Exploring Embodied and Bioenergetic Approaches in Trauma Therapy: Observing Somatic Experience and Olfactory Memory.

Recent studies highlight how body psychotherapy is becoming highly cited, especially in connection with studies on trauma-related disorders. This review highlights the theoretical assumptions and recent points in common with embodied simulation and new sensory theories by integrating bioenergetic analysis, embodiment, and olfactory memory in trauma and post-traumatic stress disorder (PTSD) therapy. Embodied memory, rooted in sensorimotor experiences, shapes cognitive functions and emotional responses. Trauma, embodied in somatic experiences, disrupts these processes, leading to symptoms such as chronic pain and dissociation. The literature discussed highlights the impact of burning odors on individuals with PTSD and those who have experienced childhood maltreatment. Burning odors can increase stress and heart rate in war veterans, with sensitivity to these odors intensifying over time since the trauma. Additionally, adults who experienced childhood maltreatment exhibit faster processing of unpleasant odors and increased symptom severity. Grounding techniques, such as adopting a balanced posture, enhance breathing and sensory capabilities, potentially aiding in managing symptoms associated with trauma-related disorders such as PTSD.

Exploring neonicotinoid effects on Drosophila: insights into olfactory memory, neurotransmission, and synaptic connectivity.

Neonicotinoid insecticides, the fastest-growing class in recent decades, interfere with cholinergic neurotransmission by binding to the nicotinic acetylcholine receptor. This disruption affects both targeted and non-targeted insects, impairing cognitive functions such as olfaction and related behaviors, with a particular emphasis on olfactory memory due to its ecological impact. Despite the persistent presence of these chemicals in the environment, significant research gaps remain in understanding the intricate interplay between cognitive function, development, neuronal activity, and neonicotinoid-induced toxicity. This study focuses on the fruit fly Drosophila melanogaster, chosen for its genetic tractability, well-characterized neural circuitry, and remarkable parallels with bees in neurotransmitter systems and brain structures. Our aim is to establish the fruit fly as a valuable model organism for studying the effects of neonicotinoids on behavior and neuronal circuitry, with particular attention to olfactory memory and associated brain circuitries. To achieve this aim, we conducted experiments to investigate the effects of short-term exposure to sublethal doses of the neonicotinoid imidacloprid, mimicking realistic environmental insecticide exposure, on the formation of odor memories. Additionally, we evaluated synaptic contacts and cholinergic neurotransmission within the mushroom body, the primary memory network of insects. Our results showed significant impairments in odor memory formation in flies exposed to imidacloprid, with exposure during the adult stage showing more pronounced effects than exposure during the larval stage. Additionally, functional studies revealed a decrease in synaptic contacts within the intrinsic olfactory projection neurons and the mushroom body. Furthermore, another experiment showed an odor-dependent reduction in cholinergic neurotransmission within this network. In summary, employing Drosophila as a model organism provides a robust framework for investigating neonicotinoid effects and understanding their diverse impacts on insect physiology and behavior. Our study initiates the establishment of the fruit fly as a pivotal model for exploring neonicotinoid influences, shedding light on their effects on olfactory memory, neuronal integrity, and synaptic transmission.

Probiotic Lactobacillus spp. improves Drosophila memory by increasing lactate dehydrogenase levels in the brain mushroom body neurons.

Probiotics are live microorganisms that offer potential benefits to their hosts and can occasionally influence behavioral responses. However, the detailed mechanisms by which probiotics affect the behavior of their hosts and the underlying biogenic effects remain unclear. Lactic acid bacteria, specifically Lactobacillus spp. are known probiotics. Drosophila melanogaster, commonly known as the fruit fly, is a well-established model organism for investigating the interaction between the host and gut microbiota in translational research. Herein, we showed that 5-day administration of Lactobacillus acidophilus (termed GMNL-185) or Lacticaseibacillus rhamnosus (termed GMNL-680) enhances olfactory-associative memory in Drosophila. Moreover, a combined diet of GMNL-185 and GMNL-680 demonstrated synergistic effects on memory functions. Live brain imaging revealed a significant increase in calcium responses to the training odor in the mushroom body ß and γ lobes of flies that underwent mixed feeding with GMNL-185 and GMNL-680. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and whole-mount brain immunohistochemistry revealed significant upregulation of lactate dehydrogenase (LDH) expression in the fly brain following the mixed feeding. Notably, the genetic knockdown of Ldh in neurons, specifically in mushroom body, ameliorated the beneficial effects of mixed feeding with GMNL-185 and GMNL-680 on memory improvement. Altogether, our results demonstrate that supplementation with L. acidophilus and L. rhamnosus enhances memory functions in flies by increasing brain LDH levels.

Vertigoheel promotes rodent cognitive performance in multiple memory tests.

Introduction: Cognitive impairment associated with old age or various brain disorders may be very disabling for affected individuals, placing their carers and public health services under considerable stress. The standard-of-care drugs produce only transient improvement of cognitive impairment in older people, so the search for novel, safe and effective therapeutics that would help to reverse or delay cognitive impairment is warranted. Repurposing pharmacological therapies with well-established safety record for additional indications is a promising recent trend in drug development. Vertigoheel (VH-04), a multicomponent drug made of Ambra grisea, Anamirta cocculus L., Conium maculatum, and Petroleum rectificatum, has been successfully used for several decades in the treatment of vertigo. Here, we investigated effects of VH-04 on cognitive performance in standard behavioral tests assessing different types of memory and explored cellular and molecular underpinnings of VH-04's biological activity. Methods: In the majority of behavioral experiments, namely in the spontaneous and rewarded alternation tests, passive avoidance test, contextual/cued fear conditioning, and social transmission of food preference, we examined the ability of single and repeated intraperitoneal administrations of VH-04 to improve cognitive parameters of mice and rats disrupted by the application of the muscarinic antagonist scopolamine. In addition, we also assessed how VH-04 affected novel object recognition and influenced performance of aged animals in Morris water maze. Furthermore, we also studied the effects of VH-04 on primary hippocampal neurons in vitro and mRNA expression of synaptophysin in the hippocampus. Results: Administration of VH-04 positively influenced visual recognition memory in the novel object recognition test and alleviated the impairments in spatial working memory and olfactory memory caused by the muscarinic antagonist scopolamine in the spontaneous alternation and social transmission of food preference tests. In addition, VH-04 improved retention of the spatial orientation memory of old rats in the Morris water maze. In contrast, VH-04 did not have significant effects on scopolamine-induced impairments in tests of fear-aggravated memory or rewarded alternation. Experiments in vitro showed that VH-04 stimulated neurite growth and possibly reversed the age-dependent decrease in hippocampal synaptophysin mRNA expression, which implies that VH-04 may preserve synaptic integrity in the aging brain. Discussion: Our findings allow a cautious conclusion that in addition to its ability to alleviate manifestations of vertigo, VH-04 may be also used as a cognitive enhancer.

Olfactory memory in mild cognitive impairment and Alzheimer's disease.

Introduction: Olfaction is impaired in Alzheimer's disease (AD). However, olfactory memory has rarely been examined. As the pathogenesis of AD remains largely unknown, collecting more data regarding the occurrence and progression of its symptoms would help gain more insight into the disease. Objective: To investigate olfactory memory and its relationship with verbal memory and other clinical features in patients with early-stage AD. Methods: Three groups of participants were enrolled in this study: patients with mild dementia due to AD (MD-AD, N = 30), patients with mild cognitive impairment due to AD (MCI-AD, N = 30), and cognitively normal older participants (CN, N = 30). All participants underwent cognitive evaluation (Clinical Dementia Rating scale, Mini Mental State Examination, Alzheimer's Disease Assessment Scale-Cognitive Subscale, delayed verbal recall, and verbal fluency tests) and assessment of olfactory immediate and delayed recognition memory. Results: Olfactory immediate and delayed recognition memory scores were significantly lower in the MD-AD group than in the MCI-AD and CN groups. The MCI-AD and CN groups did not differ significantly [in both cases, Kruskal-Wallis test, p < 0.05; post hoc analysis revealed significant differences between the MD-AD and MCI-AD groups and between the MD-AD and CN groups (p < 0.05), and no significant difference between the MCI-AD and CN groups (p > 0.05)]. Verbal immediate recall, delayed recall after 5 min, and delayed recall after 30 min scores were significantly worse in the MD-AD and MCI-AD groups than in the CN group. MD-AD and MCI-AD groups did not differ significantly [in all cases Kruskal-Wallis test, p < 0.05; post hoc analysis revealed significant differences between MD-AD and CN groups, and MCI-AD and CN groups (p < 0.05) and no significant difference between MD-AD and MCI-AD groups (p > 0.05)]. Duration of AD symptoms was a strong predictor of both immediate and delayed olfactory recognition memory scores. Conclusion: Olfactory memory impairment was observed in patients with AD. The changes progress during the course of the disease. However, unlike verbal memory, olfactory memory is not significantly impaired in the prodromal stage of AD.

Sex and Age-Dependent Olfactory Memory Dysfunction in ADHD Model Mice.

ADHD is a typical neurodevelopmental disorder with a high prevalence rate. NSCs in the subventricular zone (SVZ) are closely related to neurodevelopmental disorder and can affect olfactory function by neurogenesis and migratory route. Although olfactory dysfunction is one of the symptoms of ADHD, the relevance of cells in the olfactory bulb derived from NSCs has not been studied. Therefore, we investigated olfactory memory and NSCs in Git1-deficient mice, under the ADHD model. Interestingly, only adult male G protein-coupled receptor kinase-interacting protein-1 (GIT1)-deficient (+/-, HE) mice showed impaired olfactory memory, suggesting sex and age dependence. We performed adult NSCs culture from the SVZ and observed distinct cell population in both sex and genotype. Taken together, our study suggests that the altered differentiation of NSCs in GIT1+/- mice can contribute to olfactory dysfunction in ADHD.

Development and validation of the Olfactory Memory Test Battery (OMTB) based on odors with high- and low-verbalizability.

BACKGROUND: Odor memory is an important field of clinical research for its distinctive characteristics, which differ from those of other sensory systems. To date, several tests have been implemented for the assessment of odor memory. Despite a range of studies demonstrating the importance of verbal mediation in odor memory, few have distinguished odor memory performance in different odor verbalization levels. NEW METHOD: We aimed to develop a standardized odor memory test toolbox with one group of odors that are easily verbally identified and the other group of odors that are difficult to identify. The test contained two odor categories (high- and low-verbalizability odors), each consisting of three subtests (short- and long-term memory and working memory). RESULTS: Satisfactory test-retest reliability and solid validity of the odor recognition and working memory test were shown in both odor categories. Moreover, people scored significantly better with high- than low-verbalizability odors. A negative age effect on odor memory performance was also found. COMPARISON WITH EXISTING METHODS: No previous odor memory test distinguished odor memory performance in different odor verbalization levels, while the Olfactory Memory Test Battery (OMTB) contains high- and low-verbalizability odors and each category has three subtests. CONCLUSIONS: The present study indicated the OMTB is a comprehensive assessment of odor memory with good reliability and validity. All subtests can be used separately or in combination with each other according to the clinical and research needs.

Intentional and incidental odour-colour binding in working memory.

The question of how features are bound together in working memory has become a topic of much research in recent years. However, this is typically focused on visual and/or auditory stimuli. The purpose of this study is to apply established feature binding procedures to investigate odour binding in working memory. Across three experiments, memory for intentionally and incidentally formed odour-colour pairings was tested. Experiment 1 showed that following explicit instruction to remember the odour-colour combinations, young adults can recall lists of 3-pairings at levels above that of chance and exhibit a recency advantage for the last pairing. In Experiment 2 participants were asked to prioritise the first pairing in the list or treat all pairings equally. We observed only limited evidence of prioritisation affecting the serial position function. Experiment 3 explored incidental odour-colour binding. Using a yes/no recognition procedure, accuracy did not differ for positive test probes presented in the same (bound) or different (unbound) colour to encoding. This study is one of the first to examine odour-colour binding in working memory and, taking the evidence together, suggests that odour-colour bindings can be formed in working memory; however, functionality may be limited compared to that of visual feature binding.

Olfactory metacognition and memory in individuals with different subjective odor imagery abilities.

Imagery vividness is one of the key indicators to evaluate the ability to generate mental images. There is large inter-individual variability in olfactory imagery (OI) abilities, however, little is known about the underlying factors for individual OI abilities. Using a word cueing imagery paradigm and the trial-by-trial imagery vividness rating method, participants with high or low OI abilities (differentiated by the Vividness of Olfactory Imagery Questionnaire) completed two OI tasks with either shorter (2 s) or longer (8 s) image generation time. Participants' olfactory function, olfactory-related working memory and episodic recognition memory were measured using validated methods. Moreover, olfactory metacognition was assessed using the Odor Awareness Scale (OAS) and the Importance of Olfaction Questionnaire (IOQ). Compared to participants with high OI abilities, those with low OI abilities reported less vivid odor images during OI tasks. For participants with low OI abilities, the imagery vividness significantly improved as the image generation time increased. There was no difference regarding olfactory perception or olfactory-related memory performances between the high and the low OI ability groups. However, participants with higher OI abilities had significant higher scores on the OAS and the IOQ, indicating a superior olfactory-related metacognition. These results provide evidences supporting the underlying factors that related to variances of subjective ability of generating vivid odor mental images.

Are you my mummy? Long-term olfactory memory of mother's body odour by offspring in the domestic cat.

Longevity of odour memories, particularly those acquired during early development, has been documented in a wide range of taxa. Here, we report that kittens of the domestic cat retained a memory into adult life of their mother´s body odour experienced before weaning. Kittens from 15 litters were tested when permanently separated from their mother at weaning on postnatal week 8, and tested again when 4 and 6 months and over 1 year of age. When presented with a simultaneous three-way choice between body odour of their own mother, of an unknown female of similar reproductive condition and a blank stimulus, weaning-age kittens sniffed the cotton swab with the odour of an unknown female longer. This preference, however, changed when as adults the subjects sniffed the cotton swab with their own mother's odour longer. We conclude that kittens form a long-lasting memory of the body odour of their mother, and by implication, that mothers retain an individual odour signature sufficiently stable across age and changes in their reproductive state to be distinguishable by their adult offspring. What this means in functional or cognitive terms is not yet clear. Does such "recognition" have a specific biological function and a specific cognitive representation? Or is it rather part of a more general phenomenon well known in (human) olfaction of odours that are familiar generally being judged more pleasant, and that might then influence olfactory-guided behaviour in a variety of contexts?

[Smell and care: from professionalism to emotions]. / Odeurs et soins, du professionnalisme aux émotions.

The sense of smell presents itself to us as a means of forging an opinion of a situation, sending us back to our memories and giving us a form of knowledge of the patient. In addition, besides the obvious repulsion that certain wounds can engender, the sense of smell can also, conversely, lead us into an eroticization of care.

Differential conditioning produces merged long-term memory in Drosophila.

Multiple spaced trials of aversive differential conditioning can produce two independent long-term memories (LTMs) of opposite valence. One is an aversive memory for avoiding the conditioned stimulus (CS+), and the other is a safety memory for approaching the non-conditioned stimulus (CS-). Here, we show that a single trial of aversive differential conditioning yields one merged LTM (mLTM) for avoiding both CS+ and CS-. Such mLTM can be detected after sequential exposures to the shock-paired CS+ and -unpaired CS-, and be retrieved by either CS+ or CS-. The formation of mLTM relies on triggering aversive-reinforcing dopaminergic neurons and subsequent new protein synthesis. Expressing mLTM involves αß Kenyon cells and corresponding approach-directing mushroom body output neurons, in which similar-amplitude long-term depression of responses to CS+ and CS- seems to signal the mLTM. Our results suggest that animals can develop distinct strategies for occasional and repeated threatening experiences.

Olfactory impairment in men and mice related to aging and amyloid-induced pathology.

Olfaction, or the sense of smell, is one of the most ancient senses in men and mice, important for a large variety of innate and acquired behaviors. Clinical data reveal an early impairment of olfaction during normal aging and in the course of neurodegenerative diseases, but the underlying cellular/molecular mechanisms remain obscure. In the current review, we compare different aspects of the aging- and Alzheimer's disease related impairment of olfaction in men and mice, aiming at the identification of common morbidities and biomarkers, which can be analyzed in detail in the appropriate mouse models. We also identify common, often interdependent (patho)physiological pathways, including but not limited to extracellular amyloid depositions, neuroinflammation, ɛ4 allele of the apolipoprotein E, CNS insulin resistance, and the impairment of adult neurogenesis, to be targeted by basic and clinical research.

Rac1 Impairs Forgetting-Induced Cellular Plasticity in Mushroom Body Output Neurons.

Active memory forgetting is a well-regulated biological process thought to be adaptive and to allow proper cognitive functions. Recent efforts have elucidated several molecular players involved in the regulation of olfactory forgetting in Drosophila, including the small G protein Rac1, the dopamine receptor DAMB, and the scaffold protein Scribble. Similarly, we recently reported that dopaminergic neurons mediate both learning- and forgetting-induced plasticity in the mushroom body output neuron MBON-γ2α'1. Two open questions remain: how does forgetting affect plasticity in other, highly plastic, mushroom body compartments and how do genes that regulate forgetting affect this cellular plasticity? Here, we show that forgetting reverses short-term synaptic depression induced by aversive conditioning in the highly plastic mushroom body output neuron MBON-γ1pedc>α/ß. In addition, our results indicate that genetic tampering with normal forgetting by inhibition of small G protein Rac1 impairs restoration of depressed odor responses to learned odor by intrinsic forgetting through time passing and forgetting induced acutely by shock stimulation after conditioning or reversal learning. These data further indicate that some forms of forgetting truly erase physiological changes generated by memory encoding.

Pollen reinforces learning in honey bee pollen foragers but not in nectar foragers.

Searching for reward motivates and drives behaviour. In honey bees Apis mellifera, specialized pollen foragers are attracted to and learn odours with pollen. However, the role of pollen as a reward remains poorly understood. Unlike nectar, pollen is not ingested during collection. We hypothesized that pollen (but not nectar) foragers could learn pollen by sole antennal or tarsal stimulation. Then, we tested how pairing of pollen (either hand- or bee-collected) and a neutral odour during a pre-conditioning affects performance of both pollen and nectar foragers during the classical conditioning of the proboscis extension response. Secondly, we tested whether nectar and pollen foragers perceive the simultaneous presentation of pollen (on the tarsi) and sugar (on the antennae) as a better reinforcement than sucrose alone. Finally, we searched for differences in learning of the pollen and nectar foragers when they were prevented from ingesting the reward during the conditioning. Differences in pollen-reinforced learning correlate with division of labour between pollen and nectar foragers. Results show that pollen foragers performed better than nectar foragers during the conditioning phase after being pre-conditioned with pollen. Pollen foragers also performed better than nectar foragers in both the acquisition and extinction phases of the conditioning, when reinforced with the dual reward. Consistently, pollen foragers showed improved abilities to learn cues reinforced without sugar ingestion. We discussed that differences in how pollen and nectar foragers respond to a cue associated with pollen greatly contribute to the physiological mechanism that underlies foraging specialization in the honeybee.

Western Diet Accelerates the Impairment of Odor-Related Learning and Olfactory Memory in the Mouse.

Olfactory dysfunction could be an early indicator of cognitive decline in type 2 diabetes (T2D). However, whether obesity affects olfaction in people with T2D is unclear. This question needs to be addressed, because most people with T2D are obese. Importantly, whether different contributing factors leading to obesity (e.g., different components of diet or gain in weight) affect specific olfactory functions and underlying mechanisms is unknown. We examined whether two T2D-inducing obesogenic diets, one containing a high proportion of fat (HFD) and one with moderate fat and high sugar (Western diet, WD), affect odor detection/discrimination, odor-related learning, and olfactory memory in the mouse. We also investigated whether the diets impair adult neurogenesis, GABAergic interneurons, and neuroblasts in the olfactory system. Here, we further assessed olfactory cortex volume and cFos expression-based neuronal activity. The WD-fed mice showed declined odor-related learning and olfactory memory already after 3 months of diet intake (p = 0.046), although both diets induced similar hyperglycemia and weight gain compared to those of standard diet-fed mice (p = 0.0001 and p < 0.0001, respectively) at this time point. Eight months of HFD and WD diminished odor detection (p = 0.016 and p = 0.045, respectively), odor-related learning (p = 0.015 and p = 0.049, respectively), and olfactory memory. We observed no changes in the investigated cellular mechanisms. We show that the early deterioration of olfactory parameters related to learning and memory is associated with a high content of sugar in the diet rather than with hyperglycemia or weight gain. This finding could be exploited for understanding, and potentially preventing, cognitive decline/dementia in people with T2D. The mechanisms behind this finding remain to be elucidated.

From Synapse to Supper: A Food Preference Recipe with Olfactory Synaptic Ingredients.

C1ql3 protein and its receptor Bai3 are involved in synaptic organization and function. In this issue of Neuron, Wang et al. (2020) report that both are essential for synaptic function between the anterior olfactory nucleus and the olfactory bulb and for the generation, but not recall, of associative olfactory memories determining food preference in mice.

TOM-32-An extended test for the assessment of olfactory memory.

BACKGROUND: Olfactory memory testing is a promising way to explore different aspects of smell-related abilities in healthy subjects as well as to diagnose various diseases associated with disorders of this type of memory. Despite the obvious value of olfactory memory assessments, few available methods enable its diagnosis. NEW METHOD: The aim of our study was to create a standardized research tool designed for olfactory memory studies: an extended, 32-item version of an available "Test for Odor Memory". RESULTS: The study involving 222 subjects revealed satisfactory psychometric qualities of TOM-32, and - as hypothesized - subjects with depression scored significantly lower than healthy participants. We additionally showed that TOM-32 scores was associated with sex, age and olfactory awareness. Women performed significantly better than men, young people tended to have lower false alarm rate than older subjects and olfactory awareness was positively associated with olfactory memory. COMPARISON WITH EXISTING METHOD(S): TOM-32 could expand the possibilities of testing olfactory memory. It may offer additional information about cognitive and sensory abilities relative to existing research tools, and the large number of included test items may facilitate repeated and longitudinal testing. Wide range of applied odors increases the possibility to detect subtle differences between, as well as changes within, individuals. CONCLUSIONS: We present an extensive olfactory memory test with satisfactory psychometric qualities - a test that is comprehensive enough to show significant inter- and intra-individual differences, but time-efficient enough to be comfortable in daily research and clinical usage.

Older, non-demented apolipoprotein ε4 carrier males show hyperactivation and structural differences in odor memory regions: a blood-oxygen-level-dependent and structural magnetic resonance imaging study.

The current study sought to examine the interaction of sex and Apolipoprotein ε4 status on olfactory recognition memory within non-demented, older individuals. We separated 39 participants into groups based on ε4 status and sex. Each participant completed an olfactory memory recognition task during 2 functional magnetic resonance imaging scans and 1 structural scan. The ε4 carriers had greater functional recruitment of memory regions during false positives relative to ε4 non-carriers. During hits, the male ε4 carriers showed greater functional recruitment compared to female ε4 carriers. The ε4 carriers had larger bilateral putamen volumes relative to ε4 non-carriers. Neuroimaging data were significantly associated with Dementia Rating Scale scores solely in males. Results suggest differential olfactory memory processing in relation to sex and ε4 status. Male ε4 carriers in particular, demonstrated hyperactivation during recognition memory, which we suspect reflects neuronal compensation to maintain functional performance. Future studies should consider examining underlying mechanisms that contribute to these sex differences within ε4 carriers.

Drosophila Middle-Term Memory: Amnesiac is Required for PKA Activation in the Mushroom Bodies, a Function Modulated by Neprilysin 1.

In Drosophila, the mushroom bodies (MB) constitute the central brain structure for olfactory associative memory. As in mammals, the cAMP/PKA pathway plays a key role in memory formation. In the MB, Rutabaga (Rut) adenylate cyclase acts as a coincidence detector during associative conditioning to integrate calcium influx resulting from acetylcholine stimulation and G-protein activation resulting from dopaminergic stimulation. Amnesiac encodes a secreted neuropeptide required in the MB for two phases of aversive olfactory memory. Previous sequence analysis has revealed strong homology with the mammalian pituitary adenylate cyclase-activating peptide (PACAP). Here, we examined whether amnesiac is involved in cAMP/PKA dynamics in response to dopamine and acetylcholine co-stimulation in living flies. Experiments were conducted with both sexes, or with either sex. Our data show that amnesiac is necessary for the PKA activation process that results from coincidence detection in the MB. Since PACAP peptide is cleaved by the human membrane neprilysin hNEP, we searched for an interaction between Amnesiac and Neprilysin 1 (Nep1), a fly neprilysin involved in memory. We show that when Nep1 expression is acutely knocked down in adult MB, memory deficits displayed by amn hypomorphic mutants are rescued. Consistently, Nep1 inhibition also restores normal PKA activation in amn mutant flies. Taken together, the results suggest that Nep1 targets Amnesiac degradation to terminate its signaling function. Our work thus highlights a key role for Amnesiac in establishing within the MB the PKA dynamics that sustain middle-term memory (MTM) formation, a function modulated by Nep1.SIGNIFICANCE STATEMENT The Drosophila amnesiac gene encodes a secreted neuropeptide whose expression is required for specific memory phases in the mushroom bodies (MB), the olfactory memory center. Here, we show that Amnesiac is required for PKA activation resulting from coincidence detection, a mechanism by which the MB integrate two spatially distinct stimuli to encode associative memory. Furthermore, our results uncover a functional relationship between Amnesiac and Neprilysin 1 (Nep1), a membrane peptidase involved in memory and expressed in the MB. These results suggest that Nep1 modulates Amnesiac levels. We propose that on conditioning, Amnesiac release from the MB allows, via an autocrine process, the sustaining of PKA activation-mediating memory, which subsequently is inactivated by Nep1 degradation.