Experience-dependent tuning of the olfactory system.

Insects rely on their sense of smell to navigate complex environments and make decisions regarding food and reproduction. However, in natural settings, the odors that convey this information may come mixed with environmental odors that can obscure their perception. Therefore, recognizing the presence of informative odors involves generalization and discrimination processes, which can be facilitated when there is a high contrast between stimuli, or the internal representation of the odors of interest outcompetes that of concurrent ones. The first two layers of the olfactory system, which involve the detection of odorants by olfactory receptor neurons and their encoding by the first postsynaptic partners in the antennal lobe, are critical for achieving such optimal representation. In this review, we summarize evidence indicating that experience-dependent changes adjust these two levels of the olfactory system. These changes are discussed in the context of the advantages they provide for detection of informative odors.

Remembering how to run: A descriptive wheel run analysis in CF1 male and female mice.

Physical exercise is known to have beneficial effects on general health and wellbeing in humans and it is also related to neuronal plasticity, increasing neurogenesis and consequently leading to improvements in processes such as learning and memory. In this sense, wheel running performance in mice appears as an extensively used behavioral approach for neurobiological studies. Here, we explored the running patterns in CF1 male and female mice allowing voluntary wheel running for 20 min along three consecutive days. We analyzed differences in the accumulated distance traveled, instant velocity, and latency to run and breaks taken in both males and females, comparing performance between days. Results revealed that after a first experience with the wheel, animals that had learnt how to run on day 1 quickly look forward to stepping into the wheel in subsequent training days, reflected by a significant increase in daily running distance and velocity. Further, no differences were found in the running performance between males and females. In summary, in a first experience with the wheel, animals get familiarized with the wheel and grow accustomed to it.

Antennal movements can be used as behavioral readout of odor valence in honey bees.

The fact that honey bees have a relatively simple nervous system that allows complex behaviors has made them an outstanding model for studying neurobiological processes. Studies on learning and memory routinely use appetitive and aversive learning paradigms that involve recording of the proboscis or the sting extension. However, these protocols are based on all-or-none responses, which has the disadvantage of occluding intermediate and more elaborated behaviors. Nowadays, the great advances in tracking software and data analysis, combined with affordable video recording systems, have made it possible to extract very detailed information about animal behavior. Here we describe antennal movements that are elicited by odor that have no, positive or negative valence. We show that animals orient their antennae towards the source of the odor when it is positive, and orient them in the opposite direction when the odor is negative. Moreover, we found that this behavior was modified between animals that had been trained based on protocols of different strength. Since this procedure allows a more accurate description of the behavioral outcome using a relatively small number of animals, it represents a great tool for studying different cognitive processes and olfactory perception.

Honey bees can store and retrieve independent memory traces after complex experiences that combine appetitive and aversive associations.

Real-world experiences often mix appetitive and aversive events. Understanding the ability of animals to extract, store and use this information is an important issue in neurobiology. We used honey bees as model organism to study learning and memory after a differential conditioning paradigm that combines appetitive and aversive training trials. First, we used an aversive conditioning paradigm that constitutes a clear opposite of the well-known appetitive olfactory conditioning of the proboscis extension response. A neutral odour is presented paired with the bitter substance quinine. Aversive memory is evidenced later as an odour-specific impairment in appetitive conditioning. Then, we tested the effect of mixing appetitive and aversive conditioning trials distributed along the same training session. Differential conditioning protocols like this were used previously to study the ability to discriminate odours; however, they were not focused on whether appetitive and aversive memories are formed. We found that after differential conditioning, honey bees establish independent appetitive and aversive memories that do not interfere with each other during acquisition or storage. Finally, we moved the question forward to retrieval and memory expression to evaluate what happens when appetitive and the aversive learned odours are mixed during a test. Interestingly, opposite memories compete in such a way that they do not cancel each other out. Honey bees showed the ability to switch from expressing appetitive to aversive memory depending on their satiation level.

Learning a non-neutral conditioned stimulus: place preference in the crab Neohelice granulata.

In the wild, being able to recognize and remember specific locations related to food sources and the associated attributes of landmarks is a cognitive trait important for survival. In the present work, we show that the crab Neohelice granulata can be trained to associate a specific environment with an appetitive reward in a conditioned place preference task. After a single training trial, when the crabs were presented with a food pellet in the target quadrant of the training arena, they were able to form a long-term memory related to the event. This memory was evident at least 24 h after training and was protein synthesis dependent. Importantly, the target area of the arena proved to be a non-neutral environment, given that animals initially avoided the target quadrant. In the present work, we introduce for the first time an associative one-trial memory paradigm including a conditioned stimulus with a clear valence performed in a crustacean.

Learning-dependent plasticity in the antennal lobe improves discrimination and recognition of odors in the honeybee.

Honeybees are extensively used to study olfactory learning and memory processes thanks to their ability to discriminate and remember odors and because of their advantages for optophysiological recordings of the circuits involved in memory and odor perception. There are evidences that the encoding of odors in areas of primary sensory processing is not rigid, but undergoes changes caused by olfactory experience. The biological meaning of these changes is focus of intense discussions. Along this review, we present evidences of plasticity related to different forms of learning and discuss its function in the context of olfactory challenges that honeybees have to solve. So far, results in honeybees are consistent with a model in which changes in early olfactory processing contributes to the ability of an animal to recognize the presence of relevant odors and facilitates the discrimination of odors in a way adjusted to its own experience.

Parallel memory traces are built after an experience containing aversive and appetitive components in the crab Neohelice.

The neurobiology of learning and memory has been mainly studied by focusing on pure aversive or appetitive experiences. Here, we challenged this approach considering that real-life stimuli come normally associated with competing aversive and appetitive consequences and that interaction between conflicting information must be intrinsic part of the memory processes. We used Neohelice crabs, taking advantage of two well-described appetitive and aversive learning paradigms and combining them in a single training session to evaluate how this affects memory. We found that crabs build separate appetitive and aversive memories that compete during retrieval but not during acquisition. Which memory prevails depends on the balance between the strength of the unconditioned stimuli and on the motivational state of the animals. The results indicate that after a mix experience with appetitive and aversive consequences, parallel memories are established in a way that appetitive and aversive information is stored to be retrieved in an opportunistic manner.

Sickness Behavior in Honey Bees.

During an infection, animals suffer several changes in their normal physiology and behavior which may include lethargy, appetite loss, and reduction in grooming and general movements. This set of alterations is known as sickness behavior and although it has been extensively believed to be orchestrated primarily by the immune system, a relevant role for the central nervous system has also been established. The aim of the present work is to develop a simple animal model to allow studying how the immune and the nervous systems interact coordinately during an infection. We administered a bacterial lipopolysaccharide (LPS) into the thorax of honey bees to mimic a bacterial infection, and then we evaluated a set of stereotyped behaviors of the animals that might be indicative of sickness behavior. First, we show that this immune challenge reduces the locomotor activity of the animals in a narrow time window after LPS injection. Furthermore, bees exhibit a loss of appetite 60 and 90 min after injection, but not 15 h later. We also demonstrate that LPS injection reduces spontaneous antennal movements in harnessed animals, which suggests a reduction in the motivational state of the bees. Finally, we show that the LPS injection diminishes the interaction between animals, a crucial behavior in social insects. To our knowledge these results represent the first systematic description of sickness behavior in honey bees and provide important groundwork for the study of the interaction between the immune and the neural systems in an insect model.

Sweet taste and nutrient value subdivide rewarding dopaminergic neurons in Drosophila.

Dopaminergic neurons provide reward learning signals in mammals and insects [1-4]. Recent work in Drosophila has demonstrated that water-reinforcing dopaminergic neurons are different to those for nutritious sugars [5]. Here, we tested whether the sweet taste and nutrient properties of sugar reinforcement further subdivide the fly reward system. We found that dopaminergic neurons expressing the OAMB octopamine receptor [6] specifically convey the short-term reinforcing effects of sweet taste [4]. These dopaminergic neurons project to the ß'2 and γ4 regions of the mushroom body lobes. In contrast, nutrient-dependent long-term memory requires different dopaminergic neurons that project to the γ5b regions, and it can be artificially reinforced by those projecting to the ß lobe and adjacent α1 region. Surprisingly, whereas artificial implantation and expression of short-term memory occur in satiated flies, formation and expression of artificial long-term memory require flies to be hungry. These studies suggest that short-term and long-term sugar memories have different physiological constraints. They also demonstrate further functional heterogeneity within the rewarding dopaminergic neuron population.

Drosophila learn opposing components of a compound food stimulus.

Dopaminergic neurons provide value signals in mammals and insects. During Drosophila olfactory learning, distinct subsets of dopaminergic neurons appear to assign either positive or negative value to odor representations in mushroom body neurons. However, it is not known how flies evaluate substances that have mixed valence. Here we show that flies form short-lived aversive olfactory memories when trained with odors and sugars that are contaminated with the common insect repellent DEET. This DEET-aversive learning required the MB-MP1 dopaminergic neurons that are also required for shock learning. Moreover, differential conditioning with DEET versus shock suggests that formation of these distinct aversive olfactory memories relies on a common negatively reinforcing dopaminergic mechanism. Surprisingly, as time passed after training, the behavior of DEET-sugar-trained flies reversed from conditioned odor avoidance into odor approach. In addition, flies that were compromised for reward learning exhibited a more robust and longer-lived aversive-DEET memory. These data demonstrate that flies independently process the DEET and sugar components to form parallel aversive and appetitive olfactory memories, with distinct kinetics, that compete to guide learned behavior.

Dopamine interferes with appetitive long-term memory formation in honey bees.

Studies in vertebrates and invertebrates have proved the instructive role that different biogenic amines play in the neural representation of rewards and punishments during associative learning. Results from diverse arthropods and using different learning paradigms initially agreed that dopamine (DA) is needed for aversive learning and octopamine (OA) is needed for appetitive learning. However, the notion that both amines constitute separate pathways for appetitive and aversive learning is changing. Here, we asked whether DA, so far only involved in aversive memory formation in honey bees, does also modulate appetitive memory. Using the well characterized appetitive olfactory conditioning of the proboscis extension reflex (PER), we show that DA impairs appetitive memory consolidation. In addition, we found that blocking DA receptors enhances appetitive memory. These results are consistent with the view that aversive and appetitive components interact during learning and memory formation to ensure adaptive behavior.

Opposite actions of dopamine on aversive and appetitive memories in the crab.

The understanding of how the reinforcement is represented in the central nervous system during memory formation is a current issue in neurobiology. Several studies in insects provide evidence of the instructive role of biogenic amines during the learning and memory process. In insects it was widely accepted that dopamine (DA) mediates aversive reinforcements. However, the idea of DA being exclusively involved in aversive memory has been challenged in recent studies. Here, we study the involvement of DA during aversive and appetitive memories in the crab Chasmagnathus. We found that DA-receptor antagonists impair aversive memory consolidation, in agreement with previous reports in insects, while administration of DA facilitates memory formation after a weak training protocol. In contrast, DA treatment during appetitive training was found to impair formation of long-term appetitive memory. In addition, as a first step in elucidating the neuroanatomical correlates of DA action on memory, we mapped dopaminergic neurons in the central nervous system of the crab. Results of the current study, together with those obtained in a previous work about the role of octopamine (OA), suggest that both amines (DA and OA) play a dual action in memory processes. On the one hand, DA and OA mediate the aversive and the appetitive signals, respectively, throughout training, while on the other hand, they interfere with the formation of memory of the opposite sign (DA in appetitive and OA in aversive). Our results support a new understanding about the way appetitive and aversive stimuli are processed during memory formation to ensure adaptive behavior.

Dissecting mechanisms of reconsolidation: octopamine reveals differences between appetitive and aversive memories in the crab Chasmagnathus.

Ample evidence suggests that, when reactivated by a reminder, a consolidated memory may return to a labile state and needs to be stabilized again in order to persist, a process known as reconsolidation. In a previous study, performed in the crab Chasmagnathus, we found a dual role for the biogenic amine octopamine (OA) during memory consolidation. On the one hand, it was necessary for appetitive memory formation and, on the other, it had a deleterious effect on aversive memory consolidation. Thus, OA could be a good candidate to dissect the neurochemical mechanisms of appetitive and aversive reconsolidation. Here, we initially characterized the reconsolidation of an appetitive memory. Then, we compared appetitive reconsolidation with its aversive counterpart regarding the implication of OA in these processes, and contrasted them with previous findings obtained in the consolidation phase. Our results demonstrate that appetitive reconsolidation takes place when animals are re-exposed to the training context, as shown by the amnesic effect of cycloheximide when applied before the reminder. In addition, the no-reinforcement during the reminder is a necessary condition for appetitive reconsolidation to occur. Remarkably, appetitive reconsolidation is neither impaired by OA receptor antagonists nor facilitated by exogenous OA, whereas aversive reconsolidation can be interfered with by OA administration. Thus, our results indicate that appetitive reconsolidation does not involve OA signaling, while aversive reconsolidation is negatively modulated by OA. All in all, these results could constitute a step towards the identification of particular features of appetitive and aversive reconsolidation.