Context-dependent olfactory learning monitored by activities of salivary neurons in cockroaches

Context-dependent discrimination learning, a sophisticated form of nonelemental associative learning, has been found in many animals, including insects. The major purpose of this research is to establish a method for monitoring this form of nonelemental learning in rigidly restrained insects for investigation of underlying neural mechanisms. We report context-dependent olfactory learning (occasion-setting problem solving) of salivation, which can be monitored as activity changes of salivary neurons in immobilized cockroaches, Periplaneta americana. A group of cockroaches was trained to associate peppermint odor (conditioned stimulus, CS) with sucrose solution reward (unconditioned stimulus, US) while vanilla odor was presented alone without pairing with the US under a flickering light condition (1.0 Hz) and also trained to associate vanilla odor with sucrose reward while peppermint odor was presented alone under a steady light condition. After training, the responses of salivary neurons to the rewarded peppermint odor were significantly greater than those to the unrewarded vanilla odor under steady illumination and those to the rewarded vanilla odor was significantly greater than those to the unrewarded peppermint odor in the presence of flickering light. Similar context-dependent responses were observed in another group of cockroaches trained with the opposite stimulus arrangement. This study demonstrates context-dependent olfactory learning of salivation for the first time in any vertebrate and invertebrate species, which can be monitored by activity changes of salivary neurons in restrained cockroaches.     

Critical roles of mecamylamine-sensitive mushroom body neurons in insect olfactory learning

In insects, cholinergic neurons are thought to transmit olfactory conditioned stimulus (CS) to the sites for associating the CS with unconditioned stimulus (US), but the types of acetylcholine (ACh) receptor used by neurons participating in the association have not been determined. In cockroaches, a type of nicotinic ACh receptor specifically antagonized by mecamylamine (MEC) has been characterized. Here we investigated the roles of neurons possessing MEC-sensitive ACh receptors (MEC-sensitive neurons) in olfactory conditioning of salivation, monitored by changes in activities of salivary neurons, in cockroaches. Local and bilateral microinjection of MEC into each of the three olfactory centers, antennal lobes, calyces of the mushroom bodies and lateral protocerebra, impaired olfactory responses of salivary neurons, indicating that MEC-sensitive neurons in all olfactory centers participate in pathways mediating olfactory responses of salivary neurons. Conditioning of olfactory CS with sucrose US was impaired by injection of MEC into the antennal lobes or calyces, i.e., conditioned responses were absent even after recovery from MEC injection, suggesting that the CS-US association occurs in MEC-sensitive neurons in calyces (most probably Kenyon cells) or in neurons in downstream pathways. In contrast, conditioned responses appeared after recovery from MEC injection into the lateral protocerebra, suggesting that MEC-sensitive neurons in the lateral protocerebra are downstream of the association sites. Since lateral protocerebra are major termination areas of mushroom body efferent neurons, we suggest that input synapses of MEC-sensitive Kenyon cells, or their output synapses upon mushroom body efferent neurons, are the sites for CS-US association for conditioning of salivation.     

Context-dependent savings in procedural category learning

Environmental context can have a profound influence on the efficacy of intervention protocols designed to eliminate undesirable behaviors. This is clearly seen in drug rehabilitation clinics where patients often relapse soon after leaving the context of the treatment facility. A similar pattern is commonly observed in controlled laboratory studies of context-dependent savings in instrumental conditioning, where simply placing an animal back into the original conditioning chamber can renew an extinguished instrumental response. Surprisingly, context-dependent savings in human procedural learning has not been carefully examined in the laboratory. Here, we provide the first known empirical demonstration of context-dependent savings in a perceptual categorization task known to recruit procedural learning. We also present a computational account of these savings using a biologically detailed model in which a key role is played by cholinergic interneurons in the striatum.     

Context-dependent latent inhibition in taste aversion learning

The effects of taste stimulus preexposure, either in the presence or in the absence of a specific contextual cue consisting of a specific noise-producing bottle, upon the conditioning and testing of conditioned taste aversions to the taste (saccharin) plus context (noisy-bottle) compound stimulus were investigated. Four groups of rats were given preexposure trials (latent inhibition) to either: (1) novel saccharin in novel noisy bottles, (2) novel saccharin in familiar silent bottles, (3) familiar water in novel noisy bottles, (4) familiar water in familiar silent bottles, in six trials. During conditioning, saccharin was presented in the noisy bottles followed by lithium chloride for all the groups. At testing, saccharin was presented in the noisy bottles for both one-bottle and two-bottle tests of aversion. It was indicated that the conditioning decrement produced after both saccharin and noisy bottle preexposure was overwhelmingly greater than any produced after preexposure to the elements. These results, discussed in relation to current theories of latent inhibition and perceptual learning, further underline the overwhelming significance of exteroceptive contextual elements in conditioned taste aversions.     

Color modulates olfactory learning in honeybees by an occasion-setting mechanism

A sophisticated form of nonelemental learning is provided by occasion setting. In this paradigm, animals learn to disambiguate an uncertain conditioned stimulus using alternative stimuli that do not enter into direct association with the unconditioned stimulus. For instance, animals may learn to discriminate odor rewarded from odor nonrewarded trials if these two situations are indicated by different colors that do not themselves become associated with the reward. Despite a growing interest in nonelemental learning in insects, no study has so far attempted to study occasion setting in restrained honeybees, although this would allow direct access to the neural basis of nonelemental learning. Here we asked whether colors can modulate olfactory conditioning of the proboscis extension reflex (PER) via an occasion-setting mechanism. We show that intact, harnessed bees are not capable of learning a direct association between color and sucrose. Despite this incapacity, bees solved an occasion-setting discrimination in which colors set the occasion for appropriate responding to an odor that was rewarded or nonrewarded depending on the color. We therefore provide the first controlled demonstration of bimodal (color-odor) occasion setting in harnessed honeybees, which opens the door for studying the neural basis of such bimodal, nonelemental discriminations in insects.     

Embryonic olfactory learning in frogs

We examined the effect of embryonic olfactory experiences on behaviour after hatching in two species of anuran amphibians, Rana temporaria and Rana sylvatica. Odorants (orange and citral) injected into the egg were “learned” by the embryo and subsequently preferred by tadpoles after hatching. The observed preferences were specific to the odours experienced prior to hatching, and tadpoles discriminated between the odour experienced as embryos and a novel odour. The acquired preference was maintained after metamorphosis for those individuals that exhibited a preference as tadpoles. Preferences of tadpoles were also influenced “naturally” by odours present in the water surrounding the developing embryos. The experimentally induced olfactory preferences did not appear to influence the exhibition of sibling recognition abilities. The study represents the first demonstration of “embryonic” learning in amphibians, and the functions and importance of early olfactory learning for amphibians are discussed.     

Context-dependent use of olfactory cues by foragers of Vespula germanica social wasps

Animal Cognition - Food search is guided by cues from different sensory modalities, such as olfactory and visual. In social wasps, olfaction plays a key role in locating new resources. However,...     

Embryonic olfactory learning in frogs.

We examined the effect of embryonic olfactory experiences on behaviour after hatching in two species of anuran amphibians, Rana temporaria and Rana sylvatica. Odorants (orange and citral) injected into the egg were "learned" by the embryo and subsequently preferred by tadpoles after hatching. The observed preferences were specific to the odours experienced prior to hatching, and tadpoles discriminated between the odour experienced as embryos and a novel odour. The acquired preference was maintained after metamorphosis for those individuals that exhibited a preference as tadpoles. Preferences of tadpoles were also influenced "naturally" by odours present in the water surrounding the developing embryos. The experimentally induced olfactory preferences did not appear to influence the exhibition of sibling recognition abilities. The study represents the first demonstration of "embryonic" learning in amphibians, and the functions and importance of early olfactory learning for amphibians are discussed.     

Successive olfactory reversal learning in honeybees

Honeybees Apis mellifera can associate an originally neutral odor with a reinforcement of sucrose solution. Forward pairings of odor and reinforcement enable the odor to release the proboscis extension reflex in consecutive tests. Bees can also be conditioned differentially: They learn to respond to a reinforced odor and not to a nonreinforced one. They can also learn to reverse their choice. Here we ask whether honeybees can learn successive olfactory differential conditioning tasks involving different overlapping pairs of odors. The conditioning schedules were established in order to train the animals with 3, 2, 1, or 0 reversals previous to a last differential conditioning phase in which two additional reversals were present. We studied whether or not successive reversal learning is possible and whether or not learning olfactory discrimination reversals affects the solving of subsequent discrimination reversals. Therefore we compared the responses of bees that had experienced reversals with those of bees that had not experienced such reversals when both are confronted with a new reversal situation. In experiment 1 we showed that bees that had experienced three previous reversals were better in solving the final reversal task than bees with no previous reversal experience. In experiment 2, we showed that one reversal learning is enough for bees to perform better in the final reversal task. The successive different reversals trained in our experiments resemble the natural foraging situation in which a honeybee forager has to switch successively from an initial floral species to different ones. The fact that experiencing such changes seems to improve a bee's performance in dealing with further new exploited food sources has therefore an adaptive impact for the individual and for the colony as a whole.     

Acetylcholine and olfactory perceptual learning

Olfactory perceptual learning is a relatively long-term, learned increase in perceptual acuity, and has been described in both humans and animals. Data from recent electrophysiological studies have indicated that olfactory perceptual learning may be correlated with changes in odorant receptive fields of neurons in the olfactory bulb and piriform cortex. These changes include enhanced representation of the molecular features of familiar odors by mitral cells in the olfactory bulb, and synthetic coding of multiple coincident odorant features into odor objects by cortical neurons. In this paper, data are reviewed that show the critical role of acetylcholine (Ach) in olfactory system function and plasticity, and cholinergic modulation of olfactory perceptual learning at both the behavioral and cortical level.     

Peak shift in honey bee olfactory learning

If animals are trained with two similar stimuli such that one is rewarding (S+) and one punishing (S?), then following training animals show a greatest preference not for the S+, but for a novel stimulus that is slightly more different from the S? than the S+ is. This peak shift phenomenon has been widely reported for vertebrates and has recently been demonstrated for bumblebees and honey bees. To explore the nature of peak shift in invertebrates further, here we examined the properties of peak shift in honey bees trained in a free-flight olfactory learning assay. Hexanal and heptanol were mixed in different ratios to create a continuum of odour stimuli. Bees were trained to artificial flowers such that one odour mixture was rewarded with 2 molar sucrose (S+), and one punished with distasteful quinine (S?). After training, bees were given a non-rewarded preference test with five different mixtures of hexanal and heptanol. Following training bees’ maximal preference was for an odour mixture slightly more distinct from the S? than the trained S+. This effect was not seen if bees were initially trained with two distinct odours, replicating the classic features of peak shift reported for vertebrates. We propose a conceptual model of how peak shift might occur in honey bees. We argue that peak shift does not require any higher level of processing than the known olfactory learning circuitry of the bee brain and suggest that peak shift is a very general feature of discrimination learning.     

Elemental and non-elemental olfactory learning in Drosophila

Brain complexity varies across many orders of magnitude between animals, and it is often assumed that complexity underpins cognition. It is thus important to explore the cognitive capacity of widely used model organisms such as Drosophila. We systematically investigated the fly’s ability to learn discriminations involving compound olfactory stimuli associated with shock. Flies could distinguish binary mixtures (AB+ CD-), including overlapping mixtures (AB+ BC-). They could learn positive patterning (AB+ A- B-) but could not learn negative patterning (A+ B+ AB-) or solve a biconditional discrimination task (AB+ CD+ AC- BD-). Learning about the elements of a compound (AB+) was not affected by prior conditioning of one of the elements (A+ AB+): flies do not exhibit blocking in this task. We compare these results with the predictions from simulation of several well-known theoretical models of learning, and find none are fully consistent with the overall pattern of observed behaviour.     

Brain composition and olfactory learning in honey bees

Correlations between brain or brain component size and behavioral measures are frequently studied by comparing different animal species, which sometimes introduces variables that complicate interpretation in terms of brain function. Here, we have analyzed the brain composition of honey bees (Apis mellifera) that have been individually tested in an olfactory learning paradigm. We found that the total brain size correlated with the bees’ learning performance. Among different brain components, only the mushroom body, a structure known to be involved in learning and memory, showed a positive correlation with learning performance. In contrast, visual neuropils were relatively smaller in bees that performed better in the olfactory learning task, suggesting modality-specific behavioral specialization of individual bees. This idea is also supported by inter-individual differences in brain composition. Some slight yet statistically significant differences in the brain composition of European and Africanized honey bees are reported. Larger bees had larger brains, and by comparing brains of different sizes, we report isometric correlations for all brain components except for a small structure, the central body.     

Matching- and nonmatching-to-sample concept learning in rats using olfactory stimuli

Previous research has shown that rats can learn matching-to-sample relations with olfactory stimuli; however, the specific characteristics of this relational control are unclear. In Experiment 1, 6 rats were trained to either match or nonmatch to sample in a modified operant chamber using common household spices as olfactory stimuli. After matching or nonmatching training with 10 exemplars, the contingencies were reversed with five new stimuli such that subjects trained on matching were shifted to nonmatching and vice versa. Following these reversed contingencies, the effects of the original training persisted for many trials with new exemplars. In Experiment 2, 9 rats were trained with matching procedures in an arena that provided for 18 different spatial locations for comparison stimuli. Five subjects were trained with differential reinforcement outcomes and 4 with only one type of reinforcer. Differential outcomes and multiple exemplars facilitated learning, and there was strong evidence for generalization to new stimuli for most rats that acquired several conditional discriminations. Performances with novel samples were generally above chance, but rarely reached the high levels obtained during baseline with well-trained stimulus relations. However, taken together, the data from the two experiments extend previous work, show that rats can learn both match and nonmatch relations with different experimental protocols, and demonstrate generalization to novel sample stimuli.     

Context-dependent migrations in visual word perception

In the present study we investigate the effect of context--that is, a prime--on migration errors. Migration errors, or migrations, are caused by perceptual interactions between two words in a visual display, such as line-lace. After postcuing, subjects sometimes report lane or lice, instead of the requested word line. This phenomenon has been demonstrated by previous studies. In the experiments reported here, we replicate this phenomenon. We also find that more migrations of the lane type occur (compared with lice type), when the display line-lace is preceded by a prime related to lane (but not to lice). This shows that higher order word knowledge, in the form of semantic relations between words, influences the migration phenomenon. Further, we show that the number of migration errors are not a result of only a summation of activation from the letters in the display. The method we developed to isolate context-dependent migrations also gives valuable information about the effect of contextual information on word perception.     

Context-dependent Abduction and Relevance

Based on the premise that what is relevant, consistent, or true may change from context to context, a formal framework of relevance and context is proposed in which

Roles for Drosophila mushroom body neurons in olfactory learning and memory

Olfactory learning assays in Drosophila have revealed that distinct brain structures known as mushroom bodies (MBs) are critical for the associative learning and memory of olfactory stimuli. However, the precise roles of the different neurons comprising the MBs are still under debate. The confusion surrounding the roles of the different neurons may be due, in part, to the use of different odors as conditioned stimuli in previous studies. We investigated the requirements for the different MB neurons, specifically the alpha/beta versus the gamma neurons, and whether olfactory learning is supported by different subsets of MB neurons irrespective of the odors used as conditioned stimuli. We expressed the rutabaga (rut)-encoded adenylyl cyclase in either the gamma or alpha/beta neurons and examined the effects on restoring olfactory associative learning and memory of rut mutant flies. We also expressed a temperature-sensitive shibire (shi) transgene in these neuron sets and examined the effects of disrupting synaptic vesicle recycling on Drosophila olfactory learning. Our results indicate that although we did not detect odor-pair-specific learning using GAL4 drivers that primarily express in gamma neurons, expression of the transgenes in a subset of alpha/beta neurons resulted in both odor-pair-specific rescue of the rut defect as well as odor-pair-specific disruption of learning using shi(ts1).