Action of ammonium acetate on central primary afferent depolarization

Experiments on anesthetized spinal cats showed that ammonium acetate, injected intravenously (2–4 mmoles/kg) inhibits the depolarization of the central endings of primary afferent fibers activated by stimulation of afferent nerves. Inhibition of primary afferent depolarization is transient in character and develops parallel with depression of postsynaptic inhibition of monosynaptic reflexes. The depression produced by the action of ammonium was not due to blocking of negative postsynaptic potentials of the dorsal surface of the spinal cord or blocking of reflex electrical discharges in the ventral spinal roots. It is suggested that depression of primary afferent depolarization is due to a decrease in the emf for synaptic ion currents producing depolarization.Allergologic Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 9, No. 1, pp. 52–60, January–February, 1977.     

Depolarization of primary afferents during fictitious scratching in thalamic cats

The dorsal cord and dorsal root potentials were recorded in immobilized thalamic cats during fictitious scratching evoked by mechanical stimulation of the ear. Depolarization of primary afferents was shown to be simulated by the central scratching generator. Antidromic spike discharges appeared at the peak of the primary afferent depolarization waves in certain afferent fibers. Similar discharges arise in the resting state in response to stimulation of limb mechanoreceptors. It is suggested that during real scratching primary afferent depolarization and antidromic spikes evoked by it may effectively modulate the level of the afferent flow to spinal neurons.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 10, No. 2, pp. 173–176, March–April, 1978.     

New evidence of depression of depolarization of primary afferents with ammonium ions]

Perfusion of the central canal of the lumbar segment of the spinal cord with artificial cerebrospinal fluid (CSF) supplemented with ammonium ions induced in cats depression of slow negative electrotonic potentials of dorsal roots (PDR). PDR depression developed simultaneously with postsynaptic depression of the inhibition of monosynaptic reflexes, however it was not related to the depression of mono- or polysynaptic reflex discharges. Further perfusion of the central canal with normal CSF resulted in a complete restoration of the PDR as well as inhibition of control reflexes. It is suggested that PDR depression with ammonium ions could be the result of the blocking of the chloric pump acting the afferent terminals and creating electromotive power for outward transmembranous chloric current producing depolarization of the afferent fibres.     

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of primary afferent fibers mediated by GABAergic axo-axonal synapses (primary afferent depolarization). The strength of primary afferent depolarization can be measured by recording of volume-conducted potentials at the dorsal root (dorsal root potentials, DRP). Pathological changes of presynaptic inhibition are crucial in the abnormal central processing of certain pain conditions and in some disorders of motor hyperexcitability. Here, we describe a method of recording DRP in vivo in mice. The preparation of spinal cord dorsal roots in the anesthetized animal and the recording procedure using suction electrodes are explained. This method allows measuring GABAergic DRP and thereby estimating spinal presynaptic inhibition in the living mouse. In combination with transgenic mouse models, DRP recording may serve as a powerful tool to investigate disease-associated spinal pathophysiology. In vivo recording has several advantages compared to ex vivo isolated spinal cord preparations, e.g. the possibility of simultaneous recording or manipulation of supraspinal networks and induction of DRP by stimulation of peripheral nerves.     

Primary afferent depolarization due to selective activation of propriospinal pathways

Depolarization of primary afferent terminals in the lumbosacral portion of the spinal cord evoked by selective activation of propriospinal pathways was investigated in anesthetized cats. The strongest depolarization developed as a result of activation of short (two to five segments) propriospinal pathways in the lateral funiculus; stimulation of the long propriospinal pathways of this funiculus also induced depolarization, but of lower amplitude. Stimulation of propriospinal pathways of the ventral funiculi was ineffective. Significant primary afferent depolarization developed only following the use of a series of stimuli and strong stimulation of the propriospinal pathways. Excitation of these pathways caused depolarization of afferent terminals of both cutaneous and muscular nerves, including muscular sensory fibers of group Ia, although in the latter case its intensity was low. Neuronal mechanisms involved in the generation of this depolarization and its possible functional role are dicussed.     

The actions of serotonin on frog primary afferent terminals and cell bodies

The actions of serotonin (5-HT) were studied in the isolated frog spinal cord and dorsal root ganglion preparations. In the spinal cord, 5-HT increased the spontaneous activity recorded from dorsal roots, facilitated evoked spinal reflexes and produced fast and slow primary afferent depolarization (PAD). A direct action of 5-HT on primary afferent terminals is likely since 5-HT induced PAD remained in the presence of 1 microM tetrodotoxin and 2 mM Mn2+. The direct action of 5-HT on primary afferent terminals was blocked by methysergide and attenuated by concentrations of Mn2+ in excess of that required to block transmitter release. Cell bodies of the dorsal root ganglion were also depolarized by 5-HT. A slow hyperpolarization occasionally followed the initial depolarization. The depolarizing action of 5-HT in the dorsal root ganglion was also attenuated by treatment with Mn2+. It is concluded that 5-HT acts directly on frog primary afferents and that this influence may involve a calcium sensitive process. The dorsal root ganglion response to 5-HT appears to be a suitable model of the afferent terminal response.     

Primary afferent depolarization induced by activation of intersegmental connections of the substantia gelatinosa in the cat spinal cord

Depolarization of primary afferent terminals induced by selective activation of intersegmental connections of the substantia gelatinosa was investigated in cats anesthetized with pentobarbital. Depolarization was found to develop most rapidly in fibers of high-threshold muscular and cutaneous sensory nerves, but it was present only to a very slight degree in fibers of group Ia muscular afferents. It is suggested that the spread of activity inducing depolarization takes place in the substantia gelatinosa along a chain of excessively excited neurons. The possible role of primary afferent depolarization as a factor stabilizing coordinated activity of spinal neurons is discussed.     

Polarization of primary afferent terminals in the lumbar spinal cord during fictitious locomotion

Changes in electrical polarization of primary afferent terminals in the lumbosacral portion of the spinal cord were investigated during fictitious locomotion in immobilized decorticated and spinal cats. Fictitious locomotion was accompanied by stable hyperpolarization of the afferent terminals, against the background of which they were periodically depolarized in rhythm with efferent activity. These tonic and phasic changes were observed in terminals of all groups of afferent fibers tested: cutaneous and muscular (Ia and Ib). Periodic in-phase depolarization was carried out in different ipsilateral segments of the lumbosacral enlargement. During ficitious galloping changes in depolarization of the primary efferents were in phase on different sides; during fictitious walking, these periodic changes were out of phase. On the basis of these results the physiological importance of changes in electrical polarization of primary afferent terminals of the spinal locomotor generator is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 481–489, September–October, 1980.     

Study of tonic and evoked electrical activity in efferent fibers of the sympathetic nerve in white rats

In anaesthetised Wistar rats, electrical sympathetic activity and a somatosympathetic reflex in the cervical sympathetic trunk elicited by a single electrical shock to forelimb or hindlimb afferent nerves, were recorded. The spontaneous activity was shown to conform with the pulse and respiratory waves of arterial pressure. Somatosympathetic reflex consists of early and late discharges evoked by somatic myelinated afferent fibres stimulation, and C-response elicited by stimulation of unmyelinated afferent fibres in spinal nerves.     

Changes in antidromic discharges in peripheral nerves during polarization of the cat spinal cord by a steady current

The effect of a steady current passed through the spinal cord on antidromic discharges in primary afferent groups of Agb cutaneous nerves of the hind limb, evoked by single and paired stimulation of the terminals of these fibers, was investigated by Wall's technique in acute experiments on spinal and anesthetized cats. A current of up to 50–100 µA, flowing in the dorso-ventral direction, led to an increase in amplitude of antidromic dischanges evoked by single stimulation of afferent terminals; if the current flowed in the opposite direction, the opposite effect was observed. The relative degree of facilitation of antidromic discharges caused by conditioning stimulation of these same fibers was reduced by a polarizing current in either direction. It is suggested that the effects of the action of a steady current flowing through the spinal cord observed in these experiments are due mainly to shifts of membrane potential in primary afferent terminals.Dnepropetrovskii State University. Translated from Neirofiziologiya, Vol. 14, No. 4, pp. 386–391, July–August, 1982.     

Depolarization of different groups of lumbar afferent terminals during fictitious scratching

The kinetics of primary afferent depolarization (PAD) in the lumbar spinal cord was studied in immobilized decerebrate and decapitated cats during fictitious scratching. Fictitious scratching was accompanied by tonic and periodic PAD. Periodic PAD was cophased in different segments of the lumbosacral enlargement. Tonic depolarization was observed in terminals of all groups of afferent fibers studied (low-threshold cutaneous and of groups Ia and Ib). Periodic changes were observed only in endings of low-threshold cutaneous fibers and group Ib fibers. The physiological role of modulation of PAD by the spinal scratch generator is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 11, No. 6, pp. 569–577, November–December, 1979.     

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.     

Activity-dependent role of NMDA receptors in transmission of cardiac mechanoreceptor input to the NTS

Evidence suggests that transmission of barosensitive input from arterial baroreceptors and cardiac mechanoreceptors at nucleus tractus solitarius (NTS) neurons involves non-N-methyl-d-aspartate (NMDA) glutamate receptors, but there is a possibility that the contribution of NMDA receptors might increase during periods of increased afferent input, when enhanced neuronal depolarization could increase the activation of NMDA receptors by removal of a Mg(2+) block. Thus the effects of NMDA on cardiac mechanoreceptor-modulated NTS neuronal discharges were examined at different levels of arterial pressure used to change cardiac mechanoreceptor afferent input. To determine whether the response was specific to NMDA, (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA) was also administered at different levels of neuronal discharge. In anesthetized dogs, neuronal activity was recorded from the NTS while NMDA or AMPA was picoejected at high versus low arterial stimulating pressures. NMDA, but not AMPA, produced a significantly greater discharge of mechanoreceptor-driven NTS neurons at higher versus lower levels of stimulating pressure. These data suggest that the role played by NMDA receptors is greater during periods of enhanced neuronal depolarization, which could be produced by increases in afferent barosensitive input.     

Regional specificity of functional sensory connections in developing spinal cord cultures varies with the incidence of spontaneous bioelectric activity

Summary Cultured spinal cord explants in which little spontaneous bioelectric activity was present showed, when monitored using sensory ganglion-evoked monosynaptic action potentials, diffuse innervation by ingrowing afferent fibers at 3–4 weeks in vitro. In contrast, highly active cultures of the same age showed a strong tendency for functional sensory connections to be made within the dorsal half of the cord. Regional specificity was present in mature cultures (4–5 weeks in vitro), however, even when their spontaneous activity level was low. The results support earlier results using tetrodotoxin, and make it appear likely that centrally generated neuronal discharges can influence the topography of afferent terminals within the developing spinal cord.     

Active selection of afferent information as the basic principle of peripheral correction of the activity of spinal rhythmic movement generators

This paper summarizes information obtained in the experimental study of the dynamics of polarization of central primary afferent endings and modifications of segmental responses to afferent stimuli during fictitious locomotion and fictitious scratching in immobilized, decorticated, decerebrate, and spinal cats. Fictitious locomotion was accompanied by tonic hyperpolarization, fictitious scratching by tonic depolarization of central primary afferent endings. Against the background of these long-lasting changes in primary afferent depolarization, it exhibited periodic changes in the rhythm of efferent activity. Periodic changes of depolarization were virtually in phase in different ipsilateral segments of the lumbosacral enlargement. Data on groups of afferent fibers in whose central endings tonic and phasic changes of polarization took place. The appearance of fictitious locomotion was accompanied by a tonic increase, and of fictitious scratching by tonic inhibition of several evoked segmental responses. These tonic changes were a background against which segmental responses were modulated in step with the working rhythm of the locomotion and scratching generators. Many of the changes in evoked segmental responses were shown to be based on modulation of polarization of central endings of primary afferents by locomotion and scratching generators. It is concluded that active tonic and phase-dependent selection of incoming afferent information is effected through modulation of presynaptic inhibition of the generator. The role of this selection in peripheral collection of activity of locomotion and scratching generators is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 16, No. 3, pp. 343–353, May–June, 1984.     

Depolarizing effects of dopamine on the primary afferent fibers of a segment isolated from the spinal cord of newborn rats

Effects of dopamine on dorsal root potentials were investigated during experiments on a segment of spinal cord isolated from 12- to 18-day-old rats. Applying dopamine to the brain was found to produce a slow, reversible, dose-dependent depolarization at primary afferent fiber terminals. This dopamine-induced depolarization was retained during complete blockade of synaptic transmission brought about by exchanging calcium ions in the perfusing fluid by magnesium or manganese ions. Minimum dopamine concentration required to produce this effect was 1·10^–10–1·10^–9 M. Peak amplitude of depolarization equaled 1.5 mV. Duration of this reaction ranged from 5.5 to 36.7 min, depending on the duration and concentration of dopamine application. Depolarizing response to dopamine differed considerably from GABA-induced dorsal root depolarization in amplitude and rate of rise. Haloperidol, a dopamine antagonist, reduced dopamine-induced dorsal root depolarization. Findings indicate that dopamine acts directly on the membrane of primary afferent fiber terminals, shifting membrane potential toward depolarization. This raises the possibility that dopaminergic brainstem-spinal pathways may exert an effect on sensory information transmission in segmental reflex arcs already traveling to the spinal cord.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 19, No. 6, pp. 741–748, November–December, 1987.     

Integrative Synaptic Mechanisms in the Caudal Ganglion of the Crayfish

A study of activity recorded with intracellular micropipettes was undertaken in the caudal abdominal ganglion of the crayfish in order to gain information about central fiber to fiber synaptic mechanisms. This synaptic system has well developed integrative properties. Excitatory post-synaptic potentials can be graded, and synaptic potentials from different inputs can sum to initiate spike discharge. In most impaled units, the spike discharge fails to destroy the synaptic potential, thereby allowing sustained depolarization and multiple spike discharge following single pulse stimulation to an afferent input. Some units had characteristics which suggest a graded threshold for spike generation along the post-synaptic fiber membrane. Other impaled units responded to afferent stimulation with spike discharges of two distinct amplitudes. The smaller or "abortive" spikes in such units may represent non-invading activity in branches of the post-synaptic axon. On a few occasions one afferent input was shown to inhibit the spike discharge initiated by another presynaptic input.     

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.     

Monoaminergic Modulation of Spinal Viscero-Sympathetic Function in the Neonatal Mouse Thoracic Spinal Cord

Descending serotonergic, noradrenergic, and dopaminergic systems project diffusely to sensory, motor and autonomic spinal cord regions. Using neonatal mice, this study examined monoaminergic modulation of visceral sensory input and sympathetic preganglionic output. Whole-cell recordings from sympathetic preganglionic neurons (SPNs) in spinal cord slice demonstrated that serotonin, noradrenaline, and dopamine modulated SPN excitability. Serotonin depolarized all, while noradrenaline and dopamine depolarized most SPNs. Serotonin and noradrenaline also increased SPN current-evoked firing frequency, while both increases and decreases were seen with dopamine. In an in vitro thoracolumbar spinal cord/sympathetic chain preparation, stimulation of splanchnic nerve visceral afferents evoked reflexes and subthreshold population synaptic potentials in thoracic ventral roots that were dose-dependently depressed by the monoamines. Visceral afferent stimulation also evoked bicuculline-sensitive dorsal root potentials thought to reflect presynaptic inhibition via primary afferent depolarization. These dorsal root potentials were likewise dose-dependently depressed by the monoamines. Concomitant monoaminergic depression of population afferent synaptic transmission recorded as dorsal horn field potentials was also seen. Collectively, serotonin, norepinephrine and dopamine were shown to exert broad and comparable modulatory regulation of viscero-sympathetic function. The general facilitation of SPN efferent excitability with simultaneous depression of visceral afferent-evoked motor output suggests that descending monoaminergic systems reconfigure spinal cord autonomic function away from visceral sensory influence. Coincident monoaminergic reductions in dorsal horn responses support a multifaceted modulatory shift in the encoding of spinal visceral afferent activity. Similar monoamine-induced changes have been observed for somatic sensorimotor function, suggesting an integrative modulatory response on spinal autonomic and somatic function.     

Capsaicin and potassium evoked substance P release from the nucleus tractus solitarius and spinal trigeminal nucleus invitro

The nucleus tractus solitarius and the spinal trigeminal nucleus receive peripheral sensory input from substance P containing afferent nerves. This study demonstrates that $\text{in}$$\text{vitro}$ depolarization of these nuclei in tissue slices evokes a calcium-dependent efflux of substance P immunoreactivity. Capsaicin (33μM) also elicits substance P release from the nucleus tractus solitarius and spinal trigeminal nucleus but not from the hypothalamus. The occurrence of potassium-stimulated SP release from the two medullary nuclei fulfills one of the criteria for neurotransmitter status. The capsaicin data support the contention that this agent elicits release of substance P from nuclear regions receiving peripheral afferent information in substance P nerves independent of the particular sensory modality served but is ineffective in nonsensory areas.