Activity-dependent metaplasticity of inhibitory and excitatory synaptic transmission in the lamprey spinal cord locomotor network.

Paired intracellular recordings have been used to examine the activity-dependent plasticity and neuromodulator-induced metaplasticity of synaptic inputs from identified inhibitory and excitatory interneurons in the lamprey spinal cord. Trains of spikes at 5-20 Hz were used to mimic the frequency of spiking that occurs in network interneurons during NMDA or brainstem-evoked locomotor activity. Inputs from inhibitory and excitatory interneurons exhibited similar activity-dependent changes, with synaptic depression developing during the spike train. The level of depression reached was greater with lower stimulation frequencies. Significant activity-dependent depression of inputs from excitatory interneurons and inhibitory crossed caudal interneurons, which are central elements in the patterning of network activity, usually developed between the fifth and tenth spikes in the train. Because these interneurons typically fire bursts of up to five spikes during locomotor activity, this activity-dependent plasticity will presumably not contribute to the patterning of network activity. However, in the presence of the neuromodulators substance P and 5-HT, significant activity-dependent metaplasticity of these inputs developed over the first five spikes in the train. Substance P induced significant activity-dependent depression of inhibitory but potentiation of excitatory interneuron inputs, whereas 5-HT induced significant activity-dependent potentiation of both inhibitory and excitatory interneuron inputs. Because these metaplastic effects are consistent with the substance P and 5-HT-induced modulation of the network output, activity-dependent metaplasticity could be a potential mechanism underlying the coordination and modulation of rhythmic network activity.     

Endogenous activation of glycine and NMDA receptors in lamprey spinal cord during fictive locomotion

Strychnine is shown to abolish left-right alternation in fictive locomotion induced by sensory stimulation. Robust rhythmic activity, characterized by left-right coactivation at each segmental level, is seen in the presence of strychnine at all doses used (0.5-20 microM). The proportion of the cycle occupied by the ventral root bursts and the rostral-caudal coordination is similar to that seen in the absence of strychnine. Furthermore, the rhythm is abolished by cis-2,3-piperidine dicarboxylic acid (PDA), 2-amino-5-phosphonovalerate (APV), or the removal of Mg2+ from the perfusate, as in the absence of strychnine. Voltage clamp was applied to ventral horn neurons during stimulation in the presence of strychnine, with holding potentials negative to the plateau potential associated with a ventral root burst but positive to the potential in the interburst. Inward current was seen during the ventral root burst, but no outward current was seen at burst termination or during the interburst. The results indicate that in fictive locomotion induced by endogenous release of NMDA receptor agonists, left-right alternation is dependent on glycinergic transmission. Furthermore, evidence is provided that in the absence of glycinergic transmission, burst termination may depend on NMDA receptor-linked voltage-sensitive processes.     

The activity-dependent plasticity of segmental and intersegmental synaptic connections in the lamprey spinal cord

Activity-dependent synaptic plasticity has been proposed as a contributory factor in the patterning of rhythmic network activity. However, its role has not been examined in detail. Here, paired or triple intracellular recordings have been made from identified neurons in the lamprey locomotor network to examine the potential relevance of activity-dependent synaptic plasticity in segmental and intersegmental spinal networks. Segmental inputs from glutamatergic excitatory interneurons (EIN) to ipsilateral glycinergic crossed caudal (CC) interneurons were facilitated or depressed during spike trains at 5-20 Hz. Connections between EINs were depressed. Glycinergic inputs from small ipsilateral inhibitory interneurons were depressed in motor neurons, but were facilitated in CC interneurons. Excitatory inputs from small crossing interneurons to motor neurons were depressed, whereas inhibitory inputs were unaffected. With the exception of connections between EINs, significant effects occurred with stimulation that mimicked interneuron spiking during network activity. Intersegmental EIN synaptic properties were also investigated. EIN inputs did not differ significantly when examined from zero to four segments rostral to motor neurons or CC interneurons. However, caudally located EINs evoked greater activity-dependent facilitation than did rostral EINs. Whilst the amplitude or plasticity of EIN inputs in the rostral or mid-trunk regions of the spinal cord did not differ, EINs in the caudal trunk region evoked greater facilitation. Synaptic transmission between locomotor network neurons thus exhibits activity-dependent plasticity in response to physiologically relevant stimulation. Activity-dependent plasticity could thus contribute to the patterning of the rhythmic network output.     

Different forms of locomotion in the spinal lamprey

Forward locomotion has been extensively studied in different vertebrate animals, and the principal role of spinal mechanisms in the generation of this form of locomotion has been demonstrated. Vertebrate animals, however, are capable of other forms of locomotion, such as backward walking and swimming, sideward walking, and crawling. Do the spinal mechanisms play a principal role in the generation of these forms of locomotion? We addressed this question in lampreys, which are capable of five different forms of locomotion – fast forward swimming, slow forward swimming, backward swimming, forward crawling, and backward crawling. To induce locomotion in lampreys spinalised at the second gill level, we used either electrical stimulation of the spinal cord at different rostrocaudal levels, or tactile stimulation of specific cutaneous receptive fields from which a given form of locomotion could be evoked in intact lampreys. We found that any of the five forms of locomotion could be evoked in the spinal lamprey by electrical stimulation of the spinal cord, and some of them by tactile stimulation. These results suggest that spinal mechanisms in the lamprey, in the absence of phasic supraspinal commands, are capable of generating the basic pattern for all five forms of locomotion observed in intact lampreys. In spinal lampreys, the direction of swimming did not depend on the site of spinal cord stimulation, but on the stimulation strength. The direction of crawling strongly depended on the body configuration. The spinal structures presumably activated by spinal cord stimulation and causing different forms of locomotion are discussed.     

Activation of NMDA receptors elicits fictive locomotion and bistable membrane properties in the lamprey spinal cord

The motor pattern underlying locomotion in the lamprey can be elicited in the spinal cord in vitro by applying excitatory amino acids that activate NMDA receptors. When this is done oscillatory membrane potentials phase-linked with the locomotory rhythm can be recorded in different types of neurones. In some spinal neurones large amplitude oscillation continues after elimination of synaptic input with application of TTX. This oscillatory pacemaker-like activity is dependent on an activation of NMDA receptors, and is probably important in the generation of locomotion.     

Activation of NMDA receptors elecits fictive locomotion and bistable membrane properties in the lamprey spinal cord

The motor pattern underlying locomotion in the lamprey can be elicited in the spinal cord in vitro by applying excitatory amino acids that activate NMDA receptors. When this is done oscillatory membrane potentials phase-linked with the locomotory rhythm can be recorded in different types of neurones. In some spinal neurones large amplitude oscillation continues after elimination of synaptic input with application of TTX. This oscillatory pacemaker-like activity is dependent on an activation of NMDA receptors, and is probably important in the generation of locomotion.     

Axonal regeneration in lamprey spinal cord

Spinal cords of sea lamprey larvae were transected at one of two levels: (a) rostral, at the last gill, or (b) caudal, at the cloaca. Following various recovery times, regeneration of the posteriorly projecting giant reticulospinal axons (RAs) was demonstrated by intra-axonal injection of horseradish peroxidase (HRP). Regeneration of axons of anteriorly projecting dorsal cells (DCs) and giant interneurons (GIs) was demonstrated by intrasomatic HRP injection into cells located just below the transection scar. After 40 days of recovery, 55% of proximally transected RAs (rostral cut) regenerated at least as far as the center of the scar, whereas only 15% of distally transected RAs (caudal cut) did so. Maximum distance of regeneration was 5.3 mm beyond the scar for proximally transected RAs but only 38 u for distally transected RAs. Proximally transected RAs also branched more profusely than distally transected ones. These data (when combined with others in the literature) suggest that the regenerative capacity of RAs may decrease with distance of axotomy from the cell body. Distance of regeneration and degree of branching of proximally transected RAs peaked between 40 and 100 days. Thereafter, there appeared to be a tendency toward neurite retraction. Of axotomized GIs, 76% regenerated anteriorly at least as far as the center of a caudal transection scar (GIs are located only in the caudal part of the cord). The maximum distance of regeneration was 1.3 mm beyond the scar. Of DC axons, 56% regenerated anteriorly at least as far as the transection site. The maximum distance was 1.1 mm beyond the scar. DCs located just below a caudal transection regenerated at least as well as those located below a rostral transection. Axonal regeneration was also demonstrated for a few lateral cells, edge cells, and crossed caudally projecting interneurons.     

Heat production associated with synaptic transmission in the bullfrog spinal cord

By using heat detectors made with pyroelectric film, rapid heat production by the bullfrog spinal cord in response to dorsal root stimulation has been demonstrated. The heat production rises to its peak in about 100 ms after the arrival of afferent impulses and falls slowly with a time course comparable to that of the dorsal root potential. Stimulation of the ventral roots produces no detectable heat. The heat production was reversibly suppressed by immersion of the cord in a low Ca2+, high Mg2+ salt solution, indicating that the underlying exothermic process is associated with intraspinal synaptic transmission. The source of this 'synaptic heat' is located near the boundary between the dorsal column and the substantia gelatinosa in the vicinity of the stimulated dorsal roots.     

Autoradiographic distribution of binding sites for the non-NMDA receptor antagonist [H]CNQX in human motor cortex, brainstem and spinal cord

The distribution of non-NMDA receptors in the normal human motor cortex, brainstem and spinal cord has been investigated using [³H]CNQX. In the motor and premotor cortex, specific [³H]CNQX binding was present in all cortical laminae with the highest density of binding sites in laminae I, II and the upper part of III. In the normal brainstem, non-NMDA receptors labelled by [³H]CNQX had a heterogenous distribution. Brainstem motor nuclei subserving eye movements, which tend to be spared in motor neuron disease (MND), had a higher density of [³H]CNQX binding sites compared to other cranial nerve motor nuclei (VII, X, XII) which tend to be affected. Specific [³H]CNQX binding was present throughout the spinal grey matter, the greatest density of binding being found in the substantia gelatinosa. Excitotoxicity at non-NMDA receptors has been implicated in chronic neurodegenerative diseases such as motor neuron disease. This study suggests that the density of non-NMDA receptors, labelled by [³H]CNQX, does not account for selective vulnerability of motor neurons in this disorder.     

5-hydroxytryptamine immunoreactive varicosities in the lamprey spinal cord have no synaptic specializations--an ultrastructural study

The distribution and fine structure of 5-hydroxytryptamine (5-HT) immunoreactive cell bodies and axonal varicosities have been studied in the lamprey spinal cord, using the peroxidase-antiperoxidase (PAP) immunohistochemical technique and subsequent analysis of ultrathin serial sections. Immunostained cell bodies were found in the ventral spinal cord close to the central canal. Immunostained varicosities were found throughout the spinal cord with the highest density in the ventromedial plexus and the dorsal horn. Only large granular vesicles could be clearly distinguished in immunostained cell bodies and varicosities, but it was concluded based on a comparison with unstained normal tissue that these boutons also contained small, pleomorphic agranular vesicles. Immunoreactive varicosities were studied in the ventromedial plexus, the dorsal horn, the dorsal column, the dorsolateral and ventrolateral funiculi and the grey matter. No morphological differences could be observed between varicosities in the different loci. The varicosities were in no case seen to make synaptic contact with surrounding neuronal elements, even when followed through serial sections. Consequently, 5-HT released from boutons in all parts of the spinal cord could be expected to act on 5-HT receptors located on nearby as well as distant receptors.     

Neuromodulator interactions and spinal cord injury in lamprey

正Neuromodulation is mediated by neurotransmitters that typically act on G-protein-coupled receptors.It can confer behavioural flexibility by modifying the functional properties of anatomically hard-wired neural circuits.Single neuromodulators generally have divergent cellular and synaptic effects(Harris-Warrick and Johnson,2010),and different modulators,of     

Effects of 4-aminopyridine on synaptic transmission in the cat spinal cord

The accumulation of [3H] catecholamines from [3H] tyrosine in frontal cortical, septal, striatal and hippocampal slices was examined following intracerebroventricular (i.c.v.) injections of ACTH 1-24, lysine vasopressin (LVP) and saline. Both ACTH 1-24 and LVP (1 microgram) selectively increased the accumulation of [3H] dopamine (DA) in frontal cortical slices, but did not affect that of [3H] norepinephrine (NE). LVP but not ACTH 1-24 also inhibited the accumulation of [3H] DA in striatal slices. ACTH 1-24 did not alter the accumulation of [3H] NE in hippocampal slices, nor did LVP alter the accumulation of either catecholamine (CA) in septal slices. In vitro incubations with ACTH analogs of LVP failed to alter the rate of accumulation of [3H] CAs in striatal, substantia nigral and frontal cortical slices, except for an inhibitory effect at high doses. This effect is believed to be an artifact of precursor dilution caused by release of tyrosine following degradation of the peptides. Neither peptide modified the increased [3H] CA accumulation stimulated by 26 mM K+, nor did ACTH 1-24 modify the inhibition of [3H] CA accumulation caused by 3 X 10 -6 M haloperidol or 3 X 10 -7 M apomorphine. Selective activation of the mesocortical DA system has also been reported ot occur in response to footshock, suggesting the possibility that endogenous ACTH and/or LVP might mediate the stress-induced activation of mesocortical DA synthesis. Alternatively, i.c.v. injections of these peptides may themselves be stressful and thus indirectly elicit the response.     

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.     

Excitatory synaptic transmission between interneurons and motoneurons in chick spinal cord cell cultures

We have examined the development of synaptic transmission between interneurons and motoneurons in spinal cord cell cultures. Unitary excitatory synaptic currents and complex bursts of excitatory currents develop rapidly: EPSCs (excitatory postsynaptic currents) were detected in 100% of the motoneurons by the 4th day after plating. Inhibitory synaptic currents develop more slowly: IPSCs (inhibitory postsynaptic currents) were detected in only 10% of the motoneurons on day 5 and 40% on day 8. During the 1st and 2nd days in vitro, 24% of the motoneurons tested were dye (Lucifer Yellow) coupled to nearby interneurons. The incidence of dye coupling declined during the first week in culture. No coupling was observed between motoneurons. Our data imply that both G1 and G2 receptors are activated at each synapse. The amplitude of spontaneous excitatory synaptic currents did not change when the motoneuron was hyperpolarized from -50 to -80 mV. This behavior is similar to that of currents induced by glutamate, an agonist that activates 2 types of receptors (G1 and G2) on motoneurons. In addition, a concentration of 2-amino-5-phosphonovaleric acid sufficient to inhibit all G1 receptors only partially inhibited the excitatory synaptic currents. Given the conductance of G1 and G2 channels and the ratio of channels activated during unitary EPSCs, we estimate that as few as 25 G1 channels and 5 G2 channels may mediate excitatory interaction between interneurons and motoneurons during the first week in culture.     

Nicotinic modulation of GABAergic synaptic transmission in the spinal cord dorsal horn

While the mechanisms underlying nicotinic acetylcholine receptor (nAChR)-mediated analgesia remain unresolved, one process that is almost certainly involved is the recently-described nicotinic enhancement of inhibitory synaptic transmission in the spinal cord dorsal horn. Despite these observations, the prototypical nicotinic analgesic (epibatidine) has not yet been shown to modulate inhibitory transmission in the spinal cord. Furthermore, while nAChRs have been implicated in short-term modulation, no studies have investigated the role of nAChRs in the modulation of long-term synaptic plasticity of inhibitory transmission in dorsal horn. Whole-cell patch clamp recordings from dorsal horn neurons of neonatal rat spinal cord slices were therefore conducted to investigate the short- and long-term effects of nicotinic agonists on GABAergic transmission. GABAergic synaptic transmission was enhanced in 86% of neurons during applications of 1 microM nicotine (mean increased spontaneous GABAergic inhibitory postsynaptic current (sIPSC) frequency was approximately 500% of baseline). Epibatidine (100 nM) induced an increase to an average of approximately 3000% of baseline, and this effect was concentration dependent (EC50=43 nM). Nicotinic enhancement was inhibited by mecamylamine and DHbetaE, suggesting an important role for non-alpha7 nAChRs. Tetrodotoxin (TTX) did not alter the prevalence or magnitude of the effect of nicotine, but the responses had a shorter duration. Nicotine did not alter evoked GABAergic IPSC amplitude, yet the long-term depression (LTD) induced by strong stimulation of inhibitory inputs was reduced when paired with nicotine. These results provide support for a mechanism of nicotinic analgesia dependent on both short and long-term modulation of GABAergic synaptic transmission in the spinal cord dorsal horn.     

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.     

Ethanol reduces neuronal excitability and excitatory synaptic transmission in the developing rat spinal cord

Effects of acute ethanol (EtOH) exposure on motoneuron excitability and properties of synaptic transmission were examined in spinal cords of postnatal rats. Whole-cell patch clamp recordings and intracellular recordings with high-resistance electrodes were carried out in motoneurons of 1- to 4-day-old postnatal rats. To determine the effects of extracellular EtOH on action potential waveform, properties of current-evoked soma action potentials and motoneuron ability to generate repetitive action potential firing were examined. During a brief EtOH (70 mM) exposure, larger depolarizing current was required for action potential generation, the duration of the after hyperpolarizing potential increased, and fewer action potentials were produced during a prolonged intracellular current injection. These effects were reversed within 20 min of EtOH removal from the extracellular solution. To determine whether the reduced probability of action potential generation was associated with changes in synaptic transmission, properties of evoked synaptic potentials and spontaneous synaptic currents were investigated. In the presence of EtOH, the amplitude of dorsal root-evoked synaptic potentials was reduced, the frequency of spontaneous excitatory postsynaptic currents decreased, while the frequency of inhibitory postsynaptic currents increased. Our data suggested that acute EtOH exposure suppressed motoneuron electrical activity by decreasing motoneuron excitability and shifting the balance between excitatory and inhibitory synaptic transmission toward inhibition.     

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.     

Adenosine inhibits excitatory but not inhibitory synaptic transmission in the hippocampus.

We examined the effects of adenosine and baclofen on inhibitory (IPSC) and excitatory (EPSC) synaptic currents in dissociated rat hippocampal neurons. Adenosine dramatically reduced monosynaptic EPSCs but failed to diminish IPSCs. This selective effect on EPSCs is likely due to inhibition of excitatory transmitter release because adenosine did not directly alter any properties of postsynaptic neurons. Baclofen depressed both EPSCs and IPSCs to approximately the same extent. These experiments indicate that the presynaptic effects of adenosine and baclofen are clearly separable and that transmitter sensitivities of inhibitory and excitatory neurons can differ. These differences could be exploited in the design of antiepileptic drugs that act at adenosine receptors to limit excitatory neurotransmission without blocking tonic inhibition.