Effects of Active Conductance Distribution over Dendrites on the Synaptic Integration in an Identified Nonspiking Interneuron

The synaptic integration in individual central neuron is critically affected by how active conductances are distributed over dendrites. It has been well known that the dendrites of central neurons are richly endowed with voltage- and ligand-regulated ion conductances. Nonspiking interneurons (NSIs), almost exclusively characteristic to arthropod central nervous systems, do not generate action potentials and hence lack voltage-regulated sodium channels, yet having a variety of voltage-regulated potassium conductances on their dendritic membrane including the one similar to the delayed-rectifier type potassium conductance. It remains unknown, however, how the active conductances are distributed over dendrites and how the synaptic integration is affected by those conductances in NSIs and other invertebrate neurons where the cell body is not included in the signal pathway from input synapses to output sites. In the present study, we quantitatively investigated the functional significance of active conductance distribution pattern in the spatio-temporal spread of synaptic potentials over dendrites of an identified NSI in the crayfish central nervous system by computer simulation. We systematically changed the distribution pattern of active conductances in the neuron's multicompartment model and examined how the synaptic potential waveform was affected by each distribution pattern. It was revealed that specific patterns of nonuniform distribution of potassium conductances were consistent, while other patterns were not, with the waveform of compound synaptic potentials recorded physiologically in the major input-output pathway of the cell, suggesting that the possibility of nonuniform distribution of potassium conductances over the dendrite cannot be excluded as well as the possibility of uniform distribution. Local synaptic circuits involving input and output synapses on the same branch or on the same side were found to be potentially affected under the condition of nonuniform distribution while operation of the major input-output pathway from the soma side to the one on the opposite side remained the same under both conditions of uniform and nonuniform distribution of potassium conductances over the NSI dendrite

A behavioral analysis of force-controlled operant tasks in American lobster

Operant conditioning is a common tool for studying cognitive aspects of brain functions. As the first step toward understanding those functions in simple invertebrate microbrains, we tested whether operant conditioning could be applied to train American lobster Homarus americanus that has been extensively adopted as an animal model for neurophysiological analyses of nervous system functions and behavioral control. The animal was trained by food rewarding for gripping of a sensor bar as the operant behavior. Lobsters were first reinforced when they acted on the bar with a stronger grip than a pre-set value. After this reinforcement, the animal learnt to grip the bar for food pellets. The yoked control experiment in which the animal received action-independent reinforcement excluded the possibility of pseudoconditioning that the food simply drove the animal to frequent gripping of the sensor bar. The association of the bar grip with food was extinguished by rewarding nothing to the operant behavior, and was restored by repeating the reinforcement process as before. In addition, lobsters successfully carried out differential reinforcement regarding the gripping force: their gripping force changed depending on the increased force threshold for food reward. These data demonstrate that lobsters can be trained by operant conditioning paradigms involving acquisition and extinction procedures with the precise claw gripping even under the force control

Readiness Discharge for Spontaneous Initiation of Walking in Crayfish

Animals initiate behavior not only reflexively but also spontaneously in the absence of external stimuli. In vertebrates, electrophysiological data on the neuronal activity associated with the self-initiated voluntary behavior have accumulated extensively. In invertebrates, however, little is known about the neuronal basis of the spontaneous initiation of behavior. We investigated the spike activity of brain neurons at the time of spontaneous initiation of walking in the crayfish Procambarus clarkii and found neuronal signals indicative of readiness or preparatory activities in the vertebrate brain that precede the onset of voluntary actions. Those readiness discharge neurons became active >1 s before the initiation of walking regardless of stepping direction. They remained inactive at the onset of mechanical stimulus-evoked walking in which other descending units were recruited. These results suggest that the parallel descending mechanisms from the brain separately subserve the spontaneous and stimulus-evoked walking. Electrical stimulation of these different classes of neurons caused different types of walking. In addition, we found other descending units that represented different aspects of walking, including those units that showed a sustained activity increase throughout the walking bout depending on its stepping direction, as well as one veto unit for canceling out the output effect of the readiness discharge and three termination units for stopping the walking behavior. These findings suggest that the descending activities are modularized in parallel for spontaneous initiation, continuation, and termination of walking, constituting a sequentially hierarchical control

Discrimination learning with light stimuli in restrained American lobster

Operant discrimination learning has been extensively utilized in the study on the perceptual ability of animals and their higher-order brain functions. We tested in this study whether American lobster Homarus americanus, which was previously found to possess ability of operant learning with claw gripping, could be trained to discriminate light stimuli of different intensities. For the current purpose, we newly developed a PC-controlled operant chamber that allowed the animal under a body-fixed condition to perform operant reward learning with claw gripping. Lobsters were first reinforced when they gripped the sensor bar upon presentation of a light cue. Then they were trained to grip the bar only when the light stimulus of a specific intensity was presented to obtain food reward while the stimuli of three different intensities including the reinforced one were presented in a random order. Finally, they were re-trained to grip the bar only when the light stimulus of another intensity that was not rewarded in the preceding training to obtain food while other intensities including the one that was rewarded previously were not rewarded any more. In these training procedures, the operant behavior occurred more frequently in response to the rewarded cue than to the non-rewarded one. The action latency for the reinforced stimuli showed a significant decrease in the course of training. These data demonstrate that lobsters can be trained with the light cues of different intensity as discriminative stimuli under a restrained condition that would allow application of electrophysiological techniques to the behaving subjects

Physiological changes of premotor nonspiking interneurons in the central compensation of eyestalk posture following unilateral sensory ablation in crayfish

We investigated how the physiological characteristics and synaptic activities of nonspiking giant interneurons (NGIs), which integrate sensory inputs in the brain and send synaptic outputs to oculomotor neurons innervating eyestalk muscles, changed after unilateral ablation of the statocyst in order to clarify neuronal mechanisms underlying the central compensation process in crayfish. The input resistance and membrane time constant in recovered animals that restored the original symmetrical eyestalk posture 2 weeks after operation were significantly greater than those immediately after operation on the operated side whereas in non-recovered animals only the membrane time constant showed a significant increase. On the intact side, both recovered and non-recovered animals showed no difference. The frequency of synaptic activity showed a complex pattern of change on both sides depending on the polarity of the synaptic potential. The synaptic activity returned to the bilaterally symmetrical level in recovered animals while bilateral asymmetry remained in non-recovered ones. These results suggest that the central compensation of eyestalk posture following unilateral impairment of the statocyst is subserved by not only changes in the physiological characteristics of the NGI membrane but also the activity of neuronal circuits presynaptic to NGIs

An optical telemetry system for underwater recording of electromyogram and neuronal activity from non-tethered crayfish

We have developed an optical telemetry system for recording electrical signals associated with muscle and neuronal activities from freely walking crayfish under water. The device was made from conventional electronic parts which are commercially available, utilizing infrared light (880 nm) for signal transmission. Two or four channels of biological signals were multiplexed, the voltage of each data point modulated to the duration of subcarrier pulses and further to the interval of narrower carrier pulses that directly drove the infrared light emission diode (IRLED) under water. The light-pulse modulated signals were received by photodiodes and demodulated to restore the original two or four channel signals. Electrical recordings using wired electrodes and conventional amplifiers revealed that the optically transmitted signals were consistent with the wire-transmitted ones. In order to test the performance of this system, we recorded electromyograms (EMGs) from the second and third walking legs on each side of crayfish together with the neuronal activity in the ventral nerve cord. The results confirmed our previous observation in tethered crayfish that the background tonus of leg muscles showed an increase preceding their rhythmic activation

Electromyographic Characterization of Walking Behavior Initiated Spontaneously in Crayfish

Crayfish initiate walking behavior not only reflexively in response to external stimuli but also spontaneously in the absence of any specific stimulus. In order to analyze the initiation mechanism underlying these different types of walking, we made simultaneous electromyographic (EMG) recordings from thoracic legs when animals initiated walking, either reflexively or spontaneously, and video recorded their movements synchronously with the EMG recording. Two different stimuli, mechanical and chemical, were used to reflexively induce walking. A non-rhythmic, sustained activation of leg muscles was found to precede the behavioral initiation of either type of walking. The duration of this non-rhythmic muscle activation was significantly longer in the spontaneously initiated walking than in the mechanical stimulus-evoked walking, although no difference was observed between the spontaneous and chemical stimulus-evoked walking. EMG recordings from all eight legs revealed that their non-rhythmic muscle activation occurred almost simultaneously prior to initiation of rhythmical stepping movements. When an animal was suspended without a leg substratum, the timing of muscle activation was more variable among the legs than in the free condition on the substratum. When the circumesophageal commissures were both severed to eliminate signals descending from the brain to the thoracic ganglia, the bilaterally coordinated rhythmic burst activity was not observed in the walking legs. These findings suggest that the spontaneous initiation of walking behavior requires sensory feedback signals from leg proprioceptors, subserved by a different descending activation mechanism from that for stimulus-driven initiation of walking

Chronic electromyographic analysis of circadian locomotor activity in crayfish

Animals generally exhibit circadian rhythms of locomotor activity. They initiate locomotor behavior not only reflexively in response to external stimuli but also spontaneously in the absence of any specific stimulus. The neuronal mechanisms underlying circadian locomotor activity can, therefore, be based on the rhythmic changes in either reflexive efficacy or endogenous activity. In crayfish Procambarus clarkii, it can be determined by analyzing electromyographic (EMG) patterns of walking legs whether the walking behavior is initiated reflexively or spontaneously. In this study, we examined quantitatively the leg muscle activity that underlies the locomotor behavior showing circadian rhythms in crayfish. We newly developed a chronic EMG recording system that allowed the animal to freely behave under a tethered condition for more than 10 days. In the LD condition in which the animals exhibited LD entrainment, the rhythmic burst activity of leg muscles for stepping behavior was preceded by non-rhythmic tonic activation that lasted for 1323 +/- 488 ms when the animal initiated walking. In DD and LL free-running conditions, the pre-burst activation lasted for 1779 +/- 31 and 1517 +/- 39 ms respectively. In the mechanical stimulus-evoked walking, the pre-burst activation ended within 79 +/- 6 ms. These data suggest that periodic changes in the crayfish locomotor activity under the condition of LD entrainment or free-running are based on activity changes in the spontaneous initiation mechanism of walking behavior rather than those in the sensori-motor pathway connecting mechanoreceptors with leg movements. (c) 2013 Elsevier B.V. All rights reserved

Static electric field detection and behavioural avoidance in cockroaches

Electric fields are pervasively present in the environment and occur both as a result of man-made activities and through natural occurrence. We have analysed the behaviour of cockroaches to static electric fields and determined the physiological mechanisms that underlie their behavioural responses. The behaviour of animals in response to electric fields was tested using a Y-choice chamber with an electric field generated in one arm of the chamber. Locomotory behaviour and avoidance were affected by the magnitude of the electric fields with up to 85% of individuals avoiding the charged arm when the static electric field at the entrance to the arm was above 8–10kVm–1. Electric fields were found to cause a deflection of the antennae but when the antennae were surgically ablated, the ability of cockroaches to avoid electric fields was abolished. Fixation of various joints of the antennae indicated that hair plate sensory receptors at the base of the scape were primarily responsible for the detection of electric fields, and when antennal movements about the head–scape joint were prevented cockroaches failed to avoid electric fields. To overcome the technical problem of not being able to carry out electrophysiological analysis in the presence of electric fields, we developed a procedure using magnetic fields combined with the application of iron particles to the antennae to deflect the antennae and analyse the role of thoracic interneurones in signalling this deflection. The avoidance of electric fields in the context of high voltage power lines is discussed