Testing for concordance between predicted species richness, past prioritization, and marine protected area designations in the western Indian Ocean.

Scientific advances in environmental data coverage and machine learning algorithms have improved the ability to make large-scale predictions where data are missing. These advances allowed us to develop a spatially resolved proxy for predicting numbers of tropical nearshore marine taxa. A diverse marine environmental spatial database was used to model numbers of taxa from ∼1000 field sites, and the predictions were applied to all 7039 6.25-km2 reef cells in 9 ecoregions and 11 nations of the western Indian Ocean. Our proxy for total numbers of taxa was based on the positive correlation (r2 = 0.24) of numbers of taxa of hard corals and 5 highly diverse reef fish families. Environmental relationships indicated that the number of fish species was largely influenced by biomass, nearness to people, governance, connectivity, and productivity and that coral taxa were influenced mostly by physicochemical environmental variability. At spatial delineations of province, ecoregion, nation, and strength of spatial clustering, we compared areas of conservation priority based on our total species proxy with those identified in 3 previous priority-setting reports and with the protected area database. Our method identified 119 locations that fit 3 numbers of taxa (hard coral, fish, and their combination) and 4 spatial delineations (nation, ecoregion, province, and reef clustering) criteria. Previous publications on priority setting identified 91 priority locations of which 6 were identified by all reports. We identified 12 locations that fit our 12 criteria and corresponded with 3 previously identified locations, 65 that aligned with at least 1 past report, and 28 that were new locations. Only 34% of the 208 marine protected areas in this province overlapped with identified locations with high numbers of predicted taxa. Differences occurred because past priorities were frequently based on unquantified perceptions of remoteness and preselected priority taxa. Our environment-species proxy and modeling approach can be considered among other important criteria for making conservation decisions.

Evaluación de la concordancia entre la riqueza de especies pronosticada, priorizaciones pasadas y la designación de áreas marinas protegidas en el oeste del Océano Índico Resumen Los avances científicos en la cobertura de datos ambientales y los algoritmos de aprendizaje automatizado han mejorado la capacidad de predecir a gran escala cuando hacen falta datos. Estos avances nos permiten desarrollar un representante con resolución espacial para predecir la cantidad de taxones marinos en las costas tropicales. Usamos una base de datos espaciales de diversos ambientes marinos para modelar la cantidad de taxones a partir de ∼1000 sitios de campo y aplicamos las predicciones a las 7039 celdas arrecifales de 6.25­km2 en nueve ecorregiones y once países del oeste del Océano Índico. Nuestro representante para la cantidad total de taxones se basó en la correlación positiva (r2=0.24) de la cantidad de taxones de corales duros y cinco familias de peces arrecifales con diversidad alta. Las relaciones ambientales indicaron que el número de especies de peces estuvo influenciado principalmente por la biomasa, la cercanía a las personas, la gestión, la conectividad y la productividad y que los taxones de coral estuvieron influenciados principalmente por la variabilidad ambiental fisicoquímica. Comparamos la prioridad de las áreas de conservación a nivel de las delimitaciones espaciales de provincia, ecorregión, nación y fuerza del agrupamiento espacial basado en nuestro total de especies representantes con aquellas especies identificadas en tres reportes previos de establecimiento de prioridades y con la base de datos de áreas protegidas. Con nuestro método identificamos 119 localidades aptas para tres cantidades de taxones (corales duros, peces y su combinación) y cuatro criterios de delimitación espacial (nación, ecorregión, provincia y grupo de arrecifes). Las publicaciones previas sobre el establecimiento de prioridades identificaron 91 localidades prioritarias de las cuales seis fueron identificadas por todos los reportes. Identificamos doce localidades que se ajustan a nuestros doce criterios y se correspondieron con tres localidades identificadas previamente, 65 que se alinearon con al menos un reporte anterior y 28 que eran nuevas localidades. Sólo 34% de las 208 áreas marinas protegidas en esta provincia se traslaparon con localidades identificadas con un gran número de taxones pronosticados. Hubo diferencias porque en el pasado se priorizaba frecuentemente con base en las percepciones no cuantificadas de lo remoto y prioritario de los taxones preseleccionados. Nuestra especie representante del ambiente y nuestra estrategia de modelo pueden considerarse entre otros criterios importantes para tomar decisiones de conservación.

IR76b-expressing neurons in Drosophila melanogaster are necessary for associative reward learning of an amino acid mixture.

Learning where to find nutrients while at the same time avoiding toxic food is essential for survival of any animal. Using Drosophila melanogaster larvae as a study case, we investigate the role of gustatory sensory neurons expressing IR76b for associative learning of amino acids, the building blocks of proteins. We found surprising complexity in the neuronal underpinnings of sensing amino acids, and a functional division of sensory neurons. We found that the IR76b receptor is dispensable for amino acid learning, whereas the neurons expressing IR76b are specifically required for the rewarding but not the punishing effect of amino acids. This unexpected dissociation in neuronal processing of amino acids for different behavioural functions provides a study case for functional divisions of labour in gustatory systems.

Prediction error drives associative learning and conditioned behavior in a spiking model of Drosophila larva.

Predicting reinforcement from sensory cues is beneficial for goal-directed behavior. In insect brains, underlying associations between cues and reinforcement, encoded by dopaminergic neurons, are formed in the mushroom body. We propose a spiking model of the Drosophila larva mushroom body. It includes a feedback motif conveying learned reinforcement expectation to dopaminergic neurons, which can compute prediction error as the difference between expected and present reinforcement. We demonstrate that this can serve as a driving force in learning. When combined with synaptic homeostasis, our model accounts for theoretically derived features of acquisition and loss of associations that depend on the intensity of the reinforcement and its temporal proximity to the cue. From modeling olfactory learning over the time course of behavioral experiments and simulating the locomotion of individual larvae toward or away from odor sources in a virtual environment, we conclude that learning driven by prediction errors can explain larval behavior.

Rewarding Capacity of Optogenetically Activating a Giant GABAergic Central-Brain Interneuron in Larval Drosophila.

Larvae of the fruit fly Drosophila melanogaster are a powerful study case for understanding the neural circuits underlying behavior. Indeed, the numerical simplicity of the larval brain has permitted the reconstruction of its synaptic connectome, and genetic tools for manipulating single, identified neurons allow neural circuit function to be investigated with relative ease and precision. We focus on one of the most complex neurons in the brain of the larva (of either sex), the GABAergic anterior paired lateral neuron (APL). Using behavioral and connectomic analyses, optogenetics, Ca2+ imaging, and pharmacology, we study how APL affects associative olfactory memory. We first provide a detailed account of the structure, regional polarity, connectivity, and metamorphic development of APL, and further confirm that optogenetic activation of APL has an inhibiting effect on its main targets, the mushroom body Kenyon cells. All these findings are consistent with the previously identified function of APL in the sparsening of sensory representations. To our surprise, however, we found that optogenetically activating APL can also have a strong rewarding effect. Specifically, APL activation together with odor presentation establishes an odor-specific, appetitive, associative short-term memory, whereas naive olfactory behavior remains unaffected. An acute, systemic inhibition of dopamine synthesis as well as an ablation of the dopaminergic pPAM neurons impair reward learning through APL activation. Our findings provide a study case of complex circuit function in a numerically simple brain, and suggest a previously unrecognized capacity of central-brain GABAergic neurons to engage in dopaminergic reinforcement.SIGNIFICANCE STATEMENT The single, identified giant anterior paired lateral (APL) neuron is one of the most complex neurons in the insect brain. It is GABAergic and contributes to the sparsening of neuronal activity in the mushroom body, the memory center of insects. We provide the most detailed account yet of the structure of APL in larval Drosophila as a neurogenetically accessible study case. We further reveal that, contrary to expectations, the experimental activation of APL can exert a rewarding effect, likely via dopaminergic reward pathways. The present study both provides an example of unexpected circuit complexity in a numerically simple brain, and reports an unexpected effect of activity in central-brain GABAergic circuits.

High-resolution analysis of individual Drosophila melanogaster larvae uncovers individual variability in locomotion and its neurogenetic modulation.

Neuronally orchestrated muscular movement and locomotion are defining faculties of multicellular animals. Due to its simple brain and genetic accessibility, the larva of the fruit fly Drosophila melanogaster allows one to study these processes at tractable levels of complexity. However, although the faculty of locomotion clearly pertains to the individual, most studies of locomotion in larvae use measurements aggregated across animals, or animals tested one by one, an extravagance for larger-scale analyses. This prevents grasping the inter- and intra-individual variability in locomotion and its neurogenetic determinants. Here, we present the IMBA (individual maggot behaviour analyser) for analysing the behaviour of individual larvae within groups, reliably resolving individual identity across collisions. We use the IMBA to systematically describe the inter- and intra-individual variability in locomotion of wild-type animals, and how the variability is reduced by associative learning. We then report a novel locomotion phenotype of an adhesion GPCR mutant. We further investigated the modulation of locomotion across repeated activations of dopamine neurons in individual animals, and the transient backward locomotion induced by brief optogenetic activation of the brain-descending 'mooncrawler' neurons. In summary, the IMBA is an easy-to-use toolbox allowing an unprecedentedly rich view of the behaviour and its variability of individual larvae, with utility in multiple biomedical research contexts.

Optogenetically induced reward and 'frustration' memory in larval Drosophila melanogaster.

Animals, including humans, form oppositely valenced memories for stimuli that predict the occurrence versus the termination of a reward: appetitive 'reward' memory for stimuli associated with the occurrence of a reward and aversive 'frustration' memory for stimuli that are associated with its termination. We characterized these memories in larval Drosophila melanogaster using a combination of Pavlovian conditioning, optogenetic activation of the dopaminergic central-brain DAN-i1864 neuron, and high-resolution video-tracking. This reveals their dependency on the number of training trials and the duration of DAN-i1864 activation, their temporal stability, and the parameters of locomotion that are modulated during memory expression. Together with previous results on 'punishment' versus 'relief' learning by DAN-f1 neuron activation, this reveals a 2×2 matrix of timing-dependent memory valence for the occurrence/termination of reward/punishment. These findings should aid the understanding and modelling of how brains decipher the predictive, causal structure of events around a target reinforcing occurrence.

Accumulation of commonly used agricultural herbicides in coral reef organisms from iSimangaliso Wetland Park, South Africa.

Coral reefs are amongst the most biodiverse ecosystems on earth, but are significantly impacted by agricultural runoff. Despite herbicides being commonly detected in coastal waters, the possibility of herbicide accumulation in coral reef species has largely been overlooked. We investigate the accumulation of several herbicides in five species of coral reef invertebrates collected from ten sites along the Maputaland coast, South Africa. Multiple herbicide residues were detected in 95% of the samples, with total average concentrations across sites ranging between 25.2 ng g-1 to 51.3 ng g-1 dw. Acetochlor, alachlor and hexazinone were the predominant herbicides detected at all sites, with atrazine and simazine detected less frequently. Significant interactive effects were detected between sites nested in reef complex crossed with species, based on multiple and total herbicide concentrations. In general, multivariate herbicide concentrations varied significantly between species within and across most sites. Contrastingly, the concentrations of the different herbicides and that of total herbicide did not differ between conspecifics at most sites nested in their respective reef complexes. On average, highest total herbicide concentrations were measured in soft coral (Sarcophyton glaucum; 90.4 ± 60 ng g-1 and Sinularia gravis; 42.7 ± 25 ng g-1) and sponge (Theonela swinhoei; 39.0 ± 40 ng g-1) species, while significantly lower concentrations were detected in hard corals (Echinopora hirsutissima; 10.5 ± 5.9 ng g-1 and Acropora austera; 5.20 ± 4.5 ng g-1) at most sites. Agricultural runoff entering the ocean via the uMfolozi-St Lucia Estuary and Maputo Bay are likely sources of herbicide contamination to coral reefs in the region. There is an urgent need to assess the long-term effects of herbicide exposure on coral reef communities.

Associative learning in larval and adult Drosophila is impaired by the dopamine-synthesis inhibitor 3-Iodo-L-tyrosine.

Across the animal kingdom, dopamine plays a crucial role in conferring reinforcement signals that teach animals about the causal structure of the world. In the fruit fly Drosophila melanogaster, dopaminergic reinforcement has largely been studied using genetics, whereas pharmacological approaches have received less attention. Here, we apply the dopamine-synthesis inhibitor 3-Iodo-L-tyrosine (3IY), which causes acute systemic inhibition of dopamine signaling, and investigate its effects on Pavlovian conditioning. We find that 3IY feeding impairs sugar-reward learning in larvae while leaving task-relevant behavioral faculties intact, and that additional feeding of a precursor of dopamine (L-3,4-dihydroxyphenylalanine, L-DOPA), rescues this impairment. Concerning a different developmental stage and for the aversive valence domain. Moreover, we demonstrate that punishment learning by activating the dopaminergic neuron PPL1-γ1pedc in adult flies is also impaired by 3IY feeding, and can likewise be rescued by L-DOPA. Our findings exemplify the advantages of using a pharmacological approach in combination with the genetic techniques available in D. melanogaster to manipulate neuronal and behavioral function.

Aversive teaching signals from individual dopamine neurons in larval Drosophila show qualitative differences in their temporal "fingerprint".

Dopamine serves many functions, and dopamine neurons are correspondingly diverse. We use a combination of optogenetics, behavioral experiments, and high-resolution video-tracking to probe for the functional capacities of two single, identified dopamine neurons in larval Drosophila. The DAN-f1 and the DAN-d1 neuron were recently found to carry aversive teaching signals during Pavlovian olfactory learning. We enquire into a fundamental feature of these teaching signals, namely their temporal "fingerprint". That is, receiving punishment feels bad, whereas being relieved from it feels good, and animals and humans alike learn with opposite valence about the occurrence and the termination of punishment (the same principle applies in the appetitive domain, with opposite sign). We find that DAN-f1 but not DAN-d1 can mediate such timing-dependent valence reversal: presenting an odor before DAN-f1 activation leads to learned avoidance of the odor (punishment memory), whereas presenting the odor upon termination of DAN-f1 activation leads to learned approach (relief memory). In contrast, DAN-d1 confers punishment memory only. These effects are further characterized in terms of the impact of the duration of optogenetic activation, the temporal stability of the memories thus established, and the specific microbehavioral patterns of locomotion through which they are expressed. Together with recent findings in the appetitive domain and from adult Drosophila, our results suggest that heterogeneity in the temporal fingerprint of teaching signals might be a more general principle of reinforcement processing through dopamine neurons.

Identification of Dopaminergic Neurons That Can Both Establish Associative Memory and Acutely Terminate Its Behavioral Expression.

An adaptive transition from exploring the environment in search of vital resources to exploiting these resources once the search was successful is important to all animals. Here we study the neuronal circuitry that allows larval Drosophila melanogaster of either sex to negotiate this exploration-exploitation transition. We do so by combining Pavlovian conditioning with high-resolution behavioral tracking, optogenetic manipulation of individually identified neurons, and EM data-based analyses of synaptic organization. We find that optogenetic activation of the dopaminergic neuron DAN-i1 can both establish memory during training and acutely terminate learned search behavior in a subsequent recall test. Its activation leaves innate behavior unaffected, however. Specifically, DAN-i1 activation can establish associative memories of opposite valence after paired and unpaired training with odor, and its activation during the recall test can terminate the search behavior resulting from either of these memories. Our results further suggest that in its behavioral significance DAN-i1 activation resembles, but does not equal, sugar reward. Dendrogram analyses of all the synaptic connections between DAN-i1 and its two main targets, the Kenyon cells and the mushroom body output neuron MBON-i1, further suggest that the DAN-i1 signals during training and during the recall test could be delivered to the Kenyon cells and to MBON-i1, respectively, within previously unrecognized, locally confined branching structures. This would provide an elegant circuit motif to terminate search on its successful completion.SIGNIFICANCE STATEMENT In the struggle for survival, animals have to explore their environment in search of food. Once food is found, however, it is adaptive to prioritize exploiting it over continuing a search that would now be as pointless as searching for the glasses you are wearing. This exploration-exploitation trade-off is important for animals and humans, as well as for technical search devices. We investigate which of the only 10,000 neurons of a fruit fly larva can tip the balance in this trade-off, and identify a single dopamine neuron called DAN-i1 that can do so. Given the similarities in dopamine neuron function across the animal kingdom, this may reflect a general principle of how search is terminated once it is successful.

Reversal learning in Drosophila larvae.

Adjusting behavior to changed environmental contingencies is critical for survival, and reversal learning provides an experimental handle on such cognitive flexibility. Here, we investigate reversal learning in larval Drosophila Using odor-taste associations, we establish olfactory reversal learning in the appetitive and the aversive domain, using either fructose as a reward or high-concentration sodium chloride as a punishment, respectively. Reversal learning is demonstrated both in differential and in absolute conditioning, in either valence domain. In differential conditioning, the animals are first trained such that an odor A is paired, for example, with the reward whereas odor B is not (A+/B); this is followed by a second training phase with reversed contingencies (A/B+). In absolute conditioning, odor B is omitted, such that the animals are first trained with paired presentations of A and reward, followed by unpaired training in the second training phase. Our results reveal "true" reversal learning in that the opposite associative effects of both the first and the second training phase are detectable after reversed-contingency training. In what is a surprisingly quick, one-trial contingency adjustment in the Drosophila larva, the present study establishes a simple and genetically easy accessible study case of cognitive flexibility.

Modulations of microbehaviour by associative memory strength in Drosophila larvae.

Finding food is a vital skill and a constant task for any animal, and associative learning of food-predicting cues gives an advantage in this daily struggle. The strength of the associations between cues and food depends on a number of parameters, such as the salience of the cue, the strength of the food reward and the number of joint cue-food experiences. We investigate what impact the strength of an associative odour-sugar memory has on the microbehaviour of Drosophila melanogaster larvae. We find that larvae form stronger memories with increasing concentrations of sugar or odour, and that these stronger memories manifest themselves in stronger modulations of two aspects of larval microbehaviour, the rate and the direction of lateral reorientation manoeuvres (so-called head casts). These two modulations of larval behaviour are found to be correlated to each other in every experiment performed, which is in line with a model that assumes that both modulations are controlled by a common motor output. Given that the Drosophila larva is a genetically tractable model organism that is well suited to the study of simple circuits at the single-cell level, these analyses can guide future research into the neuronal circuits underlying the translation of associative memories of different strength into behaviour, and may help to understand how these processes are organised in more complex systems.

Neuronal processing of amino acids in Drosophila: from taste sensing to behavioural regulation.

Finding and feeding on appropriate food are crucial for all animals. Carbohydrates and amino acids are both essential nutrients, albeit with distinct roles: the former are the main energy source whereas the latter are the building blocks of proteins and are used as neurotransmitters. Despite their crucial role, neither the sensing nor the neuronal processing of amino acids is well understood. Studies in Drosophila melanogaster have only recently gained momentum in shedding new light on the molecular and neuronal mechanisms of peripheral and internal amino acid sensing, as well as the organization of amino acid feeding behaviour. Furthermore, amino acids have been shown to act as rewards in associative learning. Focusing on recent studies in Drosophila, we summarize what is known so far about the perception of, and the behavioural responses to, amino acids in insects, and try to identify key questions for future research.

One-trial learning in larval Drosophila.

Animals of many species are capable of "small data" learning, that is, of learning without repetition. Here we introduce larval Drosophila melanogaster as a relatively simple study case for such one-trial learning. Using odor-food associative conditioning, we first show that a sugar that is both sweet and nutritious (fructose) and sugars that are only sweet (arabinose) or only nutritious (sorbitol) all support appetitive one-trial learning. The same is the case for the optogenetic activation of a subset of dopaminergic neurons innervating the mushroom body, the memory center of the insects. In contrast, no one-trial learning is observed for an amino acid reward (aspartic acid). As regards the aversive domain, one-trial learning is demonstrated for high-concentration sodium chloride, but is not observed for a bitter tastant (quinine). Second, we provide follow-up, parametric analyses of odor-fructose learning. Specifically, we ascertain its dependency on the number and duration of training trials, the requirements for the behavioral expression of one-trial odor-fructose memory, its temporal stability, and the feasibility of one-trial differential conditioning. Our results set the stage for a neurogenetic analysis of one-trial learning and define the requirements for modeling mnemonic processes in the larva.

Accumulation of organochlorine pesticides in reef organisms from marginal coral reefs in South Africa and links with coastal groundwater.

Coral reefs support rich levels of biodiversity, but are globally threatened by a multitude of factors, including land-sourced pollutants. Concentrations of organochlorine pesticides (OCPs) in three species of coral reef invertebrate at five sites along the Maputaland coast, South Africa were quantified. We aimed to assess spatial and interspecies variations in pesticide accumulation. Markedly high levels of a range of OCP residues were detected within tissues, with total concentrations (ng g-1 ww) ranging from 460 to 1200 (Sarcophyton glaucum), 1100-3000 (Sinularia gravis) and 450-1500 (Theonella swinhoei), respectively. A decreasing gradient in total pesticide concentrations was detected southward from Regal Reef, opposite Lake Sibaya, the hypothesised source of the pollutants. Observed gradients in pesticide concentrations and nitrogen isotope signatures indicated coastal groundwater to be the likely source of the pollutants. Further studies are required to assess the potential ecotoxicological impacts of these contaminants at the organismal and ecosystem level.

What can South African reefs tell us about the future of high-latitude coral systems?

Coral communities are found at high latitude on the East Coast subtropical reefs of South Africa. They are biodiverse, economically important, and afforded World Heritage Site status in the iSimangaliso Wetland Park where some are subjected to recreational use. While the Park's unique coral reefs have, to date, suffered little bleaching from climate change, they are susceptible to the phenomenon and provide a natural laboratory for the study of its effects at high latitude. This review covers recent advances in the regional oceanography; coral community dynamics and the underpinning reef processes, including minor bleaching events; the incidence of coral disease; and coral genetic connectivity. The effects of human activity (SCUBA diving, recreational fishing, pesticide use) were assessed, as well as the nursery benefits of Acropora austera, a coral which provides the reefs with much structure and is vulnerable to damage and climate change. The reefs were valued in terms of human use as well as services such as sediment generation and retention. The results have provided valuable information on relatively pristine, high-latitude reefs, their socio-economic benefits, and the anticipated effects of climate change.

Drivers of Soft and Stony Coral Community Distribution on the High-Latitude Coral Reefs of South Africa.

The role of abiotic parameters in determining the distribution of coral communities was assessed on the relatively pristine Maputaland reefs of South Africa from comprehensive reef survey data. The reefs, on which 42 communities could be defined, occur within three geographically separate complexes. Patterns in benthic distribution could be partially explained by latitude and depth, in particular, with slope, turbulence and reef aspect playing far less role in that order. A few species were associated exclusively or in high abundance with some of the communities; in most, it was the ratio of otherwise cosmopolitan species within all of the reef complexes that distinguished them. Complex biotic variables were also considered but not quantified and it is likely that low genetic connectivity and high levels of self-seeding result in a measure of isolation of the communities within a latitudinal gradient.

Associative Learning of Stimuli Paired and Unpaired With Reinforcement: Evaluating Evidence From Maggots, Flies, Bees, and Rats.

Finding rewards and avoiding punishments are powerful goals of behavior. To maximize reward and minimize punishment, it is beneficial to learn about the stimuli that predict their occurrence, and decades of research have provided insight into the brain processes underlying such associative reinforcement learning. In addition, it is well known in experimental psychology, yet often unacknowledged in neighboring scientific disciplines, that subjects also learn about the stimuli that predict the absence of reinforcement. Here we evaluate evidence for both these learning processes. We focus on two study cases that both provide a baseline level of behavior against which the effects of associative learning can be assessed. Firstly, we report pertinent evidence from Drosophila larvae. A re-analysis of the literature reveals that through paired presentations of an odor A and a sugar reward (A+) the animals learn that the reward can be found where the odor is, and therefore show an above-baseline preference for the odor. In contrast, through unpaired training (A/+) the animals learn that the reward can be found precisely where the odor is not, and accordingly these larvae show a below-baseline preference for it (the same is the case, with inverted signs, for learning through taste punishment). In addition, we present previously unpublished data demonstrating that also during a two-odor, differential conditioning protocol (A+/B) both these learning processes take place in larvae, i.e., learning about both the rewarded stimulus A and the non-rewarded stimulus B (again, this is likewise the case for differential conditioning with taste punishment). Secondly, after briefly discussing published evidence from adult Drosophila, honeybees, and rats, we report an unpublished data set showing that relative to baseline behavior after truly random presentations of a visual stimulus A and punishment, rats exhibit memories of opposite valence upon paired and unpaired training. Collectively, the evidence conforms to classical findings in experimental psychology and suggests that across species animals associatively learn both through paired and through unpaired presentations of stimuli with reinforcement - with opposite valence. While the brain mechanisms of unpaired learning for the most part still need to be uncovered, the immediate implication is that using unpaired procedures as a mnemonically neutral control for associative reinforcement learning may be leading analyses astray.

Functional architecture of reward learning in mushroom body extrinsic neurons of larval Drosophila.

The brain adaptively integrates present sensory input, past experience, and options for future action. The insect mushroom body exemplifies how a central brain structure brings about such integration. Here we use a combination of systematic single-cell labeling, connectomics, transgenic silencing, and activation experiments to study the mushroom body at single-cell resolution, focusing on the behavioral architecture of its input and output neurons (MBINs and MBONs), and of the mushroom body intrinsic APL neuron. Our results reveal the identity and morphology of almost all of these 44 neurons in stage 3 Drosophila larvae. Upon an initial screen, functional analyses focusing on the mushroom body medial lobe uncover sparse and specific functions of its dopaminergic MBINs, its MBONs, and of the GABAergic APL neuron across three behavioral tasks, namely odor preference, taste preference, and associative learning between odor and taste. Our results thus provide a cellular-resolution study case of how brains organize behavior.

The Ol1mpiad: concordance of behavioural faculties of stage 1 and stage 3 Drosophila larvae.

Mapping brain function to brain structure is a fundamental task for neuroscience. For such an endeavour, the Drosophila larva is simple enough to be tractable, yet complex enough to be interesting. It features about 10,000 neurons and is capable of various taxes, kineses and Pavlovian conditioning. All its neurons are currently being mapped into a light-microscopical atlas, and Gal4 strains are being generated to experimentally access neurons one at a time. In addition, an electron microscopic reconstruction of its nervous system seems within reach. Notably, this electron microscope-based connectome is being drafted for a stage 1 larva - because stage 1 larvae are much smaller than stage 3 larvae. However, most behaviour analyses have been performed for stage 3 larvae because their larger size makes them easier to handle and observe. It is therefore warranted to either redo the electron microscopic reconstruction for a stage 3 larva or to survey the behavioural faculties of stage 1 larvae. We provide the latter. In a community-based approach we called the Ol1mpiad, we probed stage 1 Drosophila larvae for free locomotion, feeding, responsiveness to substrate vibration, gentle and nociceptive touch, burrowing, olfactory preference and thermotaxis, light avoidance, gustatory choice of various tastants plus odour-taste associative learning, as well as light/dark-electric shock associative learning. Quantitatively, stage 1 larvae show lower scores in most tasks, arguably because of their smaller size and lower speed. Qualitatively, however, stage 1 larvae perform strikingly similar to stage 3 larvae in almost all cases. These results bolster confidence in mapping brain structure and behaviour across developmental stages.