Visual processing of moving single objects and wide-field patterns in flies: Behavioural analysis after laser-surgical removal of interneurons

A laser micro-beam unit was used to reproducibly and selectively eliminate the large horizontal and vertical motion sensitive neurons (H- and V-cells) of the lobula plate on one side of the brain of house fliesMusca domestica. This was achieved by ablating the precursors of these cells deep in the larval brain without damaging other cells in the brain or other tissues. The individually reared flies were tested for their behaviour. Three tests were performed: (i) visual fixation of a single stripe, (ii) the optomotor turning and thrust response to a stripe moving clockwise and counterclockwise around the fly, (iii) the monocular turning response to a moving grating. The responses to a moving single object were normal on both sides, the control side and the one lacking the H- and V-cells. However, the responses to a moving grating were reduced on the side lacking H- and V-cells for progressive (front to back) and regressive (back to front) motion. From this we conclude that the response to single objects is controlled mainly by cells other than the H- and V-cells. We also suggest two separate pathways for the processing of single object motion and wide field pattern motion respectively (Fig. 8). Furthermore, the H- and V-cells might function as visual stabilizers and background motion processors.     

Neural correlates of the optomotor response in the fly

Summary Studies of the optomotor response, the tendency to turn in response to a moving pattern, have yielded some understanding of the motion detection capabilities of the fly. We present data from extracellular microelectrode recordings from the optic lobes of the housefly, Musca domestica and the blowflies Eucalliphora lilaea and Calliphora phaenicia. Directionally selective and directionally nonselective motion sensitive units were observed in the region between the medulla and the lobula of all three species. Employing similar stimulus conditions to those used in the optomotor reaction studies, it was found that the response of the directionally selective units exhibited most of the characteristics of the optomotor response torque measurements. It is concluded that these units code the information prerequisite to the optomotor response and hence, that much data processing is achieved in the first few synaptic layers of the insect visual nervous system.     

The activity of pleurodorsal muscles during flight and at rest in the moth Manduca sexta (L.)

In the moth Manduca sexta, the paired mesothoracic flight steering muscle II PD2m takes part in the generation of the flight rhythm and is spontaneously active in the non-flying animal. This spontaneous activity is modulated by optomotor stimuli and directionally selective. The directional response characteristics are analyzed. Another spontaneously active steering muscle pair, the III PD2c, is situated in the metathorax. The activities of this pair and of a third muscle pair, the III PD3 are also influenced by visual stimulation.The responses of all 6 muscles to optomotor stimuli which simulate the flight situations yaw, roll, thrust and lift are analyzed. Each situation elicits a unique pattern of activation/deactivation within this set of muscles. The activity pattern in non-flying animals allows the prediction of flight steering mechanisms such as changes of wing area in flight turns and provides a useful basis for the analysis of visuo-motor pathways.     

Optomotor control of wing beat and body posture in drosophila

Continuous movement of striped patterns was presented on either side of a tethered fruitfly, Drosophila melanogaster, in order to simulate the displacement of stationary landmarks within the visual field of the freely moving fly. The horizontal components of the stimulus elicit, predominantly, yaw-torque responses during flight, or turning responses on the ground, which counteract involuntary deviations from a streight course in the corresponding mode of locomotion. The vertical components elicit, predominantly, covariant responses of lift and thrust which enable the fly to maintain a given level of flight. Monocular stimulation is sufficient to produce antagonistic responses, if the direction of the stimulus is reversed. The following constituents of the responses were derived mainly from properties of wing beat and body posture on photographs of fixed flight under visual stimulation. Wing stroke modulation (W. S. M.): The difference, and the sum, of the stroke amplitudes on either side are independently controlled by horizontal and vertical movement components, respectively. The maximum range of modulation per wing (12.3°) is equivalent to a 63% change in thrust on the corresponding side. Leg stroke modulation (L.S.M.): In the walking fly each pair of legs is under control of visual stimulation. The details of leg articulation are still unknown. Abdominal deflection (A.D.): An actively induced posture effect. Facilitates steering during free flight at increased air speed. Hind leg deflection (H.L.D.): Same as before. On most of the photographs the hind legs were deflected simultaneously and in the same direction as the abdomen. Hitch inhibition (H.I.): The term hitch denotes a transient reduction of stroke amplitude which seems to occur spontaneously and independently on either side of the fly. The hitch angle (12.2±3.8° S.D.) is most probably invariant to visual stimulation. Hitches are comparatively frequent in the absence of pattern movement. Their inhibition under visual stimulation is equivalent to an increase of the average thrust of the corresponding wing. The different constituents contribute to the optomotor responses according to the following tentative scheme (Fig. 7). The torque response is essentially due to the effects of W.S.M., A.D., H.L.D. and H.I., and the turning response to L.S.M. and possibly H.L.D., if the landmarks drift from anterior to posterior. So far, H.I. seems to be the only source of the torque response, and L.S.M. the only source of the turning response, if the landmarks drift in the opposite direction. The lift/thrust response results essentially from the effects of W.S.M. and H.I., no matter whether the landmarks drift from inferior to superior or in the opposite direction. The results obtained so far suggest that the optomotor control of course and altitude in Drosophila requires at least eight independent input channels or equivalent means for the separation of the descending signals from the visual centres. Further extension and refinement of the wiring scheme is required in order to improve the identification of the sensory inputs of the motor system and the classification of optomotor defective mutants.     

Comparison between the movement detection systems underlying the optomotor and the landing response in the housefly

Flies evaluate movement within their visual field in order to control the course of flight and to elicit landing manoeuvres. Although the motor output of the two types of responses is quite different, both systems can be compared with respect to the underlying movement detection systems. For a quantitative comparison, both responses were measured during tethered flight under identical conditions. The stimulus was a sinusoidal periodic pattern of vertical stripes presented bilaterally in the fronto-lateral eye region of the fly. To release the landing response, the pattern was moved on either side from front to back. The latency of the response depends on the stimulus conditions and was measured by means of an infrared light-beam that was interrupted whenever the fly lifted its forelegs to assume a preprogrammed landing posture (Borst and Bahde 1986). As an optomotor stimulus the pattern moved on one side from front to back and on the other side in the opposite direction. The induced turning tendency was measured by a torque meter (Götz 1964). The response values which will be compared are the inverse latencies of the landing response and the amplitude of the yaw torque.

1. Optomotor course-control is more sensitive to pattern movement at small spatial wavelengths (10° and 20°) than the landing response (Fig. 1a and b). This suggests that elementary movement detectors (EMDs, Buchner 1976) with large detection base (the distance between interacting visual elements) contribute more strongly to the landing than to the optomotor system.
2. The optimum contrast frequencies of the different responses obtained at a comparatively high pattern contrast of about 0.6 was found to be between 1 and 10 Hz for the optomotor response, and around 20 Hz for the landing response (Fig. 2a and b). This discrepancy can be explained by the fact that the optomotor response was tested under stationary conditions (several seconds of stimulation) while for the landing response transient response characteristics of the movement detectors have to be taken into account (landing occurs under these conditions within less than 100 ms after onset of the movement stimulus). To test the landing system under more stationary conditions, the pattern contrast had to be reduced to low values. This led to latencies of several seconds. Then the optimum of the landing response is around 4 Hz. This is in the optimum range of the optomotor course-control response. The result suggests the same filter time constants for the movement detectors of both systems.
3. The dependence of both responses on the position and the size of the pattern was examined. The landing response has its optimum sensitivity more ventrally than the optomotor response (Fig. 3a and b). Both response amplitudes increase with the size of the pattern in a similar progression (Fig. 3c and d).

In first approximation, the present results are compatible with the assumption of a common set of movement detectors for both the optomotor course-control and the landing system. Movement detectors with different sampling bases and at different positions in the visual field seem to contribute with different gain to both responses. Accordingly, the control systems underlying both behaviors are likely to be independent already at the level of spatial integration of the detector output.     

Basic organization of operant behavior as revealed in Drosophila flight orientation

Summary Operant behavior is studied in tethered Drosophila flies using visual motion, heat or odour as operandum and yaw torque, thrust or direction of flight as operans in various combinations (Fig. 1). On the basis of these results a conceptual framework of operant behavior is proposed: (1) It requires a goal (desired state) of which the actual state deviates. (2) To attain the goal a range of motor programs is activated (initiating activity, see Fig. 7). (3) Efference copies of the motor programs are compared to the sensory input referring to the deviation from the desired state (e.g. by cross-correlation). (4) In case of a significant coincidence the respective motor program is used to modify the sensory input in the direction towards the goal. (5) Consistent control of a sensory stimulus by a behavior may lead to a more permanent behavioral change (conditioning). In this scheme operant activity (1–4) and operant conditioning (1–5) are distinguished.Abbreviations ALU arbitrary length unit - d horizontal angular width of visual pattern - IR infrared - SEM standard error of the means - T yaw torque - Th thrust - performance index - horizontal angle between visual pattern position and longitudinal body axis of the fly - vertical angular extension of visual pattern     

Are the large monopolar cells of the insect lamina on the optomotor pathway?

Evidence is presented here from experiments on the visual system of the fly that questions participation of the large monopolar cells (LMCs) in the optomotor response.

Reafferent control of optomotor yaw torque inDrosophila melanogaster

Summary In the flight simulator the optomotor response ofDrosophila melanogaster does not operate as a simple feedback loop. Reafferent and exafferent motion stimuli are processed differently. Under open-loop conditions responses to motion are weaker than under closed-loop conditions. It takes the fly less than 100 ms to distinguish reafferent from exafferent motion. In closed-loop conditions, flies constantly generate torque fluctuations leading to small-angle oscillations of the panorama. This reafferent motion stimulus facilitates the response to exafferent motion but does not itself elicit optomotor responses. Reafference control appears to be directionally selective: while a displacement of the patternm by as little as 0.1° against the expected direction leads to a fast syndirectional torque response, displacements in the expected direction have no comparable effect. Based on the behavior of the mutantrol sol, which under open-loop conditions is directionally motion-blind but in closed-loop conditions still performs optomotor balance, a model is proposed in which the fly's endogenous torque fluctuations are an essential part of the course control process. It is argued that the model may also account for wild type optomotor balance in the flight simulator.     

Deoxyglucose mapping of nervous activity induced inDrosophila brain by visual movement

Summary Autoradiographs of the brains of the visual mutantsouter rhabdomeres absent ^JK84 (ora),small optic lobes ^KS58 (KS58) andno object fixation E ^B12 (B12) have been obtained by the deoxyglucose method. The patterns of metabolic activity in the optic lobes of the visually stimulated mutants is compared with that of similarly stimulated wildtype (WT) flies which was described in Part I of this work (Buchner et al. 1984b).In the mutantKS58 the optomotor following response to movement is nearly normal despite a 40–45% reduction of volume in the visual neuropils, medulla and lobula complex. InB12 flies the volume of these neuropils and the optomotor response are reduced. In autoradiographs of both mutants the pattern of neuronal activity induced by stimulation with moving gratings does not differ substantially from that in the WT. It suggests that only neurons irrelevant to movement detection are affected by the mutation. However, in the lobula plate of someKS58 flies and in the second chiasma of allB12 flies, the pattern of metabolic activity differs from that observed in WT flies. Up to now no causal relation has been found between the modifications described in behaviour or anatomy and those observed in the labelling of these mutants.In the ommatidia ofora flies the outer rhabdomeres are lacking while the central photoreceptors appear to be normal. Stimulus-specific labelling is absent in the visual neuropil of these mutants stimulated with movement or flicker. This result underlines the importance of the outer rhabdomeres for visual tasks, especially for movement detection.Abbreviations DG deoxyglucose - KS58 small optic lobes^KS58 - B12 no object fixation E^B12 - JK84 ora outer rhabdomeres absent ^JK84 - WT wildtype     

The vertical-horizontal neurone (VH) in the lobula plate of the blowfly,Phaenicia

Summary The anatomy and physiology of a motion-sensitive neurone, the vertical-horizontal (VH-) cell in the third visual neuropil (lobula plate) of the blowfly,Phaenicia was studied by intracellular recordings combined with dye injection. The cell possesses two dendritic fields in different layers of the lobula plate. The axon runs jointly with those of the vertical cells along the caudal surface of the lobula plate and terminates in the central protocerebrum lateral to the esophageal canal. The receptive field of the VH-cell is subdivided into two physiologically different parts which correspond to the two dendritic fields: if the input reaches the dendritic field residing in a more caudal layer (V-layer), the cell responds maximally to vertical pattern motion; whereas if the input reaches the dendritic field residing in a more rostral layer (H-layer), the cell responds maximally to horizontal pattern motion. The VH-neurone responds maximally to a contrast frequency of approximately / 1.8 Hz which coincides with the contrast frequency dependence of optomotor (following) responses. It is, therefore, considered to be a likely candidate mediating the pitch response (Blondeau and Heisenberg 1982) in flies.     

Deoxyglucose mapping of nervous activity induced inDrosophila brain by visual movement

Summary The pattern of visually induced local metabolic activity in the optic lobes of two structural mutants ofDrosophila melanogaster is compared with the corresponding wildtype pattern which has been reported in Part I of this work (Buchner et al. 1984b). Individualoptomotor-blind ^H31 (omb) flies lacking normal giant HS-neurons were tested behaviourally, and those with strongly reduced responses to visual movement were processed for 3H-deoxyglucose autoradiography. The distribution of metabolic activity in the optic lobes ofomb apparently does not differ substantially from that found in wildtype. In the mutantlobula plate-less ^N684 (lop) the small rudiment of the lobula plate which lacks many small-field input neurons does not show any stimulus-specific labelling. The data provide further support for the hypothesis that small-field input neurons to the lobula plate are the cellular substrate of the direction-specific labelling inDrosophila (see Buchner et al. 1984b).Abbreviations DG deoxyglucose - omb optomotor blind^H31 - lop lobula plate-less^N684 - WT wildtype     

Convergence of visual,haltere, and prosternai inputs at neck motor neurons of Calliphora erythrocephala

Summary Neck muscles of Calliphora erythrocephala, situated in the anterior prothorax, are innervated on each side by 8 motor neurons arising in the brain (cervical nerve neurons, CN1–8) and at least 13 motor neurons arising in the prothoracic ganglion (anterior dorsal and frontal nerve neurons, ADN1,2 and FN1-11). Three prominent motor neurons (CN6 and FN1,2) are described in detail with special emphasis on their relationships with giant visual interneurons from the lobula plate, haltere interneurons, and primary afferents from the prosternal organs and halteres. These sensory organs detect head movement and body yaw, respectively. Neuronal relationships indicate that head movement is under multimodal sensory control that includes giant motion-sensitive neurons previously supposed to mediate the optomotor response in flying flies. The described pathways provide anatomical substrates for the control of optokinetic and yaw-incurred head movements that behavioural studies have shown must exist.     

Evidence for one-way movement detection in the visual system of Drosophila

Optomotor control of course and altitude in the fruitfly, Drosophila melanogaster, requires dense networks of elementary movement detectors (EMD's) which cover most if not all of the visual field. The predominant types of EMD's in these networks represent interactions between neighbouring visual elements along the three main directions of the hexagonal array in the compound eye. — Course control in the walking fly is achieved mainly by pairs of equivalent EMD's which occupy 2 o'clock and 4 o'clock positions with respect to the right eye (Buchner, 1976). Comparison of the turning response and the torque response in the present account confirms the particular properties of this network, and proves the presumed bidirectional sensitivity of its EMD's for the course control responses of legs and wings in the corresponding modes of locomotion. — Altitude control during flight is achieved by a less homogeneous network of EMD's which modifies lift and thrust simultaneously by the appropriate control of the wing beat amplitudes. The predominant types of EMD's in the lateral eye regions occupy 12 o'clock and 2 o'clock positions with respect to the right eye (Buchner et al., 1978). The present evaluation of the optomotor responses of thrust and wing beat confirms the preferred orientation of these EMD's and discloses a pecularity of their internal structure. The movement detectors of this network lack the bidirectional sensitivity of the EMD's in the course control system. At least the fronto-lateral network of the altitude control system seems to consist mainly of pairs of equivalent unidirectional EMD's. The detectors in 12 o'clock position increase wing beat in response to movement of the visual surroundings from inferior to superior. The opposite effect is produced by the detectors in 2 o'clock position which respond to movement from anterior-superior to posterior-inferior. These properties qualify unidirectional EMD's as the functional units of the optomotor control system in the fruitfly. Pairs of unidirectional antagonists would be sufficient to establish the bidirectional sensitivity found in the movement detectors of the course control system.     

Is there a motion-independent position computation of an object in the visual system of the housefly?

A single vertical stripe (long or short) was moved clockwise, with constant speed, around a tethered femaleMusca domestica fly. The yaw torque response of the fly was analyzed as a function of the position of the object. After an interval of 8 s the stripe was moved counterclockwise and a similar analysis of the torque was made. This procedure was repeated a few times and averaged to each direction separately and for all the flies tested. The results suggested that: a) There are at least two mechanisms for computing the optomotor response in the lower part of the fly's eye, one performing a position-dependent velocity computation and the other depending on the position but not on the direction of motion of an object. b) These two components are parametrized over the position on the lower part of the eye. The results also show that: c) There is a significant difference in the response between the upper and the lower part of the eye. The position-dependent component cannot be detected in the upper part of the eye. In addition: d) Two different control mechanisms are proposed, one responding to progressive (from front to back) and one to regressive (from back to front) movement of objects.     

How the two eyes add together: monocular properties of the visually guided orientation behaviour of flies

In this account fixation and the torque response to a transient moving stripe of flying femaleMusca domestica with monocular sight was tested. This was made by either covering one eye of the fly with opaque paint or by placing a screen in front of one side of the fly's visual field. A stripe was moved with constant speed once around the fly clockwise and, after a pause, counterclockwise. The torque response of the fly was measured during the motion of the stripe and shortly beforehand. The results demonstrated that the monocular torque response to progressive (from front to back) motion and regressive (from back to front) motion essentially do not differ from the binocular response, except for the region of bionocular overlap. The beginning of the response of a fly with monocular vision to progressive motion is 11 ° (on average) before the direction of flight (0°), which means that the maximal functional binocular overlap of femaleMusca domestica is stretched at least 15° to each side (3.1). In addition, the shape of the monocular torque response to a progressively moving stripe was determined (see Figs. 5Ia and 5IIb). In other experiments similar to the ones described above, a screen was placed on one side of the fly's visual field or then on the other, (instead of covering one eye) and the torque response to the moving stripe was measured. Using this method, a delay response of 90 ms was measured. We suggest that this is the delay of the direction-sensitive component of the torque response, and therefore an additional argument for the existence of two components for the optomotor torque response. Flies with a covered eye or with a screen placed in front of one side of the visual field were able to fixate a single narrow long black stripe. This, however, was possible only when an additional offset signal was added, in order to give the stripe a constant velocity component. As a result there was a shift of the fixation towards the unobscured eye. The shift was small for the monocular flies, and it was larger (13° on average) when the screen was on one side of the fly. A new type of laser torquethrust transducer was developed and is described.     

How does lateral abdomen deflection contribute to flight control ofDrosophila melanogaster?

Summary Tethered flyingDrosophila melanogaster change the posture of their caudal body appendages in response to visual stimuli. In the present paper the relevance of lateral abdomen deflections for flight control is analysed. During abdomen deflections the line of action of the gravitational force is shifted with the fly's centre of mass. The line of action of aerodynamic drag forces is displaced accordingly, because friction is increased on the side of the body to which the abdomen is deflected. These two passive forces, together with the average flight forces generated actively by the wings, induce a yaw moment. In still air, the axis of this torque is tilted about 30° backwards relative to the vertical body axis. It will be called yaw axis of the flight mechanics. Two sets of observations support the notion of a combined yaw motor output. (a) The elementary motion detectors mediating the lateral abdomen deflection and the dynamics of the response resemble that of the optomotor response measured as yaw torque or as variation of wing beat amplitudes. (b) The asymmetric directional selectivity of the motion detecting system mediating the abdomen deflection corresponds to the orientation of the yaw axis of the flight mechanics. To explain the asymmetry, a nonlinear transfer characteristic is assumed in the motion detecting system.Abbreviations EMD elementary motion detector - MDF motion detector field     

Distinct Acute Zones for Visual Stimuli in Different Visual Tasks in Drosophila

The fruit fly Drosophila melanogaster has a sophisticated visual system and exhibits complex visual behaviors. Visual responses, vision processing and higher cognitive processes in Drosophila have been studied extensively. However, little is known about whether the retinal location of visual stimuli can affect fruit fly performance in various visual tasks. We tested the response of wild-type Berlin flies to visual stimuli at several vertical locations. Three paradigms were used in our study: visual operant conditioning, visual object fixation and optomotor response. We observed an acute zone for visual feature memorization in the upper visual field when visual patterns were presented with a black background. However, when a white background was used, the acute zone was in the lower visual field. Similar to visual feature memorization, the best locations for visual object fixation and optomotor response to a single moving stripe were in the lower visual field with a white background and the upper visual field with a black background. The preferred location for the optomotor response to moving gratings was around the equator of the visual field. Our results suggest that different visual processing pathways are involved in different visual tasks and that there is a certain degree of overlap between the pathways for visual feature memorization, visual object fixation and optomotor response.     

A synergetic theory of environmentally-specified and learned patterns of movement coordination

By combining neuropharmacology and electrophysiology, we tried to determine whether the main neuronal mechanism responsible for direction-selective motion detection in the fly is based on an excitatory or an inhibitory synaptic interaction. By blocking inhibitory interactions with picrotoxinin, an antagonist of the inhibitory neurotransmitter GABA, we could abolish most of the directional selectivity of a large-field movement-sensitive neuron (H1-cell) in the lobula plate of the blowfly Calliphora erythrocephala. These modifications are similar to changes observed in the optomotor response of the fruitfly Drosophila melanogaster after application of picrotoxinin (Bülthoff and Bülthoff 1987a, b). Assuming a simplified logical model, these results are compatible with inhibitory synaptic interactions at the level of the elementary movement detectors. The picrotoxinin-induced changes in direction selectivity are not due to modifications of the peripheral visual processing in the retina and lamina. This was shown by simultaneous recordings of the electroretinogram and the H1-cell. The latencies between drug injections into various parts of the brain and their first effects on the H1-cell suggest that the inhibitory mechanism for motion detection is located in the medulla rather than in the lobula plate.     

Forewing movements and motor activity during roll manoeuvers in flying desert locusts

Desert locusts, tethered on a roll torque meter and flying in a wind tunnel are surrounded by an artificial horizon (Fig. 1). Flight motor activity and movement of forewings are monitored continuously. Movements of the artificial horizon elicit roll manoeuvers of the animal with latencies of several seconds; concomitant changes in flight motor pattern and wing movement can be correlated with the animal's roll angle and roll torque. Flight sequences with constant torque and roll angle (steady state) have been analysed with the following results. Wing Kinematics. A phase difference between the movements of the forewings on either side is correlated with roll angle (Fig. 3). Pronation of a forewing is always greater on the side to which the animal rolls, i.e. on the side that produces less lift (Fig. 5). In some experiments the slope of the wing tip path is also decreased (Fig. 5). In both cases, the aerodynamic angle of attack is decreased and the forewing on this side produces less lift. In most experiments, changes in pronation are less pronounced in the contralateral wing (Fig. 11). All factors contribute to a net roll torque that sustains the animal's roll angle. Other kinematic parameters of forewing movement, e.g. wing stroke amplitude, were not found to be correlated with roll angle and torque (Fig. 4). Motor Pattern. Activity of several flight muscles (depressors M97, M98, M99, and M129; elevators M83, M84, and M90) was investigated for changes in burst length and temporal coordination in response to roll stimuli. Most flight muscles fired only once per wing beat cycle in our preparations. Thus, burst length was not found to be correlated with roll angle. Time intervals of firing between all muscle pairs investigated change in correlation with the torque and roll angle (Fig. 9).All mesothoracic muscles are active earlier-relative to the ipsilateral metathoracic subalar muscle M129-during roll to the ipsilateral side than during roll to the contralateral side. Correlations Between Motor and Movement Pattern. The phase of muscle firing within the wing beat cycle varies with roll angle (roll torque). The first basalar M97 and second tergosternal M84 muscles, when referenced e.g. to the upper (M97) or lower (M84) reversal point of the wing tip trajectory, are active earlier on the side the animal turns to (Fig. 10). The onset of the first basalar M97 relative to the beginning of downstroke is correlated with maximum pronation and roll angle (Fig. 11). Mechanisms of Lift Control. Wing pronation, which is very important for lift production is controlled by the central nervous system by altering the phase of muscle activity within the wing beat cycle.     

Columnar cells necessary for motion responses of wide-field visual interneurons in <Emphasis Type="Italic">Drosophila</Emphasis>

Wide-field motion-sensitive neurons in the lobula plate (lobula plate tangential cells, LPTCs) of the fly have been studied for decades. However, it has never been conclusively shown which cells constitute their major presynaptic elements. LPTCs are supposed to be rendered directionally selective by integrating excitatory as well as inhibitory input from many local motion detectors. Based on their stratification in the different layers of the lobula plate, the columnar cells T4 and T5 are likely candidates to provide some of this input. To study their role in motion detection, we performed whole-cell recordings from LPTCs in Drosophila with T4 and T5 cells blocked using two different genetically encoded tools. In these flies, motion responses were abolished, while flicker responses largely remained. We thus demonstrate that T4 and T5 cells indeed represent those columnar cells that provide directionally selective motion information to LPTCs. Contrary to previous assumptions, flicker responses seem to be largely mediated by a third, independent pathway. This work thus represents a further step towards elucidating the complete motion detection circuitry of the fly.