Joshua van Kleef

The mapping of visual space by identified large second-order neurons in the dragonfly median ocellus

In adult dragonflies, the compound eyes are augmented by three simple eyes known as the dorsal ocelli. The outputs of ocellar photoreceptors converge on relatively few second-order neurons with large axonal diameters (L-neurons). We determine L-neuron morphology by iontophoretic dye injection combined with three-dimensional reconstructions. Using intracellular recording and white noise analysis, we also determine the physiological receptive fields of the L-neurons, in order to identify the extent to which they preserve spatial information. We find a total of 11 median ocellar L-neurons, consisting of five symmetrical pairs and one unpaired neuron. L-neurons are distinguishable by the extent and location of their terminations within the ocellar plexus and brain. In the horizontal dimension, L-neurons project to different regions of the ocellar plexus, in close correlation with their receptive fields. In the vertical dimension, dendritic arborizations overlap widely, paralleled by receptive fields that are narrow and do not differ between different neurons. These results provide the first evidence for the preservation of spatial information by the second-order neurons of any dorsal ocellus. The system essentially forms a one-dimensional image of the equator over a wide azimuthal area, possibly forming an internal representation of the horizon. Potential behavioural roles for the system are discussed.

The mapping of visual space by dragonfly lateral ocelli

We study the extent to which the lateral ocelli of dragonflies are able to resolve and map spatial information, following the recent finding that the median ocellus is adapted for spatial resolution around the horizon. Physiological optics are investigated by the hanging-drop technique and related to morphology as determined by sectioning and three-dimensional reconstruction. L-neuron morphology and physiology are investigated by intracellular electrophysiology, white noise analysis and iontophoretic dye injection. The lateral ocellar lens consists of a strongly curved outer surface, and two distinct inner surfaces that separate the retina into dorsal and ventral components. The focal plane lies within the dorsal retina but proximal to the ventral retina. Three identified L-neurons innervate the dorsal retina and extend the one-dimensional mapping arrangement of median ocellar L-neurons, with fields of view that are directed at the horizon. One further L-neuron innervates the ventral retina and is adapted for wide-field intensity summation. In both median and lateral ocelli, a distinct subclass of descending L-neuron carries multi-sensory information via graded and regenerative potentials. Dragonfly ocelli are adapted for high sensitivity as well as a modicum of resolution, especially in elevation, suggesting a role for attitude stabilisation by localization of the horizon.

A Spatiotemporal White Noise Analysis of Photoreceptor Responses to UV and Green Light in the Dragonfly Median Ocellus

Adult dragonflies augment their compound eyes with three simple eyes known as the dorsal ocelli. While the ocellar system is known to mediate stabilizing head reflexes during flight, the ability of the ocellar retina to dynamically resolve the environment is unknown. For the first time, we directly measured the angular sensitivities of the photoreceptors of the dragonfly median (middle) ocellus. We performed a second-order Wiener Kernel analysis of intracellular recordings of light-adapted photoreceptors. These were stimulated with one-dimensional horizontal or vertical patterns of concurrent UV and green light with different contrast levels and at different ambient temperatures. The photoreceptors were found to have anisotropic receptive fields with vertical and horizontal acceptance angles of 15° and 28°, respectively. The first-order (linear) temporal kernels contained significant undershoots whose amplitudes are invariant under changes in the contrast of the stimulus but significantly reduced at higher temperatures. The second-order kernels showed evidence of two distinct nonlinear components: a fast acting self-facilitation, which is dominant in the UV, followed by delayed self- and cross-inhibition of UV and green light responses. No facilitatory interactions between the UV and green light were found, indicating that facilitation of the green and UV responses occurs in isolated compartments. Inhibition between UV and green stimuli was present, indicating that inhibition occurs at a common point in the UV and green response pathways. We present a nonlinear cascade model (NLN) with initial stages consisting of separate UV and green pathways. Each pathway contains a fast facilitating nonlinearity coupled to a linear response. The linear response is described by an extended log-normal model, accounting for the phasic component. The final nonlinearity is composed of self-inhibition in the UV and green pathways and inhibition between these pathways. The model can largely predict the response of the photoreceptors to UV and green light.

Directional Selectivity in the Simple Eye of an Insect

Among other sensory modalities, flight stabilization in insects is performed with the aid of visual feedback from three simple eyes (ocelli). It is thought that each ocellus acts as a single wide-field sensor that detects changes in light intensity. We challenge this notion by providing evidence that, when light-adapted, the large retinal L-neurons in the median ocellus of the dragonfly respond in a directional way to upward moving bars and gratings. This ability is pronounced under UV illumination but weak or nonexistent in green light and is optimal at angular velocities of ∼750° s−1. Using a reverse-correlation technique, we analyze the functional organization of the receptive fields of the L-neurons. Our results reveal that L-neurons alter the structure of their linear spatiotemporal receptive fields with changes in the illuminating wavelength, becoming more inseparable and directional in UV light than in green. For moving bars and gratings, the strength of directionality predicted from the receptive fields is consistent with the measured values. Our results strongly suggest that, during the day, the retinal circuitry of the dragonfly median ocellus performs an early linear stage of motion processing. The likely advantage of this computation is to enhance pitch control.

Deciphering the Role of a Coleopteran Steering Muscle via Free Flight Stimulation

Testing hypotheses of neuromuscular function during locomotion ideally requires the ability to record cellular responses and to stimulate the cells being investigated to observe downstream behaviors [1]. The inability to stimulate in free flight has been a long-standing hurdle for insect flight studies. The miniaturization of computation and communication technologies has delivered ultra-small, radio-enabled neuromuscular recorders and stimulators for untethered insects [2-8]. Published stimulation targets include the areas in brain potentially responsible for pattern generation in locomotion [5], the nerve chord for abdominal flexion [9], antennal muscles [2, 10], and the flight muscles (or their excitatory junctions) [7, 11-13]. However, neither fine nor graded control of turning has been demonstrated in free flight, and responses to the stimulation vary widely [2, 5, 7, 9]. Technological limitations have precluded hypotheses of function validation requiring exogenous stimulation during flight. We investigated the role of a muscle involved in wing articulation during flight in a coleopteran. We set out to identify muscles whose stimulation produced a graded turning in free flight, a feat that would enable fine steering control not previously demonstrated. We anticipated that gradation might arise either as a function of the phase of muscle firing relative to the wing stroke (as in the classic fly b1 muscle [14, 15] or the dorsal longitudinal and ventral muscles of moth [16]), or due to regulated tonic control, in which phase-independent summation of twitch responses produces varying amounts of force delivered to the wing linkages [15, 17, 18].

Complex cell receptive fields: evidence for a hierarchical mechanism

Simple cells in the primary visual cortex have segregated ON and OFF subregions in their receptive fields, while complex cells have overlapping ON and OFF subregions. These two cell types form the extremes at each end of a continuum of receptive field types. Hubel and Wiesel in 1962 suggested a hierarchical scheme of processing whereby spatially offset simple cells drive complex cells. Simple and complex cells are often classified by their responses to moving sine wave gratings: simple cells have oscillatory responses while complex cells produce unmodulated responses. Here, using moving gratings as stimuli, we show that a significant number of cells that display low levels of response modulation at high contrasts demonstrate high levels of response modulation at low contrasts. Most often a drifting low contrast grating generates a large phasic response at the fundamental frequency of the grating (F1) and a smaller but significant phasic response that is approximately 180 deg out‐of‐phase with the F1 component. We present several models capable of capturing the effects of stimulus contrast on complex cell responses. The model that best reproduces our experimental results is a variation of the classical hierarchical model. In our model several spatially offset simple cells provide input to a complex cell, with each simple cell exhibiting a different contrast response function. At low contrasts only one of these simple cells is sufficiently excited to reveal its receptive field properties. As contrast is increased additional spatially offset simple cells with higher contrast thresholds add their responses to the overall spiking activity.

Spectral inputs and ocellar contributions to a pitch-sensitive descending neuron in the honeybee

By measuring insect compensatory optomotor reflexes to visual motion, researchers have examined the computational mechanisms of the motion processing system. However, establishing the spectral sensitivity of the neural pathways that underlie this motion behavior has been difficult, and the contribution of the simple eyes (ocelli) has been rarely examined. In this study we investigate the spectral response properties and ocellar inputs of an anatomically identified descending neuron (DNII(2)) in the honeybee optomotor pathway. Using a panoramic stimulus, we show that it responds selectively to optic flow associated with pitch rotations. The neuron is also stimulated with a custom-built light-emitting diode array that presented moving bars that were either all-green (spectrum 500-600 nm, peak 530 nm) or all-short wavelength (spectrum 350-430 nm, peak 380 nm). Although the optomotor response is thought to be dominated by green-sensitive inputs, we show that DNII(2) is equally responsive to, and direction selective to, both green- and short-wavelength stimuli. The color of the background image also influences the spontaneous spiking behavior of the cell: a green background produces significantly higher spontaneous spiking rates. Stimulating the ocelli produces strong modulatory effects on DNII(2), significantly increasing the amplitude of its responses in the preferred motion direction and decreasing the response latency by adding a directional, short-latency response component. Our results suggest that the spectral sensitivity of the optomotor response in honeybees may be more complicated than previously thought and that ocelli play a significant role in shaping the timing of motion signals.

Applicability of White-Noise Techniques to Analyzing Motion Responses

Motion processing in visual neurons is often understood in terms of how they integrate light stimuli in space and time. These integrative properties, known as the spatiotemporal receptive fields (STRFs), are sometimes obtained using white-noise techniques where a continuous random contrast sequence is delivered to each spatial location within the cells field of view. In contrast, motion stimuli such as moving bars are usually presented intermittently. Here we compare the STRF prediction of a neurons response to a moving bar with the measured response in second-order interneurons (L-neurons) of dragonfly ocelli (simple eyes). These low-latency neurons transmit sudden changes in intensity and motion information to mediate flight and gaze stabilization reflexes. A white-noise analysis is made of the responses of L-neurons to random bar stimuli delivered either every frame (densely) or intermittently (sparsely) with a temporal sequence matched to the bar motion stimulus. Linear STRFs estimated using the sparse stimulus were significantly better at predicting the responses to moving bars than the STRFs estimated using a traditional dense white-noise stimulus, even when second-order nonlinear terms were added. Our results strongly suggest that visual adaptation significantly modifies the linear STRF properties of L-neurons in dragonfly ocelli during dense white-noise stimulation. We discuss the ability to predict the responses of visual neurons to arbitrary stimuli based on white-noise analysis. We also discuss the likely functional advantages that adaptive receptive field structures provide for stabilizing attitude during hover and forward flight in dragonflies.