Quantitative Predictions Orchestrate Visual Signaling in Drosophila.

Vision influences behavior, but ongoing behavior also modulates vision in animals ranging from insects to primates. The function and biophysical mechanisms of most such modulations remain unresolved. Here, we combine behavioral genetics, electrophysiology, and high-speed videography to advance a function for behavioral modulations of visual processing in Drosophila. We argue that a set of motion-sensitive visual neurons regulate gaze-stabilizing head movements. We describe how, during flight turns, Drosophila perform a set of head movements that require silencing their gaze-stability reflexes along the primary rotation axis of the turn. Consistent with this behavioral requirement, we find pervasive motor-related inputs to the visual neurons, which quantitatively silence their predicted visual responses to rotations around the relevant axis while preserving sensitivity around other axes. This work proposes a function for a behavioral modulation of visual processing and illustrates how the brain can remove one sensory signal from a circuit carrying multiple related signals.

Muscles that move the retina augment compound eye vision in Drosophila.

Most animals have compound eyes, with tens to thousands of lenses attached rigidly to the exoskeleton. A natural assumption is that all of these species must resort to moving either their head or their body to actively change their visual input. However, classic anatomy has revealed that flies have muscles poised to move their retinas under the stable lenses of each compound eye1-3. Here we show that Drosophila use their retinal muscles to smoothly track visual motion, which helps to stabilize the retinal image, and also to perform small saccades when viewing a stationary scene. We show that when the retina moves, visual receptive fields shift accordingly, and that even the smallest retinal saccades activate visual neurons. Using a head-fixed behavioural paradigm, we find that Drosophila perform binocular, vergence movements of their retinas-which could enhance depth perception-when crossing gaps, and impairing the physiology of retinal motor neurons alters gap-crossing trajectories during free behaviour. That flies evolved an ability to actuate their retinas suggests that moving the eye independently of the head is broadly paramount for animals. The similarities of smooth and saccadic movements of the Drosophila retina and the vertebrate eye highlight a notable example of convergent evolution.

Recording physiological history of cells with chemical labeling.

Recordings of the physiological history of cells provide insights into biological processes, yet obtaining such recordings is a challenge. To address this, we introduce a method to record transient cellular events for later analysis. We designed proteins that become labeled in the presence of both a specific cellular activity and a fluorescent substrate. The recording period is set by the presence of the substrate, whereas the cellular activity controls the degree of the labeling. The use of distinguishable substrates enabled the recording of successive periods of activity. We recorded protein-protein interactions, G protein-coupled receptor activation, and increases in intracellular calcium. Recordings of elevated calcium levels allowed selections of cells from heterogeneous populations for transcriptomic analysis and tracking of neuronal activities in flies and zebrafish.

Flicker-induced eye movements and the behavioural temporal cut-off frequency in a nocturnal spider.

We investigated changes in the eye muscle activity in the spider Cupiennius salei as a response to temporal intensity modulations. These spiders are known to enhance eye muscle activity in their principal eyes when moving stimuli are detected in the secondary eyes. We measured the activity of the dorsal eye muscle using a small telemetric unit attached to the spiders' prosoma and confronted the animals to flicker stimuli presented on a cathode ray tube monitor. We registered a significant increase in eye muscle activity as response to temporal light intensity modulations, which implies that no directed motion is required to trigger the spiders' response. This allowed the determination of the behavioural temporal cut-off frequency. None of the frequencies higher than 8.6 cycles s(-1) and all of the frequencies lower than 4.3 cycles s(-1) elicited a significant increase in eye muscle activity. A behavioural cut-off frequency of only a few cycles per second is well in line with the temporal properties of the photoreceptor cells determined using intracellular recordings. A relatively low temporal resolution and a relatively high spatial resolution suit well C. salei's lifestyle as a nocturnal sit-and-wait hunter.

Suppression of motion vision during course-changing, but not course-stabilizing, navigational turns.

From mammals to insects, locomotion has been shown to strongly modulate visual-system physiology. Does the manner in which a locomotor act is initiated change the modulation observed? We performed patch-clamp recordings from motion-sensitive visual neurons in tethered, flying Drosophila. We observed motor-related signals in flies performing flight turns in rapid response to looming discs and also during spontaneous turns, but motor-related signals were weak or non-existent in the context of turns made in response to brief pulses of unidirectional visual motion (i.e., optomotor responses). Thus, the act of a locomotor turn is variably associated with modulation of visual processing. These results can be understood via the following principle: suppress visual responses during course-changing, but not course-stabilizing, navigational turns. This principle is likely to apply broadly-even to mammals-whenever visual cells whose activity helps to stabilize a locomotor trajectory or the visual gaze angle are targeted for motor modulation.

The orientation-dependent visual spatial cut-off frequency in a spider.

Cupiennius salei (Araneae, Ctenidae) has, like most spiders, eight camera-type eyes. The anterior median eyes are called principal eyes and have a movable retina; all of the other eyes are referred to as secondary eyes and are equipped with a reflecting tapetum. The photoreceptors in the secondary eyes are arranged in rows on the tapetum and the inter-receptor angle along such a row is smaller than normal to it. In this study, the vertical and horizontal spatial cut-off frequencies of moving gratings were measured for the posterior median (PM) eyes, and the data were then compared with the anatomical data reported in the literature. Detection of moving objects in the secondary eyes enhances the eye muscle potential frequency in the principal eyes. We thus recorded the eye muscle activity with a telemetric unit as a monitor for motion detection while moving stimuli - sinusoidally modulated bright and dark stripes - were presented to the PM eyes on a computer screen. A significant increase in the eye muscle activity was measured for gratings at an angular wavelength of 2.0 deg in the vertical orientation and of 2.7 deg in the horizontal direction. In the vertical orientation the critical wavelength is twice the inter-receptor angle; in the horizontal orientation the spiders responded to wavelengths that are smaller than twice the corresponding inter-receptor angle. The cut-off frequency seems thus to be limited by the visual field of the photoreceptors rather than the inter-receptor angle. The relative intensity modulations modelled for the two different grating orientations in single photoreceptor cells were in line with our data.

Asymmetric processing of visual motion for simultaneous object and background responses.

Visual object fixation and figure-ground discrimination in Drosophila are robust behaviors requiring sophisticated computation by the visual system, yet the neural substrates remain unknown. Recent experiments in walking flies revealed object fixation behavior mediated by circuitry independent from the motion-sensitive T4-T5 cells required for wide-field motion responses. In tethered flight experiments under closed-loop conditions, we found similar results for one feedback gain, whereas intact T4-T5 cells were necessary for robust object fixation at a higher feedback gain and in figure-ground discrimination tasks. We implemented dynamical models (available at http://strawlab.org/asymmetric-motion/) based on neurons downstream of T4-T5 cells­one a simple phenomenological model and another, physiologically more realistic model­and found that both predict key features of stripe fixation and figure-ground discrimination and are consistent with a classical formulation. Fundamental to both models is motion asymmetry in the responses of model neurons, whereby front-to-back motion elicits stronger responses than back-to-front motion. When a bilateral pair of such model neurons, based on well-understood horizontal system cells, downstream of T4-T5, is coupled to turning behavior, asymmetry leads to object fixation and figure-ground discrimination in the presence of noise. Furthermore, the models also predict fixation in front of a moving background, a behavior previously suggested to require an additional pathway. Thus, the models predict several aspects of object responses on the basis of neurons that are also thought to serve a key role in background stabilization.

Pupil size in spider eyes is linked to post-ecdysal lens growth.

In this study we describe a distinctive pigment ring that appears in spider eyes after ecdysis and successively decreases in size in the days thereafter. Although pigment stops in spider eyes are well known, size variability is, to our knowledge, reported here for the first time. Representative species from three families (Ctenidae, Sparassidae and Lycosidae) are investigated and, for one of these species (Cupiennius salei, Ctenidae), the progressive increase in pupil diameter is monitored. In this species the pupil occupies only a fourth of the total projected lens surface after ecdysis and reaches its final size after approximately ten days. MicroCT images suggest that the decrease of the pigment ring is linked to the growth of the corneal lens after ecdysis. The pigment rings might improve vision in the immature eye by shielding light rays that would otherwise enter the eye via peripheral regions of the cornea, beside the growing crystalline lens.