Light-induced activation of distinct modulatory neurons triggers appetitive or aversive learning in Drosophila larvae.

Many animals navigate using a combination of visual landmarks and path integration. In mammalian brains, head direction cells integrate these two streams of information by representing an animal's heading relative to landmarks, yet maintaining their directional tuning in darkness based on self-motion cues. Here we use two-photon calcium imaging in head-fixed Drosophila melanogaster walking on a ball in a virtual reality arena to demonstrate that landmark-based orientation and angular path integration are combined in the population responses of neurons whose dendrites tile the ellipsoid body, a toroidal structure in the centre of the fly brain. The neural population encodes the fly's azimuth relative to its environment, tracking visual landmarks when available and relying on self-motion cues in darkness. When both visual and self-motion cues are absent, a representation of the animal's orientation is maintained in this network through persistent activity, a potential substrate for short-term memory. Several features of the population dynamics of these neurons and their circular anatomical arrangement are suggestive of ring attractors, network structures that have been proposed to support the function of navigational brain circuits. Calcium imaging of the brain of tethered flies walking in a virtual reality arena showed that a population of neurons with dendrites that tile the ‘ellipsoid body’ use information from visual landmarks and the fly's own rotation to compute heading; this suggests insects possess neurons with similarities to ‘head direction cells’ known to contribute to spatial navigation in mammalian brains. How insect brains combine visual landmarks and path integration during navigation has been unknown. Johannes Seelig and Vivek Jayaraman perform calcium imaging of the brain of tethered flies walking in a virtual reality arena and show that a population of neurons with dendrites that tile the 'ellipsoid body' uses information from visual landmarks and the fly's own rotation to compute heading. This is the first evidence for an invertebrate equivalent of the 'head direction' neurons known to contribute to spatial navigation in mammalian brains.

Neural dynamics for landmark orientation and angular path integration

Drosophila neuropeptides in regulation of physiology and behavior

Learning and memory, defined as the acquisition and retention of neuronal representations of new information, are ubiquitous among insects. Recent research indicates that a variety of insects rely extensively on learning for all major life activities including feeding, predator avoidance, aggression, social interactions, and sexual behavior. There is good evidence that individuals within an insect species exhibit genetically based variation in learning abilities and indirect evidence linking insect learning to fitness. Although insects rely on innate behavior to successfully manage many types of variation and unpredictability, learning may be superior to innate behavior when dealing with features unique to time, place, or individuals. Among insects, social learning, which can promote the rapid spread of novel behaviors, is currently known only from a few well-studied examples in social Hymenoptera. The prevalence and importance of social learning in insects are still unknown. Similarly, we know little abo...

Evolutionary biology of insect learning

A central goal of neuroscience is to understand how neural circuits encode memory and guide behaviour. Studying simple, genetically tractable organisms, such as Drosophila melanogaster, can illuminate principles of neural circuit organization and function. Early genetic dissection of D. melanogaster olfactory memory focused on individual genes and molecules. These molecular tags subsequently revealed key neural circuits for memory. Recent advances in genetic technology have allowed us to manipulate and observe activity in these circuits, and even individual neurons, in live animals. The studies have transformed D. melanogaster from a useful organism for gene discovery to an ideal model to understand neural circuit function in memory.

Drosophila olfactory memory: single genes to complex neural circuits

Flexible goal-driven orientation requires that the position of a target be stored, especially in case the target moves out of sight. The capability to retain, recall and integrate such positional information into guiding behaviour has been summarized under the term spatial working memory. This kind of memory contains specific details of the presence that are not necessarily part of a long-term memory. Neurophysiological studies in primates indicate that sustained activity of neurons encodes the sensory information even though the object is no longer present. Furthermore they suggest that dopamine transmits the respective input to the prefrontal cortex, and simultaneous suppression by GABA spatially restricts this neuronal activity. Here we show that Drosophila melanogaster possesses a similar spatial memory during locomotion. Using a new detour setup, we show that flies can remember the position of an object for several seconds after it has been removed from their environment. In this setup, flies are temporarily lured away from the direction towards their hidden target, yet they are thereafter able to aim for their former target. Furthermore, we find that the GABAergic (stainable with antibodies against GABA) ring neurons of the ellipsoid body in the central brain are necessary and their plasticity is sufficient for a functional spatial orientation memory in flies. We also find that the protein kinase S6KII (ignorant) is required in a distinct subset of ring neurons to display this memory. Conditional expression of S6KII in these neurons only in adults can restore the loss of the orientation memory of the ignorant mutant. The S6KII signalling pathway therefore seems to be acutely required in the ring neurons for spatial orientation memory in flies.

Analysis of a spatial orientation memory in Drosophila

The neuromodulatory function of dopamine (DA) is an inherent feature of nervous systems of all animals. To learn more about the function of neural DA in Drosophila, we generated mutant flies that lack tyrosine hydroxylase, and thus DA biosynthesis, selectively in the nervous system. We found that DA is absent or below detection limits in the adult brain of these flies. Despite this, they have a lifespan similar to WT flies. These mutants show reduced activity, extended sleep time, locomotor deficits that increase with age, and they are hypophagic. Whereas odor and electrical shock avoidance are not affected, aversive olfactory learning is abolished. Instead, DA-deficient flies have an apparently “masochistic” tendency to prefer the shock-associated odor 2 h after conditioning. Similarly, sugar preference is absent, whereas sugar stimulation of foreleg taste neurons induces normal proboscis extension. Feeding the DA precursor l-DOPA to adults substantially rescues the learning deficit as well as other impaired behaviors that were tested. DA-deficient flies are also defective in positive phototaxis, without alteration in visual perception and optomotor response. Surprisingly, visual tracking is largely maintained, and these mutants still possess an efficient spatial orientation memory. Our findings show that flies can perform complex brain functions in the absence of neural DA, whereas specific behaviors involving, in particular, arousal and choice require normal levels of this neuromodulator.

http://www.pnas.org/content/pnas/108/2/834.full.pdf

Behavioral consequences of dopamine deficiency in the Drosophila central nervous system

Visual Targeting of Motor Actions in Climbing Drosophila

An understanding of associative learning is facilitated if it can be analyzed in a simple animal like the fruit fly Drosophila . Here, we introduce the first visual associative learning paradigm for larval Drosophila ; this is remarkable as larvae have an order of magnitude fewer neurons than adult flies. Larvae were subjected to either of two reciprocal training regimes: Light+/Dark- or Light-/Dark+. Subsequently, all larvae were individually tested for their preference between Light versus Dark. The difference between training regimes was therefore exclusively which visual situation was associated with which reinforcer; differences observed during the test thus reflected exclusively associative learning. For positive reinforcement (+) we used fructose (FRU), and for negative reinforcement (-) either quinine or sodium chloride (QUI, NaCl). Under these conditions, associative learning could be reproducibly observed in both wild-type strains tested. We then compared the effectiveness of training using differential conditioning, with both positive and negative reinforcement, to that using only positive or only negative reinforcement. We found that FRU only, but neither QUI nor NaCl, was in itself effective as a reinforcer. This is the first demonstration of appetitive learning in larval Drosophila . It is now possible to investigate the behavioral and neuronal organization of appetitive visual learning in this simple and genetically easy-to-manipulate experimental system.

/pdf/visual-learning-in-individually-assayed-drosophila-larvae-50rlo0nvzd.pdf

Visual learning in individually assayed Drosophila larvae.

Several aspects of locomotor control have been ascribed to the central complex of the insect brain; however, the role of distinct substructures of this complex is not well known. The tay bridge1 (tay1) mutant of Drosophila melanogaster was originally isolated on the basis of reduced walking speed and activity. In addition, tay1 is defective in the compensation of rotatory stimuli during walking and histologically, tay1 causes a mid-sagittal constriction of the protocerebral bridge, a constituent of the central complex. Cloning of the tay gene revealed that it encodes a novel protein with no significant homology to any known protein. To associate the behavioral phenotypes with the anatomical defect in the protocerebral bridge, we used different driver lines to express the tay cDNA in various neuronal subpopulations of the central brain in tay1-mutant flies. These experiments showed an association of the aberrant walking speed and activity with the structural defect in the protocerebral bridge. In contrast, the compensation of rotatory stimuli during walking was rescued without a restoration of the protocerebral bridge. The results of our differential rescue approach are supported by neuronal silencing experiments using conditional tetanus toxin expression in the same subset of neurons. These findings show for the first time that the walking speed and activity is controlled by different substructures of the central brain than the compensatory locomotion for rotatory stimuli. © 2008 Wiley Periodicals, Inc. Develop Neurobiol, 2008.

Kirsa Neuser

Papers

Analysis of a spatial orientation memory in Drosophila

Behavioral consequences of dopamine deficiency in the Drosophila central nervous system

Visual Targeting of Motor Actions in Climbing Drosophila

Visual learning in individually assayed Drosophila larvae.

Locomotor control by the central complex in Drosophila—An analysis of the tay bridge mutant