Threatening stimuli elicit a sequential cardiac pattern in arthropods.

In order to cope with the challenges of living in dynamic environments, animals rapidly adjust their behaviors in coordination with different physiological responses. Here, we studied whether threatening visual stimuli evoke different heart rate patterns in arthropods and whether these patterns are related with defensive behaviors. We identified two sequential phases of crab's cardiac response that occur with a similar timescale to that of the motor arrest and later escape response. The first phase was modulated by low salience stimuli and persisted throughout spaced stimulus presentation. The second phase was modulated by high-contrast stimuli and reduced by repetitive stimulus presentation. The overall correspondence between cardiac and motor responses suggests that the first cardiac response phase might be related to motor arrest while the second to the escape response. We show that in the face of threat arthropods coordinate their behavior and cardiac activity in a rapid and flexible manner.

A New Behavioral Paradigm for Visual Classical Conditioning in Drosophila.

Visual learning in animals is a remarkable cognitive ability that plays a crucial role in their survival and adaptation. Therefore, the ability to learn is highly conserved among animals. Despite lacking a centralized nervous system like vertebrates, invertebrates have demonstrated remarkable learning abilities. Here, we describe a simple behavioral assay that allows the analysis of visual associative learning in individually traceable freely walking adult fruit flies. The setup is based on the simple and widely used behavioral assay to study orientation behavior in flies. A single wing-clipped fly that has been starved for 21 h is placed on a platform where two unreachable opposite visual sets are displayed. This visual learning protocol was initially developed to study the cognitive ability of fruit flies to process numerical information. Through the application of the protocol, flies are able to associate a specific visual set with an appetitive reward. This association is revealed 2 h later during the testing session where we observed a change in their preference upon learning (i.e., change in their spontaneous preference). Moreover, this protocol could potentially be used to associate any other visual object/property to the reward, expanding the opportunities of studying visual learning in freely walking fruit flies at individual level.

Numerical discrimination in Drosophila melanogaster.

Sensitivity to numbers is a crucial cognitive ability. The lack of experimental models amenable to systematic genetic and neural manipulation has precluded discovering neural circuits required for numerical cognition. Here, we demonstrate that Drosophila flies spontaneously prefer sets containing larger numbers of objects. This preference is determined by the ratio between the two numerical quantities tested, a characteristic signature of numerical cognition across species. Individual flies maintained their numerical choice over consecutive days. Using a numerical visual conditioning paradigm, we found that flies are capable of associating sucrose with numerical quantities and can be trained to reverse their spontaneous preference for large quantities. Finally, we show that silencing lobula columnar neurons (LC11) reduces the preference for more objects, thus identifying a neuronal substrate for numerical cognition in invertebrates. This discovery paves the way for the systematic analysis of the behavioral and neural mechanisms underlying the evolutionary conserved sensitivity to numerosity.

Developmental transcriptional control of mitochondrial homeostasis is required for activity-dependent synaptic connectivity.

During neuronal circuit formation, local control of axonal organelles ensures proper synaptic connectivity. Whether this process is genetically encoded is unclear and if so, its developmental regulatory mechanisms remain to be identified. We hypothesized that developmental transcription factors regulate critical parameters of organelle homeostasis that contribute to circuit wiring. We combined cell type-specific transcriptomics with a genetic screen to discover such factors. We identified Telomeric Zinc finger-Associated Protein (TZAP) as a temporal developmental regulator of neuronal mitochondrial homeostasis genes, including Pink1 . In Drosophila , loss of dTzap function during visual circuit development leads to loss of activity-dependent synaptic connectivity, that can be rescued by Pink1 expression. At the cellular level, loss of dTzap/TZAP leads to defects in mitochondrial morphology, attenuated calcium uptake and reduced synaptic vesicle release in fly and mammalian neurons. Our findings highlight developmental transcriptional regulation of mitochondrial homeostasis as a key factor in activity-dependent synaptic connectivity.

Numerosity as a visual property: Evidence from two highly evolutionary distant species.

Most animals, from humans to invertebrates, possess an ability to estimate numbers. This evolutionary advantage facilitates animals' choice of environments with more food sources, more conspecifics to increase mating success, and/or reduced predation risk among others. However, how the brain processes numerical information remains largely unknown. There are currently two lines of research interested in how numerosity of visual objects is perceived and analyzed in the brain. The first argues that numerosity is an advanced cognitive ability processed in high-order brain areas, while the second proposes that "numbers" are attributes of the visual scene and thus numerosity is processed in the visual sensory system. Recent evidence points to a sensory involvement in estimating magnitudes. In this Perspective, we highlight this evidence in two highly evolutionary distant species: humans and flies. We also discuss the advantages of studying numerical processing in fruit flies in order to dissect the neural circuits involved in and required for numerical processing. Based on experimental manipulation and the fly connectome, we propose a plausible neural network for number sense in invertebrates.

A neurodevelopmental origin of behavioral individuality in the Drosophila visual system.

The genome versus experience dichotomy has dominated understanding of behavioral individuality. By contrast, the role of nonheritable noise during brain development in behavioral variation is understudied. Using Drosophila melanogaster, we demonstrate a link between stochastic variation in brain wiring and behavioral individuality. A visual system circuit called the dorsal cluster neurons (DCN) shows nonheritable, interindividual variation in right/left wiring asymmetry and controls object orientation in freely walking flies. We show that DCN wiring asymmetry instructs an individual's object responses: The greater the asymmetry, the better the individual orients toward a visual object. Silencing DCNs abolishes correlations between anatomy and behavior, whereas inducing DCN asymmetry suffices to improve object responses.

A simple computer vision pipeline reveals the effects of isolation on social interaction dynamics in Drosophila.

Isolation profoundly influences social behavior in all animals. In humans, isolation has serious effects on health. Drosophila melanogaster is a powerful model to study small-scale, temporally-transient social behavior. However, longer-term analysis of large groups of flies is hampered by the lack of effective and reliable tools. We built a new imaging arena and improved the existing tracking algorithm to reliably follow a large number of flies simultaneously. Next, based on the automatic classification of touch and graph-based social network analysis, we designed an algorithm to quantify changes in the social network in response to prior social isolation. We observed that isolation significantly and swiftly enhanced individual and local social network parameters depicting near-neighbor relationships. We explored the genome-wide molecular correlates of these behavioral changes and found that whereas behavior changed throughout the six days of isolation, gene expression alterations occurred largely on day one. These changes occurred mostly in metabolic genes, and we verified the metabolic changes by showing an increase of lipid content in isolated flies. In summary, we describe a highly reliable tracking and analysis pipeline for large groups of flies that we use to unravel the behavioral, molecular and physiological impact of isolation on social network dynamics in Drosophila.

A crustacean lobula plate: Morphology, connections, and retinotopic organization.

The lobula plate is part of the lobula complex, the third optic neuropil, in the optic lobes of insects. It has been extensively studied in dipterous insects, where its role in processing flow-field motion information used for controlling optomotor responses was discovered early. Recently, a lobula plate was also found in malacostracan crustaceans. Here, we provide the first detailed description of the neuroarchitecture, the input and output connections and the retinotopic organization of the lobula plate in a crustacean, the crab Neohelice granulata using a variety of histological methods that include silver reduced staining and mass staining with dextran-conjugated dyes. The lobula plate of this crab is a small elongated neuropil. It receives separated retinotopic inputs from columnar neurons of the medulla and the lobula. In the anteroposterior plane, the neuropil possesses four layers defined by the arborizations of such columnar inputs. Medulla projecting neurons arborize mainly in two of these layers, one on each side, while input neurons arriving from the lobula branch only in one. The neuropil contains at least two classes of tangential elements, one connecting with the lateral protocerebrum and the other that exits the optic lobes toward the supraesophageal ganglion. The number of layers in the crab's lobula plate, the retinotopic connections received from the medulla and from the lobula, and the presence of large tangential neurons exiting the neuropil, reflect the general structure of the insect lobula plate and, hence, provide support to the notion of an evolutionary conserved function for this neuropil.

Organization of columnar inputs in the third optic ganglion of a highly visual crab.

Motion information provides essential cues for a wide variety of animal behaviors such as mate, prey, or predator detection. In decapod crustaceans and pterygote insects, visual codification of object motion is associated with visual processing in the third optic neuropile, the lobula. In this neuropile, tangential neurons collect motion information from small field columnar neurons and relay it to the midbrain where behavioral responses would be finally shaped. In highly ordered structures, detailed knowledge of the neuroanatomy can give insight into their function. In spite of the relevance of the lobula in processing motion information, studies on the neuroarchitecture of this neuropile are scant. Here, by applying dextran-conjugated dyes in the second optic neuropile (the medulla) of the crab Neohelice, we mass stained the columnar neurons that convey visual information into the lobula. We found that the arborizations of these afferent columnar neurons lie at four main lobula depths. A detailed examination of serial optical sections of the lobula revealed that these input strata are composed of different number of substrata and that the strata are thicker in the centre of the neuropile. Finally, by staining the different lobula layers composed of tangential processes we combined the present characterization of lobula input strata with the previous characterization of the neuroarchitecture of the crab's lobula based on reduced-silver preparations. We found that the third lobula input stratum overlaps with the dendrites of lobula giant tangential neurons. This suggests that columnar neurons projecting from the medulla can directly provide visual input to the crab's lobula giant neurons.

Behaviorally related neural plasticity in the arthropod optic lobes.

BACKGROUND: Due to the complexity and variability of natural environments, the ability to adaptively modify behavior is of fundamental biological importance. Motion vision provides essential cues for guiding critical behaviors such as prey, predator, or mate detection. However, when confronted with the repeated sight of a moving object that turns out to be irrelevant, most animals will learn to ignore it. The neural mechanisms by which moving objects can be ignored are unknown. Although many arthropods exhibit behavioral adaptation to repetitive moving objects, the underlying neural mechanisms have been difficult to study, due to the difficulty of recording activity from the small columnar neurons in peripheral motion detection circuits. RESULTS: We developed an experimental approach in an arthropod to record the calcium responses of visual neurons in vivo. We show that peripheral columnar neurons that convey visual information into the second optic neuropil persist in responding to the repeated presentation of an innocuous moving object. However, activity in the columnar neurons that convey the visual information from the second to the third optic neuropil is suppressed during high-frequency stimulus repetitions. In accordance with the animal's behavioral changes, the suppression of neural activity is fast but short lasting and restricted to the retina's trained area. CONCLUSIONS: Columnar neurons from the second optic neuropil are likely the main plastic locus responsible for the modifications in animal behavior when confronted with rapidly repeated object motion. Our results demonstrate that visually guided behaviors can be determined by neural plasticity that occurs surprisingly early in the visual pathway.

Regionalization in the eye of the grapsid crab Neohelice granulata (=Chasmagnathus granulatus): variation of resolution and facet diameters.

Crabs have panoramic compound eyes, which can show marked regional specializations of visual acuity. These specializations are thought to be related to the particular features of the animal's ecological environment. Modern knowledge on the neuroanatomy and neurophysiology of the crabs' visual system mainly derives from studies performed in the grapsid crab Neohelice granulata (=Chasmagnathus granulatus). However, the organization of the visual sampling elements across the eye surface of this animal had not yet been addressed. We analyzed the sampling resolution across the eye of Neohelice by measuring the pseudopupil displacement with a goniometer. In addition, we measured the facet sizes in the different regions of the eye. We found that Neohelice possesses an acute band of high vertical resolution around the eye equator and an increase in horizontal sampling resolution and lenses diameter towards the lateral side of the eye. Therefore, the analysis of the optical apparatus indicates that this crab possesses greater visual acuity around the equator and at the lateral side of the eye. These specializations are compared with those found in different species of crabs and are discussed in connection to the particular ecological features of Neohelice's habitat.