Olfactory learning in individually assayed Drosophila larvae

Insect and mammalian olfactory systems are strikingly similar. Therefore, Drosophila can be used as a simple model for olfaction and olfactory learning. The brain of adult Drosophila, however, is still complex. We therefore chose to work on the larva with its yet simpler but adult-like olfactory system and provide evidence for olfactory learning in individually assayed Drosophila larvae. We developed a differential conditioning paradigm in which odorants are paired with positive (“+” fructose) or negative (“-” quinine or sodium chloride) gustatory reinforcers. Test performance of individuals from two treatment conditions is compared—one received odorant A with the positive reinforcer and odorant B with a negative reinforcer (A+/B-); animals from the other treatment condition were trained reciprocally (A-/B+). During test, differences in choice between A and B of individuals having undergone either A+/B- or A-/B+ training therefore indicate associative learning. We provide such evidence for both combinations of reinforcers; this was replicable across repetitions, laboratories, and experimenters. We further show that breaks improve performance, in accord with basic principles of associative learning. The present individual assay will facilitate electrophysiological studies, which necessarily use individuals. As such approaches are established for the larval neuromuscular synapse, but not in adults, an individual larval learning paradigm will serve to link behavioral levels of analysis to synaptic physiology.     

A neutral odor may become a sexual incentive through classical conditioning in male rats

A neutral olfactory stimulus was employed as CS in a series of experiments with a sexually receptive female as UCS and the execution of an intromission as the UCR. Each experimental session lasted until the male ejaculated. The time the experimental subject spent in a zone adjacent to the source of the olfactory stimulus during the 10 s of CS presentation and the latency to enter that area were employed as measures of conditioning. In Experiment 1, the CS was always presented at the same location in the conditioning arena, bordering the site where the female became accessible. Conditioning resulted in a gradual increase in the time spent in the CS zone as well as in a reduction of the latency to enter it. In Experiment 2, the site of presentation of the CS was varied randomly from one side of the conditioning arena to another in order to assure that the subjects indeed associated the CS and not a particular location with access to a female. Again, the time spent in the CS zone increased and the latency to enter it was reduced by conditioning. Furthermore, after conditioning the CS was approached more than a neutral odor in a different context. In a third experiment one group of animals was randomly exposed to the CS to control for non-associative effects on performance. Another group was conditioned as in Experiment 2. The random presentation of the CS failed to enhance approach, while the paired UCS-CS had effects identical to those found in Experiment 2. In the last experiment, an olfactory CS⁻ was included. The CS⁺ and CS⁻ were presented in random order at random locations. Both the time spent in the CS⁺ and CS⁻ zones was increased, while the latency to enter the zone was significantly reduced only for the CS⁺. These data suggest that the subjects had partially learned to discriminate between the CS⁺ and the CS⁻, at most. The procedure employed in Experiments 2 and 3 seems to be the most promising for studies of sexual conditioning in male rats.     

Pavlovian processes do mediate control of the “advance” response strategy

Two parallel experiments used pigeons in a transfer of control design to determine the basis of the transfer of the use of the “advance strategy” across successive discriminations (W. K. Honig & H. Lindsay, Learning and Motivation, 1975, 6, 157–178). Pigeons were first trained under instrumental contingencies to use an “advance” response to maximize their exposure to S^D and minimize their exposure to S^Δ. Then orthogonal stimuli (key lights in Experiment 1 and diffuse illuminations in Experiment 2) were independently established through separate discriminative Pavlovian procedures as CS+ and CS? or as CS^o's. Later in special tests, it was demonstrated that the associative values of the Pavlovian CS's were a major factor governing the use of the advance strategy: only pigeons exposed to $\text{CS+}\text{CS?}$ used the advance response to regulate their exposure to either CS. Although these experiments showed that the necessary and sufficient conditions for utilization of the advance strategy are the Pavlovian associative values of the discriminative stimuli, opportunity for differential “operant” responding during the discriminative stimuli seems to be a contributor to the optimal use of the advance response across successive discriminations.     

Learning in honey bees with brain lesions: how partial mushroom-body ablations affect sucrose responsiveness and tactile antennal learning

Honey bees are ideal organisms for studying associative learning, because they can rapidly learn different sensory cues, even under laboratory conditions. Classical olfactory learning experiments have shown that the mushroom bodies (MBs), a prominent neuropil of the central nervous system of the bee, are involved in olfactory learning and memory formation. We tested whether the MBs are also involved in tactile antennal learning. As in olfactory learning, bees use the antennae during tactile learning to sense tactile cues. We produced specific MB ablations by applying the mitotic blocker hydroxyurea (HU). In Drosophila, HU-induced brain lesions of the MBs strongly impaired olfactory memory formation. As treatment with HU might also interfere with the processing of the reward stimulus, sucrose, we measured the responsiveness to sucrose stimuli in these bees. Treatment with HU led to partial ablations of the median MB sub-units on one or both sides of the brain. We analysed side-specific effects in double-blind tests, testing sucrose responsiveness separately for each antenna, and conditioning first one antenna and then the other in a reversal learning assay. HU-treated bees without detectable ablations were less responsive to sucrose and had a poorer learning performance than untreated controls. Partial MB ablation did not additionally affect responsiveness to sucrose or tactile antennal learning. Interestingly, bees with bilateral MB ablations did not differ from untreated controls in their learning performance during the first learning phase. During reversal learning, acquisition in these bees was significantly lower than that in untreated controls. It is concluded that HU treatment affects sucrose responsiveness and tactile learning even without detectable ablation of neuropils. The effects of MB ablations on tactile learning are not side-specific and not correlated with the volume of the ablated neuropil. Accepted after revision: 15 January 2001 ❚ Electronic Publication     

The Organization of Extrinsic Neurons and Their Implications in the Functional Roles of the Mushroom Bodies in Drosophila melanogaster Meigen

Although the importance of the Drosophila mushroom body in olfactory learning and memory has been stressed, virtually nothing is known about the brain regions to which it is connected. Using Golgi and GAL4–UAS techniques, we performed the first systematic attempt to reveal the anatomy of its extrinsic neurons. A novel presynaptic reporter construct, UAS-neuronal synaptobrevin–green fluorescent protein (n-syb–GFP), was used to reveal the direction of information in the GAL4-labeled neurons. Our results showed that the main target of the output neurons from the mushroom body lobes is the anterior part of the inferior medial, superior medial, and superior lateral protocerebrum. The lobes also receive afferents from these neuropils. The lack of major output projections directly to the deutocerebrum’s premotor pathways discourages the view that the role of the mushroom body may be that of an immediate modifier of behavior. Our data, as well as a critical evaluation of the literature, suggest that the mushroom body may not by itself be a “center” for learning and memory, but that it can equally be considered as a preprocessor of olfactory signals en route to “higher” protocerebral regions.     

Pavlovian conditioned stimulus effects upon instrumental choice behavior are reinforcer specific

In a transfer-of-control experiment with rats, Pavlovian CSs were tested for the specificity of their effects. The instrumental behavior consisted of a discriminative, conditional two-lever choice task in which qualitatively different appetitive reinforcers were contingent upon the two correct choices. In a Pavlovian phase, subjects experienced conditioning to establish either a CS⁺ or CS^? for one reinforcer or a CS⁺ or CS^? for the other reinforcer. Finally, in a test, these CSs were presented when there was the opportunity to make choice responses. The CS⁺s evoked choices of the lever which had eventuated in the reinforcer that had served as the Pavlovian US, while the CS^?s showed only a slight tendency to evoke the other choice responses. When the CSs were compounded with the original S^Ds, the CS⁺s had little effect upon the vigor of responding while the CS^?s reduced the vigor of responding to the S^D for the reinforcer that was the same as the US used in establishing the CS^?. The results are discussed in terms of associative mediational theory and the reinforcer specificity of Pavlovian conditioned excitation and inhibition.     

Octopaminergic neurons have multiple targets in Drosophila larval mushroom body calyx and can modulate behavioral odor discrimination

Discrimination of sensory signals is essential for an organism to form and retrieve memories of relevance in a given behavioral context. Sensory representations are modified dynamically by changes in behavioral state, facilitating context-dependent selection of behavior, through signals carried by noradrenergic input in mammals, or octopamine (OA) in insects. To understand the circuit mechanisms of this signaling, we characterized the function of two OA neurons, sVUM1 neurons, that originate in the subesophageal zone (SEZ) and target the input region of the memory center, the mushroom body (MB) calyx, in larval Drosophila. We found that sVUM1 neurons target multiple neurons, including olfactory projection neurons (PNs), the inhibitory neuron APL, and a pair of extrinsic output neurons, but relatively few mushroom body intrinsic neurons, Kenyon cells. PN terminals carried the OA receptor Oamb, a Drosophila α1-adrenergic receptor ortholog. Using an odor discrimination learning paradigm, we showed that optogenetic activation of OA neurons compromised discrimination of similar odors but not learning ability. Our results suggest that sVUM1 neurons modify odor representations via multiple extrinsic inputs at the sensory input area to the MB olfactory learning circuit.

Behavioral choices depend on discrimination among “sensory objects,” which are neural representations of multiple coincident sensory inputs, across a range of sensory modalities. For example, “odor objects” (Gottfried 2009; Wilson and Sullivan 2011; Gire et al. 2013) are represented in sparse ensembles of neurons, that are coincidence detectors of multiple parallel inputs from odor quality channels. This principle is used widely in animals, including in mushroom bodies (MBs), the insect center for associative memory (Masuda-Nakagawa et al. 2005; Honegger et al. 2011), and in the piriform cortex (PCx) of mammals (Stettler and Axel 2009; Davison and Ehlers 2011).The selectivity of sensory representations can be modulated dynamically by changes in behavioral state, allowing an animal to learn and respond according to perceptual task. In mammals, the noradrenergic system originating in the locus coeruleus (LC) is implicated in signaling behavioral states such as attention, arousal and expectation (Aston-Jones and Cohen 2005; Sara and Bouret 2012).In insects, octopamine (OA), structurally and functionally similar to noradrenalin (NA) in mammals (Roeder 2005), can mediate changes in behavioral state that often promote activity; for example, sensitization of reflex actions in locusts (Sombati and Hoyle 1984), aggressive state in crickets (Stevenson et al. 2005), initiation and maintenance of flight state (Brembs et al. 2007; Suver et al. 2012), and enhanced excitability of Drosophila motion detection neurons during flight (Strother et al. 2018). Another role of OA is as a reward signal: A single OA neuron, VUMmx1, mediates the reinforcing function of unconditioned stimulus in the honeybee proboscis extension reflex (Hammer 1993; Hammer and Menzel 1998; Menzel 2012). In Drosophila, acquisition of appetitive memory is impaired in TβH mutants, unable to synthesize OA (Schwaerzel et al. 2003), and activation of OA neurons can substitute reinforcing stimulus in appetitive learning (Schroll et al. 2006). Moreover, OA receptors are necessary for reward learning in Drosophila (Burke et al. 2012) and crickets (Matsumoto et al. 2015).To understand the neural mechanisms of OA in higher order sensory discrimination, we used the simple sensory “cortex” of larval Drosophila, the calyx, which is the sensory input region of the mushroom bodies (MBs), the insect memory center. Here, each MB neuron (Kenyon cell [KC]) typically arborizes in several glomeruli, most of which are organized around the terminus of an olfactory projection neuron (PN); KCs thus combinatorially integrate multiple sensory input channels (Masuda-Nakagawa et al. 2005) and are coincidence detectors of multiple inputs. The APL provides inhibitory feedback (Lin et al. 2014; Masuda-Nakagawa et al. 2014) and helps to maintain KC sparse responses and odor selectivity (Honegger et al. 2011), analogous to inhibition in the mammalian PCx (Poo and Isaacson 2009; Stettler and Axel 2009; Gire et al. 2013). Thus, odors are represented as a sparse ensemble of KCs that are highly odor selective, a property beneficial for memory (Olshausen and Field 2004).In addition, the larval MB calyx is innervated by two OA neurons, sVUMmd1 and sVUMmx1, ventral unpaired medial neurons with dendritic fields originating in the mandibular and maxillary neuromeres, respectively, of the SEZ in the third instar larva (Selcho et al. 2014). sVUMmd1 and sVUMmx1 are named as OANa-1 and OANa-2, respectively, in the EM connectomic analysis of a 6-h first instar larva (Eichler et al. 2017; Supplemental Fig. 3 of Saumweber et al. 2018). These sVUM1 neurons also innervate the first olfactory neuropile of the antennal lobe (AL). This pattern of innervation is conserved in other insects, for example, the dorsal unpaired median (DUM) neurons in locusts (for review, see Bräunig and Pflüger 2001), the VUMmx1 neuron in honeybees (Hammer 1993; Schröter et al. 2007), and OA-VUMa2 neurons in adult Drosophila (Busch et al. 2009). In adult Drosophila, OA-VUMa2 neurons also show a dense innervation of the lateral horn, implicated in innate behaviors (Busch et al. 2009). The widespread innervation of the insect olfactory neuropiles also resembles the widespread NA innervation of mammalian olfactory processing areas, such as the olfactory bulb, and piriform cortex, by LC neurons originating in the brainstem.We characterized the innervation pattern and synaptic targets of sVUM1 neurons in the calyx, with MB intrinsic and also extrinsic neurons, the localization of the OA receptor Oamb in the calyx circuit, and the impact of sVUM1 neuron activation on behavioral odor discrimination. For this we used an appetitive conditioning paradigm, and tested the ability of larvae to discriminate between similar odors, as opposed to dissimilar odors. Since the larval connectome is based on a single brain, at first instar stage before octopaminergic connections have become as extensive as at third instar, and to obtain a comprehensive understanding of the synaptic targets of sVUM1s in the third-instar larval calyx, we extended our analysis to previously unanalyzed connectivity of VUM1s, to APL and PNs. Further, we combined light microscopy of third-instar larvae with the connectome described by Eichler et al. (2017).We find that sVUM1 neurons in third-instar larvae contact all the major classes of calyx neuron to some degree, consistent with EM synaptic analysis of the 6-h larva (Eichler et al. 2017). A GFP fusion of the OA receptor Oamb is localized in the terminals of PNs, and activating a subset of five SEZ neurons, including sVUM1 neurons, can affect discrimination of similar odors, without affecting underlying olfactory learning and memory ability. We suggest a broad modulatory effect of sVUM1 neurons in the calyx, including a potential role in modulating PN input at the second synapse in the olfactory pathway.     

Drosophila Mushroom Bodies Are Dispensable for Visual, Tactile, and Motor Learning

A total of 18 associative learning/memory tests have been applied to Drosophila melanogaster flies lacking mushroom bodies. Only in paradigms involving chemosensory cues as conditioned stimuli have flies been found to be compromised by a block in the mushroom body pathway. Among the learning tasks not requiring these structures are a case of motor learning (yaw torque/heat), a test of the fly’s spatial orientation in total darkness, conditioned courtship suppression by mated females, and nine different examples of visual learning. The latter used the reinforcers of heat, visual oscillations, mechanical shaking, or sucrose, and as conditioned stimuli, color, intensity contrast, as well as stationary and moving visual patterns. No forms of consolidated memory have been tested in mushroom body-less flies. With respect to short-term memory the mushroom bodies of Drosophila are specially required for chemosensory learning tasks, but not for associative learning and memory in general.     

Neurogenetic approaches to habituation and dishabituation in Drosophila

We review work in the major model systems for habituation in Drosophila melanogaster, encompassing several sensory modalities and behavioral contexts: visual (giant fiber escape response, landing response); chemical (proboscis extension reflex, olfactory jump response, locomotory startle response, odor-induced leg response, experience-dependent courtship modification); electric (shock avoidance); and mechanical (leg resistance reflex, cleaning reflex). Each model system shows several of Thompson and Spencer’s [Thompson, R. F., & Spencer, W. A. (1966). Habituation: A model phenomenon for the study of neuronal substrates of behavior. Psychological Review, 73, 16–43] parametric criteria for habituation: spontaneous recovery and dishabituation have been described in almost all of them and dependence of habituation upon stimulus frequency and stimulus intensity in the majority. Stimulus generalization (and conversely, the delineation of stimulus specificity) has given insights into the localization of habituation or the neural architecture underlying sensory processing.The strength of Drosophila for studying habituation is the range of genetic approaches available. Mutations have been used to modify specific neuroanatomical structures, ion channels, elements of synaptic transmission, and second-messenger pathways. rutabaga and dunce, genes of the cAMP signal pathway that have been studied most often in the reviewed experiments, have also been implicated in synaptic plasticity and associative conditioning in Drosophila and other species including mammals. The use of the Gal4/UAS system for targeting gene expression has enabled genetic perturbation of defined sets of neurons. One clear lesson is that a gene may affect habituation differently in different behaviors, depending on the expression, processing, and localization of the gene product in specific circuits. Mutations of specific genes not only provide links between physiology and behavior in the same circuit, but also reveal common mechanisms in different paradigms of behavioral plasticity. The rich repertoire of models for habituation in the fly is an asset for combining a genetic approach with behavioral, anatomical and physiological methods with the promise of a more complete understanding.     

Cellular site and molecular mode of synapsin action in associative learning

Synapsin is an evolutionarily conserved, presynaptic vesicular phosphoprotein. Here, we ask where and how synapsin functions in associative behavioral plasticity. Upon loss or reduction of synapsin in a deletion mutant or via RNAi, respectively, Drosophila larvae are impaired in odor-sugar associative learning. Acute global expression of synapsin and local expression in only the mushroom body, a third-order "cortical" brain region, fully restores associative ability in the mutant. No rescue is found by synapsin expression in mushroom body input neurons or by expression excluding the mushroom bodies. On the molecular level, we find that a transgenically expressed synapsin with dysfunctional PKA-consensus sites cannot rescue the defect of the mutant in associative function, thus assigning synapsin as a behaviorally relevant effector of the AC-cAMP-PKA cascade. We therefore suggest that synapsin acts in associative memory trace formation in the mushroom bodies, as a downstream element of AC-cAMP-PKA signaling. These analyses provide a comprehensive chain of explanation from the molecular level to an associative behavioral change.     

Second-order ‘fear’ conditioning in humans: Persistence of CR2 following extinction of CR1

Two groups of subjects were given pairings of a slide of a triangle (CS₁) with an aversive loud noise (US). After first-order conditioning a different figure (CS₂⁺) was paired with CS₁, whilst a third figure acted as a non-paired control (CS₂⁰). Conditioned second-order skin conductance responses (SCRs) were established to CS₂⁺ for all subjects who had undergone first-order conditioning. Following second-order conditioning, SCRs to CS₁ were extinguished and CS₂⁺ was subsequently tested for retention of its SCR evoking properties. For the group of subjects who believed they had an avoidance response available during the second-order conditioning phase, CR₂ failed to survive extinction of CR₁. However, for the group of subjects who had no perceived control over US presentation, extinction of cr₁ did not eliminate CR₂. This latter finding is consistent with associative analyses of second-order conditioning in animals and has clinical implications for the treatment of fear-related problems.     

Associative representational plasticity in the auditory cortex: a synthesis of two disciplines

Historically, sensory systems have been largely ignored as potential loci of information storage in the neurobiology of learning and memory. They continued to be relegated to the role of "sensory analyzers" despite consistent findings of associatively induced enhancement of responses in primary sensory cortices to behaviorally important signal stimuli, such as conditioned stimuli (CS), during classical conditioning. This disregard may have been promoted by the fact that the brain was interrogated using only one or two stimuli, e.g., a CS(+) sometimes with a CS(-), providing little insight into the specificity of neural plasticity. This review describes a novel approach that synthesizes the basic experimental designs of the experimental psychology of learning with that of sensory neurophysiology. By probing the brain with a large stimulus set before and after learning, this unified method has revealed that associative processes produce highly specific changes in the receptive fields of cells in the primary auditory cortex (A1). This associative representational plasticity (ARP) selectively facilitates responses to tonal CSs at the expense of other frequencies, producing tuning shifts toward and to the CS and expanded representation of CS frequencies in the tonotopic map of A1. ARPs have the major characteristics of associative memory: They are highly specific, discriminative, rapidly acquired, exhibit consolidation over hours and days, and can be retained indefinitely. Evidence to date suggests that ARPs encode the level of acquired behavioral importance of stimuli. The nucleus basalis cholinergic system is sufficient both for the induction of ARPs and the induction of specific auditory memory. Investigation of ARPs has attracted workers with diverse backgrounds, often resulting in behavioral approaches that yield data that are difficult to interpret. The advantages of studying associative representational plasticity are emphasized, as is the need for greater behavioral sophistication.     

Translocation and activation of Rac in the hippocampus during associative contextual fear learning

Small G proteins including Rac are mediators of changes in neuronal morphology associated with synaptic plasticity. Previous studies in our laboratory showed that Rac is highly expressed in the adult mouse hippocampus, a brain area that exhibits robust synaptic plasticity and is crucial for the acquisition of memories. In this study, we investigated whether Rac was involved in NMDA receptor-dependent associative fear learning in the area CA1 of adult mouse hippocampus. We found that Rac translocation and activation was increased in the hippocampus following associative fear conditioning in mice, and that these increases are blocked by intraperitoneal injection of the NMDA receptor channel blocker MK801 at the acquisition stage. Our data indicate that NMDA receptor-dependent associative fear learning alters Rac localization and function in the mouse hippocampus.     

Color and intensity discrimination in <Emphasis Type="Italic">Xenopus laevis</Emphasis> tadpoles

Investigations into the physiology of Xenopus laevis have the potential to greatly accelerate biomedical research, especially concerning neural plasticity and sensory systems, but are limited by the lack of available information on behavioral learning in this species. Here, we attempt to lay the foundations for a behavioral assay in Xenopus that can be used in conjunction with biological manipulations. We tested cohorts of Xenopus tadpoles across four light-mediated active-avoidance experiments, using either wavelength or intensity as the salient discriminative cue. In the wavelength task, we determine a baseline learning rate and characterize retention of learning, identifying active extinction effects as far more potent than the passage of time in the loss of behavior. In the intensity task, we examine the effects of varying differences between the discriminative stimuli on acquisition and extinction and identify a critical range of intensity differences where learning changes. The results of our experiments demonstrate that Xenopus is a tractable model organism for cognitive research and can learn a variety of associative tasks in the laboratory settings. These data reveal new aspects of the Xenopus larval visual processing system and facilitate future research between cognitive methods and biological/chemical manipulations to study mechanisms of brain structure and function.     

Mapping of the Anatomical Circuit of CaM Kinase-Dependent Courtship Conditioning in Drosophila

Globally inhibiting CaM kinase activity in Drosophila, using a variety of genetic techniques, disrupts associative memory yet leaves visual and chemosensory perception intact. These studies implicate CaM kinase in the plastic processes underlying learning and memory but do not identify the neural circuitry that specifies the behavior. In this study, we use the GAL4/UAS binary expression system to define areas of the brain that require CaM kinase for modulation of courtship conditioning. The CaM kinase-dependent neurons that determine the response to the mated female during conditioning and those involved in formation and expression of memory were found to be located in distinct areas of the brain. This supports the idea that courtship conditioning results in two independent behavioral modifications: a decrement in courtship during the conditioning period and an associative memory of conditioning. This study has allowed us for the first time to genetically determine the circuit of information flow for a memory process in Drosophila. The map we have generated dissects the behavior into multiple components and will provide tools that allow both molecular and electrophysiological access to this circuit.     

Reelin supplementation enhances cognitive ability, synaptic plasticity, and dendritic spine density

Apolipoprotein receptors belong to an evolutionarily conserved surface receptor family that has intimate roles in the modulation of synaptic plasticity and is necessary for proper hippocampal-dependent memory formation. The known lipoprotein receptor ligand Reelin is important for normal synaptic plasticity, dendritic morphology, and cognitive function; however, the in vivo effect of enhanced Reelin signaling on cognitive function and synaptic plasticity in wild-type mice is unknown. The present studies test the hypothesis that in vivo enhancement of Reelin signaling can alter synaptic plasticity and ultimately influence processes of learning and memory. Purified recombinant Reelin was injected bilaterally into the ventricles of wild-type mice. We demonstrate that a single in vivo injection of Reelin increased activation of adaptor protein Disabled-1 and cAMP-response element binding protein after 15 min. These changes correlated with increased dendritic spine density, increased hippocampal CA1 long-term potentiation (LTP), and enhanced performance in associative and spatial learning and memory. The present study suggests that an acute elevation of in vivo Reelin can have long-term effects on synaptic function and cognitive ability in wild-type mice.     

Learning and memory deficits upon TAU accumulation in Drosophila mushroom body neurons

Mutations in the neuronal-specific microtubule-binding protein TAU are associated with several dementias and neurodegenerative diseases. However, the effects of elevated TAU accumulation on behavioral plasticity are unknown. We report that directed expression of wild-type vertebrate and Drosophila TAU in adult mushroom body neurons, centers for olfactory learning and memory in Drosophila, strongly compromised associative olfactory learning and memory, but olfactory conditioning-relevant osmotactic and mechanosensory responses remained intact. In addition, TAU accumulation in mushroom body neurons did not result in detectable neurodegeneration or premature death. Therefore, TAU-mediated structural or functional perturbation of the microtubular cytoskeleton in mushroom body neurons is likely causal of the behavioral deficit. These results indicate that behavioral plasticity decrements may be the earliest detectable manifestations of tauopathies.     

Enhanced cholinergic suppression of previously strengthened synapses enables the formation of self-organized representations in olfactory cortex

Computational modeling assists in analyzing the specific functional role of the cellular effects of acetylcholine within cortical structures. In particular, acetylcholine may regulate the dynamics of encoding and retrieval of information by regulating the magnitude of synaptic transmission at excitatory recurrent connections. Many abstract models of associative memory function ignore the influence of changes in synaptic strength during the storage process and apply the effect of these changes only during a so-called recall-phase. Efforts to ensure stable activity with more realistic, continuous updating of the synaptic strength during the storage process have shown that the memory capacity of a realistic cortical network can be greatly enhanced if cholinergic modulation blocks transmission at synaptic connections of the association fibers during the learning process. We here present experimental data from an olfactory cortex brain slice preparation showing that previously potentiated fibers show significantly greater suppression (presynaptic inhibition) by the cholinergic agonist carbachol than unpotentiated fibers. We conclude that low suppression of non-potentiated fibers during the learning process ensures the formation of self-organized representations in the neural network while the higher suppression of previously potentiated fibers minimizes interference between overlapping patterns. We show in a computational model of olfactory cortex, that, together, these two phenomena reduce the overlap between patterns that are stored within the same neural network structure. These results further demonstrate the contribution of acetylcholine to mechanisms of cortical plasticity. The results are consistent with the extensive evidence supporting a role for acetylcholine in encoding of new memories and enhancement of response to salient sensory stimuli.     

A cGMP-dependent protein kinase gene, foraging, modifies habituation-like response decrement of the giant fiber escape circuit in Drosophila

The Drosophila giant fiber jump-and-flight escape response is a model for genetic analysis of both the physiology and the plasticity of a sensorimotor behavioral pathway. We previously established the electrically induced giant fiber response in intact tethered flies as a model for habituation, a form of nonassociative learning. Here, we show that the rate of stimulus-dependent response decrement of this neural pathway in a habituation protocol is correlated with PKG (cGMP-Dependent Protein Kinase) activity and foraging behavior. We assayed response decrement for natural and mutant rover and sitter alleles of the foraging (for) gene that encodes a Drosophila PKG. Rover larvae and adults, which have higher PKG activities, travel significantly farther while foraging than sitters with lower PKG activities. Response decrement was most rapid in genotypes previously shown to have low PKG activities and sitter-like foraging behavior. We also found differences in spontaneous recovery (the reversal of response decrement during a rest from stimulation) and a dishabituation-like phenomenon (the reversal of response decrement evoked by a novel stimulus). This electrophysiological study in an intact animal preparation provides one of the first direct demonstrations that PKG can affect plasticity in a simple learning paradigm. It increases our understanding of the complex interplay of factors that can modulate the sensitivity of the giant fiber escape response, and it defines a new adult-stage phenotype of the foraging locus. Finally, these results show that behaviorally relevant neural plasticity in an identified circuit can be influenced by a single-locus genetic polymorphism existing in a natural population of Drosophila.     

Evidence for associative learning in newly emerged honey bees (Apis mellifera)

The honey bee is a model organism for studies on the neural substrates of learning and memory. Associative olfactory learning using sucrose rewards is fast and reliable in foragers and older hive bees. However, researchers have so far failed to show any significant learning in newly emerged bees. It is generally argued that in these bees only part of the brain structures important for learning are fully developed. Here we show for the first time that newly emerged honey bees are capable of associative learning, if they are sufficiently responsive to sucrose. Responsiveness to sucrose, which can be measured using the proboscis extension response (PER), increases with age. Newly emerged bees are on average very unresponsive to sucrose. We show that if newly emerged bees displaying a PER to 10% sucrose or lower sucrose concentrations are conditioned to an odour, they show significant associative learning and early long-term memory. Nevertheless, the level of acquisition is still lower than in foragers. The general assumption that newly emerged honey bees are incapable of associative learning must therefore be reconsidered. Further, our study suggests that an age-dependent increase in responsiveness to rewarding stimuli is directly related to the development of early learning abilities. The decisive influence of responsiveness to rewarding stimuli in associative learning of newly emerged bees has far reaching consequences for studies on the development of associative learning capabilities in insects and vertebrates.