Olfactory Behaviors Assayed by Computer Tracking Of <em>Drosophila</em> in a Four-quadrant Olfactometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an ...

Imperial College London

A fly or bee's responses to widefield image motion depend on two basic parameters: temporal frequency and angular speed. Rotational optic flow is monitored using temporal frequency analysers, whereas translational optic flow seems to be monitored in terms of angular speed. Here we present a possible model of an angular speed detector which processes input signals through two parallel channels. The output of the detector is taken as the ratio of the two channels' outputs. This operation amplifies angular speed sensitivity and depresses temporal frequency tuning. We analyse the behaviour of two versions of this model with different filtering properties in response to a variety of input signals. We then embody the detector in a simulated agent's visual system and explore its behaviour in experiments on speed control and odometry. The latter leads us to suggest a new algorithm for optic flow driven odometry.

A model of visual detection of angular speed for bees.

Many bees and wasps learn about the immediate surroundings of their nest during learning flights, in which they look back towards the nest and acquire visual information that guides their subsequent returns. Visual guidance to the nest is simplified by the insects' tendency to adopt similar viewing directions during learning and return flights. To understand better the factors determining the particular viewing directions that insects choose, we have recorded the learning and return flights of a ground-nesting bumblebee in two visual environments--an enclosed garden with a partly open view between north and west, and a flat roof with a more open panorama. In both places, bees left and returned to an inconspicuous nest hole in the centre of a tabletop, with the hole marked by one or more nearby cylinders. In all experiments, bees adopted similar preferred orientations on their learning and return flights. Bees faced predominantly either north or south, suggesting the existence of two attractors. The bees' selection between attractors seems to be influenced both by the distribution of light, as determined by the shape of the skyline, and by the direction of wind. In the partly enclosed garden with little or no wind, bees tended to face north throughout the day, i.e. towards the pole in the brighter half of their surroundings. When white curtains, which distributed skylight more evenly, were placed around the table, bees faced both north and south. The bees on the roof tended to face south or north when the wind came from a wide arc of directions from the south or north, respectively. We suggest that bees switch facing orientation between north and south as a compromise between maintaining a single viewing direction for efficient view-based navigation and responding to the distribution of light for the easier detection of landmarks seen against the ground or to the direction of the wind for exploiting olfactory cues.

Preferred viewing directions of bumblebees (Bombus terrestris L.) when learning and approaching their nest site.

The courtship behavior of Drosophilid flies has served as a long-standing model for studying the bases of animal communication. During courtship, male flies flap their wings to send a complex pattern of airborne vibrations to the antennal ears of the females. These "courtship songs" differ in their spectrotemporal composition across species and are considered a crucial component of the flies' premating barrier. However, whether the species-specific differences in song structure are also reflected in the receivers of this communication system, i.e., the flies' antennal ears, has remained unexplored. Here we show for seven members of the melanogaster species group that (1) their ears are mechanically tuned to different best frequencies, (2) the ears' best frequencies correlate with high-frequency pulses of the conspecific courtship songs, and (3) the species-specific tuning relies on amplificatory mechanical feedback from the flies' auditory neurons. As a result of its level-dependent nature, the active mechanical feedback amplification is particularly useful for the detection of small stimuli, such as conspecific song pulses, and becomes negligible for sensing larger stimuli, such as the flies' own wingbeat during flight.

Active process mediates species-specific tuning of Drosophila ears.

The visually guided foraging routes of some formicine ants are individually stereotyped, suggesting the importance of visual learning in maintaining these routes. We ask here whether the wood ant Formica rufa learns a sequence of visual features encountered at different stages along a route, as reported for honeybees. We trained ants in several simple mazes to follow two alternative routes. Along each two-stage route, the ants first encountered one of two priming stimuli. The identity of the priming stimulus determined which of two choice stimuli was rewarded in the second stage of the route. As stimuli we used ultraviolet and yellow/green light panels, and two black-and-white patterns. Did ants learn to pair each colour with the appropriate black-and-white pattern? Ants learnt readily to discriminate between the two coloured stimuli or between the two black-and-white patterns. They could also pair coloured and black-and-white patterns, provided that the two were presented simultaneously. The ants' behaviour with sequential stimuli varied according to whether the priming stimulus was a coloured stimulus or a black-and-white pattern. When the priming stimulus was coloured, ants seemed to learn the two sequences, but tests showed that their success was probably caused by the after-effects of colour adaptation. With a black-and-white priming stimulus and a coloured second stage stimulus, robust sequential learning could not be demonstrated, although under certain experimental conditions a tiny proportion of ants did acquire the sequence. Thus, ants perform conditional discriminations reliably when priming and choice stimuli are simultaneous, but they usually fail when the stimuli are sequential.

Do wood ants learn sequences of visual stimuli?

Many wasps and bees learn the position of their nest relative to nearby visual features during elaborate 'learning' flights that they perform on leaving the nest. Return flights to the nest are thought to be patterned so that insects can reach their nest by matching their current view to views of their surroundings stored during learning flights. To understand how ground-nesting bumblebees might implement such a matching process, we have video-recorded the bees' learning and return flights and analysed the similarities and differences between the principal motifs of their flights. Loops that take bees away from and bring them back towards the nest are common during learning flights and less so in return flights. Zigzags are more prominent on return flights. Both motifs tend to be nest based. Bees often both fly towards and face the nest in the middle of loops and at the turns of zigzags. Before and after flight direction and body orientation are aligned, the two diverge from each other so that the nest is held within the bees' fronto-lateral visual field while flight direction relative to the nest can fluctuate more widely. These and other parallels between loops and zigzags suggest that they are stable variations of an underlying pattern, which enable bees to store and reacquire similar nest-focused views during learning and return flights.

Bumblebee calligraphy: the design and control of flight motifs in the learning and return flights of Bombus terrestris.

Bumblebees tend to face their nest over a limited range of compass directions when learning the nest's location on departure and finding it on their approach after foraging. They thus obtain similar views of the nest and its surroundings on their learning and return flights. How do bees coordinate their flights relative to nest-based and compass-based reference frames to get such similar views? We show, first, that learning and return flights contain straight segments that are directed along particular compass bearings, which are independent of the orientation of a bee's body. Bees are thus free within limits to adjust their viewing direction relative to the nest, without disturbing flight direction. Second, we examine the coordination of nest-based and compass-based control during likely information gathering segments of these flights: loops during learning flights and zigzags on return flights. We find that bees tend to start a loop or zigzag when flying within a restricted range of compass directions and to fly towards the nest and face it after a fixed change in compass direction, without continuous interactions between their nest-based and compass-based directions of flight. A preferred trajectory of compass-based flight over the course of a motif, combined with the tendency of the bees to keep their body oriented towards the nest automatically narrows the range of compass directions over which bees view the nest. Additionally, the absence of interactions between the two reference frames allows loops and zigzags to have a stereotyped form that can generate informative visual feedback.

Coordinating compass-based and nest-based flight directions during bumblebee learning and return flights.

Insects inform themselves about the 3D structure of their surroundings through motion parallax. During flight, they often simplify this task by minimising rotational image movement. Coordinated head and body movements generate rapid shifts of gaze separated by periods of almost zero rotational movement, during which the distance of objects from the insect can be estimated through pure translational optic flow. This saccadic strategy is less appropriate for assessing the distance between objects. Bees and wasps face this problem when learning the position of their nest-hole relative to objects close to it. They acquire the necessary information during specialised flights performed on leaving the nest. Here, we show that the bumblebee's saccadic strategy differs from other reported cases. In the fixations between saccades, a bumblebee's head continues to turn slowly, generating rotational flow. At specific points in learning flights these imperfect fixations generate a form of 'pivoting parallax', which is centred on the nest and enhances the visibility of features near the nest. Bumblebees may thus utilize an alternative form of motion parallax to that delivered by the standard 'saccade and fixate' strategy in which residual rotational flow plays a role in assessing the distances of objects from a focal point of interest.

Head movements and the optic flow generated during the learning flights of bumblebees.

The Q system is a repressible binary expression system for transgenic manipulations in living organisms. Through protein engineering and in vivo functional tests, we report here variants of the Q-system transcriptional activator, including QF2, for driving strong and ubiquitous expression in all Drosophila tissues. Our QF2, Gal4QF and LexAQF chimeric transcriptional activators substantially enrich the toolkit available for transgenic regulation in Drosophila melanogaster.

Improved and expanded Q-system reagents for genetic manipulations.

Intracellular Ca(2+) is a widely used neuronal activity indicator. Here we describe a transcriptional reporter of intracellular Ca(2+) (TRIC) in Drosophila that uses a binary expression system to report Ca(2+)-dependent interactions between calmodulin and its target peptide. We found that in vitro assays predicted in vivo properties of TRIC and that TRIC signals in sensory systems depend on neuronal activity. TRIC was able to quantitatively monitor neuronal responses that changed slowly, such as those of neuropeptide F-expressing neurons to sexual deprivation and neuroendocrine pars intercerebralis cells to food and arousal. Furthermore, TRIC-induced expression of a neuronal silencer in nutrient-activated cells enhanced stress resistance, providing a proof of principle that TRIC can be used for circuit manipulation. Thus, TRIC facilitates the monitoring and manipulation of neuronal activity, especially those reflecting slow changes in physiological states that are poorly captured by existing methods. TRIC's modular design should enable optimization and adaptation to other organisms.

Olena Riabinina

MRC Clinical Sciences Center

Imperial College London

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

Olena Riabinina

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}MRC Clinical Sciences Center

Imperial College London

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

MRC Clinical Sciences Center