Possibility of different effects of primary afferent depolarization on spinal reflexes

The development of intensive depolarization of terminals through stimulation of the afferent nerve or dorsal root of sufficient strength causes the appearance of antidromic discharges recordable as a dorsal-root reflex. The discharges were shown to spread simultaneously in the dorso-ventral direction, with consequent facilitation of spinal reflexes. The results suggest the possible existence of two types of effect of presynaptic depolarization on spinal afferent pathways.     

Role of propriospinal neurons in inhibition of the afferent flow

The effects of stimulation of the dorsal funiculus on dorsal surface potentials (DSPs) of the spinal cord evoked by stimulation of a peripheral nerve and on antidromic action potentials (AAPs) evoked by stimulation of terminal branches of primary afferent fibers and recorded from the afferent nerve or dorsal root, were investigated in acute experiments on spinal cats and on cats anesthetized with pentobarbital and chloralose. Stimulation of the dorsal funiculus led to biphasic inhibition of the N₁-component of the DSP with maxima at the 15th–30th and 60th–80th milliseconds between the conditioning and testing stimuli. Maximal reinforcement of the AAP was found with these intervals. Bilateral division of the dorsal funiculi between the point of application of the conditioning stimuli and the point of recording the DSP abolished the first wave of inhibition of the DSP and the reinforcement of the AAP. After total transection of the cord above the site of conditioning stimulation the picture was unchanged. It is concluded that the initial changes in DSP and AAP are due to activation of the presynaptic inhibition mechanism by antidromic impulses traveling along nerve fibers running in the dorsal funiculus. Repeated inhibition of the DSP, like reinforcement of the AAP, can possibly be attributed to activation of similar inhibitory mechanisms through the propriospinal neurons of the spinal cord.Dnepropetrovsk State University. Translated from Neirofiziologiya, Vol. 5, No. 4, pp. 401–405, July–August, 1973.     

Inhibition of spinal reflex responses in the last period of embryogenesis

Some characteristics of spinal reflex reaction inhibition were studied in cat fetuses during the last three weeks of antenatal development. The experiments were conducted on fetuses with intact placental circulation. Restoration of the excitability of the spinal reflex arcs was very slow after stimulation of the dorsal root by a single stimulus. In embryos studied 20 days before birth the full inhibition of reflex responses lasted about 500 msec. Even 2–3 sec after a single stimulation of the afferent fibers the amplitude of the reflex response to the second stimulus was only 30–40% of the control value. It was determined that such long postactivation depression is unrelated to refractoriness or antidromic inhibition. The presence of a prolonged intense depolarization of afferent terminal fibers at these stages suggests a presynaptic inhibition as one of the most probable reasons for the prolonged postactivation depression. Another important factor in the appearance of postactivation depression is probably the morphologic and functional immaturity of synaptic structures. A reciprocal inhibition was observed in cat fetuses on the 10–12th antenatal day. On the basis of these data it is suggested that in embryogenesis presynaptic inhibition considerably precedes that of postsynaptic fibers.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 3, No. 1, pp. 68–75, January–February, 1971.     

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.     

Antidromic dorsal root impulses during naturally occurring locomotion in rats

Antidromic dorsal root activity during naturally occurring locomotion (swimming and treadmill walking) was investigated during experiments on white rats. The activity observed consisted of phaselinked and tonic components of antidromic action potentials (APP). A strong correlation was found between intensity of AAP and that of afferent input during actual locomotion; AAP correlated less well with degree of electromyographic activity. Possible sources of the initiation of antidromic activity and the part played in spinal reflex control by presynaptic depolarization are discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 20, No. 5, pp. 579–585, September–October, 1988.     

Segmental inhibitory response in the frog spinal cord during orthodromic motoneuron activation

Different types of reflex discharges were produced in various preparations by stimulating the dorsal root of isolated frog spinal cord. These ranged from multiphasic low-amplitude waves to distinctly synchronized monosynaptic response. The discharges were followed by facilitation in the former and deep, protracted inhibition of response to test dorsal root stimulation in the latter. When interstimulus intervals measured 40–50 msec, inhibitory action was less pronounced than at shorter (15–30 msec) or longer (60–100 msec) intervals, thus indicating that at least two types of inhibition were at work, one at an earlier and the other at a later stage. Strychnine at a concentration of 10^–5 M effectively reinforced the former and blocked the latter, while 10^–4 M d-tubocurarine attenuated both types of inhibition substantially. It is concluded that inhibition of response occurs mainly as a result of recurrent activation of inhibitory systems via recurrent motoneuron axon collaterals when frog spinal cord afferents are excited. Intensity of the later (presynaptic) and earlier (postsynaptic) inhibition of reflex transmission is determined by the degree of synchrony in motoneuronal discharge in response to orthodromic stimulation.Institute of Medical Radiology, Academy of Medical Sciences of the USSR, Obninsk, Kaluga Oblast. Translated from Neirofiziologiya, Vol. 19, No. 3, pp. 343–350, May–June, 1987.     

Electrophysiology of the Fetal Spinal Cord : II. Interaction among peripheral inputs and recurrent inhibition

Interactions of peripheral inputs to the motoneuron of the kitten fetus as young as 3 weeks prenatal were studied by reflex discharge from the ventral root as well as by recording from single motoneurons. Facilitation was found between two synergists in fetuses 1 to 2 weeks before birth. Intracellular recording showed that the facilitation could be explained by summation of excitatory postsynaptic potentials. Inhibition was found between antagonists in the fetuses 2 to 3 weeks before birth and was accompanied by inhibitory postsynaptic potentials. Recurrent inhibition was very powerful in the fetal spinal cord as shown by large motoneuron hyperpolarization by antidromic stimulation. Cells presumed to be "Renshaw cells" and which responded to both ortho- and antidromic stimulation with repetitive firing were shown in the 2 weeks prenatal fetus. These results lead to the conclusion that there is considerable effective synaptic connection of afferent collaterals already established by the later stage of intrauterine life and that this may be achieved independently of external stimuli.     

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.     

The effect of stimulation of Golgi tendon organs and spindle receptors from hindlimb extensor muscles on supraspinal descending inhibitory mechanisms

Experiments were performed in precollicular decerebrate cats to investigate whether proprioceptive volleys originating from Golgi tendon organs and muscle spindles may activate supraspinal descending inhibitory mechanisms. Conditioning stimulation of the distal stump of ventral root filaments of L7 or S1 leading to isometric contraction of the gastrocnemius-soleus (GS) muscle inhibited the monosynaptic reflex elicited by stimulation of the ipsilateral plantaris-flexor digitorum and hallucis longus (Pl-FDHL) nerve. The amount and the time course of this Golgi inhibition were greatly increased by direct cross-excitation of the intramuscular branches of the group Ia afferents due to ephaptic stimulation of the sensory fibers, which occurred when a large number of a fibers had been synchronously activated. The postsynaptic and the presynaptic nature of these inhibitory effects, as well as their segmental origin, have been discussed. In no instance, however, did the stimulation of Golgi tendon organs elicit any late inhibition of the test monosynaptic reflex, which could be attributed to a spino-bulbo-spinal (SBS) reflex. Conditioning stimulation of both primary and secondary endings of muscle spindles, induced by dynamic stretch of the lateral gastrocnemius-soleus (LGS) muscle, was unable to elicit any late inhibition of the medial gastrocnemius (MG) monosynaptic reflex. The only changes observed in this experimental condition were a facilitation of the test reflex during the dynamic stretch of the LGS, followed at the end of the stimulus by a prolonged depression. These effects however were due to segmental interactions, since they persisted after postbrachial section of the spinal cord. Intravenous injection of an anticholinesterase, at a dose which greatly potentiated the SBS reflex inhibition produced by conditioning stimulation of the dorsal root L6, did not alter the changes in time course of the test reflex induced either by muscle contraction or by dynamic muscle stretch. Conditioning stimulation of a muscle nerve activated the supraspinal descending mechanism responsible for the inhibitory phase of the SBS reflex only when the high threshold group III muscle afferents (innervating pressure-pain receptors) had been recruited by the electric stimulus. This finding contrasts with the great availability of the system to the low threshold cutaneous afferents. The proprioceptive afferent volleys originating from Golgi tendon organs as well as from both primary and secondary endings of muscle spindles, contrary to the cutaneous and the high threshold muscle afferent volleys, were apparently unable to elicit not only a SBS reflex inhibition, but also any delayed facilitation of monosynaptic extensor reflexes attributable to inhibition of the cerebellar Purkinje cells.     

Contribution of antidromic efferent-fiber excitation to mass spinal potentials in the cat

The contribution of antidromic excitation of motoneurons to cord dorsum potentials (CDP) was studied in the spinal cord of anesthetized cats. It was shown that stimulation of ventral roots (VR) or peripheral nerves following deafferentiation of a number of segments by crosscutting of dorsal roots on the dorsal surface evokes appreciable positive-negative CDP (VR-CDP). Under intact conditions, VR effects of antidromic stimulation of efferent fibers brings appreciable input to the initial "fast" CDP component (the "afferent" peak); input values for the main mixed nerves of the hindlimb are presented. After conditioning stimulation of a mixed nerve, VR-CDP undergo inhibition with two maximums, associated with blocking of the effects of antidromic excitation of efferents by orthodromic mono- and polysynaptic reflex discharges of motoneurons. The hypothesis that intactness of efferents in nerves under stimulation can be determined from an analysis of initial CDP components is stated.Scientific-Research Institute of Biology, Dnepropetrovsk State University, Dnepropetrovsk. Translated from Neirofiziologiya, Vol. 23, No. 6, pp. 655–661, November–December, 1991.     

Intracellular recording of the frog dorsal root single fibres' responses mediated by the GABAA and 5-HT-receptors

The effects of GABA, bicuculline and 5-HT on primary afferents in the isolated spinal cord of the frog Rana ridibunda were studied. Bath application of GABA (1 mM) reduced the primary afferent depolarisation (PAD) in IX segment of the spinal cord evoked by X dorsal root stimulation (57 +/- 8% of initial level, n = 5, p < 0.05). The action potentials (AP) recorded in dorsal root afferents was also suppressed under the GABA action (74 +/- 9%, p < 0.05). Bath application of bicuculline (50 microM) reduced the PAD (21 +/- 7%), n = 6, p < 0.05), meanwhile the AP in dorsal root afferents was resistant against the bicuculline action. Bath application of 5-HT (25 microM) depressed the PAD (34 +/- 7%, n = 7, p < 0.05) and the amplitude of the AP recorded from the single afferent fibre in dorsal column (76 +/- 6%, n = 7, p < 0.05). In contrast to GABA, 5-HT more effectively suppressed the late phase of the PAD evoked by X dorsal root stimulation and caused (76 +/- 6%, n = 7, p < 0.05) an alteration of the AP shape. All effects induced by these drugs were reversible. The mechanisms of GABA and 5-HT modulation of spinal cord afferent income are discussed.     

A possible coordinating role for presynaptic inhibition

Conditioning stimuli were applied to the common peroneal or superficial peroneal nerve in acute experiments on anesthetized cats. Changes in the N₁-component of the dorsal cord potential evoked by stimulation of one of these nerves or of other nerves (tibial, deep peroneal) and changes in the amplitude of antidromic action potentials in the afferent fibers of these nerves were investigated. The degree of reinforcement of antidromic action potentials, reflecting the degree of depolarization of the afferent terminals, was found to be greater for the passive nerve than for the active to which the conditioning stimulus was applied. Inhibition of the N₁-component of the dorsal cord potential was deeper when a pair of stimuli was applied to two different nerves (under these conditions only the mechanism of presynaptic inhibition was activated) than when they were applied to the same nerve. It is concluded that presynaptic inhibition, by selectively controlling afferent volleys, can evidently play a coordinating role.     

Extrapyramidal descending influences on neurons of the spinal cord in a superreflexia state

The spinal superreflexia state was modeled in experiments on rats using preliminary transection of the spinal cord and injection (in the course of the acute experiment) of 4-aminopyridine. An extremely high (reaching 15–20 mV) amplitude of monosynaptic reflex discharges (MRs) evoked by stimulation of the dorsal root and recorded from the ventral root (VR) L ₄ and the presence of an additional component in the above discharges were phenomena indicative of the development of the above state. Under such conditions, the amplitudes of the discharges evoked in the VR by electrical stimulation of the round window of the labyrinth (vestibular stimulation) and of the discharges elicited by stimulation of the motor cortex under conditions of bilateral transection of the pyramids increased several times. Thresholds of the VR responses to vestibular and cortical stimulations demonstrated an about threefold drop; latencies of the mass responses and responses of single spinal moto-and interneurons decreased about twofold, on average. The pattern of vestibular conditioning effects on the VR MRs changed: in intact animals vestibular stimulation induced inhibition of the VR MRs, while in animals with superreflexia such stimulation led to facilitation of the MRs. Cortical stimulation under conditions of pyramidotomy in both intact animals and animals with superreflexia resulted in facilitation of the VR MRs of a nearly the same intensity. The levels of convergence of the segmental and supraspinal effects on interneurons and motoneurons of the rat spinal cord dramatically increased under superreflexia conditions. The possible mechanisms of augmentation of the descending influences on spinal neuronal systems under the above conditions are discussed. Neirofiziologiya/Neurophysiology, Vol. 38, No. 2, pp. 140–149, March–April, 2006.     

Presynaptic inhibition and antidromic discharges in crayfish primary afferents.

The mechanisms of presynaptic inhibition have been studied in sensory afferents of a stretch receptor in an in vitro preparation of the crayfish. Axon terminals of these sensory afferents display primary afferent depolarisations (PADs) mediated by the activation of GABA receptors that open chloride channels. Intracellular labeling of sensory axons by Lucifer yellow combined with GABA immunohistochemistry revealed the presence of close appositions between GABA-immunoreactive boutons and sensory axons close to their first branching point within the ganglion. Electrophysiological studies showed that GABA inputs mediating PADs appear to occur around the first axonal branching point, which corresponds to the area of transition between active and passive propagation of spikes. Moreover, this study demonstrated that whilst shunting appeared to be the sole mechanism involved during small amplitude PADs, sodium channel inactivation occurred with larger amplitude PADs. However, when the largest PADs (>25 mV) are produced, the threshold for spike generation is reached and antidromic action potentials are elicited. The mechanisms involved in the initiation of antidromic discharges were analyzed by combining electrophysiological and simulation studies. Three mechanisms act together to ensure that PAD-mediated spikes are not conveyed distally: 1) the lack of active propagation in distal regions of the sensory axons; 2) the inactivation of the sodium channels around the site where PADs are produced; and 3) a massive shunting through the opening of chloride channels associated with the activation of GABA receptors. The centrally generated spikes are, however, conveyed antidromically in the sensory nerve up to the proprioceptive organ, where they inhibit the activity of the sensory neurons for several hundreds of milliseconds.     

The dorsal root reflex in isolated mammalian spinal cord

1. The dorsal root reflex has been investigated in an isolated preparation of adult mammalian spinal cord. 2. Both evoked and spontaneous activity can be recorded from the cord in the dorsal spinal roots. 3. The spontaneous activity has a characteristic pattern of firing in bursts of action potentials. Spontaneous and evoked activity are optimum at temperatures between 25 and 27 degrees C; little activity can be detected above 35 degrees C. 4. The spontaneous dorsal root activity has been shown to be correlated with negative potentials in the dorsal horn of the cord, and intracellular recordings made from primary afferent fibres have shown spontaneous primary afferent depolarizations (PAD) which underlie the generation of the spontaneous dorsal root activity. 5. The evoked dorsal root reflex has been shown to spread up to 16 spinal segments both rostrally and caudally from the stimulated dorsal root, and to the contralateral side of the cord. 6. The spontaneous dorsal root activity in widely separated segments has been shown by cross-correlation analysis to be linked both ipsi- and contra-laterally. 7. The significance of such a widespread system for the generation of PAD is discussed.     

Modeling the spinal pudendo-vesical reflex for bladder control by pudendal afferent stimulation

Electrical stimulation of the pudendal nerve (PN) is a promising approach to restore continence and micturition following bladder dysfunction resulting from neurological disease or injury. Although the pudendo-vesical reflex and its physiological properties are well established, there is limited understanding of the specific neural mechanisms that mediate this reflex. We sought to develop a computational model of the spinal neural network that governs the reflex bladder response to PN stimulation. We implemented and validated a neural network architecture based on previous neuroanatomical and electrophysiological studies. Using synaptically-connected integrate and fire model neurons, we created a network model with realistic spiking behavior. The model produced expected sacral parasympathetic nucleus (SPN) neuron firing rates from prescribed neural inputs and predicted bladder activation and inhibition with different frequencies of pudendal afferent stimulation. In addition, the model matched experimental results from previous studies of temporal patterns of pudendal afferent stimulation and selective pharmacological blockade of inhibitory neurons. The frequency- and pattern-dependent effects of pudendal afferent stimulation were determined by changes in firing rate of spinal interneurons, suggesting that neural network interactions at the lumbosacral level can mediate the bladder response to different frequencies or temporal patterns of pudendal afferent stimulation. Further, the anatomical structure of excitatory and inhibitory interneurons in the network model was necessary and sufficient to reproduce the critical features of the pudendo-vesical reflex, and this model may prove useful to guide development of novel, more effective electrical stimulation techniques for bladder control.     

Functional expression of AMPA receptors on central terminals of rat dorsal root ganglion neurons and presynaptic inhibition of glutamate release

No direct evidence has been found for expression of functional AMPA receptors by dorsal root ganglion neurons despite immunocytochemical evidence suggesting they are present. Here we report evidence for expression of functional AMPA receptors by a subpopulation of dorsal root ganglion neurons. The AMPA receptors are most prominently located near central terminals of primary afferent fibers. AMPA and kainate receptors were detected by recording receptor-mediated depolarization of the central terminals under selective pharmacological conditions. We demonstrate that activation of presynaptic AMPA receptors by exogenous agonists causes inhibition of glutamate release from the terminals, possibly via primary afferent depolarization (PAD). These results challenge the traditional view that GABA and GABA(A) receptors exclusively mediate PAD, and indicate that PAD is also mediated by glutamate acting on presynaptically localized AMPA and kainate receptors.     

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.     

Effects of tractotomy on reflex activity in the lumbar segment of the rat spinal cord previously disrupted by severing the sciatic nerve

Evoked electrical discharges in the spinal cord roots and dorsal surface ipsilateral to the previously severed sciatic nerve (as well as on the contralateral side) were investigated in rats one, three, seven, and 14 days after tractotomy. Monosynaptic reflex discharges in the ventral roots were found to return to 20–40% of the level of this parameter as measured on the contralateral side within seven and 14 days after tractotomy. Mean amplitude of antidromic dorsal root discharges, afferent peak, and the N₁ component of potential(s) at the dorsal surface ipsilateral to the severed nerve barely altered, remaining significantly lower than on the contralateral side. Mechanisms are suggested for the increase in monosynaptic reflex ventral root discharges ipsilateral to the severed nerve following tractotomy — thought to be largely due to raised sensitivity to transmitter at the motoneuronal membrane resulting from degeneration of synapses of descending pathways.Medical Institute of the Ukrainian Ministry of Health, Dnepropetrovsk. Translated from Neirofiziologiya, Vol. 21, No. 3, pp. 366–371, May–June, 1989.     

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.