Involvement of calcium in rhythmic activity induced by disinhibition in cultured spinal cord networks

Disinhibition of rat spinal networks induces a spontaneous rhythmic bursting activity. The major mechanisms involved in the generation of such a bursting are intrinsic neuronal firing of a subpopulation of interneurons, recruitment of the network by recurrent excitation, and autoregulation of neuronal excitability. We have combined whole cell recording with calcium imaging and flash photolysis of caged-calcium to investigate the contribution of [Ca(2+)](i) to rhythmogenesis. We found that calcium mainly enters the neurons through voltage-activated calcium channels and N-methyl-D-aspartate (NMDA) channels as a consequence of the depolarization during the bursts. However, [Ca(2+)](i) could neither predict the start nor the termination of bursts and is therefore not critically involved in rhythmogenesis. Also calcium-induced calcium release is not involved as a primary mechanism in bursting activity. From these findings, we conclude that in the rhythmic activity induced by disinhibition of spinal cord networks, the loading of the cells with calcium is a consequence of bursting and does not functionally contribute to rhythm generation.     

Activity-dependent modification of inhibitory synapses in models of rhythmic neural networks

The faithful production of rhythms by many neural circuits depends critically on the strengths of inhibitory synaptic connections. We propose a model in which the strengths of inhibitory synapses in a central pattern-generating circuit are subject to activity-dependent plasticity. The strength of each synapse is modified as a function of the global activity of the postsynaptic neuron and by correlated activity of the pre- and postsynaptic neurons. This allows the self-assembly, from random initial synaptic strengths, of two cells into reciprocal oscillation and three cells into a rhythmic triphasic motor pattern. This self-assembly illustrates that complex oscillatory circuits that depend on multiple inhibitory synaptic connections can be tuned via simple activity-dependent rules.     

Ontogeny of rhythmic motor patterns generated in the embryonic rat spinal cord

Patterned spontaneous activity is generated in developing neuronal circuits throughout the CNS including the spinal cord. This activity is thought to be important for activity-dependent neuronal growth, synapse formation, and the establishment of neuronal networks. In this study, we examine the spatiotemporal distribution of motor patterns generated by rat spinal cord and medullary circuits from the time of initial axon outgrowth through to the inception of organized respiratory and locomotor rhythmogenesis during late gestation. This includes an analysis of the neuropharmacological control of spontaneous rhythms generated within the spinal cord at different developmental stages. In vitro spinal cord and medullary-spinal cord preparations isolated from rats at embryonic ages (E)13.5-E21.5 were studied. We found age-dependent changes in the spatiotemporal pattern, neurotransmitter control, and propensity for the generation of spontaneous rhythmic motor discharge during the prenatal period. The developmental profile of the neuropharmacological control of rhythmic bursting can be divided into three periods. At E13.5-E15.5, the spinal networks comprising cholinergic and glycinergic synaptic interconnections are capable of generating rhythmic activity, while GABAergic synapses play a role in supporting the spontaneous activity. At late stages (E18.5-E21.5), glutamate drive acting via non- N-methyl-d-aspartate (non-NMDA) receptors is primarily responsible for the rhythmic activity. During the middle stage (E16.5-E17.5), the spontaneous activity results from the combination of synaptic drive acting via non-NMDA glutamatergic, nicotinic acetylcholine, glycine, and GABA(A) receptors. The modulatory actions of chloride-mediated conductances shifts from predominantly excitatory to inhibitory late in gestation.     

Muscles express motor patterns of non-innervating neural networks by filtering broad-band input

We describe three slow muscles that responded to low-frequency modulation of a high-frequency neuronal input and, consequently, could express the motor patterns of neural networks whose neurons did not directly innervate the muscles. Two of these muscles responded to different frequency components present in the same input, and as a result each muscle expressed the motor pattern of a different, non-innervating, neural network. In an analogous manner, the distinct dynamics of the multiple intracellular processes that most cells possess may allow each process to respond to, and hence differentiate among, specific frequency ranges present in broad-band input.     

Multiple spontaneous rhythmic activity patterns generated by the embryonic mouse spinal cord occur within a specific developmental time window

Spontaneous rhythmic activity is a ubiquitous phenomenon in developing neural networks and is assumed to play an important role in the elaboration of mature circuitry. Here we describe the day-by-day evolution of spontaneous activity in the embryonic mouse spinal cord and show that, at a specific developmental stage, 2 distinct rhythms coexist. On embryonic days E12.5 and E13.5, we observed a single type of regularly recurring short spike-episodes synchronized across cervical, thoracic, and lumbar levels. By E14.5, in addition to this motor rhythm, another type of spontaneous synchronous activity appeared, characterized by much longer lasting episodes separated by longer time intervals. On E15.5, these long episodes disappeared. Short episodes were less numerous and more irregular except at the cervical level where a rhythm was occasionally observed. By E16.5, this cervical rhythm became more robust, whereas the lumbar level fell almost silent. Surprisingly, at E17.5, spontaneous activity resumed at caudal levels, now characterized by numerous erratic short episodes. A striking ontogenetic feature of spontaneous activity was the occurrence of long episodes only at E14.5. Although concomitant at all levels of the spinal cord, long episodes displayed different patterns along the spinal cord, with tonic firing at the thoracic level and rhythmic discharge with occasional sequences of left/right alternation at the lumbar level. Thus at E14.5, the originally synchronized network has started to segregate into more specialized subnetworks. In conclusion, this work suggests that ongoing spontaneous rhythms do not follow a smooth evolution during maturation, but rather undergo profound changes at very specific stages.     

The motor output and behavior produced by rhythmogenic sacrocaudal networks in spinal cords of neonatal rats

The characteristics of the rhythmic motor output and behavior produced by intrinsic sacrocaudal networks were studied in isolated tail-spinal cord preparations of neonatal rats. An alternating left-right rhythm could be induced in the sacral cord by stimulus trains applied to sacrocaudal afferents at various intensities. Strengthening the stimulation intensity enhanced the rhythmic efferent firing and accelerated the rhythm by < or =30%. High stimulation intensities induced tonic excitation or inhibition and thereby perturbed the rhythm. Increasing the stimulation frequency from 1 to 10 Hz decreased the cycle time of the rhythm by 36%. The rhythm was blocked during prolonged afferent stimulation but could be restored by stimulation of contralateral afferents. Sacrocaudal afferent activation produced ventroflexion accompanied by either low- or high-amplitude rhythmic abduction of the tail. The low-amplitude abductions were produced by alternating flexor bursts during long stimulus trains. The activity of abductors and extensors was substantially reduced during these trains, their recruitment lagged after that of the flexors, and their activity bursts were much shorter. It is suggested that tail extensor/abductor motoneurons were suppressed during the stimulus train by inhibitory afferent projections. The high-amplitude abductions appeared after cessation of stimulus trains. Alternating left-right activation of the tail muscles, and coactivation of the principal muscles on each side of the tail were observed during these abductions. It is suggested that flexors and extensors assist the abductors to produce the high-amplitude abductions. This suggestion is supported by the finding that tail abduction could be produced by direct unilateral stimulation of any of the principal tail muscles. The relevance of the findings described in the preceding text to the use of regional sacral circuits in generation of stereotypic motor behaviors and to future studies of rhythmogenic sacrocaudal networks is discussed.     

Cortical response amplitude changes produced by rhythmic acoustic stimulation in cats

Summary Under a short rhythmic acoustic stimulation lasting for several minutes and a prolonged rhythmic stimulation a decrease and increase was observable in the amplitude of the individual components of cortical evoked responses registered in the cat in the anterior portion of the medial suprasylvian gyrus and in the primary region. These changes, however, are not depending upon the duration of the rhythmic acoustic stimulation at all but they reflect changes in the amplitude of the EEG of the corresponding region. An increase in the amplitude integral of the EEG of the cortical acoustic region is connected with a parallel increase in the magnitude of the first positivity and negativity of the cortical response registered in the same region. The first positivity and negativity registered during acoustic stimulation in the suprasylvian gyrus vary in dependence on changes in the value of EEG amplitude integral of this region in the same way as those of the primary acoustic response. The changes of the second positive wave registered during rhythmic stimulation in the primary region are somewhat more complicated, however.     

Short-term plasticity of spinal reflex excitability induced by rhythmic arm movement

Rhythmic arm movement reduces Hoffmann (H)-reflex amplitudes in leg muscles by modulation of presynaptic inhibition in group Ia transmission. To date only the acute effect occurring during arm movement has been studied. We hypothesized that the excitability of soleus H-reflexes would remain suppressed beyond a period of arm cycling conditioning. Subjects used a customized arm ergometer to perform rhythmic 1-Hz arm cycling for 30 min. H-reflexes were evoked before, during, and after arm cycling via stimulation of the tibial nerve in the popliteal fossa. The most important finding was that the H-reflex amplitudes were significantly suppressed during and 相似文献   

Nature of rhythmic discharges in ventral spinal roots

After recording rhythmic discharges in the ventral spinal roots Perret and co-workers postulated the existence of a special “locomotor pacemaker”. The present writers showed that the “locomotor discharges” discovered by Perret and co-workers increase during asphyxia and disappear during apnea. Rhythmic discharges in the ventral roots thus depend on the irradiation of excitation from the respiratory center and they cannot be evidence of the existence of a special locomotor pacemaker.     

Detection of ECG waveforms by neural networks

In this study, ECG waveform detection was performed by using artificial neural networks (ANNs). Initially, the R peak of the QRS complex is detected, and then feature vectors are formed by using the amplitudes of the significant frequency components of the DFT spectrum. Grow and Learn (GAL) and Kohonen networks are comparatively investigated to detect four different ECG waveforms. The comparative performance results of GAL and Kohonen networks are reported.     

Respiratory-like rhythmic activity can be produced by an excitatory network of non-pacemaker neuron models

It is still unclear whether the respiratory-like rhythm observed in slice preparations containing the pre-B?tzinger complex is of pacemaker or network origin. The rhythm persists in the absence of inhibition, but blocking pacemaker activity did not always result in rhythm abolition. We developed a computational model of the slice to show that respiratory-like rhythm can emerge as a network property without pacemakers or synaptic inhibition. The key currents of our model cell are the low- and high-threshold calcium currents and the calcium-dependent potassium current. Depolarization of a single unit by current steps or by raising the external potassium concentration can induce periodic bursting activity. Gaussian stimulation increased the excitability of the model without evoking oscillatory activity, as indicated by autocorrelation analysis. In response to hyperpolarizing pulses, the model produces prolonged relative refractory periods. At the network level, an increase of external potassium concentration triggers rhythmic activity that can be attributed to cellular periodic bursting, network properties, or both, depending on different parameters. Gaussian stimulation also induces rhythmic activity that depends solely on network properties. In all cases, the calcium-dependent potassium current has a central role in burst termination and interburst duration. However, when periodic inhibition is considered, the activation of this current is responsible for the characteristic amplification ramp of the emerged rhythm. Our results may explain controversial results from studies blocking pacemakers in vitro and show a shift in the role of the calcium-dependent potassium current in the presence of network inhibition.     

Independent neural control of rhythmic sequences--behavioral and fMRI evidence

Can the temporal structure of movement sequences can be represented and learned independently of their ordinal structure? Are some brain regions particularly important for temporal sequence performance? We addressed these questions in behavioral and functional magnetic resonance imaging (fMRI) experiments. Using a learning transfer design, we found evidence for independent temporal representations: learning a spatiotemporal sequence facilitated learning its temporal and ordinal structure alone; learning a temporal and an ordinal structure facilitated learning of a sequence where the two were coupled. Secondly, learning of temporal structures was found during reproduction of sequential stimuli with random ordinal structure, suggesting independent mechanisms for temporal learning. We then used fMRI to investigate the neural control of sequences during well-learned performance. The temporal and ordinal structure of the sequences were varied in a 2x2 factorial design. A dissociation was found between brain regions involved in ordinal and temporal control, the latter mainly involving the pre-supplementary motor area, the inferior frontal gyrus, the precentral sulcus, and the superior temporal gyri. In a second fMRI experiment, temporal sequences were performed with the left or right index fingers, or using rhythmic speech. The overlap in brain activity during performance with the different effectors included a similar set of brain regions to that found in the first fMRI experiment: the supplementary motor area and the superior temporal and inferior frontal cortices. We suggest that these regions are important for abstract, movement-independent temporal sequence control. This organization may be important for flexibility in voluntarily timed motor tasks.     

Dynamics of spinal cord changes produced by prenatal asphyxia

Changes in neurons and nerve fibers of the spinal cord at the level C4–C5 were studied in rats exposed to asphyxiain utero and killed at different times. In the first 10 days the changes in the nerve cells progressed gradually. By the age of 1–2 months the state of most neurons is back to normal, but in each hundred fields of vision pathologically changed neurons are found, mainly in zones of the collateral circulation. Changes also are observed in the nerve fibers.Laboratory for the Study of Brain Development, Scientific-Research Institute of Pediatrics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR M. Ya. Studenikin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 3, pp. 361–364, March, 1977.     

Properties of rhythmic activity generated by the isolated spinal cord of the neonatal mouse

We examined the ability of the isolated lumbosacral spinal cord of the neonatal mouse (P0-7) to generate rhythmic motor activity under several different conditions. In the absence of electrical or pharmacological stimulation, we recorded several patterns of spontaneous ventral root depolarization and discharge. Spontaneous, alternating discharge between contralateral ventral roots could occur two to three times over a 10-min interval. We also observed other patterns, including left-right synchrony and rhythmic activity restricted to one side of the cord. Trains of stimuli delivered to the lumbar/coccygeal dorsal roots or the sural nerve reliably evoked episodes of rhythmic activity. During these evoked episodes, rhythmic ventral root discharges could occur on one side of the cord or could alternate from side to side. Bath application of a combination of N-methyl-D,L-aspartate (NMA), serotonin, and dopamine produced rhythmic activity that could last for several hours. Under these conditions, the discharge recorded from the left and right L(1)-L(3) ventral roots alternated. In the L(4)-L(5) segments, the discharge had two peaks in each cycle, coincident with discharge of the ipsilateral and contralateral L(1)-L(3) roots. The L(6) ventral root discharge alternated with that recorded from the ipsilateral L(1)-L(3) roots. We established that the drug-induced rhythm was locomotor-like by recording an alternating pattern of discharge between ipsilateral flexor and extensor hindlimb muscle nerves. In addition, by recording simultaneously from ventral roots and muscle nerves, we established that ankle flexor discharge was in phase with ipsilateral L(1)/L(2) ventral root discharge, while extensor discharge was in phase with ipsilateral L(6) ventral root discharge. Rhythmic patterns of ventral root discharge were preserved following mid-sagittal section of the spinal cord, demonstrating that reciprocal inhibitory connections between the left and right sides of the cord are not essential for rhythmogenesis in the neonatal mouse cord. Blocking N-methyl-D-aspartate receptors, in both the intact and the hemisected preparation, revealed that these receptors contribute to but are not essential for rhythmogenesis. In contrast, the rhythm was abolished following blockade of kainate/AMPA receptors with 6-cyano-7-nitroquinoxalene-2,3-dione. These findings demonstrate that the isolated mouse spinal cord can produce a variety of coordinated activities, including locomotor-like activity. The ability to study these behaviors under a variety of different conditions offers promise for future studies of rhythmogenic mechanisms in this preparation.     

Role of group II and III metabotropic glutamate receptors in rhythmic patterns of the neonatal rat spinal cord in vitro

Electrophysiological recordings were used to explore the role of group II and III metabotropic glutamate receptors (mGluRs) in oscillatory patterns generated by the neonatal rat spinal cord in vitro. Neither the group II agonist DCG-IV (and the selective antagonist EGLU), nor the group III agonist l-AP4 (and its selective antagonist CPPG) had any effect on lumbar motoneuron membrane potential or input resistance. This observation suggests that motoneurons expressed no functional group II and III mGluRs and received no network-based, tonic influence mediated by them. DCG-IV or l-AP4 strongly depressed synaptic responses evoked by single dorsal root (DR) stimuli, an effect counteracted by their respective antagonist. EGLU or CPPG per se had no effect on synaptic responses, indicating no mGluR autoreceptor-dependent control of transmitter release. l-AP4 largely depressed cumulative depolarization, windup and associated oscillations, whereas synaptic depression induced by DCG-IV waned with repeated stimuli. l-AP4 slowed down fictive locomotor patterns and arrested disinhibited bursting, which could, however, be promptly restored by DR electrical stimulation. DCG-IV had no significant effect on fictive locomotion, but it blocked disinhibited bursting. EGLU facilitated bursting, suggesting that burst termination was partly controlled by group II mGluRs. All these effects were reversible on washout. It is concluded that activation of group II and III mGluRs differentially modulated rhythmic patterns recorded from motoneurons via network-dependent actions, which probably included decrease in the release of neurotransmitters at key circuit points.G. Taccola and C. Marchetti contributed equally to the work     

Extracellular magnesium enhances the damage to locomotor networks produced by metabolic perturbation mimicking spinal injury in the neonatal rat spinal cord in vitro

An acute injury to brain or spinal cord produces profound metabolic perturbation that extends and exacerbates tissue damage. Recent clinical interventions to treat this condition with i.v. Mg²⁺ to stabilize its extracellular concentration provided disappointing results. The present study used an in vitro spinal cord model from the neonatal rat to investigate the role of extracellular Mg²⁺ in the lesion evoked by a pathological medium mimicking the metabolic perturbation (hypoxia, aglycemia, oxidative stress, and acid pH) occurring in vivo. Damage was measured by taking as outcome locomotor network activity for up to 24 h after the primary insult. Pathological medium in 1 mM Mg²⁺ solution (1 h) largely depressed spinal reflexes and suppressed fictive locomotion on the same and the following day. Conversely, pathological medium in either Mg²⁺-free or 5 mM Mg²⁺ solution evoked temporary network depression and enabled fictive locomotion the day after. While global cell death was similar regardless of extracellular Mg²⁺ solution, white matter was particularly affected. In ventral horn the number of surviving neurons was the highest in Mg²⁺ free solution and the lowest in 1 mM Mg²⁺, while motoneurons were unaffected. Although the excitotoxic damage elicited by kainate was insensitive to extracellular Mg²⁺, 1 mM Mg²⁺ potentiated the effect of combining pathological medium with kainate at low concentrations. These results indicate that preserving Mg²⁺ homeostasis rendered experimental spinal injury more severe. Furthermore, analyzing ventral horn neuron numbers in relation to fictive locomotion expression might provide a first estimate of the minimal size of the functional locomotor network.