Further evidence for excitatory amino acid transmission in lamprey reticulospinal neurons: selective retrograde labeling with (3H)D-aspartate

The distribution of radiolabeled neurons in the brain stem of Lampetra fluviatilis was studied following unilateral injections of (3H)D-aspartate in the rostral spinal cord. After survival periods of 1-3 days, labeled perikarya were present within and nearby the posterior, middle, and anterior rhombencephalic reticular nuclei and in the mesencephalic reticular nucleus. The highest number of (3H)D-aspartate labeled cell bodies were present in the posterior rhombencephalic reticular nucleus. The labeled reticulospinal neurons were distributed mainly ipsilateral to the injection site and included the giant Müller cells as well as medium-sized and small neurons. Contralateral labeling occurred in cell bodies scattered along the lateral margin of the rhombencephalic reticular formation, the most rostral of these contralaterally projecting neurons being the Mauthner cell. The (3H)D-aspartate labeling correlates with previous electrophysiological studies showing that lamprey reticulospinal neurons utilize excitatory amino acid transmission.     

The role of spinal cord inputs in modulating the activity of reticulospinal neurons during fictive locomotion in the lamprey

Lamprey reticulospinal neurons are rhythmically modulated during fictive swimming. The present study examines the possibility that this modulation may originate from the spinal cord locomotor networks rather than from the brainstem. To test this, the in vitro preparation of the lamprey brainstem-spinal cord was separated into two compartments which could be exposed to different chemical environments. Locomotor activity was induced pharmacologically in the caudal spinal cord compartment and reticulospinal (RS) neurons from the posterior rhombencephalic reticular nucleus (PRRN) were recorded intracellularly in the rostral compartment containing normal lamprey Ringer. Under these conditions, the membrane potential of RS neurons showed clear rhythmic oscillations which are correlated with the ongoing locomotor activity in the caudal spinal cord bath, although no locomotor discharges were present in the ventral roots of the rostral bath. Such oscillations were not present in the absence of locomotion. These results indicate that the spinal cord locomotor networks can contribute to the rhythmic oscillations which occur in RS neurons during fictive locomotion. Moreover, the latter oscillations of membrane potential are due to both phasic excitation and Cl- -dependent inhibition in the opposite phase.     

Phasic modulation of transmission from vestibular inputs to reticulospinal neurons during fictive locomotion in lampreys

The aim of this study was to determine whether the transmission from sensory inputs to reticulospinal neurons is modulated during fictive locomotion in lampreys. Reticulospinal neurons play a key role in the control of locomotion; modulation of sensory transmission to these neurons might be of importance for the adaptation of the control they exert during locomotion. In this series of experiments, intracellular synaptic responses of reticulospinal neurons of the posterior rhombencephalic reticular nucleus elicited by electrical stimulation of vestibular nerves on each side were studied during fictive locomotion induced by 50 microM N-methyl-D-aspartate (NMDA). Interestingly, shortly after NMDA had reached the bath and much before locomotor discharges were apparent in the recorded ventral roots, there was a significant depression of the synaptic transmission from vestibular nerves. The effect was reversed by washing out the NMDA and persisted in the isolated brainstem after spinal transection at the first segmental level. As locomotor discharges appeared in the ventral roots, synaptic responses elicited by vestibular nerve stimulation showed a clear phasic modulation of their amplitude during the locomotor cycle. Responses to stimulation of the ipsilateral vestibular nerve were smaller during the ipsilateral burst discharge than during the contralateral activity, whilst responses to stimulation of the contralateral vestibular nerve were minimal during contralateral activity and maximal during ipsilateral activity. This opposite pattern of modulation observed in the same reticulospinal neuron suggests that the phasic modulation of vestibular transmission is not due to changes in the membrane properties of the reticulospinal cell but is produced at a pre-reticular level.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of stimulating the reticular formation during fictive locomotion in lampreys

The effects of stimulating the reticular formation were studied during fictive locomotion in lampreys (Ichthyomyzon unicuspis). The in vitro isolated preparation of the brainstem and spinal cord was used and fictive locomotion was induced by bath application of N-methyl-

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-aspartate (NMDA; 50–100 μM). During different phases of the locomotor cycle, short trains of stimuli (10 pulses at 80–100 Hz; 10 μA) were delivered through glass-coated tungsten microelectrodes positioned within the middle rhombencephalic reticular nucleus (MRRN) and their effects were studied on ipsi- and contralateral ventral root locomotor discharges. Irrespective of the locomotor phase during which the stimulation train was delivered, a resetting effect occurred. It was characterized by a re-synchronization of the locomotor discharges with a constant latency for each ventral root on the ipsilateral side. The latency increased as the recorded root was located further caudally. This increase in latency was in the range of the phase lag observed between roots during control bouts of locomotion. These results suggest that reticulospinal neurones exert strong resetting effects on spinal locomotor networks. These effects may play a significant role with respect to changes of direction during swimming.     

Descending brain neurons in larval lamprey: Spinal projection patterns and initiation of locomotion

In larval lamprey, partial lesions were made in the rostral spinal cord to determine which spinal tracts are important for descending activation of locomotion and to identify descending brain neurons that project in these tracts. In whole animals and in vitro brain/spinal cord preparations, brain-initiated spinal locomotor activity was present when the lateral or intermediate spinal tracts were spared but usually was abolished when the medial tracts were spared. We previously showed that descending brain neurons are located in eleven cell groups, including reticulospinal (RS) neurons in the mesenecephalic reticular nucleus (MRN) as well as the anterior (ARRN), middle (MRRN), and posterior (PRRN) rhombencephalic reticular nuclei. Other descending brain neurons are located in the diencephalic (Di) as well as the anterolateral (ALV), dorsolateral (DLV), and posterolateral (PLV) vagal groups. In the present study, the Mauthner and auxillary Mauthner cells, most neurons in the Di, ALV, DLV, and PLV cell groups, and some neurons in the ARRN and PRRN had crossed descending axons. The majority of neurons projecting in medial spinal tracts included large identified Müller cells and neurons in the Di, MRN, ALV, and DLV. Axons of individual descending brain neurons usually did not switch spinal tracts, have branches in multiple tracts, or cross the midline within the rostral cord. Most neurons that projected in the lateral/intermediate spinal tracts were in the ARRN, MRRN, and PRRN. Thus, output neurons of the locomotor command system are distributed in several reticular nuclei, whose neurons project in relatively wide areas of the cord.     

Reticulospinal neurones activate excitatory amino acid receptors

Paired intracellular recordings were used to study the monosynaptic excitatory postsynaptic potentials (EPSP) in lamprey motoneurones evoked by stimulation of single reticulospinal Müller and Mauthner cells. The chemical component of the synaptic potentials was depressed by both application of the non-selective excitatory amino acid antagonists kynurenic acid and cis-2,3-piperidine dicarboxylate. The N-methyl-D-aspartate (NMDA) antagonists Mg2+ and 2-amino-5-phosphonovalerate caused a selective depression of a late component of the EPSP. Thus, fast-conducting reticulospinal neurones appear to release an excitatory amino acid acting at both NMDA and non-NMDA receptors.     

Axonal regeneration in the adult lamprey spinal cord

Larval sea lampreys recover from complete spinal transection by a process involving directionally specific axonal regeneration. In order to determine whether this is also true of adults, 14 adult lampreys were transected at the level of the 5th gill and allowed to recover for 10 weeks. Müller and Mauthner cells and their giant reticulospinal axons (GRAs) were impaled with microelectrodes and injected with horseradish peroxidase (HRP). The tissue was processed for HRP histochemistry and wholemounts of brain and spinal cord were prepared. All animals recovered coordinated swimming; 61 of 121 (50%) neurites emanating from 30 axons regenerated caudal to the scar into the distal stump. Of the neurites which had grown beyond the scar, 92% were correctly oriented, i.e., caudalward and ipsilateral to the parent axon. Retransection in two additional animals eliminated the recovered swimming. Thus, behavioral recovery in adult sea lampreys is accompanied by directionally specific axonal regeneration.     

Origins of the descending spinal projections in petromyzontid and myxinoid agnathans

The origins of the descending spinal pathways in sea lampreys (Petromyzon marinus), silver lampreys (Ichthyomyzon unicuspis), and Pacific hagfish (Eptatretus stouti) were identified by the retrograde transport of horseradish peroxidase (HRP) placed in the rostral spinal cord. In lampreys, the majority of HRP-labeled cells were located along the length of the brainstem reticular formation in the inferior, middle, and superior reticular nuclei of the medulla, mesencephalic tegmentum, and nucleus of the medial longitudinal fasciculus. Labeled reticular cells included the Mauthner and Müller cells. Horseradish-peroxidase-filled cells were also present in the descending trigeminal tract, intermediate and posterior octavomotor nuclei, and a diencephalic cell group, the nucleus of the posterior tubercle. As in lampreys, the reticular formation of the Pacific hagfish was the largest source of descending afferents to the spinal cord. Labeled cells were found in the dorsolateral and ventromedial reticular nuclei, the dorsal tegmentum at the juncture of the medulla and midbrain, and the nucleus of the medial longitudinal fasciculus. Additional medullary cells projecting to the cord were located in the perivagal nucleus, the central gray, and the anterior and posterior magnocellular octavolateralis nuclei. The existence of reticulospinal and possible vestibulo-, trigemino-, and solitary spinal projections in lampreys and hagfishes and the wide distribution of these pathways in jawed vertebrates suggest that they evolved in the common ancestor of gnathostomes and both groups of jawless fishes. However, descending spinal pathways from the cerebellum, red nucleus, and telencephalon appear to be gnathostome characters.     

Effects of bicuculline and strychnine on synaptic inputs to edge cells during fictive locomotion

Edge cells are mechanoreceptive neurones located in the lateral tracts of the lamprey spinal cord. Phasic activation of these cells by lateral bending can entrain the activity of the locomotor central pattern generator. During fictive locomotion induced by bath-applied NMDLA (N-methyl-D,L-aspartate) or sensory stimulation, edge cells receive synaptic input. In this paper we provide evidence that the tonic inhibition received during sensory-induced fictive locomotion is glycinergic. We also show that during fictive locomotion induced by application of NMDLA to the spinal cord, bicuculline unveils phasic synaptic activity in edge cells. During sensory-evoked fictive locomotion, by contrast, only tonic synaptic activity is apparent in the presence of bicuculline.     

Descending pathways to the spinal cord in the himé salmon (landlocked red salmon, Oncorhynchus nerka)

Distribution and morphology of the cells of origin of the descending spinal pathways and their axonal courses were studied in the himé salmon, using retrograde labelling with cobaltic lysine and horseradish peroxidase (HRP). Following application of the tracers to the cut end of the spinal cord or injection of the tracers at the 10th to 15th spinal segment, neurons mainly labelled via the axons of passage were distributed in the mesencephalon and the rhombencephalon. Mesencephalic cell groups consisted of the nucleus pretectalis, the nucleus fasciculi longitudinalis medialis, and the nucleus ruber. The former two cell groups sent their axons to the fasciculus longitudinalis medialis. The axons of the nucleus ruber formed a separate loose bundle, the "tractus rubrospinalis." The rhombencephalic cell groups consisted of the rhombencephalic reticular formation, the Mauthner cells (one cell for each side), and the octavolateral area. The rhombencephalic reticular formation could be further subdivided into the nucleus reticularis superior, nucleus reticularis medius, and nucleus reticularis inferior. The axons of these cell groups joined the fasciculus longitudinalis medialis and the "tractus bulbospinalis." The Mauthner cell had two main gigantic dendrites, and its giant axons formed a conspicuous fiber of Mauthner throughout the rhombencephalon down to the spinal cord. The octavolateral area could be subdivided into the nucleus vestibularis magnocellularis, nucleus tangentialis, nucleus vestibularis descendens and nucleus intermedius. The axons of the nucleus vestibularis magnocellularis and nucleus intermedius entered the fasciculus longitudinalis medialis and/or the tractus bulbospinalis. Those of the nucleus vestibularis descendens and nucleus tangentialis formed the "tractus vestibulospinalis". The descending spinal pathways of the himé salmon were compared with those of other fishes and other vertebrates. The significance of these descending spinal pathways in the control of locomotion and sexual behavior is also discussed.     

Structural and functional properties of reticulospinal neurons in the early-swimming stage Xenopus embryo

This study presents direct evidence that in Xenopus laevis embryos ipsi- and contralaterally descending reticulospinal fibers from the caudal brain stem project to the spinal cord, where they directly contact primary motoneurons. At stage 30, occasional contacts between primary motoneurons and descending axons are present. These contacts are possibly already functional since presynaptic vesicles were sometimes observed. Furthermore, the physiological data obtained in this study suggest that reticulospinal neurons in the caudal brain stem are involved in the central generation of early swimming. The first ingrowth of reticulospinal axons was observed in the rostral spinal cord after application of HRP to the caudal brain stem of stage 27/28 embryos. By stage 32, many supraspinal axons could be found in the spinal cord at the level of the 12/13th myotome, near the time of the first rhythmic swimming. Both lamellipodial and varicose growth cones were found. Intracellular recordings from the brain stem and extracellular recordings from the myotomal muscles in curarized embryos around stage 30 revealed neurons in the caudal brain stem which were active during early fictive swimming. After intracellular staining with Lucifer yellow neurons with descending axons were found in the brain-stem reticular formation. These reticulospinal neurons showed "motoneuron-like" phasic activity, producing one spike each swimming cycle. Rhythmically occurring spikes with swimming periodicity were superimposed on a sustained depolarization level of some 5-30 mV. Reticulospinal neurons in the brain stem resemble descending interneurons in the spinal cord by their morphology, projection pattern, and activity during early swimming. Reticulospinal neurons and descending interneurons might therefore form one continuous population of projecting interneurons with a different location but a similar function. In support of this we propose that the embryonic brain-stem reticular formation forms part of the swimming pattern generator.     

Rostro-caudal distribution of reticulospinal projections from different brainstem nuclei in the lamprey

The reticulospinal (RS) system in the lamprey is responsible for the control of locomotion, postural corrections and steering. To perform these functions, the RS system has to affect different muscular compartments along the body axis selectively. In this study, the possibility that RS neurones in different nuclei may project to different parts of the spinal cord, was investigated. The rostro-caudal extent of single RS axons was defined by stimulating them antidromically while recording from their cell body. All recorded mesencephalic RS neurones projected to the caudal tip of the spinal cord. Of the rhombencephalic RS neurones, 26% of the recorded neurones did not reach the caudalmost fourth of the spinal cord and this proportion varied between the anterior (18%), middle (17%) and posterior (36%) rhombencephalic reticular nuclei. For these RS axons, the level of termination covered the whole rostro-caudal extent of the spinal cord. No correlation was found between the length of an axon and its conduction velocity or between the length of an axon and the rostro-caudal position of its cell body in the nuclei.     

Development of early brainstem projections to the tail spinal cord of xenopus

Horseradish peroxidase (HRP) was used to determine the sequence in which axons from different brain neurons reach the tail spinal cord during embryonic and early larval development of Xenopus laevis. Brainstem cells of several classes project to the tail at these stages: mesencephalic reticulospinal neurons of the nucleus of the medial longitudinal fasciculus, a variety of other reticulospinal neurons, vestibulospinal neurons, and a group of median basal cells which may be raphe neurons. Among the reticulospinal neurons the paired Mauthner cells are the most prominent. They and caudally situated reticular neurons are the first to label with HRP applied to the tail spinal cord (stage 37). Vestibulospinal and other reticular neurons begin to label next (stage 39), followed by mesencephalic and then median basal neurons (stage 41). Except for the Mauthner cells, the number of labeled cells belonging to each neuron class increases gradually as development proceeds.     

Locomotion evoked by stimulation of the brain stem in the Atlantic stingray, Dasyatis sabina

The primary pathway descending to the spinal cord to initiate locomotion in the stingray is located in the intermediate to ventral portion of the lateral funiculus; a second pathway is located in the dorsolateral funiculus. The goal of this study was to identify the origins of these pathways in the rhombencephalic reticular formation (RF). To do this we used microstimulation of the RF in conjunction with selective lesions of the brain stem and spinal cord. In some animals microinjections of excitatory amino acids were used to avoid stimulating axons of passage. Locomotion in the contralateral pectoral fin was evoked by microstimulation of the dorsal and ventral reticular nuclei, the middle and superior RF, and the ventral portion of the lateral RF. The regions from which locomotion was evoked by chemical stimulation were more restricted and included the rostral dorsal reticular nucleus, the middle RF, and the adjacent ventral lateral RF. This area encompasses the magnocellular RF and coincides with the distribution of numerous reticulospinal cells that project ipsilaterally into the ventral half of the lateral funiculus. Our results indicate, then, that locomotion in the stingray is mediated primarily by a pathway originating in the magnocellular RF that descends ipsilaterally in the ventral half of the lateral funiculus to elicit swimming in the contralateral pectoral fin. We suggest that this primary pathway is specifically associated with the control of locomotion. We also demonstrated that locomotion can be evoked independently from the lateral RF, but is probably mediated by an indirect pathway relaying near the spinomedullary junction or in the rostral spinal cord.     

Initiation of locomotion in lampreys

The spinal circuitry underlying the generation of basic locomotor synergies has been described in substantial detail in lampreys and the cellular mechanisms have been identified. The initiation of locomotion, on the other hand, relies on supraspinal networks and the cellular mechanisms involved are only beginning to be understood. This review examines some of the findings relative to the neural mechanisms involved in the initiation of locomotion of lampreys. Locomotion can be elicited by sensory stimulation or by internal cues associated with fundamental needs of the animal such as food seeking, exploration, and mating. We have described mechanisms by which escape swimming is elicited in lampreys in response to mechanical skin stimulation. A rather simple neural connectivity is involved, including sensory and relay neurons, as well as the brainstem rhombencephalic reticulospinal cells, which act as command neurons. We have shown that reticulospinal cells have intrinsic membrane properties that allow them to transform a short duration sensory input into a long-lasting excitatory command that activates the spinal locomotor networks. These mechanisms constitute an important feature for the activation of escape swimming. Other sensory inputs can also elicit locomotion in lampreys. For instance, we have recently shown that olfactory signals evoke sustained depolarizations in reticulospinal neurons and chemical activation of the olfactory bulbs with local injections of glutamate induces fictive locomotion. The mechanisms by which internal cues initiate locomotion are less understood. Our research has focused on one particular locomotor center in the brainstem, the mesencephalic locomotor region (MLR). The MLR is believed to channel inputs from many brain regions to generate goal-directed locomotion. It activates reticulospinal cells to elicit locomotor output in a graded fashion contrary to escape locomotor bouts, which are all-or-none. MLR inputs to reticulospinal cells use both glutamatergic and cholinergic transmission; nicotinic receptors on reticulospinal cells are involved. MLR excitatory inputs to reticulospinal cells in the middle (MRRN) are larger than those in the posterior rhombencephalic reticular nucleus (PRRN). Moreover at low stimulation strength, reticulospinal cells in the MRRN are activated first, whereas those in the PRRN require stronger stimulation strengths. The output from the MLR on one side activates reticulospinal neurons on both sides in a highly symmetrical fashion. This could account for the symmetrical bilateral locomotor output evoked during unilateral stimulation of the MLR in all animal species tested to date. Interestingly, muscarinic receptor activation reduces sensory inputs to reticulospinal neurons and, under natural conditions, the activation of MLR cholinergic neurons will likely reduce sensory inflow. Moreover, exposing the brainstem to muscarinic agonists generates sustained recurring depolarizations in reticulospinal neurons through pre-reticular effects. Cells in the caudal half of the rhombencephalon appear to be involved and we propose that the activation of these muscarinoceptive cells could provide additional excitation to reticulospinal cells when the MLR is activated under natural conditions. One important question relates to sources of inputs to the MLR. We found that substance P excites the MLR, whereas GABA inputs tonically maintain the MLR inhibited and removal of this inhibition initiates locomotion. Other locomotor centers exist such as a region in the ventral thalamus projecting directly to reticulospinal cells. This region, referred to as the diencephalic locomotor region, receives inputs from several areas in the forebrain and is likely important for goal-directed locomotion. In summary, this review focuses on the most recent findings relative to initiation of lamprey locomotion in response to sensory and internal cues in lampreys.     

Spino-bulbar neurons convey information to the brainstem about different phases of the locomotor cycle in the lamprey

Lamprey retriculospinal neurons show phasic oscillations of their membrane potential during fictive locomotion. This modulation originates from the spinal cord locomotor networks. The aim of the present study was to elucidate the pattern of discharge of the spino-bulbar axons responsible for this modulation. Experiments were performed on in vitro brainstem/spinal cord preparations. Two baths were formed in the recording chamber. The caudal one was perfused with 150 microM N-methyl-D-aspartate (NMDA) solution to induce fictive locomotion. The rostral bath containing the brain and the first 3-5 segments of the spinal cord was exposed to a 0 Ca2+ + 2.6 mM Mn2+ solution to block synaptic transmission and therefore to abolish any rhythmic descending activity. Spinobulbar axons were recorded intracellularly at the level of the brain/spinal cord junction. They exhibited phasic discharges correlated with the ongoing motor activity in the caudal pool. Some discharged in phase with either the ipsilateral or the contralateral ventral root bursts, others with either of the transition phases between these two bursts. These spinal cells with ascending axons, running in the ventrolateral spinal cord, may be important for modulating the activity of supraspinal neurons to match the ongoing locomotor activity.     

Spinal inputs from lateral columns to reticulospinal neurons in lampreys

This study characterizes the inputs from the lateral columns of the spinal cord to reticulospinal neurons in the lampreys, using the in vitro isolated brainstem and spinal cord preparation. Synaptic responses to the electrical stimulation of the lateral columns were recorded in reticulospinal neurons of the posterior and middle rhombencephalic reticular nuclei. The responses consisted of a mixture of excitation and inhibition. They were markedly potentiated when using trains of two to five pulses, suggesting that the larger part of these synaptic responses was mediated via an oligosynaptic pathway. An early component, however, persisted when using twin pulses at 10–20 Hz on the ipsilateral side, suggesting the presence of an early mono- or disynaptic component. When increasing the stimulation strength, an early fast rising excitatory component appeared. It most likely resulted from an antidromic activation of vestibulospinal axons in the lateral tracts, which make en passant synaptic contacts with reticulospinal neurons. Responses were practically abolished by adding CNQX and AP5 to the Ringer's solution. The late component of excitatory responses was decreased by AP5, suggesting that NMDA receptors were activated. The NMDA receptor-mediated component was larger when using trains of stimuli or in Mg²⁺-free Ringer's. The application of NMDA depolarized reticulospinal neurons. The glycinergic inhibitory component was markedly increased in Mg²⁺-free Ringer's. Moreover, GABA_B-receptor activation with (−)-baclofen abolished both excitatory and inhibitory responses. Taken together, the present results indicate that ascending lateral column axons generate large excitatory and inhibitory synaptic potentials in reticulospinal neurons. The possible role of these inputs in modulating the activity of reticulospinal neurons during locomotion is discussed.     

Spinal interneuronal networks in the cat: elementary components

This review summarises features of networks of commissural interneurones co-ordinating muscle activity on both sides of the body as an example of feline elementary spinal interneuronal networks. The main feature of these elementary networks is that they are interconnected and incorporated into more complex networks as their building blocks. Links between networks of commissural interneurones and other networks are quite direct, with mono- and disynaptic input from the reticulospinal and vestibulospinal neurones, disynaptic from the contralateral and ipsilateral corticospinal neurones and fastigial neurones, di- or oligosynaptic from the mesencephalic locomotor region and mono-, di- or oligosynaptic from muscle afferents. The most direct links between commissural interneurones and motoneurones are likewise simple: monosynaptic and disynaptic via premotor interneurones with input from muscle afferents. By such connections, a particular elementary interneuronal network may subserve a wide range of movements, from simple reflex and postural adjustments to complex centrally initiated phasic and rhythmic movements, including voluntary movements and locomotion. Other common features of the commissural and other interneuronal networks investigated so far is that input from several sources is distributed to their constituent neurones in a semi-random fashion and that there are several possibilities of interactions between neurones both within and between various populations. Neurones of a particular elementary network are located at well-defined sites but intermixed with neurones of other networks and distributed over considerable lengths of the spinal cord, which precludes the topography to be used as their distinguishing feature.     

Stimulation of the mesencephalic locomotor region elicits controlled swimming in semi-intact lampreys

The role of the mesencephalic locomotor region (MLR) in initiating and controlling the power of swimming was studied in semi-intact preparations of larval and adult sea lampreys. The brain and the rostral portion of the spinal cord were exposed in vitro, while the intact caudal two-thirds of the body swam freely in the Ringer's-containing chamber. Electrical microstimulation (2-10 Hz; 0. 1-5.0 microA) within a small periventricular region in the caudal mesencephalon elicited well-coordinated and controlled swimming that began within a few seconds after the onset of stimulation and lasted throughout the stimulation period. Swimming stopped several seconds after the end of stimulation. The power of swimming, expressed by the strength of the muscle contractions and the frequency and the amplitude of the lateral displacement of the body or tail, increased as the intensity or frequency of the stimulating current were increased. Micro-injection of AMPA, an excitatory amino acid agonist, into the MLR also elicited active swimming. Electrical stimulation of the MLR elicited large EPSPs in reticulospinal neurons (RS) of the middle rhombencephalic reticular nucleus (MRRN), which also displayed rhythmic activity during swimming. The retrograde tracer cobalt-lysine was injected into the MRRN and neurons (dia. 10-20 microm) were labelled in the MLR, indicating that this region projects to the rhombencephalic reticular formation. Taken together, the present results indicate that, as higher vertebrates, lampreys possess a specific mesencephalic region that controls locomotion, and the effects onto the spinal cord are relayed by brainstem RS neurons.     

Vibration-evoked startle behavior in larval lampreys

Larval lampreys (ammocoetes) exhibit a rapid vibration-evoked startle response involving a bilateral activation of musculature along the length of the body. The resulting movement is variable, contingent on the animal's prestimulus posture: lateral curves along the trunk and tail contract more on the inner side of the curve than on the outer side. Thus, the startle response increases preexisting body curvature. Because ammocoetes are burrowing filter feeders, this startle behavior results in rapid withdrawal of the head into the burrow. A vibratory pulse to the otic capsules in a semi-intact preparation evokes simultaneous action potentials in both primary Mauthner neurons. Vibration also excites several Müller cells. Intracellular stimulation of one primary Mauthner axon (eliciting one action potential) produces bilateral trunk electromyographic potentials that are smaller than those evoked by vibration; simultaneous stimulation of both Mauthner axons (one action potential each) reproduces the vibration-evoked electromyographic amplitudes. The Mauthner cell's sensitivity to vestibular input is centrally modulated during changes in behavioral state. Mauthner action potentials are most easily elicited by vibratory or electrical stimulation of vestibular afferents while an intact animal is at rest; the same stimuli become subthreshold for Mauthner activity while the animal is swimming. A similar depression of Mauthner excitability is observed in semi-intact preparations during arousal. 'Arousal' was defined by the occurrence of tonic, descending spinal cord discharge. Mauthner cells are tonically depolarized during arousal and exhibit an increased membrane conductance; excitatory postsynaptic potentials evoked by vibratory or electrical stimulation of vestibular afferents are greatly attenuated. Modulated sensory transmission to the Mauthner cell may help to prevent inappropriate activation of the startle circuit.