Automated Charting of the Visual Space of Housefly Compound Eyes.

This paper describes the automatic measurement of the spatial organization of the visual axes of insect compound eyes, which consist of several thousands of visual units called ommatidia. Each ommatidium samples the optical information from a small solid angle, with an approximate Gaussian-distributed sensitivity (half-width on the order of 1˚) centered around a visual axis. Together, the ommatidia gather the visual information from a nearly panoramic field of view. The spatial distribution of the visual axes thus determines the eye's spatial resolution. Knowledge of the optical organization of a compound eye and its visual acuity is crucial for quantitative studies of neural processing of the visual information. Here we present an automated procedure for mapping a compound eye's visual axes, using an intrinsic, in vivo optical phenomenon, the pseudopupil, and the pupil mechanism of the photoreceptor cells. We outline the optomechanical setup for scanning insect eyes and use experimental results obtained from a housefly, Musca domestica, to illustrate the steps in the measurement procedure.

Rapid mapping of compound eye visual sampling parameters with FACETS, a highly automated wide-field goniometer.

A highly automated goniometer instrument (called FACETS) has been developed to facilitate rapid mapping of compound eye parameters for investigating regional visual field specializations. The instrument demonstrates the feasibility of analyzing the complete field of view of an insect eye in a fraction of the time required if using non-motorized, non-computerized methods. Faster eye mapping makes it practical for the first time to employ sample sizes appropriate for testing hypotheses about the visual significance of interspecific differences in regional specializations. Example maps of facet sizes are presented from four dipteran insects representing the Asilidae, Calliphoridae, and Stratiomyidae. These maps provide the first quantitative documentation of the frontal enlarged-facet zones (EFZs) that typify asilid eyes, which, together with the EFZs in male Calliphoridae, are likely to be correlated with high-spatial-resolution acute zones. The presence of EFZs contrasts sharply with the almost homogeneous distribution of facet sizes in the stratiomyid. Moreover, the shapes of EFZs differ among species, suggesting functional specializations that may reflect differences in visual ecology. Surveys of this nature can help identify species that should be targeted for additional studies, which will elucidate fundamental principles and constraints that govern visual field specializations and their evolution.

The allometry of CNS size and consequences of miniaturization in orb-weaving and cleptoparasitic spiders.

Allometric studies of the gross neuroanatomy of adults from nine species of spiders from six web-weaving families (Orbicularia), and nymphs from six of these species, show that very small spiders resemble other small animals in having disproportionately larger central nervous systems (CNSs) relative to body mass when compared with large-bodied forms. Small spiderlings and minute adult spiders have similar relative CNS volumes. The relatively large CNS of a very small spider occupies up to 78% of the cephalothorax volume. The CNSs of very small spiders extend into their coxae, occupying as much as 26% of the profile area of the coxae of an Anapisona simoni spiderling (body mass < 0.005 mg). Such modifications occur both in species with minute adults, and in tiny spiderlings of species with large-bodied adults. In at least one such species, Leucauge mariana, the CNS of the spiderling extends into a prominent ventral bulge of the sternum. Tiny spiders also have reduced neuronal cell body diameters. The adults of nearly all orbicularian spiders weave prey capture webs, as do the spiderlings, beginning with second instar nymphs. Comparable allometric relations occur in adults of both orb-weaving and cleptoparasitic species, indicating that this behavioral difference is not reflected in differences in gross CNS allometry.

An inexpensive and portable microvolumeter for rapid evaluation of biological samples.

We describe an improved microvolumeter (MVM) for rapidly measuring volumes of small biological samples, including live zooplankton, embryos, and small animals and organs. Portability and low cost make this instrument suitable for widespread use, including at remote field sites. Beginning with Archimedes' principle, which states that immersing an arbitrarily shaped sample in a fluid-filled container displaces an equivalent volume, we identified procedures that maximize measurement accuracy and repeatability across a broad range of absolute volumes. Crucial steps include matching the overall configuration to the size of the sample, using reflected light to monitor fluid levels precisely, and accounting for evaporation during measurements. The resulting precision is at least 100 times higher than in previous displacement-based methods. Volumes are obtained much faster than by traditional histological or confocal methods and without shrinkage artifacts due to fixation or dehydration. Calibrations using volume standards confirmed accurate measurements of volumes as small as 0.06 microL. We validated the feasibility of evaluating soft-tissue samples by comparing volumes of freshly dissected ant brains measured with the MVM and by confocal reconstruction.

Diverse speed response properties of motion sensitive neurons in the fly's optic lobe.

Speed and acceleration are fundamental components of visual motion that animals can use to interpret the world. Behavioral studies have established that insects discriminate speed largely independently of contrast and spatial frequency, and physiological recordings suggest that a subset of premotor descending neurons is in this sense speed-selective. Neural substrates and mechanisms of speed selectivity in insects, however, are unknown. Using blow flies Phaenicia sericata, intracellular recordings and dye-fills were obtained from medulla and lobula complex neurons which, though not necessarily speed-selective themselves, are positioned to participate in circuits that produce speed-selectivity in descending neurons. Stimulation with sinusoidally varied grating motion (0-200 degrees /s) provided a range of instantaneous velocities and accelerations. The resulting speed response profiles are indicative of four distinct speed ranges, supporting the hypothesis that the spatiotemporal tuning of mid-level neurons contains sufficient diversity to account for the emergence of speed selectivity at the descending neuron level. This type of mechanism has been proposed to explain speed discrimination in both insects and mammals, but has seemed less likely for insects due to possible constraints on small brains. Two additional recordings are suggestive of acceleration-selectivity, a potentially useful visual capability that is of uncertain functional significance for arthropods.

Sign-conserving amacrine neurons in the fly's external plexiform layer.

Amacrine cells in the external plexiform layer of the fly's lamina have been intracellulary recorded and dye-filled for the first time. The recordings demonstrate that like the lamina's short photoreceptors R1-R6, type 1 lamina amacrine neurons exhibit nonspiking, "sign-conserving" sustained depolarizations in response to illumination. This contrasts with the sign-inverting responses that typify first-order retinotopic relay neurons: monopolar cells L1-L5 and the T1 efferent neuron. The contrast frequency tuning of amacrine neurons is similar to that of photoreceptors and large lamina monopolar cells. Initial observations indicate that lamina amacrine receptive fields are also photoreceptor-like, suggesting either that their inputs originate from a small number of neighboring visual sampling units (VSUs), or that locally generated potentials decay rapidly with displacement. Lamina amacrines also respond to motion, and in one recording these responses were selective for the orientation of moving edges. This functional organization corresponds to the anatomy of amacrine cells, in which postsynaptic inputs from several neighboring photoreceptor endings are linked by a network of very thin distal processes. In this way, each VSU can receive convergent inputs from a surround of amacrine processes. This arrangement is well suited for relaying responses to local intensity fluctuations from neighboring VSUs to a central VSU where amacrines are known to be presynaptic to the dendrites of the T1 efferent. The T1 terminal converges at a deeper level with that of the L2 monopolar cell relaying from the same optic cartridge. Thus, the localized spatial responses and receptor-like temporal response properties of amacrines are consistent with possible roles in lateral inhibition, motion processing, or orientation processing.

The computational basis of an identified neuronal circuit for elementary motion detection in dipterous insects.

Based on comparative anatomical studies and electrophysiological experiments, we have identified a conserved subset of neurons in the lamina, medulla, and lobula of dipterous insects that are involved in retinotopic visual motion direction selectivity. Working from the photoreceptors inward, this neuronal subset includes lamina amacrine (alpha) cells, lamina monopolar (L2) cells, the basket T-cell (T1 or beta), the transmedullary cell Tm1, and the T5 bushy T-cell. Two GABA-immunoreactive neurons, the transmedullary cell Tm9 and a local interneuron at the level of T5 dendrites, are also implicated in the motion computation. We suggest that these neurons comprise the small-field elementary motion detector circuits the outputs of which are integrated by wide-field lobula plate tangential cells. We show that a computational model based on the available data about these neurons is consistent with existing models of biological elementary motion detection, and present a comparable version of the Hassenstein-Reichardt (HR) correlation model. Further, by using the model to synthesize a generic tangential cell, we show that it can account for the responses of lobula plate tangential cells to a wide range of transient stimuli, including responses which cannot be predicted using the HR model. This computational model of elementary motion detection is the first which derives specifically from the functional organization of a subset of retinotopic neurons supplying the lobula plate. A key prediction of this model is that elementary motion detector circuits respond quite differently to small-field transient stimulation than do spatially integrated motion processing neurons as observed in the lobula plate. In addition, this model suggests that the retinotopic motion information provided to wide-field motion-sensitive cells in the lobula is derived from a less refined stage of processing than motion inputs to the lobula plate.

Anatomical organization of retinotopic motion-sensitive pathways in the optic lobes of flies.

Anatomical methods have identified conserved neuronal morphologies and synaptic relationships among small-field retinotopic neurons in insect optic lobes. These conserved cell shapes occur across many species of dipteran insects and are also shared by Lepidoptera and Hymenoptera. The suggestion that such conserved neurons should participate in motion computing circuits finds support from intracellular recordings as well as older studies that used radioactive deoxyglucose labeling to reveal strata with motion-specific activity in an achromatic neuropil called the lobula plate. While intracellular recordings provide detailed information about the motion-sensitive or motion-selective responses of identified neurons, a full understanding of how arrangements of identified neurons compute and integrate information about visual motion will come from a multidisciplinary approach that includes morphological circuit analysis, the use of genetic mutants that exhibit specific deficits in motion processing, and biomimetic models. The latter must be based on the organization and connections of real neurons, yet provide output properties similar to those of more traditional theoretical models based on behavioral observations that date from the 1950s. Microsc. Res. Tech. 62:132-150, 2003.

Retinotopic pathways providing motion-selective information to the lobula from peripheral elementary motion-detecting circuits.

Recordings from afferent channels from the medulla supplying deep neuropils of the fly's optic lobes reveal different filter properties among the three classes of afferent neurons: transmedullary cells, T2 neurons, and Y cells. Whereas transmedullary cells respond to local flicker stimuli without discriminating these from directional or oriented motion, the T2 afferent neurons show clear motion orientation selectivity, which corresponds closely with a morphological bias in the orientation of their dendrites and could also be influenced by systems of local recurrent neurons in the medulla. A Y cell having a clearly defined terminal in the lobula, but having dendrite-like processes in the medulla and, possibly, the lobula plate, discriminates the direction of motion and its orientation. These results demonstrate unambiguously that the lobula receives information about motion and that the channels carrying it are distinct from those supplying wide-field motion-selective neurons in the lobula plate. Furthermore, recordings from a newly identified recurrent neuron linking the lobula back to the inner medulla demonstrate that the lobula discriminates nondirectional edge motion from flicker, thereby reflecting a property of this neuropil that is comparable with that of primary visual cortex in cats. The present findings support the proposal that elementary motion detecting circuits supply several parallel channels through the medulla, which segregate to, but are not shared by, the lobula and the lobula plate. The results are discussed in the context of other intracellular recordings from retinotopic neurons and with analogous findings from mammalian visual systems.