The action of L-DOPA on the spontaneous activity generated by the isolated spinal cord of 16- to 20-day-old chick embryos]

Effect of bath application of L-DOPA, dopamine (DA), noradrenaline (NA) on the spontaneous activity--cyclical oscillations of electrotonic potentials in dorsal and ventral roots (DR and VR, respectively) generated by isolated spinal cord of 16-20-day-old chick embryo was studied. L-DOPA in low concentration (30-150 mumol/l) activated spinal generator: suprathreshold rhythms appeared (i.e., spike activity in DR and VR). High concentration (200-1000 mumol/l) of L-DOPA depressed generator activity, but in the course of washing of the spinal cord spontaneous activity was enhanced and suprathreshold rhythms appeared. DA and NA (50 mumol/l) did not influence the activity but in concentration 100 mumol/l they depressed spontaneous activity in DR and VR. Bath application of 2-amino-5-phosphonovaleric acid (20 mumol/l) blocked activity induced by small concentration of L-DOPA. Activity generated by the neuronal network of the isolated dorsal horn was intensified after bath-application of L-DOPA in small concentration. Rhythmic activity in the isolated ventral horn did not appear after bath-application of L-DOPA. It is concluded that excitatory action of L-DOPA on the neuronal network of dorsal horn chick embryo was direct (i.e., did not depend on catecholamines).     

The modification of the cyclic activity of the isolated spinal cord in l6- to 20-day-old chick embryos through changes in the Ca2+ and Mg2+ concentrations]

Spontaneous rhythmic activity which appeared as synchronous oscillations of electrotonic potentials in dorsal and ventral roots (DR and VR, respectively) of isolated spinal cord sections of 16-20-day-old chick embryos was completely blocked with a rise of Mg2+ concentration from the control level (1.3 mmol/l) to 5 mmol/l. The similar effect was observed when Ca2+ concentration fell from 2.6 to 1 mmol/l, but in that case spike discharges could appear in VR. Moreover, during disappearance of the above spontaneous activity (after changes in Mg2+ or Ca2+ concentration) rhythmic activity of other type was observed in 6 from 8 experiments. That rhythmic activity consisted of oscillations of electrotonic VR potentials (amplitude to 200 microV and duration to 400 ms) which arose irrespective of oscillations of DR potentials. Maximum frequency of that activity was 0.4/s. Possible functional role of the found types of rhythmic activity is discussed.     

Cholinergic modulation of synaptic transmission in the spinal cord of the frog

Superfusion of the isolated frog spinal cord by the Ringer solution containing arecoline (10 mumol/l) evoked depolarization and increase of the input resistance and PSP amplitude of motoneurons. Depolarizing electrotonic potentials and reflex discharges in the ventral roots also increased, but duration of dorsal root potentials decreased. The observed arecoline facilitation of synaptic transmission in the spinal cord has postsynaptic nature evoked by motoneuron M2-cholinoreceptor activation and bound to an increase of the cyclic nucleotide metabolism. Arecoline inhibited the synaptic transmission in the spinal cord under conditions when its postsynaptic action was eliminated. This effect was due to presynaptic M1-cholinoreceptor activation without changing the cyclic nucleotide metabolism.     

The influence of bemegride on frog spinal cord root potentials]

The effect of the convulsant bemegride (beta-ethyl-beta-methyl-glutharimide) on the electronic potentials in the frog spinal roots was investigated in situ. Intravenous injection of the bemegride subconvulsant doses (6.8 +/- 2.7 mg/kg) depressed rapidly the electrotonic dorsal root potentials (DRP) evoked by a stimulation of the adjacent dorsal root (DR) or the ventral root (VR). They were reduced by 55--67% 3-6 min after the bemegride injection. The effect of bemegride was reversible and the DRP amplitude restored its initial value within an hour. Ventral root potentials after bemegride injection revealed only greater amplitude fluctuations. A conclusion is made that bemegride is a selective and potent depressor of the primary afferent depolarization in the frog spinal cord.     

Spinal dorsal horn neurons in elevated extracellular calcium: cell properties and spontaneous discharges

Recordings were made from neurons in the dorsal horn (DH), and from dorsal and ventral roots (DRs and VRs) of isolated spinal cords of infant mice. Raising calcium concentration ([Ca2+]) in the organ bath from 1.2 to 2.4 mmol/l resulted in a slight hyperpolarization, elevation of threshold current (rheobase), and augmentation of excitatory postsynaptic potentials (EPSPs). In many cells EPSPs acquired a much prolonged late phase. Orthodromic stimulation evoked in some DH neurons an action potential that had the same threshold as, and coincided in time with, the 'dorsal horn response' (DHR) recorded from DR. In spinal cords bathed in elevated [Ca2+], DR recordings showed irregularly recurring spontaneous waves, and DH neurons generated spontaneous EPSPs, often with spikes. Some neurons fired irregularly timed spontaneous action potentials that did not appear triggered by EPSPs. In less than 50% of the neurons the spontaneous EPSPs coincided in time with the spontaneous DR waves. The action potentials that appeared without EPSP were fired independently from DR activity. These observations confirm that elevation of interstitial free calcium concentration results in strong enhancement of excitatory transmission, especially of an EPSP of much extended duration. Virtually all neurons showed increased spontaneous activity in high [Ca2+], but only a minority appeared recruited into the synchronized discharges that are detectable as spontaneous waves in DR and VR recordings.     

Inhibitory control of the urinary bladder in the neonatal rat in vitro spinal cord-bladder preparation

Urinary bladder activity of the neonatal rat is tonically inhibited by neural input from the spinal cord passing through axons in the pelvic nerve. The present study was undertaken to examine the organization of this inhibitory mechanism using in vitro spinal cord-bladder preparations of neonatal rats in which the lumbosacral dorsal roots (DRs) or ventral roots (VRs) were transected. Isovolumetric bladder contractions occurring spontaneously or induced by electrical stimulation of the bladder wall (ES-BW) were measured. In DR transected (DRT) preparations, removal of the spinal cord significantly enhanced (50-59%) the amplitude of spontaneous and ES-BW-evoked bladder contractions; whereas in VR transected (VRT) preparations removal of the spinal cord produced only a small enhancement (6.7-12%). However, in VRT preparations, electrical stimulation of the dorsal roots reduced the amplitude of spontaneous contractions, an effect blocked by a nicotinic ganglionic blocking agent, hexamethonium. In DRT preparations, MK-801 enhanced the amplitude of spontaneous and ES-BW-evoked contractions. These results demonstrate that bladder activity of the neonatal rat is tonically inhibited by input from the lumbosacral spinal cord via parasympathetic pathways in the pelvic nerve. The inhibitory outflow is not dependent upon afferent input to the cord but is facilitated by NMDA glutamatergic transmission in the spinal cord. Antidromic activation of afferent axons also appears to induce inhibition in the bladder via a mechanism involving nicotinic cholinergic receptors. These findings suggest that spinal and peripheral inhibitory mechanisms may play an important role in controlling voiding in the neonatal rat.     

The Influence of Magnesium Ions on the NMDA Mediated Responses of Ventral Rhythmic Neurons in the Spinal Cord of Xenopus Embryos

Xenopus embryos immobilized in tubocurarine respond to natural skin stimulation with fictive swimming. This can also occur in saline without Mg2+ and is blocked by NMDA antagonists. Ventral spinal cord neurons which are rhythmically active during swimming are depolarized by bath applied N-methyl-D-aspartate (NMDA) (in 1 microM tetrodotoxin (TTX) to block indirect effects). By using current clamp techniques this depolarization is shown to be partially blocked by 0.5 and 1 mM Mg2+ in a voltage-dependent manner similar to that described in cultured neurons. Mg2+ partially and reversibly reduces the slow NMDA-mediated component of excitatory post-synaptic potentials (EPSPs) in ventral neurons. However, in 1 mM Mg2+ fictive swimming can still be evoked by natural stimulation. The frequency of swimming is slightly lower than in nominally 0 mM Mg2+, but the pattern of ventral root activity and synaptic drive to ventral neurons seems little affected. Fictive swimming can also be induced by applying NMDA to spinal preparations. In 0 mM Mg2+, such rhythmic activity is unstable and transient over a narrow NMDA concentration range. In 0.5 mM Mg2+, continuous rhythmic activity is induced over a wide range of NMDA concentrations. Lower spinal preparations need higher NMDA concentrations to induce activity. We conclude that the neurons rhythmically active in swimming have NMDA receptor channels which show a voltage dependent block in the presence of Mg2+. However, while Mg2+ exerts a powerful stabilizing influence on rhythmic activity induced in spinal embryos by exogenous NMDA, its influence on 'naturally' evoked fictive swimming is less clear. The fictive swimming machinery in the brain and spinal cord can produce stable swimming with or without Mg2+ induced voltage dependency of the NMDA channels.     

Long-range oscillatory Ca2+ waves in rat spinal dorsal horn

Synchronous activity of large populations of neurons shapes neuronal networks during development. However, re-emergence of such activity at later stages of development could severely disrupt the orderly processing of sensory information, e.g. in the spinal dorsal horn. We used Ca2+ imaging in spinal cord slices of neonatal and young rats to assess under which conditions synchronous activity occurs in dorsal horn. No spontaneous synchronous Ca2+ transients were detected. However, increasing neuronal excitability by application of 4-aminopyridine after pretreatment of the slice with blockers of (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate, gamma-aminobutyric acid (GABA)(A) and glycine receptors evoked repetitive Ca2+ waves in dorsal horn. These waves spread mediolaterally with a speed of 1.0 +/- 0.1 mm/s and affected virtually every dorsal horn neuron. The Ca2+ waves were associated with large depolarizing shifts of the membrane potential of participating neurons and were most likely synaptically mediated because they were abolished by blockade of action potentials or N-methyl-D-aspartate (NMDA) receptors. They were most pronounced in the superficial dorsal horn and absent from the ventral horn. A significant proportion of the Ca2+ waves spread to the contralateral dorsal horn. This seemed to be enabled by disinhibition as primary afferent-induced dorsal horn excitation crossed the midline only when GABA(A) and glycine receptors were blocked. Interestingly, the Ca2+ waves occurred under conditions where AMPA/kainate receptors were blocked. Thus, superficial dorsal horn NMDA receptors are able to sustain synchronous neuronal excitation in the absence of functional AMPA/kainate receptors.     

Mechanisms of pattern generation in co-cultures of embryonic spinal cord and skeletal muscle

Spontaneous output patterns of embryonic spinal cord slices in vitro were investigated in order to study the formation of pattern-generating networks. Patterns of spontaneous contractions of skeletal muscle fibers were recorded in co-cultures of embryonic rat spinal cord, dorsal root ganglia and skeletal muscle. A part of these contractions was shown to be driven by spinal circuits. These neuron-driven activity patterns changed from random to rhythmic when the inhibitory synapses in the spinal cord were blocked by strychnine, bicuculline or both. Rhythmic patterns consisted of bursts of activity (tetanic contractions) followed by periods of relaxation. The transition from random to rhythmic patterns occurred during a period of heavily increased rate of activity. Presynaptic inhibition was not involved critically in the generation of rhythmic patterns. Such patterns were, however, modulated through muscarinic and α-adrenergic receptors. Neither NMDA nor glutamate nor its uptake blocker dihydrokainate induced rhythmic patterns of contraction, although NMDA in the presence of low magnesium increased moderately the rate of random activity. In order to study the size of pattern-generating networks, parts of the spinal cord slices were sectioned during rhythmic activity. Tangential cuts at the lateral or dorsal side of the slices reduced either the rate or the duration of the bursts or both. Sagittal cuts suppressed the activity almost totally. These findings suggest that the pattern generators in the slices consist of excitatory networks covering the entire slice, and that these networks reverberate following spontaneous activity of some distributed elements.     

Changes in the electrical activity of spinal cord neurons of adrenalectomized rats under the effect of adrenal cortex hormones

Effect of dexamethasone and desoxycorticosterone on the electrical activity of neurons in dorsal and ventral horn of spinal cord evoked by sciatic nerve stimulation were studied in adrenalectomized rats as well as effect of the same hormones on the background activity of single cells in the dorsal horn. The results demonstrated that both hormones (dexamethasone and desoxycorticosterone) provided enhancement of the amplitude of the field potentials recorded from the dorsal half of the spinal cord and facilitation of the background neuronal discharges of the single cells under investigation. It was stated that gluco- and mineralocorticoid hormones exerted different effects on the activity of ventral horn neurons of the spinal cord: dexamethasone++ potentiated and desoxycorticosterone depressed the amplitudes of the field potentials recorded from the region of motoneurons. The presented data have shown the modulatory effects of neurosteroids on the electrical activity of the spinal cord neurons.     

The role of putative excitatory amino acid neurotransmitters in the initiation of locomotion in the lamprey spinal cord. I. The effects of excitatory amino acid antagonists

The activation of N-methyl-D-aspartate (NMDA) and kainate receptors will evoke fictive locomotion in the appropriate motor pattern for locomotion in the isolated lamprey spinal cord, but not a selective activation of quisqualate receptors. The present experiments test whether the initiation of locomotion in response to sensory stimulation depends on these types of receptors. An in vitro preparation of the lamprey spinal cord with part of its tailfin left innervated has been used. In this preparation a sequence of fictive locomotion (i.e. alternating bursts in the segmental ventral roots with a rostrocaudal phase lag) can be elicited by continual sensory stimulation of the tailfin. The effects of excitatory amino acid antagonists were studied by recordings from ventral roots (extracellularly) and motoneurones (intracellularly). It was found that the strong initial bursts of each swimming sequence induced by sensory stimulation were depressed by combined NMDA/kainate antagonists (cis-2,3-piperidine dicarboxylate (PDA) and gamma-D-glutamylglycine (gamma-DGG] whereas the less intense burst activity, occurring particularly towards the end of each swimming sequence, was depressed by a selective NMDA antagonist, 2-amino-5-phosphonovalerate (2-APV). This condition could be mimicked in an isolated spinal cord preparation by an application of L-glutamate; the low-level fictive locomotion induced by low doses of L-Glu (less than 100 microM) was depressed by a NMDA antagonist (2-APV), and, if higher doses were applied, the activity was only depressed by PDA/gamma-DGG. The mode and time course of the depression (by excitatory amino acid antagonists) of fictive locomotion, induced by sensory stimulation, shows that the putative excitatory amino acid neurotransmitter directly or indirectly acts at the pattern generating circuitry within the spinal cord.     

Multiple-site optical recording reveals embryonic organization of synaptic networks in the chick spinal cord

We examined embryonic expression of postsynaptic potentials in stages 26-31 (E5 to E7) chick spinal cord slices. Slow optical signals related to the postsynaptic potentials which were evoked by electrical stimulation of afferent fibers were identified in the dorsal grey matter and the ventral motoneuronal area. In cervical spinal cord (C13) preparations, the dorsal slow signal appeared from stage 28 (E6), whilst the ventral slow signal was recognized from stage 29. At stages 26 and 27 (E5), no slow signal was observed in either the dorsal or ventral regions. On the other hand, in lumbosacral spinal cord (LS5) preparations, the dorsal, as well as ventral, slow signals appeared from stage 29; at stage 28 no slow signal was detected in the dorsal or ventral regions. These results suggest that there are differences in the ontogenetic expression of synaptic functions between the dorsal and ventral regions, and between the cervical and lumbosacral spinal cords. In embryos older than stage 29, removal of Mg2+ from the bathing solution markedly enhanced the amplitude and incidence of the ventral slow signal. In addition, in C13 preparations at stage 28, removal of Mg2+ elicited small slow signals in the ventral region in which no synaptic response was evoked in normal Ringer's solution. The slow signals induced in the Mg2+-free solution were blocked by 2-amino-5-phosphonovaleric acid (APV), showing that they are attributable to N-methyl- D-aspartate (NMDA) receptors. These results suggest that functional synaptic connections via polysynaptic pathways are already generated on motoneurons, but are suppressed by a Mg2+ block on the NMDA receptors at developmental stages when synaptic transmission from the primary afferents to the dorsal interneurons is initially expressed in the dorsal region.     

The effects of bicuculline on the isolated spinal cord of the frog

An investigation has been made of the effects of topically applied bicuculline, a reported gamma-aminobutyric acid (GABA) antagonist, on the isolated, hemisected frog spinal cord by recording ventral and dorsal root potentials and reflexes evoked by volleys to various spinal cord inputs. Bicuculline had potent excitatory effects causing depolarization, spontaneous potentials in ventral and dorsal roots, and an increased polysynaptic ventral root reflex. More importantly, the alkaloid blocked presynaptic inhibition of orthodromic reflex activity produced by preceding ventral root stimulation and primary afferent depolarization. These effects were attributed to a demonstrated antagonism of the direct depolarizing effects of GABA on dorsal root terminals by the alkaloid. These actions of bicuculline suggest that GABA may be the transmitter responsible for primary afferent depolarization and presynaptic inhibition in the amphibian.     

The effects of human myelin basic protein (HMBP) on the bioelectrical activity of the frog spinal cord

The effects of human myelin basic protein (HMBP) and its individual microheterogeneous components on the electrical activity of the frog spinal cord are compared with those of bovine MBP (BMBP) and glutamate (Glu). HMBP causes a dose-dependent depolarization of the ventral (VR) and dorsal (DR) roots with a reduction of the evoked root potentials. The amplitude of the depolarization induced by 0.01 mM HMBP corresponds to that observed with application of 1 mM Glu. A weaker depolarization persists during blockade of synaptic transmission by MgSO₄ or tetrodotoxin, suggesting a direct depolarizing effect on motoneurones and primary afferents. HMBP causes a long-lasting tachyphylaxis. The individual microheterogeneous components of HMBP (peaks 1, 2 and 3), and a mixture of smaller myelin basic proteins elicit similar responses, whereas BMBP and its microheterogeneous components are slightly less active.     

Spontaneous network activity transiently depresses synaptic transmission in the embryonic chick spinal cord.

We examined the effects of spontaneous or evoked episodes of rhythmic activity on synaptic transmission in several spinal pathways of embryonic day 9-12 chick embryos. We compared the amplitude of synaptic potentials evoked by stimulation of the ventrolateral funiculus (VLF), the dorsal or ventral roots, before and after episodes of activity. With the exception of the short-latency responses evoked by dorsal root stimulation, the potentials were briefly potentiated and then reduced for several minutes after an episode of rhythmic activity. Their amplitude progressively recovered in the interval between successive episodes. The lack of post-episode depression in the short-latency component of the dorsal root evoked responses is probably attributable to the absence of firing in cut muscle afferents during an episode of activity. The post-episode depression of VLF-evoked potentials was mimicked by prolonged stimulation of the VLF, subthreshold for an episode of activity. By contrast, antidromically induced motoneuron firing and the accompanying calcium entry did not depress VLF-evoked potentials recorded from the stimulated ventral root. In addition, post-episode depression of VLF-evoked synaptic currents was observed in voltage-clamped spinal neurons. Collectively, these findings suggest that somatic postsynaptic activity and calcium entry are not required for the depression. We propose instead that the mechanism may involve a form of long-lasting activity-induced synaptic depression, possibly a combination of transmitter depletion and ligand-induced changes in the postsynaptic current accompanying transmitter release. This activity-dependent depression appears to be an important mechanism underlying the occurrence of spontaneous activity in developing spinal networks.     

Network bursting by organotypic spinal slice cultures in the presence of bicuculline and/or strychnine is developmentally regulated

Organotypic cocultures of dorsal root ganglia and spinal cord from embryonic rats provides direct access to spinal interneurons in a culture system in which the cytoarchitectural organization of the spinal cord slice is maintained. This preparation was used to investigate the possible induction of rhythmic behaviour at different times of development in vitro. Spontaneous rhythmic bursts induced by coapplication of strychnine (1 μm ) and bicuculline (20 μm ) were observed with patch-clamp recordings from ventral interneurons. Ventral horn interneurons consistently developed a very regular pattern of activity which was superimposed on a background of sustained synaptic activity. The pattern of the spontaneous bursting following application of strychnine and bicuculline showed a developmentally regulated difference in frequency between two distinct stages of in vitro development.     

NR2B receptors are involved in the mediation of spinal segmental reflex potentials but not in the cumulative motoneuronal depolarization in vitro

Windup, the frequency dependent build-up of spinal neuronal responses is an electrophysiological model of the development of the central sensitization in the chronic pain states. NR2B subunit containing NMDA-type glutamate receptors are implicated in the windup of dorsal horn neurons, while their role at the motoneuronal level is controversial. The cumulative motoneuronal depolarization in hemisected rat spinal cord preparation is an in vitro model of windup. The role of NR2B receptors in this process, and in the mediation of dorsal root stimulation evoked ventral root reflex potentials was elucidated. Three selective NR2B antagonists; CP-101,606; CI-1041 and Co-101244 (1 microM) were used. They had only weak, but statistically significant inhibitory effect on the early part of ventral root response, and did not influence the cumulative depolarization. On the contrary, non-selective NMDA antagonist APV (40 microM) decreased both responses markedly. We conclude that the pharmacological sensitivities of windup at the sensory and motor levels are different. NR2B containing NMDA receptors have major role in the mediation of the windup of dorsal horn neurons, but their contribution to this phenomenon at the motor level is negligible.     

Formation of the central pattern generator for locomotion in the rat and mouse

It is well known that in the neonatal rat spinal cord preparation, alternating rhythmic bursts in the left and right ventral roots in a given lumbar segment can be induced by bath-application of N-methyl-D-aspartate or 5-hydroxytryptamine. Alternation between L2 and L5 ventral roots on the same side, representing the activity of flexor and extensor muscles, respectively, can be observed as well. In the prenatal period in the rat, alternation between the left and right ventral roots is established between embryonic day (E) 16.5 and E18.5. The alternation between the L2 and L5 ventral roots emerges at E20.5. Recent findings show that locomotor-like rhythmic activity with similar characteristics can be induced in the neonatal mouse preparation. In the lumbar spinal cord in the neonatal mouse, it is likely that the rhythm-generating network is distributed throughout the lumbar region with a rostro-caudal gradient, a situation similar to that in the neonatal and fetal rat spinal cord. With this review we hope to highlight the dramatic changes that neuronal networks generating locomotor-like activity undergo during the prenatal development of the rat. Moreover, the distribution of the neuronal network generating the locomotor rhythm in the neonatal rat and mouse spinal cord is compared.     

Influence of growth medium, age in vitro and spontaneous bioelectric activity on the distribution of sensory ganglion-evoked activity in spinal cord explants

The role of serum added to the culture medium and of spontaneous bioelectric activity in the development of sensory afferent connections was studied, employing fetal mouse spinal cord explants with attached dorsal root ganglia (DRG) as an in vitro model system. Afferent DRG terminals in the cord explants were localized on the basis of 'fixed-latency' DRG-evoked action potentials, which were anatomically verified in several experiments using horseradish peroxidase histology. In serum-supplemented medium (HSSM), but not in chemically defined medium (CDM), those DRG fibers which grew into the dorsal side of the cord terminated predominantly within the dorsal cord region, and remained there throughout the experimental period (18-33 days in vitro). In contrast, ventrally entering fibers terminated equally in both the dorsal and the ventral cord regions in young cultures (18-24 days in vitro) but were no longer observed after 27 days in vitro. Cultures grown in HSSM with the addition of xylocaine, in order to chronically suppress spontaneous bioelectric activity, essentially corresponded (at 25-32 days in vitro) to the picture seen in the control series at the same age. On the basis of polysynaptic DRG-evoked responses in the cord, developmental changes in local neuronal networks were inferred which resulted in less spread of DRG-evoked activity with age in HSSM, and more spread with age in CDM-grown cultures. It is concluded that for the formation of selective DRG connections in the spinal cord: (i) a serum-borne factor plays a role: and (ii) functional activity is not required.     

Forelimb locomotor generators and quadrupedal locomotion in the neonatal rat

The spinal localization of the forelimb locomotor generators and their interactions with other spinal segments were investigated on in vitro brainstem-spinal cord preparations of new-born rats. Superfusion of the cervicothoracic cord (C1-T4) with high K+/low Mg2+ artificial cerebrospinal fluid (aCSF) evoked rhythmic motor root activity that was limited to low cervical (C7, C8) and high thoracic (T1) spinal levels. This activity consisted of synchronous, homolateral bursts and a typical alternating bilateral pattern. Rhythmic activity with similar locomotor-like characteristics could be induced with either serotonin (5-HT, 5 microm), N-methyl-d-aspartate (NMDA, 5 microm), kainate (10 microm) or a "cocktail" of 5-HT (5 microm) and NMDA (5 microm). During 5-HT/NMDA perfusion of the cervicothoracic cord, induced bursting was no longer restricted to C7-T1 levels, but also occurred at cervical C3-C5 levels and with C5-C8 homolateral alternation. Spinal transections between C6 and C7 cervical segments did not abolish rhythmic activity in C7-T1, but suppressed locomotor-like rhythmicity at C3-C5 levels. Reduced regions comprising the C7-C8 or C8-T1 segments maintained rhythmicity. Superfusion of the whole cord with 5-HT/NMDA induced ventral root bursting with similar frequencies at all recorded segments (cervical, thoracic and lumbar). After isolation, the T3-T10 cord was unable to sustain any rhythmic activity while cervical and lumbar segmental levels continued to burst, albeit at different frequencies. We also found that the faster caudal and the slower rostral locomotor generators interact to produce coordinated locomotor-like activity in all segments of the intact spinal cord. In conclusion, C7-T1 spinal levels display a strong motor rhythmogenic ability; with the lumbar generators, they contribute to coordinated rhythmic activity along the entire spinal cord of a quadrupedal locomoting mammal.