An Analysis of Field Potentials During Different Tailflip Behaviours in Crayfish

Crayfish tailflips have been intensively studied to reveal the decision-making processes and neural organisation underlying a stereotyped escape behaviour. Three behaviours mediated by different neural pathways have been well described: medial giant, lateral giant and non-giant tailflips. It has proved difficult to distinguish between the three without invasive or restrictive experimental manipulation. We report unambiguous differences between the signals generated by the crayfish Cherax destructor during the three types of tailflip when recorded by bath electrodes placed in the holding aquarium. Using our ability to distinguish between the different behaviours in freely moving animals we examined the relationship between the type of tailflip evoked by stimulation to different parts of the body. The transition zone between medial and lateral giant tailflips is the thoracic-abdominal border but it is not absolute and some stimuli produce responses that cannot be unambiguously assigned to either behavioural category. We examined the latency between stimulation at different points down the length of the body and the appearance of the electrical signal accompanying escape for both medial and lateral tailflips. We used two methods to estimate the proportion of the latency accounted for by giant fibre conduction velocity. The results support current views of the differences between the activation sites of the two giant fibre systems and suggest why stimulation in the transition zone results in ambiguous outcomes.     

Inhibition of an abdominal proprioceptive reflex in crayfish

Summary Stimulation of the tonic muscle receptor organs inProcambarus clarkii results in reflex activation of the superficial extensors. This pathway is inhibited by activity in the lateral giant fibers and both medial giants. A labile synapse in the same pathway works synergistically with the giant fiber-mediated inhibition to prevent maladaptive activation of the superficial extensors during escape behavior. Evidence did not support the notion of giant fiber-mediated peripheral inhibition (via the accessory nerve) of MRO activity.This work was supported by NIH grant NS 02944 (to D. Kennedy). I would like to thank Drs. Kennedy, Wine, and the members of the Kennedy laboratory for their help and support.     

Loss of escape responses and giant neurons in the tailflipping circuits of slipper lobsters, Ibacus spp. (Decapoda, Palinura, Scyllaridae)

In many decapod crustaceans, escape tailflips are triggered by lateral giant (LG) and medial giant (MG) interneurons, which connect to motor giant (MoG) abdominal flexor neurons. Several decapods have lost some or all of these giant neurons, however. Because escape-related giant neurons have not been documented in palinurans, I examined tailflipping and abdominal nerve cords for giant neurons in two scyllarid lobster species, Ibacus peronii and Ibacus alticrenatus. Unlike decapods with giant neurons, Ibacus do not tailflip in response to sudden taps. Ibacus can perform non-giant tailflipping: the frequency of tailflips during swimming is adjusted by altering the gap between each individual tailflip. Abdominal nerve cord sections show no LG or MG interneurons. Backfilling nerve 3 of abdominal ganglia revealed no MoG neurons, and the fast flexor motor neuron population is otherwise identical to that described for crayfish. The loss of giant neurons in Ibacus represents an independent deletion of these cells compared to other reptantian decapods known to have lost these giant neurons. This loss is correlated with the normal posture in scyllarids, in which the last two abdominal segments are flexed, and an alternative defensive strategy, concealment by digging into sand.     

Modulation of the crayfish escape reflex--physiology and neuroethology

We review here factors that control the excitability of thegiant neuron-mediated tail-flip escape behavior in crayfish,focusing especially on recent findings concerning serotonergicmodulation. Serotonin can either facilitate or inhibit escapedepending on concentration and pattern of application. Low concentrationsfacilitate while high ones inhibit; however, if high concentrationsarise gradually they facilitate instead of inhibiting. The effectsof serotonin can also be altered by social experience, withapplication regimens that cause facilitation in social isolatescoming to produce inhibition after an extended period of livingas a subordinate. Attempts to understand both the possible physiologicalbasis of some of these complexities and their possible functionare discussed. Neuroethological investigations indicate thatgiant neuron-mediated escape is inhibited during the initialfights that establish social relationships and is facilitatedin their immediate aftermath. Once the relationship of a pairis well-established, the presence of the dominant tends to suppressgiant neuron-mediated escape (but not tail-flip escape mediatedby non-giant circuitry) in the subordinate, but the presenceof the subordinate has relatively little effect on the dominant.These patterns of modulation can be seen as consistent withthe known variations in serotonin's effect as a function ofconcentration and social experience and may provide a biologicalreason for these variations.     

Long-lasting potentiation of excitatory synaptic signaling to the crayfish lateral giant neuron

The neural circuit that underlies the lateral giant fiber (LG)-mediated reflex escape in crayfish has provided findings relating synaptic change to nonassociative learning such as sensitization and habituation. The LGs receive sensory inputs from the primary sensory afferents and a group of mechanosensory interneurons (MSIs). An increase of excitability by suprathreshold repetitive excitation of this circuit, which is similar to Hebbian long-term potentiation (LTP), has been reported [Miller et al. (1987) J Neurosci 7:1081]. This potentiation was previously thought to result from the enhancement of transmission at cholinergic synapses between primary afferents and MSIs but not the electrical synapses onto LG. In this study, we found that potentiation of synaptic signaling at the electrical synapse onto LG can also be induced when the synapse was activated with subthreshold repetitive pulses or with a few strong suprathreshold shocks. LG LTP was induced in the preparation which had received pulses at limited frequency range. Although whether this LTP is involved in the learning process of escape behavior in crayfish is not clear, the intensity and amount of sensory stimulation used here mimicked those that could easily be produced by a predator trying to catch a crayfish and could be of adaptive significance in life.     

The Tentacle Cilia of Aglantha digitale (Hydrozoa: Trachylina) and their Control

The tentacles of Aglantha have ciliary bands along the sides. Metachronal waves pass along these bands. The strong ciliary currents produced propel water past the tentacles, increasing the probability of prey capture. The ciliated cells are unusual in having many (up to about 500) cilia per cell, where most cnidarian ciliated cells have only one. The cells are also peculiar in containing numerous axonemes without membrane coverings, lying loose in the cytoplasm. Tentacles show independent, rhythmic, slow flexions in the oral direction and groups of tentacles show coordinated, slow flexions as part of a regularly repeated fishing cycle. In both cases, these slow, graded movements are mediated by a slowly conducting system, probably the network of small neurons present in the ectoderm, and are accompanied by ciliary arrests. Much faster, more powerful, coordinated contractions of the tentacles occur in the context of escape behaviour; these are mediated by giant axons which run down the tentacles and are also accompanied by ciliary arrest. Ciliary and muscle effectors evidently share a common motor innervation. Electron microscopy shows that the giant and non-giant nerves both synapse with muscle cells. The latter are joined to the ciliated cells by gap junctions, and it is suggested that whenever the muscles are excited depolarizations spread to the ciliated cells through the gap junctions and cause ciliary arrests. Neuronal control of ciliary activity has not previously been reported in the Hydrozoa.     

Effects of Removal of Muscle Receptor Organ Input on the Temporal Structure of Non-Giant Swimming Cycles in the Crayfish, Cherax Destructor

Most studies of the muscle receptor organs (MROs) of decapod crustaceans have focused on their role in local reflex loops. This may not be their only function. We examine their involvement in the regulation of non-giant swimming cycles by removing stretch receptor (SR) input from the MROs in abdominal segments 2-5 of the crayfish Cherax destructor . SR input was left intact in two control groups, one of which had sham surgery and the other no surgery at all. We recorded electromyograms (EMGs) from selected uropod muscles during tailflipping in sequences of non-giant swimming in tethered animals. The removal of SR input had a significant effect. The opener muscle period was shorter in the experimental group than in either of the control groups. This suggests that by using SR afference, crayfish sacrifice speed for increased control of the swimming movement.     

Effects of Removal of Muscle Receptor Organ Input on the Temporal Structure of Non-Giant Swimming Cycles in the Crayfish,Cherax Destructor

Most studies of the muscle receptor organs (MROs) of decapod crustaceans have focused on their role in local reflex loops. This may not be their only function. We examine their involvement in the regulation of non-giant swimming cycles by removing stretch receptor (SR) input from the MROs in abdominal segments 2-5 of the crayfish Cherax destructor. SR input was left intact in two control groups, one of which had sham surgery and the other no surgery at all. We recorded electromyograms (EMGs) from selected uropod muscles during tailflipping in sequences of non-giant swimming in tethered animals. The removal of SR input had a significant effect. The opener muscle period was shorter in the experimental group than in either of the control groups. This suggests that by using SR afference, crayfish sacrifice speed for increased control of the swimming movement.     

Neurobehavioral specializations for respiratory movements and rapid escape from predators in posterior segments of the tubificid Branchiura sowerbyi

The posterior end of the aquatic oligochaete, Branchiura sowerbyi (Tubificidae) protrudes above the sediments and is specialized to carry out several rhythmic respiratory movements. These include 1) waves of flexion by paired gill filaments on each posterior segment, 2) body undulations, and 3) rectal water pumping. Since execution of these behaviors renders the worm's posterior end vulnerable to predation, appropriate neurobehavioral mechanisms have evolved that permit extremely rapid escape of tail segments into the sediments. Some of these mechanisms include 1) highly sensitive sensory apparatus for detecting substrate vibrations, water displacements, or touch, 2) large diameter and rapidly conducting lateral giant nerve fibers, and 3) adequacy of a single lateral giant fiber impulse for evoking an all-or-none longitudinal muscle contraction. The significance of these posterior respiratory and escape reflex specializations are discussed in relation to possible predator foraging strategies.     

Behavioral plasticity and central regeneration of locomotor reflexes in the freshwater oligochaete, Lumbriculus variegatus. I: Transection studies

Abstract. Access to the ventral nerve cord in living specimens of Lumbriculus variegatus , an aquatic oligochaete, is normally impossible because surgical invasion induces segmental autotomy (self-fragmentation). We show here that nicotine is a powerful paralytic agent that reversibly immobilizes worms, blocks segmental autotomy, and allows experimental access to the nerve cord. Using nicotine-treated worms, we transected the ventral nerve cord and used non-invasive electrophysiological recordings and behavioral analyses to characterize the functional recovery of giant nerve fibers and other reflex pathways. Initially, after transection, medial giant fiber (MGF) and lateral giant fiber (LGF) spikes conducted up to, but not across, the transection site. Reestablishment of MGF and LGF through-conduction across the transection site occurred as early as 10 h (usually by 20 h) after transection. Analyses of non-giant-mediated behavioral responses (i.e., helical swimming and body reversal) were also made following nerve cord transection. Immediately after transection, functional reorganization of touch-evoked locomotor reflexes occurred, so that the two portions of the worm anterior and posterior to the transection site were independently capable of helical swimming and body reversal responses. Similar reorganization of responses occurred in amputated body fragments. Reversion back to the original whole-body pattern of swimming and reversal occurred as early as 8 h after transection. Thus, functional restoration of the non-giant central pathways appeared slightly faster than giant fiber pathways. The results demonstrate the remarkable plasticity of locomotor reflex behaviors immediately after nerve cord transection or segment amputation. They also demonstrate the exceptional speed and specificity of regeneration of the central pathways that mediate locomotor reflexes.     

A rectifying electrotonic synapse in the central nervous system of a vertebrate

The adductor muscles of the pectoral fins of the hatchetfish Gasteropelecus are innervated by bilateral pools of about 40 motoneurons which lie primarily in the first spinal segment. A pair of giant fibers on each side of the medulla send processes ventroposteriorly to the motoneuron pools. Electrophysiological evidence indicates that giant fibers are presynaptic to ipsilateral motoneurons, but not to contralateral ones. Transmission across the giant fiber, motoneuron synapse is electrically mediated as is indicated by direct measurement of electrotonic spread in either direction across the synapse, and by the extremely short latency of the giant fiber postsynaptic potentials (PSP's) in the motoneuron. The coupling resistance across the synapse was calculated from measurements of input and transfer resistance. The coupling resistance rectifies in such a way as to facilitate spread of depolarization from giant fiber to motoneuron, and to oppose transmission in the opposite direction. As a consequence of rectification, the giant fiber PSP in a motoneuron is augmented by hyperpolarization of the motoneuron. The coupling resistance calculated on the basis of this effect is in good agreement with calculations from input and transfer resistance data. Rectification at the electrotonic synapses may permit the motoneurons to act in small swimming movements as well as to fire synchronously in an extremely fast escape reflex mediated by Mauthner and giant fibers.     

Decrease in excitability of LG following habituation of the crayfish escape reaction

Crayfish escapes from threatening stimuli to the abdomen by tailflipping upwards and forwards. This lateral giant (LG)-mediated escape reaction habituates readily upon repetitive sensory stimulation. Using an isolated abdominal nerve cord preparation, we have analyzed the change in LG activity by applying additional sensory stimulation after different periods following habituation to characterize the retention of LG habituation. Results show that the LG mediated response habituates more quickly, but the retention time is shorter, as repetitive sensory stimulation is applied at progressively shorter inter-stimulus time intervals. The spike response of LG recovers quickly, within several minutes after habituation, but they fail to spike when an additional stimulus is applied after specific long periods following habituation. The critical period of the delay for this decrease in excitability of LG is dependent on the inter-stimulus time interval of the initial repetitive stimulus. As the inter-stimulus interval became longer, the delay needed for decrease in excitability became shorter. Furthermore, the local injection of 10^–6 mol l^–1 octopamine into the neuropil just following habituation promotes the achievement of decrease in excitability. No effects were observed when 10^–6 mol l^–1 serotonin and tyramine were injected. These results suggested octopamine promotes decrease in excitability of LG following habituation.     

Monosynaptic excitation of lateral giant fibres by proprioceptive afferents in the crayfish

Giant interneurones mediate a characteristic `tail flip' escape response of the crayfish, Procambarus clarkii, which move it rapidly away from the source of stimulation. We have analysed the synaptic connections of proprioceptive sensory neurones with one type of giant interneurone, the lateral giant. Spikes in sensory neurones innervating an exopodite-endopodite chordotonal organ in the tailfan, which monitors the position and movements of the exopodite, are followed at a short and constant latency by excitatory postsynaptic potentials in a lateral giant interneurone (LG) recorded in the terminal abdominal ganglion. These potentials are unaffected by manipulation of the membrane potential of LG, by bath application of saline with a low calcium concentration, or by one containing the nicotinic antagonist, curare. The potentials evoked in LG by chordotonal organ stimulation are thus thought to be monosynaptic and electrically mediated. This is the first demonstration that LG receives input from sensory receptors other than exteroceptors in the terminal abdominal ganglion. Accepted: 7 April 1997     

Synchronous firing by specific pairs of cercal giant interneurons in crickets encodes wind direction

One prominent stimulus to evoke an escape response in crickets is the detection of air movement, such as would result from an attacking predator. Wind is detected by the cercal sensory system that consists of hundreds of sensory cells at the base of filiform hairs. These sensory cells relay information to about a dozen cercal giant and non-giant interneurons. The response of cercal sensory cells depends both, on the intensity and the direction of the wind. Spike trains of cercal giant interneurons then convey the information about wind direction and intensity to the central nervous system. Extracellular recording of multiple cercal giant interneurons shows that certain interneuron pairs fire synchronously if a wind comes from a particular direction. We demonstrate here that directional tuning curves of synchronously firing pairs of interneurons are sharper than those of single interneurons. Moreover, the sum total of all synchronously firing pairs eventually covers all wind directions. The sharpness of the tuning curves in synchronously firing pairs results from excitatory and inhibitory input from the cercal sensory neurons. Our results suggest, that synchronous firing of specific pairs of cercal giant interneurons encodes the wind direction. This was further supported by behavioral analyses.     

Morphallaxis in an aquatic oligochaete, Lumbriculus variegatus: reorganization of escape reflexes in regenerating body fragments

We describe functional and anatomical correlates of the reorganization of giant nerve fiber-mediated escape reflexes in body fragments of an aquatic oligochaete, Lumbriculus variegatus, a species that reproduces asexually by fragmentation. Since fragments from any axial position always regenerate short heads (seven or eight segments long) and much longer tail sections, segments originating from posterior fragments become transposed along the longitudinal axis and acquire, by morphallaxis, features of escape reflex organization that conform to their new anterior position. Using noninvasive electrophysiological recordings we have quantified, on a day-to-day and a segment-by-segment basis, the reorganization that occurs in sensory field arrangements of the medial (MGF) and lateral (LGF) giant nerve fibers, as well as changes in giant fiber conduction velocity and morphometry. Our results show that (1) posterior fragments, originally subserved by the LGF sensory field gradually become subserved by the MGF sensory field; (2) appropriate increases in the ratio of MGF:LGF cross-sectional area, perimeter, and conduction velocity accompany the reorganization in giant fiber sensory fields; and (3) sensory field reorganization can be repeatedly reversed by additional amputations. These results demonstrate that the functional organization of escape reflexes is highly plastic and that morphallaxis may result from the counterbalance of morphogenic influences localized within the anterior and posterior ends of regenerating body fragments.     

Modulation of soleus H-reflex during dorsal and plantar flexions in the human ankle joint

Recording of the H-reflex was used to study the changes in the reflex excitability of soleus motoneurons during dorsal and plantar flexions of the ipsilateral and contralateral feet performed with different strengths by 15 healthy subjects. The dorsiflexion of the ipsilateral foot was accompanied by the “classic” reciprocal inhibition of the soleus motoneurons, the degree of the inhibition being directly proportional to the strength of the contraction of pretibial muscles and depending on the presence of foot support. The plantar flexion of the ipsilateral foot was accompanied by changes in reflex excitability, which were inversely proportional to the strength of the flexion. This was apparently related to the activation of a mechanism protecting the muscle against excessive contraction. The dorsal and plantar flexions of the contralateral foot were accompanied by similar changes in the reflex excitability of soleus motoneurons, namely, an increase in the case of weak contraction and a decrease in the case of strong contraction. However, the increase in reflex excitability during contralateral dorsiflexion was smaller and its decrease began at a weaker contraction than in the case of contralateral plantar flexion. The changes in the reflex excitability of soleus motoneurons during movements of the contralateral foot, which were also strength-dependent, confirmed the presence of cross-projections that are likely to be part of the generator of the central pattern of lower limb movement coordination.     

低氧对横断脊髓大鼠脊髓反射兴奋性的影响

本文用低压舱模拟不同海拔高度,测定横断脊髓大鼠脊髓反射兴奋性恢复曲线。断脊髓组和对照组自海拔２０００ｍ以后脊髓反射兴奋性逐渐增高,但两组出现差异有显著性的海拔高度不同,断脊髓组为海拔４０００ｍ,对照组为海拔３０００ｍ。对照组与断脊髓组在同一海拔高度上比较,除海拔３０００ｍ时差别有显著性（Ｐ＜０．０５）外,在其它各海拔高度上两组间差异无显著性（Ｐ＞０．０５）。结果表明,低氧时脊髓以上中枢和脊髓都参与脊髓反射兴奋性升高的调节     

Effect of cerebral ventricles perfusion with naloxone on trigemino-hypoglossal reflex in rats

The goal of this study was to determine whether opioid receptor antagonist naloxone abolishes the influence of periaqueductal central gray (PAG) on nociceptive evoked tongue jerks (ETJ) -- a trigemino-hypoglossal reflex induced by tooth pulp stimulation. In rats under chloralose anesthesia three series of experiments were performed. In the first two groups perfusions of lateral ventricles-cerebellomedullary cistern with McIlwain-Rodnight's solution and naloxone were carried out. In group 3 naloxone was infused through a catheter through the jugular vein. The amplitudes of tongue jerks induced by tooth pulp stimulation were recorded during subsequent 10 min perfusions. Mean amplitude of tongue movements induced by tooth pulp stimulation was regarded as the indicator of the magnitude of trigemino-hypoglossal reflex. We observed that perfusion of the cerebral ventricles with naloxone (100 nmol/ml) increased the trigemino-hypoglossal reflex up to 143%. The amplitude of ETJ was significantly reduced during PAG stimulation with a train of electrical impulses. After obtaining a significant -- 93% -- inhibition of ETJ (7% of the control), naloxone (100 nmol/ml) was added to the perfusion fluid. This led to a significant increase of the reflex up to 68%. Infusion of naloxone through the jugular vein did not affect the reflex. The above results suggest that the inhibition of ETJ due to PAG stimulation is partially reversed by naloxone and mediated via interactions with endogenous opioid systems involved in modulation of nociception.     

Raising cytosolic Cl- in cerebellar granule cells affects their excitability and vestibulo-ocular learning

Cerebellar cortical throughput involved in motor control comprises granule cells (GCs) and Purkinje cells (PCs), both of which receive inhibitory GABAergic input from interneurons. The GABAergic input to PCs is essential for learning and consolidation of the vestibulo-ocular reflex, but the role of GC excitability remains unclear. We now disrupted the Kcc2 K-Cl cotransporter specifically in either cell type to manipulate their excitability and inhibition by GABA(A)-receptor Cl(-) channels. Although Kcc2 may have a morphogenic role in synapse development, Kcc2 disruption neither changed synapse density nor spine morphology. In both GCs and PCs, disruption of Kcc2, but not Kcc3, increased [Cl(-)](i) roughly two-fold. The reduced Cl(-) gradient nearly abolished GABA-induced hyperpolarization in PCs, but in GCs it merely affected excitability by membrane depolarization. Ablation of Kcc2 from GCs impaired consolidation of long-term phase learning of the vestibulo-ocular reflex, whereas baseline performance, short-term gain-decrease learning and gain consolidation remained intact. These functions, however, were affected by disruption of Kcc2 in PCs. GC excitability plays a previously unknown, but specific role in consolidation of phase learning.     

Central neural circuitry in the jellyfish Aglantha: a model 'simple nervous system'

Like other hydrozoan medusae, Aglantha lacks a brain, but the two marginal nerve rings function together as a central nervous system. Twelve neuronal and two excitable epithelial conduction systems are described and their interactions summarized. Aglantha differs from most medusae in having giant axons. It can swim and contract its tentacles in two distinct ways (escape and slow). Escape responses are mediated primarily by giant axons but conventional interneurons are also involved in transmission of information within the nerve rings during one form of escape behavior. Surprisingly, giant axons provide the motor pathway to the swim muscles in both escape and slow swimming. This is possible because these axons can conduct calcium spikes as well as sodium spikes and do so on an either/or basis without overlap. The synaptic and ionic bases for these responses are reviewed. During feeding, the manubrium performs highly accurate flexions to points at the margin. At the same time, the oral lips flare open. The directional flexions are conducted by FMRFamide immunoreactive nerves, the lip flaring by an excitable epithelium lining the radial canals. Inhibition of swimming during feeding is due to impulses propagated centrifugally in the same epithelium. Aglantha probably evolved from an ancestor possessing a relatively simple wiring plan, as seen in other hydromedusae. Acquisition of giant axons resulted in considerable modification of this basic plan, and required novel solutions to the problems of integrating escape with non-escape circuitry.