheniramine to abolish estradiol-induced CCRs is not due to state-dependency or to the antihistaminergic properties of chlorpheniramine. It is proposed that the results of all of the experiments can be accounted for by the aversive properties of chlorpheniramine.

Peripheral vasopressin accelerates extinction of conditioned taste avoidance.

Both peripheral and central administration of vasopressin improves retention and delays extinction when given before or after acquisition of shock avoidance learning. For conditioned taste avoidance, however, vasopressin prolongs extinction when injected peripherally before acquisition tests and accelerates extinction when infused intracerebroventricularly after acquisition. The following experiments were designed to determine whether this inconsistency is based on the route of administration or timing of vasopressin treatment. Because acquisition of conditioned taste avoidance is strengthened when an agent that is capable of inducing avoidance is administered after LiCl injection, it was determined in experiment 1 that a 6 microg/kg dose of vasopressin did not induce conditioned taste avoidance when administered 50 min after consumption of a sucrose solution. In experiment 2, it was determined that this dose of vasopressin accelerated extinction of a LiCl-induced conditioned taste avoidance when given 50 min after LiCl injection. These results suggest that the inconsistency is not based on route of administration. In experiment 3, it was determined that there was a tendency for animals to show prolonged extinction when vasopressin was administered 20 min before access to a sucrose solution. All of the results taken together suggest that the differential effects of vasopressin on extinction are due to the timing of administration. It was suggested that vasopressin accelerates extinction when given after acquisition by reducing the effectiveness of LiCl and it prolongs extinction when given before acquisition by altering neural responsiveness in areas mediating conditioned taste avoidance.

Role of the lateral parabrachial nucleus in apomorphine-induced conditioned consumption reduction: cooling lesions and relationship of c-Fos-like immunoreactivity to strength of conditioning.

The following experiments were designed to determine whether the lateral parabrachial nucleus (lPBN) mediates acquisition of conditioned consumption reduction induced by apomorphine, an agent that also has reinforcing properties. Temporary cooling lesions of the PBN blocked acquisition of apomorphine-induced conditioned consumption reduction. In addition, both apomorphine and LiCl activated c-Fos-like immunoreactivity (c-FLI) in the central, external, and crescent lPBN, and there was a strong correspondence between amount of c-FLI expression and strength of conditioned consumption reduction in these subnuclei. Taken together, these results support the hypothesis that the lPBN mediates apomorphine-induced conditioned consumption reduction, as is true for LiCl. Furthermore, they raise the possibility that the specific part of the lPBN mediating this conditioning effect of apomorphine and LiCl is 1 of the 3 subnuclei.

CD40 monoclonal antibody activation of antigen-presenting cells improves therapeutic efficacy of tumor-specific T cells.

OBJECTIVE: The goal of this study was to determine whether CD40 ligation of antigen presenting cells (APCs) enhances the anti-tumor effector function of tumor draining lymph node (TDLN) T lymphocytes in an adoptive immunotherapy model. STUDY DESIGN: MCA 205 TDLNs were culture activated both in the presence and absence of a stimulatory anti-CD40 monoclonal antibody (mAb) and effector cell phenotype, cytokine secretion in vitro and therapeutic efficacy in vivo were compared. RESULTS: Anti-CD40 mAb induced upregulation of APC cell surface activation markers that promoted generation of T cells that demonstrated an increase in tumor-specific IFN-gamma secretion and a statistically significant reduction in the number of pulmonary tumors (p< 0.01) after adoptive transfer. CONCLUSION: CD40 ligation of APCs in vitro results in the generation of T cells with enhanced effector function against established pulmonary tumors in vivo. SIGNIFICANCE: These findings have direct implications in the development of effective T cell-based immunotherapy of malignant conditions in human beings.

Vasopressin content in select brain regions during extinction of a conditioned taste aversion.

Previous studies have shown that low levels of vasopressin during extinction of conditioned taste avoidance are associated with a faster extinction, that fluid deprivation differentially alters vasopressin levels in various neural areas, and that extinction of conditioned taste avoidance is accelerated in fluid deprived male rats. The following study was designed to identify areas of the brain in which vasopressin levels are different in fluid deprived and nondeprived males during extinction of conditioned taste avoidance. Arginine vasopressin content was determined by radioimmunoassay in the paraventricular nucleus (PVN), medial amygdala (AMe), bed nucleus of the stria terminalis (BNST), nucleus tractus solitarius (NTS), medial septum (MS), lateral septum (LS), and insular cortex (IC) of unconditioned nondeprived males and conditioned males that were maintained on a 23-h fluid deprivation schedule or that were nondeprived. Vasopressin content in the PVN of deprived and nondeprived males differed during extinction. Based on comparisons with unconditioned nondeprived males, this difference was due to an elevation in the vasopressin content of the nondeprived but not the deprived males. These results raise the possibility that a vasopressinergic system in the PVN plays a critical role in the differential extinction rate of fluid deprived and nondeprived males, which will need to be verified by manipulating vasopressin levels in this brain site during extinction of a conditioned taste avoidance.

The meninges contribute to the conditioned taste avoidance induced by neural cooling in male rats.

After consumption of a novel sucrose solution, temporary cooling of neural areas that mediate conditioned taste avoidance can itself induce conditioned avoidance to the sucrose. It has been suggested that this effect is either a result of inactivation of neurons in these areas or of cooling the meninges. In a series of studies, we demonstrated that cooling the outer layer of the meninges, the dura mater, does not contribute to the conditioned taste avoidance induced by cooling any of these areas. The present experiments were designed to determine whether the inner layers of the meninges are involved. If they are involved, then one would expect that cooling locations in the brain that do not mediate conditioned taste avoidance, such as the caudate putamen (CP), would induce conditioned taste avoidance as long as the meninges were cooled as well. One also would expect that cooling neural tissue without cooling the meninges would reduce the strength of the conditioned taste avoidance. Experiment 1 established that the temperature of the neural tissue and meninges around the cold probes implanted in the CP were cooled to temperatures that have been shown to block synaptic transmission. Experiment 2 demonstrated that cooling the caudate putamen and overlying cortex and meninges induced conditioned taste avoidance. In experiment 3, a circle of meninges was cut away so that the caudate putamen and overlying cortex could be cooled without cooling the meninges. The strength of the conditioned taste avoidance was substantially reduced, but it was not entirely eliminated. These data support the hypothesis that cooling the meninges contributes to the conditioned taste avoidance induced by neural cooling. They also allow the possibility that neural inactivation produces physiological changes that can induce conditioned taste avoidance.

Exposure to estradiol before but not during acquisition of LiCl-induced conditioned taste avoidance accelerates extinction.

Estradiol accelerates extinction of LiCl-induced conditioned taste avoidance when it is present continuously before and during acquisition. We have suggested that the effect of estradiol on extinction is due to its illness-associated, rather than learning-associated, properties. If this were the case, then one would expect estradiol to act before but not during acquisition. This expectation is based on previous work showing attenuation of learned taste avoidance when rats are given distal preexposure (greater than 24 h before conditioning) or proximal preexposure (less than 24 h before conditioning) to the illness-inducing agent LiCl before acquisition of a LiCl-induced conditioned taste avoidance. In three separate experiments, estradiol was administered during three different time periods via subcutaneous implantation of a 10-mm estradiol-filled capsule. In each experiment, the extinction of estradiol-treated females was compared to that of females implanted with empty capsules. In the first experiment, female rats were given distal exposure to estradiol before acquisition. Capsules were implanted 11 days before acquisition and were removed 2 days before acquisition. In the second experiment, female rats were given proximal exposure to estradiol before acquisition. Capsules were implanted 2.5 h before LiCl was paired with a sucrose solution and were removed 16.5 h later. In the third experiment, female rats were given exposure to estradiol during acquisition. Capsules were implanted at the same time as LiCl administration and were removed 18 h later. The only estradiol-treated females to show accelerated extinction were those given distal preexposure to estradiol in Experiment 1. These data do not support a learning-associated hypothesis and only partially support an illness-associated hypothesis. The failure to find accelerated extinction following proximal preexposure may reflect an inappropriate choice of the parameters used in the experiment or a difference in the stimulus properties of LiCl and estradiol that allow each to serve as conditioning and preexposure agents in conditioned taste avoidance paradigms [corrected].

Cooling lesions of the lateral parabrachial nucleus during LiCl activation block acquisition of conditioned taste avoidance in male rats.

Lesions of the lateral parabrachial nucleus (lPBN) disrupt acquisition of LiCl-induced conditioned taste avoidance. Animals with lesions in this area also fail to exhibit taste neophobia. This raises the possibility that an inability of rats to recognize the taste solution as novel contributes to the deficit in taste avoidance learning. If this is the case, then one would expect conditioned taste avoidance not to be disrupted if the lPBN is functional during taste processing but not during LiCl processing. The first three experiments demonstrated that cooling was a viable method by which to temporarily inactivate the lPBN. Measurement of neural temperature during cooling indicated that the lPBN was cooled to temperatures that have been shown to block synaptic transmission but not axonal transmission. Cooling the lPBN itself induced a conditioned avoidance to a sucrose solution but this avoidance was abolished by exposure to daily cooling for 1 week prior to acquisition. In experiment 4, all animals were preexposed to lPBN cooling for 1 week. Those rats that received cooling lesions during a period that started immediately after sucrose solution consumption and extended through the peak effectiveness of LiCl failed to acquire a taste avoidance. These results fail to support the hypothesis that the deficit in taste avoidance learning after permanent lesions of the lPBN is due to an inability of lesioned animals to recognize the taste as novel. They are consistent with the hypothesis that this neural area processes ascending information about LiCl.

Central infusion of vasopressin in male rats accelerates extinction of conditioned taste avoidance induced by LiCl.

In shock avoidance tasks, extinction is prolonged when vasopressin is infused into the lateral ventricle after an acquisition session. Experiments were performed to determine whether a dose of vasopressin that does not induce conditioned taste avoidance (CTA) could prolong extinction of a LiCl-induced CTA when it is infused into the lateral ventricle of Sprague-Dawley male rats after acquisition. The first experiment was designed to determine whether infusion of vasopressin into the lateral ventricle would induce a CTA. Consumption of a sucrose solution was paired with infusion of vasopressin or saline, and even after two pairings, none of the vasopressin-treated rats showed decreases in sucrose consumption. Therefore, in the second experiment, this same dose of vasopressin was infused into the lateral ventricle 50 min after consumption of a sucrose solution was paired with an injection of LiCl. Vasopressin increased the rate of extinction of the LiCl-induced CTA. These results are the opposite of what has been found after peripheral administration of vasopressin before acquisition and/or extinction of a LiCl-induced CTA. Possible reasons for the difference in the direction of the effect on extinction include differential effects of vasopressin depending on the route of administration, the timing of injection, and the presence of aversive effects produced by the neuropeptide.