Wasp manipulates cockroach behavior by injecting venom cocktail into prey central nervous system.

The parasitoid wasp Ampulex compressa induces behavioral changes in the cockroach prey by injecting venom into its central nervous system. In contrast to most other venomous predators, the wasp's sting does not induce paralysis. Rather, the two consecutive stings in the thoracic and head ganglia induce three stereotypic behavioral effects. The prey behavior is manipulated in a way beneficial to the wasp and its offspring by providing a living meal for its newborn larva. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis prevents the cockroach from fighting with its front legs, thereby facilitating the second sting in the head. A postsynaptic block of central synaptic transmission mediates this leg paralysis. Following the head sting, dopamine identified in the venom induces 30 minutes of intense grooming that appears to prevent the cockroach from straying until the last and third behavioral effect of hypokinesia commences. In this lethargic state that lasts about three weeks, the cockroach does not respond to various stimuli nor does it initiates movement. However, other specific behaviors of the prey are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to specific thoracic neurons.

Wasp uses venom cocktail to manipulate the behavior of its cockroach prey.

The sting of the parasitoid wasp Ampulex compressa is unusual, as it induces a transient paralysis of the front legs followed by grooming behavior and then by a long-term hypokinesia of its cockroach prey. Because the wasp's goal is to provide a living meal for its newborn larva, the behavioral changes in the prey are brought about by manipulating the host behavior in a way beneficial to the wasp and its offspring. To this end, the wasp injects its venom cocktail with two consecutive stings directly into the host's central nervous system. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis is due to the presence of a venom component that induces a postsynaptic block of central cholinergic synaptic transmission. Following the head sting, dopamine identified in the venom appears to induce 30 min of intense grooming. During the long-term hypokinesia that follows the grooming, specific behaviors of the prey are inhibited while others are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to the thoracic neurons.

Wasp venom blocks central cholinergic synapses to induce transient paralysis in cockroach prey.

The parasitoid wasp Ampulex compressa induces a set of unique behavioral effects upon stinging its prey, the cockroach. It stings into the first thoracic segment inducing 2 to 3 min of transient flaccid paralysis of the front legs. This facilitates a second sting in the cockroach's head that induces 30 min of excessive grooming followed by a 2 to 5-week long lethargic state. In the present study, we examine the immediate effect of the first sting, which is a transient paralysis of the front legs. Using radiolabeled wasps, we demonstrate that the wasp injects its venom directly into the cockroach's first thoracic ganglion. The artificial injection of milked venom into a thoracic ganglion abolishes spontaneous and evoked responses of the motoneurons associated with leg movements. To investigate the physiological mechanism of action of the venom, we injected venom into the last abdominal ganglion of the cockroach, which houses a well-characterized cholinergic synapse. Injected venom abolishes both sensory-evoked and agonist-evoked postsynaptic potentials recorded in the postsynaptic neuron for 2 to 3 min without affecting action potential propagation. Thus, the venom blocking effect has a postsynaptic component that follows the same time course as the transient paralysis induced by the thoracic sting. Finally, injection of a nicotinic antagonist in the front thoracic ganglion induces paralysis of the front legs. We conclude that the transient paralytic effect of the thoracic sting can be mainly accounted for by the presence of a venom active component that induces a postsynaptic block of central cholinergic synaptic transmission.

Are monoaminergic systems involved in the lethargy induced by a parasitoid wasp in the cockroach prey?

The venom of the parasitoid wasp Ampulex compressa induces long-lasting hypokinesia in the cockroach prey. Previous work indicates that the venom acts in the subesophageal ganglion to indirectly affect modulation of thoracic circuits for locomotion. However, the target of the venom in the subesophageal ganglion, and the mechanism by which the venom achieves its effects are as yet unknown. While the stung cockroaches appear generally lethargic, not all behaviors were affected, indicating that the venom targets specific motor systems and not behavior in general. Stung cockroaches were observed "freezing" in abnormal positions. Reserpine, which depletes monoamines, mimics the behavioral effects of the venom. We treated cockroaches with antagonists to dopamine and octopamine receptors, and found that the dopamine system is required for normal escape response. Dopamine injection induces prolonged grooming in normal cockroaches, but not in stung, suggesting that the venom is affecting dopamine receptors, or targets downstream of these receptors, in the subesophageal ganglion. This dopamine blocking effect fades slowly over the course of several weeks, similar to the time course of recovery from hypokinesia. The similarity in the time courses suggests that the mechanism underlying the hypokinesia may be the block of the dopamine receptors.

Synaptic reorganization induced by selective photoablation of an identified neuron.

The maintenance of synaptic strength and specificity in the CNS may depend on interactions among postsynaptic dendrites. We examined the effect of removing a neuron on synaptic organization. A single identified postsynaptic neuron in the adult cercal system of the cockroach was removed with photoablation. After a 30 d recovery period, the synaptic connectivity and morphology of the intact presynaptic and postsynaptic neurons were analyzed. The synaptic connectivity was reorganized in a manner that was consistent with functional plasticity. To associate anatomical changes with this reorganization, we analyzed the morphology of the presynaptic and postsynaptic neurons by quantitative morphometry. Both presynaptic and intact postsynaptic neurons maintained a stable morphology after removal of a neighboring postsynaptic neuron. Using the Hausdorff Match method (HM) (Mizrahi et al. 2000), we found that the spatial organization of the intact dendritic and axonal trees after ablation of a postsynaptic neuron remained stable. Thus, interactions with neighboring neurons were not necessary for maintaining dendritic morphology in the adult nervous system. However, adult central synapses were capable of adjusting to maintain normal function.

Presynaptic inhibition and antidromic spikes in primary afferents of the crayfish: a computational and experimental analysis.

Primary afferent depolarizations (PADs) are associated with presynaptic inhibition and antidromic discharges in both vertebrates and invertebrates. In the present study, we have elaborated a realistic compartment model of a primary afferent from the coxobasipodite chordotonal organ of the crayfish based on anatomical and electrophysiological data. The model was used to test the validity of shunting and sodium channel inactivation hypotheses to account for presynaptic inhibition. Previous studies had demonstrated that GABA activates chloride channels located on the main branch close to the first branching point. We therefore focused the analysis on the effect of GABA synapses on the propagation of action potentials in the first axonal branch. Given the large diameters of the sensory axons in the region in which PADs were likely to be produced and recorded, the model indicates that a relatively large increase in chloride conductance (up to 300 nS) is needed to significantly reduce the amplitude of sensory spikes. The role of the spatial organization of GABA synapses in the sensory arborization was analyzed, demonstrating that the most effective location for GABA synapses is in the area of transition from active to passive conduction. This transition is likely to occur on the main branch a few hundred micrometers distal to the first branching point. As a result of this spatial organization, antidromic spikes generated by large-amplitude PADs are prevented from propagating distally.

Comparative analysis of dendritic architecture of identified neurons using the Hausdorff distance metric.

Dendritic trees often are complex, three-dimensional structures. Comparative morphologic studies have not yet provided a reliable measure to analyze and compare the geometry of different dendritic trees. Therefore, it is important to develop quantitative methods for analyzing the three-dimensional geometry of these complex trees. The authors developed a comparison measure based on the Hausdorff distance for comparing quantitatively the three-dimensional structure of different neurons. This algorithm was implemented and incorporated into a new software package that the authors developed called NeuroComp. The authors tested this algorithm to study the variability in the three-dimensional structure of identified central neurons as well as measuring the structural differences between homologue neurons. They took advantage of the uniform dendritic morphology of identified interneurons of an insect, the giant interneurons of the cockroach. More specifically, after establishing a morphometric data base of these neurons, the authors found that the algorithm is a reliable tool for distinguishing between dendritic trees of different neurons, whereas conventional metric analysis often is inadequate. The authors propose to use this method as a quantitative tool for the investigation of the effects of various experimental paradigms on three-dimensional dendritic architecture.

Venom of a parasitoid wasp induces prolonged grooming in the cockroach

The parasitoid wasp Ampulex compressa hunts cockroaches Periplaneta americana, stinging them first in the thorax and then in the head, the sting penetrating towards the subesophageal ganglion. After being stung the cockroach grooms almost continuously for approximately 30 min, performing all the normal components of grooming behavior. This excessive grooming is only seen after the head sting and cannot be attributed to stress, to contamination of the body surface or to systemic or peripheral effects. This suggests that the venom is activating a neural network for grooming. We suggest that the venom induces prolonged grooming by stimulating dopamine receptors in the cockroach, for the following reasons. (1) Reserpine, which causes massive release of monoamines, induces excessive grooming. (2) Dopamine injected into the hemocoel also induces excessive grooming and is significantly more effective than octopamine or serotonin. In addition, the dopamine agonist SKF 82958 induces excessive grooming when injected directly into the subesophageal ganglion. (3) Injection of the dopamine antagonist flupenthixol greatly reduces venom-induced grooming. (4) Dopamine, or a dopamine-like substance, is present in the venom.

Independent coding of wind direction in cockroach giant interneurons.

Independent coding of wind direction in cockroach giant interneurons. J. Neurophysiol. 78: 2655-2661, 1997. In this study we examined the possible role of cell-to-cell interactions in the localization processing of a wind stimulus by the cockroach cercal system. Such sensory processing is performed primarily by pairs of giant interneurons (GIs), a group of highly directional cells. We have studied possible interactions among these GIs by comparing the wind sensitivity of a given GI before and after removing another GI with the use of photoablation. Testing various combinations of GI pairs did not reveal any suprathreshold interactions. This was true for all unilateral GI pairs on the left or right side as well as all the bilateral GI pairs (left and right homologues). Those experiments in which we were able to measure synaptic activity did not reveal subthreshold interactions between the GIs either. We conclude that the GIs code independently for a given wind direction without local GI-GI interactions. We discuss the possible implications of the absence of local interactions on information transfer in the first station of the escape circuit.

In situ visualization and photoablation of individual neurons using a low cost fiber optic based system.

This paper describes a low-cost system for in situ visualization and photoablation of single neurons. The system includes a fiber optics light source equipped with a filter port, a blue excitation filter and a yellow barrier filter, and a light guide which terminates with a focusing lens. This system is inexpensive, easy to use, and requires minimal maintenance. Given its price, this system is readily accessible and has the potential of becoming standard equipment for in situ visualization and killing of individual neurons.

Wind-evoked evasive responses in flying cockroaches.

1) A standing cockroach (Periplaneta americana) responds to the air displacement made by an approaching predator, by turning away and running. The wind receptors on the cerci, two posterior sensory appendages, excite a group of ventral giant interneurons that mediate this response. While flying, these interneurons remain silent, owing to strong inhibition; however, the dorsal giant interneurons respond strongly to wind. Using behavioral and electromyographic analysis, we sought to determine whether flying cockroaches also turn away from air displacement like that produced by an approaching flying predator; and if so, whether the cerci and dorsal giant interneurons mediate this response. 2) When presented with a wind puff from the side, a flying cockroach carries out a variety of maneuvers that would cause a rapid turn away and perhaps a dive. These are not evoked if the cerci are ablated (Figs. 4, 5,6). 3) This evasive response appears to be mediated by a circuit separate from that mediating escape when the cockroach is standing (Fig. 7). 4) The dorsal giant interneurons respond during flight in a directional manner that is suited to mediate this behavior (Fig. 8). 5) Recordings of the wind produced by a moving model predator (Fig. 9), together with measurements of the behavioral latency of tethered cockroaches, suggest that the evasive response would begin just milliseconds before a predator actually arrives. However, as explained in the Discussion section, under natural conditions, the evasive response may well begin earlier, and could indeed be useful in escaping from predators. 6) If cockroaches had a wind-mediated yaw-correcting behavior, as locusts have, this could conflict with the wind-evoked escape. In fact, cockroaches show the opposite, yaw-enhancing response, mediated by the cerci, that does not present a conflict with escape (Figs. 10-14).

Frequency as a releaser in the courtship song of two crickets, Gryllus bimaculatus (de Geer) and Teleogryllus oceanicus: a neuroethological analysis.

1. The courtship behavior of male field crickets, Gryllus bimaculatus (De Geer) and Teleogryllus oceanicus, is a complex, multimodal behavioral act that involves acoustic signals (a courtship song; Fig. 1A, B). The dominant frequency is 4.5 kHz for T. oceanicus song (Fig. 1A) and 13.5 kHz for G. bimaculatus (Fig. 1B). 2. When courting males are deprived of their courtship song by wing amputation, their courtship success declines markedly but is restored when courting is accompanied by tape-recordings of their courtship songs or a synthetic courtship song with only the dominant frequency of the natural song; other naturally occurring frequency components are ineffective for restoring mating success (Figs. 4, 5). 3. It has been suggested that an identified auditory interneuron, AN2, plays a critical role in courtship success. Chronic recordings of AN2 in an intact, tethered female show that AN2's response to the natural courtship song and synthesized songs at 4.5 and 13.5 kHz is similar in T. oceanicus. By contrast, in G. bimaculatus, AN2's response to the natural courtship song and synthesized song at 13.5 kHz, but not at 4.5 kHz, is similar (Fig. 2,3). 4. In behavioral experiments, playback of a 30 kHz synthetic courtship song in G. bimaculatus does not restore courtship success, yet this same stimulus elicits as strong a response from AN2 as does the normal courtship song (Fig. 6). Thus, contrary to earlier work by others, we conclude AN2 is not, by itself, a critical neural link in the courtship behavior of these two species of crickets.

Ultrasonic startle behavior in bushcrickets (Orthoptera; Tettigoniidae).

1. In the present work, we show that in flight, bushcrickets not previously known to respond to ultrasound alter their flight course in response to ultrasonic stimuli. Such stimuli elicit in flying Neoconocephalus ensiger an extension of the front and middle legs along the body and a rapid closure of all 4 wings (Fig. 1). This is a short latency acoustic startle response to ultrasound, consistent with acoustic startle responses of other insects. 2. The percentage of trials on which acoustic startle responses were elicited was maximum (90%) for sound frequencies ranging from 25 to at least 60 kHz. No acoustic startle response was observed at frequencies of 5 or 10 kHz (Fig. 2). The threshold for the response was roughly 76 dB between 25 to 60 kHz (Fig. 2) and the behavioral latency was 45 ms (Fig. 3). Recordings from flight muscles show that they cease discharging during the acoustic startle response (Fig. 4). 3. The characteristics of the acoustic startle response match those of an auditory interneuron called the T-neuron. The frequency sensitivity of this neuron is greatest for sound frequencies ranging from 13 to 60 kHz (Fig. 6). Moreover, we found that the neuron produces many more spikes to ultrasound (30 kHz) of increasing intensities than to a conspecific communication sound, whose dominant frequency is 14 kHz (Fig. 7).

Multiple feedback loops in the flying cockroach: excitation of the dorsal and inhibition of the ventral giant interneurons.

1. In a tethered cockroach (Periplaneta americana) whose wings have been cut back to stumps, it is possible to elicit brief sequences of flight-like activity by puffing wind on the animal's body. 2. During such brief sequences, rhythmic bursts of action potentials coming from the thorax at the wingbeat frequency, descend the abdominal nerve cord to the last abdominal ganglion (A6). This descending rhythm is often accompanied by an ascending rhythm (Fig. 2). 3. Intracellular recording during flight-like activity from identified ascending giant interneurons, and from some unidentified descending axons in the abdominal nerve cord, shows that: (a) ventral giant interneurons (vGIs) remain silent (Fig. 3); (b) dorsal giant interneurons (dGIs) are activated at the onset of the flight-like activity and remain active sporadically throughout the flight sequence (Fig.4); (c) some descending axons in the abdominal nerve cord show rhythmic activity phase-locked to the flight rhythm (Fig. 5). 4. Also during such brief sequences, the cercal nerves, running from the cerci (paired, posterior, wind sensitive appendages) to the last abdominal ganglion, show rhythmic activity at the wingbeat frequency (Fig. 6). This includes activity of some motor axons controlling vibratory cercal movements and of some sensory axons. 5. More prolonged flight sequences were elicited in cockroaches whose wings were not cut and which flew in front of a wind tunnel (Fig. 1B). 6. In these more prolonged flight sequences, the number of ascending spikes per burst was greater than that seen in the wingless preparation (Fig. 8; compare to Fig. 2). Recordings from both ventral and dorsal GIs show that: in spite of the ongoing wind from both the tunnel and the beating wings, which is far above threshold for the vGIs in a resting cockroach, the vGIs are entirely silent during flight. Moreover, the vGIs response to strong wind puffs that normally evoke maximal GI responses is reduced by a mean of 86% during flight (Fig. 9). The dGIs are active in a strong rhythm (Figs. 11 and 12). 7. Three sources appear to contribute to the ascending dGI rhythm (1) the axons carrying the rhythmic descending bursts; (2) the rhythmic sensory activity resulting from the cercal vibration; and (3) the sensory activity resulting from rhythmic wind gusts produced by the wingbeat and detected by the cerci. The contribution of each source has been tested alone while removing the other two (Figs. 13 and 14). Such experiments suggest that all 3 feedback loops are involved in rhythmically exciting the dGIs (Fig. 15).

Nonsynaptic regulation of sensory activity during movement in cockroaches.

Here we describe a nonsynaptic mechanism for filtering out potentially perturbing sensory feedback during locomotion. During flight, the cockroach moves its cerci, two abdominal sensory appendages, about their joint with the body and holds them in place. The cerci bear highly sensitive wind-receptive hairs, which would be strongly stimulated by flight wind. Such wind could cause habituation of the synaptic connections from these cercal receptors onto interneurons responsible for the running escape response to an approaching predator. We have found that the cercal displacement blocks one-third to one-half of the action potentials along the sensory nerve, possibly aiding in protection against such habituation. This block occurs if one experimentally displaces a cercus, and the block persists in the complete absence of any connections with the central nervous system. The block appears to be nonsynaptic and to result instead from mechanical pressure on the nerve near the joint. The results suggest that activity in peripheral nerves in other animals may also be affected by the position or movement of joints through which the nerves pass.

Single-unit responses and reflex effects of force-sensitive mechanoreceptors of the dactyl of the crab.

This paper examines the responses and reflex effects of force-sensitive mechanoreceptors of the most distal leg segment, the dactyl, of the leg of the crab, Carcinus maenas. The goals of these studies are to establish the potential activities and functions of these receptors in posture and locomotion. The responses of force-sensitive mechanoreceptors to imposed mechanical stimuli depended upon their location on the dactyl. A distal group of receptors is located on a specialized region, the dactyl tip, which is composed solely of epicuticle. Another group of receptors is distributed throughout more proximal regions of the dactyl where the cuticle is completely calcified. Both groups of receptors showed vigorous responses to imposed bending forces. When bending forces were applied as step functions at the dactyl, tip distal receptors showed only phasic responses to all levels of force application. Receptors located at more proximal positions on the dactyl showed phasic responses to low levels of step applied forces and phasicotonic discharges at higher levels of force. Increasing levels of force produced a sigmoid increase in the tonic firing of these units. When bending forces were applied using ramp functions, receptors of the distal group responded with an intense initial discharge followed by firing at a constant rate throughout both force application and release. This response was not related to the velocity of force application. In contrast, receptors located more proximally responded directionally to force application and release. Proximal receptors also effectively encoded the velocity of force application. Responses of these two groups of receptors also differed when vibrations were applied at the dactyl tip: proximal receptors only followed vibrational stimuli up to 50 Hz, whereas distal receptors showed 1:1 responses at vibrations as high as 95 Hz. Mechanoreceptors of the dactyl also responded intensely to bending forces resulting from resisted contractions of the animal's own muscles. No responses were obtained from unresisted movements of the leg. Stimulation of force-sensitive mechanoreceptors of the dactyl produced intra- and interleg reflex discharges in motor neurons to leg muscles. Mechanical bending of the dactyl or electrical stimulation of dactyl nerves in which force-sensitive mechanoreceptors were recorded produced strong tonic excitation of motors neurons to the levator muscles of the same leg.(ABSTRACT TRUNCATED AT 400 WORDS)

Force-sensitive mechanoreceptors of the dactyl of the crab: single-unit responses during walking and evaluation of function.

The activities of individual force-sensitive mechanoreceptors of the dactyl (terminal leg segment) of the crab, Carcinus maenas, have been recorded during free walking. These receptors have also been mechanically and electrically stimulated in freely moving animals to directly evaluate their function in locomotion. All force-sensitive mechanoreceptors fired during the stance phase of walking and were silent during swing. Receptor discharges showed regular phase relationships to bursts in motor neurons of leg muscles. Crabs walk laterally and use the legs of one side either in trailing to actively push the animal to the opposite side, or in leading, to less forcefully pull the animal in that direction. Individual force-sensitive mechanoreceptors differed in their patterns of activity during trailing or leading according to their location on the dactyl. Units of proximal receptors fired more vigorously when used in trailing than in leading. Discharges in trailing were also increased by loading of the animal. In contrast, distal receptors near the dactyl tip fired equally intensely during walking in either direction. Proximal receptors thus encode forces and loads applied to the leg. Distal receptors do not encode loads but can signal leg contact and, potentially, exteroceptive vibrations. Sensory stimulation of force-sensitive mechanoreceptors was produced during walking by a device that imposed continuous mechanical bending of the dactyl and by electrical stimulation of dactyl nerves. Intra- and inter-segmental reflexes were evaluated by myographic recordings from leg muscles. Continuous mechanical deformation of the dactyl increased the activity of the levator and decreased firing in the depressor muscles of the homonymous leg during walking. The same stimulus produced enhanced activity in depressor muscles of adjacent legs. The latter effect was not due to simple mechanical coupling resulting from reflexes in the stimulated leg. These reflexes can function to limit forces applied to a leg and provide compensatory adjustments in other legs. Brief low-threshold electrical stimuli applied to nerves in which the activities of force-sensitive mechanoreceptors were recorded produced reflex effects similar to those obtained by mechanical stimulation. These stimuli also reset the rhythm of motor neuron bursting in both homonymous and adjacent legs during walking. These studies confirm the importance of force-sensitive mechanoreceptors in adapting walking patterns and in determining leg coordination in locomotion.

Dual locomotor activity selectively controlled by force- and contact-sensitive mechanoreceptors.

The crab Carcinus maenas walks laterally; moreover, as soon as leg contact with the support is lost, it starts swimming. In free-moving animals, discharges from individual force- and contact-mechanoreceptors located in the terminal segment of the last pair of walking legs have been recorded. These receptors are active during the stance phase of walking and remain silent during swimming. Selective electrical stimulation of their afferent fibers during swimming inhibits this behaviour. The possible role of such sensory information in selecting different motor patterns is discussed.