Vertical System Research Articles

Voltage to Calcium Transformation Enhances Direction Selectivity in Drosophila T4 Neurons.

An important step in neural information processing is the transformation of membrane voltage into calcium signals leading to transmitter release. However, the effect of voltage to calcium transformation on neural responses to different sensory stimuli is not well understood. Here, we use in vivo two-photon imaging of genetically encoded voltage and calcium indicators, ArcLight and GCaMP6f, respectively, to measure responses in direction-selective T4 neurons of female Drosophila Comparison between ArcLight and GCaMP6f signals reveals calcium signals to have a significantly higher direction selectivity compared with voltage signals. Using these recordings, we build a model which transforms T4 voltage responses into calcium responses. Using a cascade of thresholding, temporal filtering and a stationary nonlinearity, the model reproduces experimentally measured calcium responses across different visual stimuli. These findings provide a mechanistic underpinning of the voltage to calcium transformation and show how this processing step, in addition to synaptic mechanisms on the dendrites of T4 cells, enhances direction selectivity in the output signal of T4 neurons. Measuring the directional tuning of postsynaptic vertical system (VS)-cells with inputs from other cells blocked, we found that, indeed, it matches the one of the calcium signal in presynaptic T4 cells.SIGNIFICANCE STATEMENT The transformation of voltage to calcium influx is an important step in the signaling cascade within a nerve cell. While this process has been intensely studied in the context of transmitter release mechanism, its consequences for information transmission and neural computation are unclear. Here, we measured both membrane voltage and cytosolic calcium levels in direction-selective cells of Drosophila in response to a large set of visual stimuli. We found direction selectivity in the calcium signal to be significantly enhanced compared with membrane voltage through a nonlinear transformation of voltage to calcium. Our findings highlight the importance of an additional step in the signaling cascade for information processing within single nerve cells.

Mathematical modelling and performance analysis of airflow distribution systems inside convection hot-air dryers

As a section on improving the energy efficiency and thermal efficiency of convection dryers, a different airflow distribution system in the convection dryer was established. The grape drying experiment was investigated by applying three types of distribution air inside a convection hot-air dryer. The vertical system (VS) in which hot air passes vertically through the tray of samples. However, the air was delivered parallel to the sample tray in two horizontal systems (HS) Types (I, II). The investigations were approved with the different air-drying temperatures of 40, 50, and 60 °C at different velocity 1.0, 1.5, and 2.0 m/s. The drying time in the horizontal dryer compared to the vertical system was significantly less. The data analysis showed that horizontal energy consumption with a type (II) dryer was low while the energy consumption was high under the vertical dryer. Moreover, applying the horizontal Type (II) for grapes drying had numerous improvements above the conventional vertical system, such as shorter processing times, less overall energy utilisation, and higher dryer thermal efficiency. According to the results, after fitting the experimental data with eleven drying kinetics models and comparing the results in terms ofthe root mean square error (RMSE), the reduced chi-square (χ2), and the correlation coefficient (R2), it was obtained that logarithmic model can give practically accurate evaluations of the drying curve for grapes drying in different convection dryers.

Non-uniform weighting of local motion inputs underlies dendritic computation in the fly visual system

The fly visual system offers a unique opportunity to explore computations performed by single neurons. Two previous studies characterized, in vivo, the receptive field (RF) of the vertical system (VS) cells of the blowfly (calliphora vicina), both intracellularly in the axon, and, independently using Ca2+ imaging, in hundreds of distal dendritic branchlets. We integrated this information into detailed passive cable and compartmental models of 3D reconstructed VS cells. Within a given VS cell type, the transfer resistance (TR) from different branchlets to the axon differs substantially, suggesting that they contribute unequally to the shaping of the axonal RF. Weighting the local RFs of all dendritic branchlets by their respective TR yielded a faithful reproduction of the axonal RF. The model also predicted that the various dendritic branchlets are electrically decoupled from each other, thus acting as independent local functional subunits. The study suggests that single neurons in the fly visual system filter dendritic noise and compute the weighted average of their inputs.

Near-optimal decoding of transient stimuli from coupled neuronal subpopulations.

Coupling between sensory neurons impacts their tuning properties and correlations in their responses. How such coupling affects sensory representations and ultimately behavior remains unclear. We investigated the role of neuronal coupling during visual processing using a realistic biophysical model of the vertical system (VS) cell network in the blow fly. These neurons are thought to encode the horizontal rotation axis during rapid free-flight maneuvers. Experimental findings suggest that neurons of the VS are strongly electrically coupled, and that several downstream neurons driving motor responses to ego-rotation receive inputs primarily from a small subset of VS cells. These downstream neurons must decode information about the axis of rotation from a partial readout of the VS population response. To investigate the role of coupling, we simulated the VS response to a variety of rotating visual scenes and computed optimal Bayesian estimates from the relevant subset of VS cells. Our analysis shows that coupling leads to near-optimal estimates from a subpopulation readout. In contrast, coupling between VS cells has no impact on the quality of encoding in the response of the full population. We conclude that coupling at one level of the fly visual system allows for near-optimal decoding from partial information at the subsequent, premotor level. Thus, electrical coupling may provide a general mechanism to achieve near-optimal information transfer from neuronal subpopulations across organisms and modalities.

From Dendritic Compartments to Neuronal Networks: A Multilevel Analysis of Motion Vision

Animals typically rely on vision to direct their locomotion through the environment. Flies, who move in three dimensions while in flight, have evolved the fastest visual system in the animal kingdom to help them stabilize their flight posture and trajectories (Autrum, 1958). Partly for this reason, they have been the subject of extensive research on the neuronal basis of motion vision, the component of visual function involved in detecting movement within a scene. Using a variety of techniques, including electrophysiology, genetic manipulation, and behavioral analysis, researchers have started to unravel the earliest stages of motion processing (Clark et al., 2011; Eichner et al., 2011). Visual motion processing in the fly begins with the elementary motion detectors (EMDs), which are units sensitive to one direction of motion over a small receptive field. The identities of the cells involved in this computation are under active research, and a complete picture has yet to emerge. For over four decades, however, the identity of one set of downstream cells that receive input from the EMDs has been known (Braitenberg, 1972). These cells, located in the lobula plate of the optic lobe of the fly, are called the horizontal system (HS) and vertical system (VS) cells.

Octopamine Neurons Mediate Flight-Induced Modulation of Visual Processing in Drosophila

Activity-dependent modulation of sensory systems has been documented in many organisms and is likely to be essential for appropriate processing of information during different behavioral states. However, the mechanisms underlying these phenomena remain poorly characterized. We investigated the role of octopamine neurons in the flight-dependent modulation observed in visual interneurons in Drosophila. The vertical system (VS) cells exhibit a boost in their response to visual motion during flight compared to quiescence. Pharmacological application of octopamine evokes responses in quiescent flies that mimic those observed during flight, and octopamine cells that project to the optic lobes increase in activity during flight. Using genetic tools to manipulate the activity of octopamine neurons, we find that they are both necessary and sufficient for the flight-induced visual boost. This study provides the first evidence that endogenous release of octopamine is involved in state-dependent modulation of visual interneurons in flies.

Robust Coding of Ego-Motion in Descending Neurons of the Fly

In many species, motion-sensitive neurons responding to optic flow at higher processing stages are well characterized; however, less is known how this representation of ego-motion is further transformed into an appropriate motor response. Here, we analyzed in the blowfly Calliphora vicina the visuomotor transformation from motion-sensitive neurons in the lobula plate [V2 and vertical system (VS) cells] onto premotor descending neurons [descending neurons of the ocellar and vertical system (DNOVS) cells] feeding into the motor circuit of the fly thoracic ganglion. We found that each of these cells is tuned to rotation of the fly around a particular body axis. Comparing the responses of presynaptic and postsynaptic cells revealed that DNOVS cells have approximately the same tuning widths as V2 and VS cells. However, DNOVS signals cells are less corrupted by fluctuations arising from the spatial structure of the visual input than their presynaptic elements. This leads to a more robust representation of ego-motion at the level of descending neurons. Thus, when moving from lobula plate cells to descending neurons, the selectivity for a particular optic flow remains unaltered, but the robustness of the representation increases.

Synaptic Organization of Lobula Plate Tangential Cells in Drosophila: Dα7 Cholinergic Receptors

: The nervous system of seeing animals derives information about optic flow in two subsequent steps. First, local motion vectors are calculated from moving retinal images, and second, the spatial distribution of these vectors is analyzed on the dendrites of large downstream neurons. In dipteran flies, this second step relies on a set of motion-sensitive lobula plate tangential cells (LPTCs), which have been studied in great detail in large fly species. Yet, studies on neurons that convey information to LPTCs and neuroanatomical investigations that enable a mechanistic understanding of the underlying dendritic computations in LPTCs are rare. We investigated the subcellular distribution of nicotinic acetylcholine receptors (nAChRs) on two sets of LPTCs: vertical system (VS) and horizontal system (HS) cells in Drosophila melanogaster. In this paper, we describe that both cell types express Dα7-type nAChR subunits specifically on higher order dendritic branches, similar to the expression of gamma aminobutyric acid (GABA) receptors. These findings support a model in which directional selectivity of LPTCs is achieved by the dendritic integration of excitatory, cholinergic, and inhibitory GABA-ergic input from local motion detectors with opposite preferred direction. Nonetheless, whole-cell recordings in mutant flies without Dα7 nAChRs revealed that direction selectivity of VS and HS cells is largely retained. In addition, mutant LPTCs were responsive to acetylcholine and remaining nAChR receptors were labeled by α-bungarotoxin. These results in LPTCs with genetically manipulated excitatory input synapses suggest a robust cellular implementation of dendritic processing that warrants direction selectivity. The underlying mechanism that ensures appropriate nAChR-mediated synaptic currents and the functional implications of separate sets or heteromultimeric nAChRs can now be addressed in this system.

Eigenanalysis of a neural network for optic flow processing

Flies gain information about self-motion during free flight by processing images of the environment moving across their retina. The visual course control center in the brain of the blowfly contains, among others, a population of ten neurons, the so-called vertical system (VS) cells that are mainly sensitive to downward motion. VS cells are assumed to encode information about rotational optic flow induced by self-motion (Krapp and Hengstenberg 1996 Nature 384 463–6). Recent evidence supports a connectivity scheme between the VS cells where neurons with neighboring receptive fields are connected to each other by electrical synapses at the axonal terminals, whereas the boundary neurons in the network are reciprocally coupled via inhibitory synapses (Haag and Borst 2004 Nat. Neurosci. 7 628–34; Farrow et al 2005 J. Neurosci. 25 3985–93; Cuntz et al 2007 Proc. Natl Acad. Sci. USA). Here, we investigate the functional properties of the VS network and its connectivity scheme by reducing a biophysically realistic network to a simplified model, where each cell is represented by a dendritic and axonal compartment only. Eigenanalysis of this model reveals that the whole population of VS cells projects the synaptic input provided from local motion detectors on to its behaviorally relevant components. The two major eigenvectors consist of a horizontal and a slanted line representing the distribution of vertical motion components across the fly's azimuth. They are, thus, ideally suited for reliably encoding translational and rotational whole-field optic flow induced by respective flight maneuvers. The dimensionality reduction compensates for the contrast and texture dependence of the local motion detectors of the correlation-type, which becomes particularly pronounced when confronted with natural images and their highly inhomogeneous contrast distribution.

Reciprocal inhibitory connections within a neural network for rotational optic-flow processing

Neurons in the visual system of the blowfly have large receptive fields that are selective for specific optic flow fields. Here, we studied the neural mechanisms underlying flow–field selectivity in proximal Vertical System (VS)-cells, a particular subset of tangential cells in the fly. These cells have local preferred directions that are distributed such as to match the flow field occurring during a rotation of the fly. However, the neural circuitry leading to this selectivity is not fully understood. Through dual intracellular recordings from proximal VS cells and other tangential cells, we characterized the specific wiring between VS cells themselves and between proximal VS cells and horizontal sensitive tangential cells. We discovered a spiking neuron (Vi) involved in this circuitry that has not been described before. This neuron turned out to be connected to proximal VS cells via gap junctions and, in addition, it was found to be inhibitory onto VS1.

Robust coding of flow-field parameters by axo-axonal gap junctions between fly visual interneurons

Complex flight maneuvers require a sophisticated system to exploit the optic flow resulting from moving images of the environment projected onto the retina. In the fly's visual course control center, the lobula plate, 10 so-called vertical system (VS) cells are thought to match, with their complex receptive fields, the optic flow resulting from rotation around different body axes. However, signals of single VS cells are unreliable indicators of such optic flow parameters in the context of their noisy, texture-dependent input from local motion measurements. Here we propose an alternative encoding scheme based on network simulations of biophysically realistic compartmental models of VS cells. The simulations incorporate recent data about the highly selective connectivity between VS cells consisting of an electrical axo-axonal coupling between adjacent cells and a reciprocal inhibition between the most distant cells. We find that this particular wiring performs a linear interpolation between the output signals of VS cells, leading to a robust representation of the axis of rotation even in the presence of textureless patches of the visual surround.

Sharing Receptive Fields with Your Neighbors: Tuning the Vertical System Cells to Wide Field Motion

In the blowfly, the direction-selective response of the 60 lobula-plate tangential cells has been ascribed to the integration of local motion information across their extensive dendritic trees. Because the lobula plate is organized retinotopically, the receptive fields of the tangential cells ought to be determined by their dendritic architecture. However, this appears not always to be the case. One compelling example is the exceptionally wide receptive fields of the vertical system (VS) tangential cells. Using dual-intracellular recordings, Haag and Borst (2004) found VS cells to be mutually coupled in such a way that each VS cell is connected exclusively to its immediate neighbors. This coupling may form the basis of the broad receptive fields of VS cells. Here, we tested this hypothesis directly by photoablating individual VS cells. The receptive field width of VS cells indeed narrowed after the ablation of single VS cells, specifically depending on whether the receptive field of the ablated cell was more frontal or more posterior to the recorded cell. In particular, the responses changed as if the neuron lost access to visual information from the ablated neuron and those VS cells more distal than it from the recorded neuron. These experiments provide strong evidence that the lateral connections among VS cells are a crucial component in the mechanism underlying their complex receptive fields, augmenting the direct columnar input to their dendrites.

Golgi analysis of tangential neurons in the lobula plate of Drosophila melanogaster.

The lobula plate (LP), which is the third order optic neuropil of flies, houses wide-field neurons which are exquisitely sensitive to motion. Among Diptera, motion-sensitive neurons of larger flies have been studied at the anatomical and physiological levels. However, the neurons of Drosophila lobula plate are relatively less explored. As Drosophila permits a genetic analysis of neural functions, we have analysed the organization of lobula plate of Drosophila melanogaster. Neurons belonging to eight anatomical classes have been observed in the present study. Three neurons of the horizontal system (HS) have been visualized. The HS north (HSN) neuron, occupying the dorsal lobula plate is stunted in its geometry compared to that of larger flies. Associated with the HS neurons, thinner horizontal elements known as h-cells have also been visualized in the present study. Five of the six known neurons of the vertical system (VS) have been visualized. Three additional neurons in the proximal LP comparable in anatomy to VS system have been stained. We have termed them as additional VS (AVS)-like neurons. Three thinner tangential cells that are comparable to VS neurons, which are elements of twin vertical system (tvs); and two cells with wide dendritic fields comparable to CH neurons of Diptera have been also observed. Neurons comparable to VS cells but with 'tufted' dendrites have been stained. The HSN and VS1-VS2 neurons are dorsally stunted. This is possibly due to the shape of the compound eye of Drosophila which is reduced in the fronto-dorsal region as compared to larger flies.

Small GTPase Cdc42 Is Required for Multiple Aspects of Dendritic Morphogenesis

The study of dendritic development in CNS neurons has been hampered by a lack of complex dendritic structures that can be studied in a tractable genetic system. In an effort to develop such a system, we recently characterized the highly complex dendrites of the vertical system (VS) neurons in the Drosophila visual system. Using VS neurons as a model system, we show here using loss-of-function mutations that endogenous Cdc42, a member of Rho family of small GTPases, is required for multiple aspects of dendritic morphogenesis. Cdc42-mutant VS neurons display normal complexity but increased dendritic length compared with wild type and have defects in dendrite caliber and stereotyped dendritic branch positions. Remarkably, Cdc42 mutant neurons also show a 50% reduction in dendritic spine density. These results demonstrate that Cdc42 is a regulator for multiple aspects of dendritic development.

Dendritic development of Drosophila high order visual system neurons is independent of sensory experience

BackgroundThe complex and characteristic structures of dendrites are a crucial part of the neuronal architecture that underlies brain function, and as such, their development has been a focal point of recent research. It is generally believed that dendritic development is controlled by a combination of endogenous genetic mechanisms and activity-dependent mechanisms. Therefore, it is of interest to test the relative contributions of these two types of mechanisms towards the construction of specific dendritic trees. In this study, we make use of the highly complex Vertical System (VS) of motion sensing neurons in the lobula plate of the Drosophila visual system to gauge the importance of visual input and synaptic activity to dendritic development.ResultsWe find that the dendrites of VS1 neurons are unchanged in dark-reared flies as compared to control flies raised on a 12 hour light, 12 hour dark cycle. The dendrites of these flies show no differences from control in dendrite complexity, spine number, spine density, or axon complexity. Flies with genetically ablated eyes show a slight but significant reduction in the complexity and overall length of VS1 dendrites, although this effect may be due to a reduction in the overall size of the dendritic field in these flies.ConclusionsOverall, our results indicate no role for visual experience in the development of VS dendrites, while spontaneous activity from photoreceptors may play at most a subtle role in the formation of fully complex dendrites in these high-order visual processing neurons.

A preparation of the blowfly ( Calliphora erythrocephala) brain for in vitro electrophysiological and pharmacological studies

We describe a method for the preparation and maintenance of the blowfly ( Calliphora erythrocephala) brain in a recording chamber under in vitro conditions in a semi-slice configuration. Large identified neurones in the posterior part of the 3rd optic lobe (lobula plate) can be penetrated easily with microelectrodes. The so-called vertical system (VS) cells which respond to vertical image motion in vivo could be encountered best because their axons are escorted individually by specific tracheae. Fluorescent stained cells show their natural shape as being in vivo. Electrophysiological properties of the cells investigated so far, i.e., resting potential (about −40 mV) and firing properties (single rebound spikes), are comparable to recordings in intact flies. Initial pharmacological experiments on VS cells in this preparation reveal that iontophoretical application of acetylcholine and carbamylcholine results in depolarization. VS cells also respond to bath-applied nicotine (1 μM) with a slow depolarization of their membrane potential in normal fly saline as well as in a Ca 2+-free saline, suggesting direct cholinergic input via nicotinic receptors. The suitability of the preparation for a wide range of electrophysiological and pharmacological studies is discussed.

The number and structure of giant vertical cells (VS) in the lobula plate of the blowflyCalliphora erythrocephala

1. The structure of one class of giant tangential neurons in the lobula plate ofCalliphora, the ‘Vertical System (VS)’ has been investigated by light microscopy. Different staining and reconstruction procedures were employed to ensure that all existing VS-neurons are revealed. 2. There are 11 VS-cells in a characteristic, and constant arrangement (Fig. 2). Each cell covers a particular area of the lobula plate, i.e., a distinct area of the retinotopic input array (Table 2), and therefore has a distinct receptive field. 3. Although VS-cells in general tend to occupy the posterior surface of the lobula plate, only three of them (VS 2-VS 5) reside exclusively in this layer. The other cells (VS1 and VS6-VS10) have bistratified dendritic arborizations (Fig. 6), whose dorsal parts are apposed to the anterior surface of the lobula plate. 4. The arrangement, territory and stratification of any given VS-cell is largely invariant in different individuals, whereas the branching pattern may vary considerably (Fig. 3). 5. The present results provide the framework for physiological studies of the role of individual VS-cells in movement perception, and their involvement in the control of particular locomotor behaviour.