N-methyl-D-aspartate effect on spontaneous activity of 16–20-day-old chick embryo spinal cord

The effects of N-methyl-D-aspartate (NMDA) and the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid (2-APV), on spontaneous activity of dorsal and ventral roots (DR and VR, respectively) generated by isolated spinal cord from 16–20-day-old chick embryo were studied. This activity was synchronous oscillations of electrotonic potentials in DR and VR. There was no impulse activity in the VR. When NMDA was applied at 2–25 µM, the amplitudes of the oscillations increased, the impulse activities in VR and DR developed, and the tonic component of electrotonic potentials appeared. At 20 µM, 2-APV decreased both, the spontaneous and NMDA induced activity. After sectioning of the spinal cord, the neuronal network of the isolated dorsal arm conserved the capacity to generate spontaneous activity in the DR which increased after NMDA application. There was no rhythm in the ventral part of the spinal cord. The localization of the NMDA-sensitive neuronal network, generator of the rhythmic (motor) activity, in the spinal cord is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 23, No. 2, pp. 205–213, March–April, 1991.     

Nature of rhythmic discharges in ventral spinal cord roots]

Rhythmic "locomotor discharges" recorded in the ventral spinal roots by Perret et al proved to increase during asphyxia and disappeared during apnea. Thus, rhythmic discharges in the ventral spinal roots depended on irradiation from the respiratory centre and could not confirm the fact of existence of a special "locomotor pacemaker".     

Dual action of intracellularly released calcium on the quantal mediator secretion

The mice diaphragm muscle and microelectrode technique were used to check the influence of ryanodine (0.5 mcM) on spontaneous and evoked mediator release under conditions of potassium depolarization (8-16 mM [K+]ex or rhythmic (4-100 Hz) stimulation of motor nerve terminals. Weak tonic calcium loading (by muscle exposition to 8 mM [K+]ex) caused a two-fold frequency increase if miniature and plate potentials (MEPPs), which was returned to the basal level by subsequent application of ryanodine. This inhibitory effect of ryanodine was blocked by apamin (500 nM) a blocker of K+(Ca)-channels. A greater calcium load of terminals (in solution with 16 mM [K+]ex) caused a 15-fold increase of MEPPs frequency. Subsequent ryanodine application caused an additional 2-3-fold increase of MEPPs frequency. During rhythmic activity of motor synapses, ryanodine was able to decrease the amplitude of EPP by 60% at plateau phase at short low frequency (4 Hz) of discharges and to increase the amplitude of EPP by 60-150% at high frequency (70-100 Hz) of discharges. It is concluded that rynodine induced calcium release from intraterminal Ca2+-stores can influence dual: excitatory or inhibitory, action on spontaneous and evoked mediator release, due to different intraterminal calcium loads and regimen of synaptic activity.     

Antidromic discharges of dorsal root afferents in the neonatal rat.

Presynaptic inhibition of primary afferents can be evoked from at least three sources in the adult animal: 1) by stimulation of several supraspinal structures; 2) by spinal reflex action from sensory inputs; or 3) by the activity of spinal locomotor networks. The depolarisation in the intraspinal afferent terminals which is due, at least partly, to the activation of GABA(A) receptors may be large enough to reach firing threshold and evoke action potentials that are antidromically conducted into peripheral nerves. Little is known about the development of presynaptic inhibition and its supraspinal control during ontogeny. This article, reviewing recent experiments performed on the in vitro brainstem/spinal cord preparation of the neonatal rat, demonstrates that a similar organisation is present, to some extent, in the new-born rat. A spontaneous activity consisting of antidromic discharges can be recorded from lumbar dorsal roots. The discharges are generated by the underlying afferent terminal depolarizations reaching firing threshold. The number of antidromic action potentials increases significantly in saline solution with chloride concentration reduced to 50% of control. Bath application of the GABA(A) receptor antagonist, bicuculline (5-10 microM) blocks the antidromic discharges almost completely. Dorsal root discharges are therefore triggered by chloride-dependent GABA(A) receptor-mediated mechanisms; 1) activation of descending pathways by stimulation delivered to the ventral funiculus (VF) of the spinal cord at the C1 level; 2) activation of sensory inputs by stimulation of a neighbouring dorsal root; or 3) pharmacological activation of the central pattern generators for locomotion evokes antidromic discharges in dorsal roots. VF stimulation also inhibited the response to dorsal root stimulation. The time course of this inhibition overlapped with that of the dorsal root discharge suggesting that part of the inhibition of the monosynaptic reflex may be exerted at a presynaptic level. The existence of GABA(A) receptor-independent mechanisms and the roles of the antidromic discharges in the neonatal rat are discussed.     

Modification of cyclical activity in the spinal cord of the 16- to 20-day-old chick embryo due to changes in Ca2+ and Mg2+ concentrations

In isolated segments of the 16- to 20-day old chick embryo, increasing the Mg²⁺ concentration from 1.3 to 5.0 mmoles/liter in the superfusate caused complete suppression of spontaneous rhythmic activity that appeared as synchronous cyclical oscillations of electrotonic potentials in the dorsal and ventral roots. A similar change was recorded when the Ca²⁺ concentration was decreased from 2.6 to 1.0 mmole/liter, but in this case tonic discharges of impulses (spikes) could occur. Further, during the disappearance of the spontaneous activity due to changing the concentration of Ca²⁺ or Mg²⁺, in six out of eight experiments another type of rhythmic activity was seen, appearing as oscillations of electrotonic potentials in the ventral roots that were independent of oscillations in the dorsal roots. The amplitude of these oscillations of potential in the ventral roots was up to 200 µV, and their duration was up to 400 msec. The highest frequency of this activity was 0.4 sec^–1. The possible functional significance of the observed patterns of activity is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 23, No. 3, pp. 333–338, May–June, 1991.     

Effect of fenibut on the GABA B receptors of the spinal motor neurons

It has been established in experiments on the isolated spinal cord of 7-14-day-old rats that the GABAB-mimetic phenibut (10(-5)--10(-4) M) elicits a slow-developing depolarization of motoneurons, suppression of spontaneous activity and polysynaptic reflex discharges of motoneurons, recorded from the ventral roots. Administered under the same conditions GABA produces de- and hyperpolarization of motoneurons. The depolarization of motoneurons elicited by phenibut and GABA is not reversed by picrotoxin in contradistinction to the GABA-induced hyperpolarization of motoneurons, being associated with a direct action of the GABA-mimetics on postsynaptic GABAB receptors of motoneurons. Diazepam (10(-9)--10(-6) M) potentiates the effects of phenibut supposedly via benzodiazepine receptors bound with GABAA receptors (an independent interaction).     

Interaction between hindbrain and spinal networks during the development of locomotion in zebrafish

Little is known about the role of the hindbrain during development of spinal network activity. We set out to identify the activity patterns of reticulospinal (RS) neurons of the hindbrain in fictively swimming (paralyzed) zebrafish larvae. Simultaneous recordings of RS neurons and spinal motoneurons revealed that these were coactive during spontaneous fictive swim episodes. We characterized four types of RS activity patterns during fictive swimming: (i) a spontaneous pattern of discharges resembling evoked high-frequency spiking during startle responses to touch stimuli, (ii) a rhythmic pattern of excitatory postsynaptic potentials (EPSPs) whose frequency was similar to the motoneuron EPSP frequency during swim episodes, (iii) an arrhythmic pattern consisting of tonic firing throughout swim episodes, and (iv) RS cell activity uncorrelated with motoneuron activity. Despite lesions to the rostral spinal cord that prevented ascending spinal axons from entering the hindbrain (normally starting at approximately 20 h), RS neurons continued to display the aforementioned activity patterns at day 3. However, removal of the caudal portion of the hindbrain prior to the descent of RS axons left the spinal cord network unable to generate the rhythmic oscillations normally elicited by application of N-methyl-d-aspartate (NMDA), but in approximately 40% of cases chronic incubation in NMDA maintained rhythmic activity. We conclude that there is an autonomous embryonic hindbrain network that is necessary for proper development of the spinal central pattern generator, and that the hindbrain network can partially develop independently of ascending input.     

Presynaptic depression of synaptic response of Renshaw cells by adenosine 5'-monophosphate

The action of AMP (adenosine 5'-monophosphate) on synaptic transmission of Renshaw cells has been studied in cats under Dial anaesthesia. AMP applied iontophoretically reversibly reduced synaptic responses of Renshaw cells evoked by stimulation of ventral roots, as well as their spontaneous firing; however, there were no marked effects on discharges of these cells caused by iontophoretic application of acetylcholine, asparatate, and glutamate. On the other hand, AMP had no comparable effect on synaptic responses of dorsal horn interneurones evoked by stimulation of dorsal roots or their spontaneous discharges.     

Effects of hydrocortisone on electrical activity of the cat spinal cord

The effect of the corticosteroid hormone hydrocortisone on electrical activity in the lumbosacral portion of the spinal cord was studied in acute experiments on cats anesthetized with urethane and chloralose and immobilized with succinylcholine. The amplitude of mono- and polysynaptic discharges arising in the ventral roots in response to stimulation of various afferents of the animal's hind limb was increased by a statistically significant degree after intravenous injection of the hormone. The potentiating action of the hormone was strongest and most stable with respect to early and late postsynaptic potentials of the spinal cord. The dorsal cord potentials were not significantly changed by hydrocortisone. Spontaneous unit activity in the intermediate nucleus of the spinal cord rose sharply after administration of hydrocortisone. Before the action of the hormone the mean frequency of spontaneous discharges of 46 neurons was 7.91/sec, rising to 20/sec after the injection. The number of neurons with a high spontaneous firing rate also was increased. Prolonged extracellular recording of the spontaneous activity of the same neuron before and after administration of hydrocortisone also revealed a marked increase in the frequency of its discharges. The results are evidence of the activating effect of hydrocortisone on spinal interneuronal activity.     

Effect of hypertonic solutions on electrical activity of the isolated frog muscle spindle

Changes in electrical activity of the isolated frog muscle spindle were investigated in hypertonic solutions obtained by adding 400 mM sucrose, glucose, or glycerol to Ringer's solution. The spontaneous firing rate in hypertonic sucrose and glucose solutions increased at first (for 3–5 min) and then fell rapidly to zero; the receptor potential and evoked spike activity diminished under these conditions and disappeared. In the hypertonic solution with glycerol a similar effect was observed but, unlike in the first two media, in this case spike activity returned after its initial increase to the normal level; a second rise in the firing rate was then observed up to a steady value which was higher than normal. After rinsing out the hypertonic sucrose and glucose solutions with ordinary Ringer's solution the spontaneous and evoked activity gradually returned to normal with a small overshoot. During the rinsing out of the hypertonic glycerol solution a sharp and considerable rise in spontaneous activity was first observed, while the changes in frequency of the evoked activity were negligible. The spike activity then returned to normal. The observed changes in electrical activity of the muscle spindle in hypertonic media are attributed to deformation of the sensory terminals and intrafusal muscle fibers (in the glycerol medium), leading to depolarization of the receptor membrane.P. K. Anokhin Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 8, No. 3, pp. 291–299, May–June, 1976.     

The Role of Calcium Ions in Modulation of Impulse Activity of the Isolated Muscle Spindle of the Frog Rana temporaria

The effect of various concentrations of Ca⁺² and Mg⁺² as well as of calcium channel blockers verapamil and nifedipin on impulse activity of frog isolated muscle spindles was studied. Removal of Ca ions from the external Ringer solution was established to increase spontaneous and evoked activity of the muscle spindle. A 4- and 8-fold increase of Ca⁺² concentration produces inhibition and complete cessation of the spontaneous and evoked activity in the muscle spindle. Replacement of Ca⁺² by Mg⁺² is observed to cause no statistically significant change of the spontaneous activity of the isolated muscle spindle; at the same time, at the dynamic spindle extension, the impulse activity rate at the dynamic and static phases of the response rises. Nifedipin and verapamil, blockers of Ca⁺² channels, suppress impulse activity both in norm and on the background of increased impulse activity evoked by removal of Ca⁺² from the external solution. An increase of muscle spindle impulse activity after the removal of Ca⁺² from the external solution is accounted for by transformation of calcium channels of the muscle spindle sensory endings into selective sodium channels.     

Peculiarities of spinal hyperreflexia after combined synchronous transection of the sciatic nerve and chordotomy in rats

Parameters of the reflex discharges evoked by spinal dorsal root stimulation were measured in rats with the sciatic nerve and spinal cord (at low thorasic level) transected five days earlier. Monosynaptic discharges in the ventral roots were found to increase after the operation; the degree of increase was significantly higher as compared with that observed after isolated transections of the spinal cord or the nerve. The combined lesion of the nerve and spinal cord could result in the appearance of high-amplitude reflex discharge components, probably of a polysynaptic nature. We concluded, from the comparison of modifications of reflex discharges, that the mechanisms underlying spinal hyperreflexia after nerve or spinal cord lesions differ considerably from each other.Neirofiziologiya/Neurophysiology, Vol. 26, No. 3, pp. 197–202, May–June, 1994.     

A comparison of experimental procedures of investigation of the dorsal root evoked ventral root reflex in the frog

Effects of the drug methyl-m-aminobenzoate (MS-222 Sandoz) on the dorsal root evoked ventral root responses were studied by electrophysiological methods in the frog spinal cord. A fairly quick and marked depression of the response was observed from which complete recovery was seen within 60 minutes. Larger doses or repeated injection of small amounts of the drug prolonged the recovery and the monosynaptic discharge component of the ventral root reflex often deteriorated irreversibly. - In further experiments, monosynaptic ventral root discharges were demonstrated in spinal cords isolated from the vertebral canal and kept in Ringer solution. The results are discussed in the light of controversial views about the occurrence of monosynaptic ventral root discharge to stimulation of the primary afferents in the amphibian spinal cord.     

Influence of homologous n-alkanoic acids on functional properties of isolated skeletal muscles. I. Muscle contraction]

1.The contractility of isolated muscles of the frog (and in some instances of the rat) was investigated at room temperature in Ringer's solutions containing homologous alkanoic acids (100 mM C4 to 0.4 mM C10). 2. Free fatty acids decrease the contraction amplitudes evoked by direct stimulation. The effects increase with concentration, exposure, and chain length of the fatty acids. In Ringer's solution the changes are totally or partly reversible. 3. The depression of contraction amplitude induced by free fatty acids is removed by small concentrations of caffeine (2--5 mM) in Ringer's solution. 4. Interactions of fatty acids with different structures of skeletal muscle (mitochondria, sarcolemma and membranes of sarcoplasmic vesicles) are discussed. The distinct effect of fatty acids on stimulated muscles and the importance of membranes in the regulation of the calcium ion concentration in the cytoplasm suggest that fatty acids interact with membrane lipids.     

The late prolonged motor discharges during stimulation of the segmentary afferents in rat pups: the effect of L-DOPA

In experiments on 5-30-day rat puppies, under slight urethane narcosis, studies have been made on the late prolonged discharges in extensor muscles evoked by stimulation of contralateral tibial nerve before and after injection of DOPA. Within the first 16 days, the drug completely abolished or significantly reduced late discharges, increasing the spontaneous motor activity. Beginning from the 16th day, periods of potentiation of late discharges were observed. The data obtained are discussed in relation to the development of mechanisms of control of the activity of spinal generators which are responsible for late discharges and spontaneous motor activity.     

Effects of barium on isolated frog spinal cord

The effects of Ba2+ were studied in vitro on the isolated frog spinal cord. Ba2+ (25 microM-5 mM) caused a concentration-dependent depolarization of ventral (VR) and dorsal (DR) roots. TTX and Mg2+ substantially reduced the depolarization suggesting that interneuronal effects were involved. Ba2+ (25-500 microM) markedly increased the frequency and duration of spontaneous VR and DR potentials and substantially enhanced the duration (and frequently the amplitude) of VR and DR potentials evoked by DR stimulation. Higher concentrations of Ba2+ (1-5 mM) reduced both spontaneous and evoked potentials. Ba2+ (25-500 microM) enhanced the amount of K+ released by a DR volley and by application of L-glutamate and L-aspartate. The cation reduced VR and DR root depolarizations produced by elevated [K+]0. VR potentials induced by L-glutamate, L-aspartate, GABA and glycine and DR depolarizations caused by GABA were reduced by Ba2+. These results show that Ba2+ has complex actions on reflex transmission, interneuronal activity, the postsynaptic actions of excitatory and inhibitory amino acids and the evoked release of K+.     

Effect of local depression of inhibition on electrical activity of the spinal cord

The dorsal cord, dorsal root, and focal potentials in response to peripheral nerve stimulation were investigated in rats with local depression of inhibition in the left or right half of the lumbar segments produced by the action of tetanus toxin. The investigation was carried out at the stage of poisoning when excitation of the neuron population with disturbed inhibition caused generalized excitation of spinal and bulbar motoneurons. Experiments on spinal animals showed that if a cutaneous nerve is stimulated on the side affected by the toxin these responses have a greater amplitude and a much longer duration than those evoked by stimulation of the opposite nerve or responses in healthy rats. The maximal increase in amplitude and duration of the negative component of the focal potential corresponding to the time of the increased P wave of the dorsal cord potential was found in the ventral quadrant on the side affected by the toxin. Besides evoked focal potentials, spontaneous rhythmic negative waves also were recorded in this area. The mechanisms of spread of seizure activity from the focus of depressed inhibition are discussed and the structures generating spreading seizure activity are identified.     

Effects of dexamethasone on the electrical activity of spinal neurons in rats with the transected sciatic nerve

Effects of the glucocorticoid hormone dexamethasone on the reflex discharges in the lumbar ventral roots and background activity (BA) of single neurons in the dorsal laminae of spinal grey were studied in rats after transection of the sciatic nerve. Administration of the hormone during early post-traumatic period (up to seven days) evoked no significant changes in the amplitude of increased (due to the postdenervation hyperreflexia) monosynaptic discharges on the side of nerve transection. At the same time, the monosynaptic discharges grew by 150–170% on the intact side. During later post-transection periods (up to 35 days), when ventral root reflex discharges were suppressed, dexamethasone facilitated reflex transmission via the polysynaptic segmental pathways on both the operated and intact sides. Nonetheless, the monosynaptic component of reflex discharges on the injured side did not recover. Dexamethasone treatment resulted in an increase in the number of BA-generating interneurons within the superficial dorsal horn laminae, and in a decrease in the proportion of units generating bursting activity (possibly of pathological nature).Neirofiziologiya/Neurophysiology, Vol. 27, No. 1, pp. 26–31, January–February, 1995.     

Cholinergic modulation of the synaptic transmission in the frog spinal cord

It was found during experiments on isolated frog spinal cord involving extracellular recording from the dorsal roots (sucrose bridging) and intracellular recording from motoneurons by microelectrodes that 10 mM of the M-cholinomimetic arecoline produces motoneuronal depolarization which is matched by depolarizing electronic ventral root potentials and a rise in motoneuronal input resistance. Arecoline changes synaptic transmission by increasing the amplitude of postsynaptic potentials during intracellular recording and that of motoneuronal reflex discharges in the ventral roots but reduces the duration of dorsal root potentials. In the presence of arecoline, L-glutamate-induced motoneuronal response increases. Facilitation of synaptic transmission produced by arecoline in the spinal cord is bound up with cholinergic M₂- activation, since it is suppressed by atropine but not by low concentrations of pirenzipine; it is also coupled with a reduction in adenylcyclase activity. When motoneuronal postsynaptic response has been suppressed, as in the case of surplus calcium or theophylline, arecoline produces an inhibitory effect on the amplitude of motoneuronal monosynaptic reflex discharges which is suppressed by pirenzipine at a concentration of 1×10^–7 M. This would indicate the presence at the primary afferent terminals of presynaptic cholinergic M₁ receptors which mediate its inhibition of impulses of transmitter release. This effect is independent of changes in cyclic nucleotide concentration.A. M. Gorkii Medical Institute, Donetsk. Translated from Neirofiziologiya, Vol. 19, No. 3, pp. 399–405, May–June, 1987.     

Stimulation of the hippocampus and its effect on electrographic manifestations of the brain in unrestrained rats.

The dorsal hippocampus was electrically stimulated in unanaesthetized, unrestrained rats with a cobalt-gelatin rod in their cortex. The significance of the hippocampus in the elicitation of both physiological spontaneous rhythmic activity (episodic activity of 8--9/sec frequency bound, in rats, to a state of quiet wakefulness, and "sleep spindles") and pathological rhythmic activity of the self-sustained after-discharge (SSAD) type was determined from the aspect of the EEG and behavioural characteristics. 1. Single electrical pulses (0.1 msec, 1--10 V, 0.3/sec) elicited an evoked potential bilaterally in the somatosensory cortex. Elicitation of rhythmic after-activity (of the type of episodes or sleep spindles) was observed only in some cases in which an adequately strong stimulus was used. 2. Repeated series of rhythmic electrical stimuli following each other at short intervals (2--3 min) led to the formation of SSAD in about one third of the cases and at all stimulation frequencies (3-15/sec), although low frequencies (3--4/sec) were the least effective. The character of the SSAD and simultaneous behavioural phenomena differed fundamentally from those evoked by electrical stimulation of the thalamus (Chocholová et al. 1977). The development of paroxysmal after-activity was signalled by responses of a more or less distinct "recruiting" character during stimulation. On the basis of a comparison of electrographic and behavioural manifestations after electrical stimulation of the thalamus and hippocampus, the possibility of both thalamic and extrathalamic projection from the hippocampus to the cortical region is considered.