Sensorimotor pathways involved in interjoint reflex action of an insect leg

Coordination of motor output between leg joints is crucial for the generation of posture and active movements in multijointed appendages of legged organisms. We investigated in the stick insect the information flow between the middle leg femoral chordotonal organ (fCO), which measures position and movement in the femur-tibia (FT) joint and the motoneuron pools supplying the next proximal leg joint, the coxa-trochanteral (CT) joint. In the inactive animal, elongation of the fCO (by flexing the FT joint) induced a depolarization in eight of nine levator trochanteris motoneurons, with a suprathreshold activation of one to three motoneurons. Motoneurons of the depressor trochanteris muscle were inhibited by fCO elongation. Relaxation signals, i.e., extension of the FT joint, activated both levator and depressor motoneurons; i.e., both antagonistic muscles were coactivated. Monosynaptic as well as polysynaptic pathways contribute to interjoint reflex actions in the stick insect leg. fCO afferents were found to induce short latency EPSPs in levator motoneurons, providing evidence for direct connections between fCO afferents and levator motoneurons. In addition, neuronal pathways via intercalated interneurons were identified that transmit sensory information from the fCO onto levator and/or depressor motoneurons. Finally, we describe two kinds of alterations in interjoint reflex action: (a) With repetitive sensory stimulation, this interjoint reflex action shows a habituation-like decrease in strength. (b) In the actively moving animal, interjoint reflex action in response to fCO elongation, mimicking joint flexion, qualitatively remained the same sign, but with a marked increase in strength, indicating an increased influence of sensory signals from the FT joint onto the adjacent CT joint in the active animal. © 1997 John Wiley & Sons, Inc. J Neurobiol 33: 891–913, 1997     

Vibration signals from the FT joint can induce phase transitions in both directions in motoneuron pools of the stick insect walking system

The influence of vibratory signals from the femoral chordotonal organ fCO on the activities of muscles and motoneurons in the three main leg joints of the stick insect leg, i.e., the thoraco-coxal (TC) joint, the coxa-trochanteral (CT) joint, and the femur-tibia (FT) joint, was investigated when the animal was in the active behavioral state. Vibration stimuli induced a switch in motor activity (phase transition), for example, in the FT joint motor activity switched from flexor tibiae to extensor tibiae or vice versa. Similarly, fCO vibration induced phase transitions in both directions between the motoneuron pools of the TC joint and the CT joint. There was no correlation between the directions of phase transition in different joints. Vibration stimuli presented during simultaneous fCO elongation terminated the reflex reversal motor pattern in the FT joint prematurely by activating extensor and inactivating flexor tibiae motoneurons. In legs with freely moving tibia, fCO vibration promoted phase transitions in tibial movement. Furthermore, ground vibration promoted stance-swing transitions as long as the leg was not close to its anterior extreme position during stepping. Our results provide evidence that, in the active behavioral state of the stick insect, vibration signals can access the rhythm generating or bistable networks of the three main leg joints and can promote phase transitions in motor activity in both directions. The results substantiate earlier findings on the modular structure of the single-leg walking pattern generator and indicate a new mechanism of how sensory influence can contribute to the synchronization of phase transitions in adjacent leg joints independent of the walking direction.     

Intersegmental transfer of sensory signals in the stick insect leg muscle control system

Intersegmental coordination during locomotion in legged animals arises from mechanical couplings and the exchange of neuronal information between legs. Here, the information flow from a single leg sense organ of the stick insect Cuniculina impigra onto motoneurons and interneurons of other legs was investigated. The femoral chordotonal organ (fCO) of the right middle leg, which measures posture and movement of the femur-tibia joint, was stimulated, and the responses of the tibial motoneuron pools of the other legs were recorded. In resting animals, fCO signals did not affect motoneuronal activity in neighboring legs. When the locomotor system was activated and antagonistic motoneurons were bursting in alternation, fCO stimuli facilitated transitions from flexor to extensor activity and vice versa in the contralateral leg. Following pharmacological treatment with picrotoxin, a blocker of GABA-ergic inhibition, the tibial motoneurons of all legs showed specific responses to signals from the middle leg fCO. For the contralateral middle leg we show that fCO signals encoding velocity and position of the tibia were processed by those identified local premotor nonspiking interneurons known to contribute to posture and movement control during standing and voluntary leg movements. Interneurons received both excitatory and inhibitory inputs, so that the response of some interneurons supported the motoneuronal output, while others opposed it. Our results demonstrate that sensory information from the fCO specifically affects the motoneuronal activity of other legs and that the layer of premotor nonspiking interneurons is a site of interaction between local proprioceptive sensory signals and proprioceptive signals from other legs.     

Octopaminergic modulation of the femoral chordotonal organ in the stick insect

The modulatory action of DL-octopamine on the multicellular femoral chordotonal organ (fCO) of the stick insect Cuniculina impigra was examined using extracellular recordings from the fCO nerve and intracellular recordings from single sensory neurons. To determine the octopaminergic effect on position, velocity and/or acceleration sensitivity of mechanoreceptors direct mechanical stimulations with defined parameters were applied to the fCO apodeme. The spontaneous activity in the fCO nerve was enhanced in a dose-dependent manner by octopamine (threshold at 5 × 10^?7 M). This was based on enhanced activity of position sensitive neurons as the fCO activity for all position stimuli was shifted to higher values. Intracellular recordings of single sensory cells showed that velocity-sensitivity of single sensory cells was not altered by octopamine. Similarly, the response of fCO afferents to ramp-and-hold stimuli revealed that acceleration sensitivity was unaffected by octopamine. The observed alterations in the fCO activity indicate that responses to static stimuli are enhanced while responses to motion stimuli are not affected by octopamine. These findings suggest that the octopaminergic modulation of the fCO may affect the animals' posture and those leg movements that rely on position information.     

Identified nonspiking interneurons in leg reflexes and during walking in the stick insect

In the stick insect Carausius morosus identified nonspiking interneurons (type E4) were investigated in the mesothoracic ganglion during intraand intersegmental reflexes and during searching and walking.In the standing and in the actively moving animal interneurons of type E4 drive the excitatory extensor tibiae motoneurons, up to four excitatory protractor coxae motoneurons, and the common inhibitor 1 motoneuron (Figs. 1–4).In the standing animal a depolarization of this type of interneuron is induced by tactile stimuli to the tarsi of the ipsilateral front, middle and hind legs (Fig. 5). This response precedes and accompanies the observed activation of the affected middle leg motoneurons. The same is true when compensatory leg placement reflexes are elicited by tactile stimuli given to the tarsi of the legs (Fig. 6).During forward walking the membrane potential of interneurons of type E4 is strongly modulated in the step-cycle (Figs.8–10). The peak depolarization occurs at the transition from stance to swing. The oscillations in membrane potential are correlated with the activity profile of the extensor motoneurons and the common inhibitor 1 (Fig. 9).The described properties of interneuron type E4 in the actively behaving animal show that these interneurons are involved in the organization and coordination of the motor output of the proximal leg joints during reflex movements and during walking.Abbreviations CLP reflex, compensatory leg placement reflex - CI1 common inhibitor I motoneuron - fCO femoral chordotonal organ - FETi fast extensor tibiae motoneuron - FT femur-tibia - SETi slow extensor tibiae motoneuron     

Role of presynaptic inputs to proprioceptive afferents in tuning sensorimotor pathways of an insect joint control network

The femur-tibia (FT) joint of insects is governed by a neuronal network that controls activity in tibial motoneurons by processing sensory information about tibial position and movement provided by afferents of the femoral chordotonal organ (fCO). We show that central arborizations of fCO afferents receive presynaptic depolarizing synaptic inputs. With an average resting potential of −71.9 ± 3.72 mV (n = 10), the reversal potential of these potentials is on average −62.8 ± 2.3 mV (n = 5). These synaptic potentials occur either spontaneously or are related to movements at the fCO. They are thus induced by signals from other fCO afferents. Therefore, the synaptic inputs to fCO afferents are specific and depend on the sensitivity of the individual afferent affected. These potentials reduce the amplitude of concurrent afferent action potentials. Bath application of picrotoxin, a noncompetitive blocker of chloride ion channels, blocks these potentials, which indicates that they are mediated by chloride ions. From these results, it is concluded that these are inhibitory synaptic potentials generated in the central terminals of fCO afferents. Pharmacologic removal of these potentials affects the tuning of the complete FT control system. Following removal, the dependence of the FT control loop on the tibia position increases relative to the dependency on the velocity of tibia movements. This is due to changes in the relative weighting of the position and velocity signals in the parallel interneuronal pathways from the fCO onto tibial motoneurons. Consequently, the FT joint is no longer able to perform twig mimesis (i.e., catalepsy), which is known to rely on a low position compared to the high-velocity dependency of the FT control system. © 1997 John Wiley & Sons, Inc. J Neurobiol 32: 359–376, 1997.     

Physiological changes in central neuronal pathways contributing to the generation of a reflex reversal

1. In the stick insect Carausius morosus the properties of the neuronal network governing the femur-tibia joint depend on the behavioral state of the animal. In the inactive animal flexion of the femur-tibia joint results in the generation of a resistance reflex, while in the active animal the same stimulus induces the so-called active reaction, the first part of which is a reflex reversal.
2. Recordings from motoneurons innervating the extensor tibiae muscle indicated that their time course of activity during the active reaction is due to inputs from intercalated pathways. We therefore investigated the role of identified nonspiking interneurons that transmit sensory information from the chordotonal organ onto the extensor motoneurons in the inactive animal. We can show that (i) the nonspiking interneurons received altered inputs whereas (ii) they provided qualitatively the same synaptic drive onto leg motoneurons.
3. From our results it is clear that (i) neuronal pathways contributing to the generation of the resistance reflex are also involved in the generation of the reflex reversal in the same control loop, (ii) thereby adopting the same principle of information processing (parliamentary principle), because both, supporting and opposing pathways contribute to the generation of the motor output.

     

Gain control in a proprioceptive feedback loop as a prerequisite for working close to instability

In the artificially closed femur-tibia control system of stick insects oscillations were induced in 3 different ways: Increasing the phase-shift by introducing an electronic delay, afference sign reversal and coupling the tibia to an inert mass. In all 3 cases the oscillations stopped after some time. The gain of the open-loop system was significantly smaller after the oscillations. Afference sign reversal by surgically crossing of the receptor apodeme of the femoral chordotonal organ for 25–85 days does not lead to altered characteristics of the control loop. When sinusoidal passive movements are forced upon the intact femur-tibia joint the forces resisting these movements do not decrease with time. In contrast to direct stimulation of the femoral chordotonal organ, these passive movements also influence the contralateral leg. The experiments show that the gain-control system of the femur-tibia control loop of stick insects consists of at least two components: A sensitization system (with inputs from many kinds of stimuli indicating some kind of disturbance) increases the gain of all reflex loops. A specific habituation-like system decreases the gain with repetitive stimulation only of one control system.Abbreviations fCO femoral chordotonal organ - SETi slow extensor tibiae motor neuron     

Variability of femoral chordotonal organ reflexes in the locust, Locusts miaratoria

Abstract. The femoral chordotonal organ of a locust front, middle or hind leg was stimulated mechanically under open-loop conditions and the forces produced by the muscles moving the tibia were measured. In nearly all cases resistance reflexes were elicited in the inactive animal. However, in rare, but reproducible cases, a positive feedback occurred. In an animal performing active movements the responses were very labile: often no response occurred, sometimes a reaction comparable to the active reaction in stick insects was found, and sometimes resistance reflexes were present.     

Mass-induced oscillations in the femur-tibia control system of stick insects

Attaching an inert mass to a freely moving tibia of an otherwise fixed stick insect Carausius morosus, induces undamped oscillations of the tibia. We describe the use of a rotational pendulum to observe these oscillations applying various amounts of inertia. The dependence of the frequency of these oscillations on the moment of inertia is similar to that of a purely mechanical system. The sequence of the oscillatory behavior can be separated into 3 distinct behavioural states. The transitions between some of these states could be elicited by external stimuli and partly showed characteristics of habituation and dishabituation. With a rotational pendulum on each middle leg, simultaneous oscillations of both legs were measured to investigate coupling effects between the neural control systems of the two legs. In some cases, significant coupling effects could be observed in phase and frequency. In many other cases, no coupling was found. The habituation and dishabituation effects were not transferred between the middle legs.     

Sensorimotor pathways processing vibratory signals from the femoral chordotonal organ of the stick insect

The femoral chordotonal organ of stick insects senses position and velocity of movements in the femur-tibia joint, as well as tibial vibration. While sensory information about large-scale tibial movements is processed by a well-known neuronal network and elicits resistance reflexes in extensor and flexor tibiae motoneurons, it is not yet known how sensory information about vibration of the tibia is processed. We investigated the transmission of vibration stimuli to tibial extensor motoneurons and their premotor interneurons. Vibration stimuli applied to the femoral chordotonal organ evoked responses in tibial extensor and flexor muscles. During ongoing vibration this response adapted rapidly. This adaptation had no effect on the motoneuronal response to large-scale tibial movements. Recording from premotor interneurons revealed that vibratory signals were processed in part by the same interneuronal pathways as (large-scale) velocity and position information. While only certain parts of the interneuronal reflex pathways showed little or no response during vibration stimuli, most neurons responded to both position or velocity stimuli and vibration at the femoral chordotonal organ. We conclude that sensory information about vibration of the tibia shares part of the interneuronal pathways that transmit sensory information about large-scale tibial movements to the motoneurons. Accepted: 25 April 1999     

Influence of aminergic and peptidergic substances on heart beat frequency in the stick insect Carausius morosus (Insecta,Phasmatodea)

The dorsal heart of the Indian stick insect, Carausius morosus, is responsible for the anterograde flow of hemolymph to the aorta and into the body cavity. The contraction frequency of the insect heart is known to be influenced by several substances of neural source. Here, a semi‐exposed heart assay was employed to study the effect of an aminergic substance (octopamine) and three neuropeptides (C. morosus hypertrehalosemic hormone [Carmo‐HrTH], crustacean cardioactive peptide [CCAP], and proctolin) on heart contraction. The contraction frequency was measured as beats per minute in adults ligated between the head and the prothorax. All three investigated neuropeptides had a stimulatory effect on heart contraction that lasted approximately 6 min, after which the normal heart beat rate was restored. Proctolin and CCAP stimulated the rate of heart beat also in unligated stick insects, whereas Carmo‐HrTH was active only in ligated insects. The latter could suggest that when the stick insect is not ligated, a competing substance may be released from the head of C. morosus; the competing substance is, apparently, not physiologically active but it binds or blocks access to the receptor of Carmo‐HrTH‐II, thereby rendering the HrTH peptide “not active.” In ligated stick insects, 6.7 × 10^?8 M Carmo‐HrTH‐II significantly increased the heart beat rate; higher doses resulted in no further increase, suggesting the saturation of the HrTH receptor. Octopamine inhibited the rate at which the heart contracted in a dose‐dependent manner; inhibition was achieved with 10^?4 M of octopamine.     

Reflex modulation in locusts walking on a treadwheel intracellular recordings from motoneurons

Summary In locusts (Locusta migratoria) walking on a treadwheel, afferents of tarsal hair sensilla were stimulated via chronically implanted hook electrodes (Fig. 1). Stimuli applied to the middle leg tarsus elicited avoidance reflexes (Fig. 2). In quiescent animals, the leg was lifted off the ground and the femur adducted. In walking locusts, the response was phase-dependent. During the stance phase, no reaction was observed except occasional, premature triggering of swing movements; stimuli applied near the end of the swing phase were able to elicit an additional, short leg protraction.Central nervous correlates of phase-dependent reflex modulation were observed by recording intracellularly from motoneuron somata in walking animals. As a rule, motoneurons recruited during the swing phase showed excitatory stimulus-related responses around the end of the swing movement, correlated to the triggering of additional leg protractions (Figs. 3, 4, 5). Motoneurons active during the stance phase were often inhibited by tarsal stimulation, some showed only weak responses (Figs. 8, 9, 10). Common inhibitory motoneuron 1 was excited by tarsal stimulation during all phases of the leg movement (Figs. 6, 7). In one type of flexor tibiae motoneuron, a complex response pattern was observed, involving the inversion of stimulus-related synaptic potentials from excitatory, recorded during rest, to inhibitory, observed during long-lasting stance phases (Figs. 11, 12).The results demonstrate how reflex modulation is represented on the level of synaptic input to motoneurons. They further suggest independent gain control in parallel, antagonistic pathways converging onto the same motoneuron as a mechanism for reflex reversal during locomotion.Abbreviations CI ₁ common inhibitory motoneuron (1) - EMG electromyogram - Feti fast extensor muscle of the tibia     

Motor pattern selection by combinatorial code of interneuronal pathways

We use a modeling approach to examine ideas derived from physiological network analyses, pertaining to the switch of a motor control network between two opposite control modes. We studied the femur–tibia joint control system of the insect leg, and its switch between resistance reflex in posture control and “active reaction” in walking, both elicited by the same sensory input. The femur–tibia network was modeled by fitting the responses of model neurons to those obtained in animals. The strengths of 16 interneuronal pathways that integrate sensory input were then assigned three different values and varied independently, generating a database of more than 43 million network variants. We demonstrate that the same neural network can produce the two different behaviors, depending on the combinatorial code of interneuronal pathways. That is, a switch between behaviors, such as standing to walking, can be brought about by altering the strengths of selected sensory integration pathways. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Interneurones participating in the “active reaction” in stick insects

The active reaction upon stimulation of the femoral chordotonal organ in stick insects is known to control velocity and endpoint of flexion movements of the femur-tibia joint (Bässler 1988). This article presents evidence that spiking interneurones in the ventral anterior median part of the adjacent ganglion participate in the generation of the active reaction as well as non-spiking interneurones in the dorsal lateral region do. Three different kinds of experiments lead to this conclusion:

Regulation of the body-substrate-distance in the stick insect: Step responses and modelling the control system

The feed back mechanism subserving the regulation of the body-substrate-distance in the stick insect Carausius morosus has been investigated by means of step-like stimuli. Based on the results obtained in open-loop experiments a model is developed which describes the results obtained under closed loop conditions. When the experimental animal is pushed or pulled in dorso-ventral direction an initial fast and a subsequent very slow change of the body height z over the substrate are observed. The late slow response is a nearly linear function of time and can last for more than one hour if the animal is pulled with moderate forces. It withstands less effectively if pushed. During the slow phase of the response sudden changes of z and of the slope of the z(t)-curves occur, presumably due to corresponding changes of the amplification within the feed back loop.     

Information processing in the femur-tibia control loop of stick insects

The complicated response characteristics of the identified nonspiking interneuron type E4 upon elongation stimuli to the femoral chordotonal organ (fCO) can be obtained by a computer simulation using the neuronal network simulator BioSim, if the following assumptions were introduced: (1) The interneurons receive direct excitatory input from position- and velocity-sensitive fCO afferents but also, in parallel delayed inhibition from the same velocity-sensitive afferents. (2) Position-sensitive afferents in part show adaptation with a rather long time-constant. A subsequent experimental analysis demonstrated that all these assumptions fit the reality: (1) Interneurons of type E4 receive direct excitatory input from fCO afferents. (2) Interneurons of type E4 are affected by velocity dependent delayed inhibitory inputs from the fCO. (3) The fCO does contain adapting position-sensitive sensory neurons, which have not been described before. The described principle of the information processing is also able to generate the response in interneurons of type E6 with less steep amplitude-velocity characteristic due to a different weighting of the direct excitation and delayed inhibition.Abbreviations EPSP excitatory postsynaptic potential - FETi fast extensor tibiae motor neuron - fCO femoral chordotonal organ - FT-control loop femur-tibia control loop - IPSP inhibitory postsynaptic potential - SETi slow extensor tibiae motor neuron     

Premotor interneurons in generation of adaptive leg reflexes and voluntary movements in stick insects

We investigated the role of local nonspiking interneurons involved in motor control of legs in the stick insect, Carausius morosus. In a preparation that allowed the animals to perform active leg movements such as adaptive tactile reflexes, proprioceptive reflexes, and walking, we gathered the following results. Almost all tested nonspiking interneurons that provide synaptic drive onto motoneurons of the proximal leg muscles contribute to all of the motor programs underlying tactile reflexes and voluntary leg movements such as walking, searching, and rocking. Most of them are also involved in the generation of proprioceptive reflexes. All motor programs for coactivation, avoidance reflexes, resistance reflexes, and voluntary leg movements result from parallel pathways including nonspiking interneurons that support and others that oppose the motoneuronal activity. The contribution of a single interneuron to the different motor programs is specific: it can be supporting for one motor program but opposing for the other. Even for the same motor program, for example, coactivation, the contribution of an individual interneuron can depend on the stimulus site from where the response is elicited. Our results support the idea that the different motor patterns for adaptive tactile reflexes, resistance reflexes, and voluntary leg movements emerge from a multifunctional neuronal circuit that is reorganized corresponding to the motor behavior performed. The actual motor pattern is then shaped by distributed information processing in parallel supporting and opposing pathways. © 1996 John Wiley & Sons, Inc.     

Modelling of the active reaction of stick insects by a network of neuromimes

Depending on the activity status of the animal, the control system of the femur-tibia joint in stick insects exhibits either a resistance reflex, or the active reaction, a totally different action pattern (Bässler 1988a). Using analog electronic neuron models, several different neuronal circuits are explored that model the active reaction with all its features. The models differ in complexity, redundance and the robustness against small variations of network parameters (e.g. coupling strengths). The circuit with the highest robustness and redundancy requires interneurons with special features, such as found in real animals. When inserted into a closed loop modeling movement and sensory feedback from the periphery, this circuit produces oscillations similar to searching movements found in the real animal. In addition to intracellular recording methods, the authors propose modeling with realistic neuromimes as a complementary method in the investigation of neuronal networks which have well documented input-output relationships.     

Nonspiking pathways antagonize the resistance reflex in the thoraco-coxal joint of stick insects

In the stick insect (Carausius morosus) imposed forward and backward movements of the coxa of the middle leg induce resistance reflexes in the retractor or protractor coxae muscles, depending on the direction of movement. The hairs of the ventral coxal hairplate (cxHPv) function as the primary transducer of the retractor part of the underlying feedback loop: bending of the hairs of the cxHPv during an imposed forward movement of the coxa leads to a reflex activation of the retractor motoneurones, whereas releasing of the hairs causes an inhibition of these motoneurones. Local nonspiking interneurones were investigated, which transmit information from the cxHPv onto the retractor motoneurones: 1) they are depolarized during bending of the hair sensilla of the cxHPv and 2) they decrease the activities of retractor motoneurones. In addition, four of the interneurones drive a protractor motoneurone, when they are depolarized. As bending stimuli at the cxHPv (mimicking an imposed forward movement of the leg) induce reflex activation of the retractor motoneurones and reflex inhibition of the protractor motoneurones, the physiology of the recorded interneurones appears to antagonize the resistance reflex in the thoraco-coxal joint. The results indicate that these nonspiking interneurones take part in the shaping of the reflex response and that furthermore these interneurones are involved in the organization of the motor output to the two antagonistic sets of motoneurones. The possible role of these interneurones might be the adjustment of the gain and of the time constant in the thoraco-coxal feedback loop.