Specific and potassium components in the depolarization of the Ia afferents in the spinal cord of the cat

In the cat spinal cord, primary afferent depolarization (PAD) of group Ia fibers of extensor muscles is produced by high-frequency stimulation (100 Hz) of group I muscle flexor afferents without significant increases in extracellular potassium. On the other hand, the PAD produced by stimulation of mixed and pure cutaneous nerves correlates well with increases in potassium ions. We conclude that the PAD produced by group I muscle afferents results from the activation of specific pathways making axo-axonic synapses with the Ia fiber terminals. The PAD of Ia fibers resulting from activation of cutaneous nerves involves instead unspecific accumulation of potassium ions.     

Persistence of PAD and presynaptic inhibition of muscle spindle afferents after peripheral nerve crush

Two to twelve weeks after crushing a muscle nerve, still before the damaged afferents reinnervate the muscle receptors, conditioning stimulation of group I fibers from flexor muscles depolarizes the damaged afferents [M. Enriquez, I. Jimenez, P. Rudomin, Changes in PAD patterns of group I muscle afferents after a peripheral nerve crush. Exp. Brain Res., 107 (1996), 405-420]. It is not known, however, if this primary afferent depolarization (PAD) is indeed related to presynaptic inhibition. We now show in the cat that 2-12 weeks after crushing the medial gastrocnemius nerve (MG), conditioning stimulation of group I fibers from flexors increases the excitability of the intraspinal terminals of both the intact lateral gastrocnemius plus soleus (LGS) and of the previously damaged MG fibers ending in the motor pool, because of PAD. The PAD is associated with the depression of the pre- and postsynaptic components of the extracellular field potentials (EFPs) evoked in the motor pool by stimulation of either the intact LGS or of the previously damaged MG nerves. These observations indicate, in contrast to what has been reported for crushed cutaneous afferents [K.W. Horch, J.W. Lisney, Changes in primary afferent depolarization of sensory neurones during peripheral nerve regeneration in the cat, J. Physiol., 313 (1981), 287-299], that shortly after damaging their peripheral axons, the synaptic efficacy of group I spindle afferents remains under central control. Presynaptic inhibitory mechanisms could be utilized to adjust the central actions of muscle afferents not fully recovered from peripheral lesions.     

Modification of primary afferent depolarization in cat group Ia afferents following high frequency intra-axonal tetanization of individual afferents

Intra-axonal tetanization of a single, functionally-identified group Ia afferent from the triceps surae muscle in the anesthetized cat produces marked enhancement and slowing of the primary afferent depolarization (PAD) generated in the Ia afferent by volleys in flexor muscle group Ia afferents, plus a pronounced transmembrane hyperpolarizing undershoot (HPU) which disappears more rapidly than the enhanced PAD. These alterations are qualitatively and quantitatively similar to those found after conditioning tetani are applied to the whole muscle nerve. The occurrence of these PAD changes after intra-axonal tetanization of a single group Ia afferent appears to rule out the participation of non-specific alterations in extracellular ionic concentrations or activation of polysynaptic pathways in their genesis.     

The role of cardiovascular and muscle afferent systems in control of body water balance

Influences of afferent inputs from cardiovascular and muscle receptors on the activities of neurosecretory neurons in the hypothalamus, which secrete vasopressin (ADH) were studied. Recordings were made from identified neurosecretory neurons in the supraoptic (SON) and paraventricular nuclei (PVN) of cats and rats. Activation of baroreceptors in the carotid sinus and aortic arch and atrial receptors inhibited SON and PVN neuron activities, while activation of chemoreceptors in the carotid sinus excited them. Repetitive electrical stimulation of the carotid sinus and aortic nerves showed that weak stimulation produced excitation and stronger stimulation produced inhibition of SON and PVN neurons. Electrical stimulation of these nerves and the nucleus tractus solitarius (NTS) by a single or short train of pulses showed that 'fast' and 'slow' pathways between the NTS and the SON existed, while these two types of pathways were not observed between the NTS and the PVN. Evidence of direct connections from the NTS to the PVN was found by means of antidromic stimulation of the PVN. Electrical stimulations of group I afferent fibers from the gastrocnemius muscle did not change SON neuron discharges, while activation of group III and IV afferent fibers excited them. Injection of chemicals (NaCl, KCl, bradykinin) into arteries supplying the muscle excited SON neurons. The excitation disappeared after section of the muscle nerves. The results indicated that activation of small afferents from the muscle excites the SON neurons, leading to an increase in vasopressin secretion. All these studies show that afferent inputs from receptors in the cardiovascular system and in the muscle have modulatory effects on neurosecretory neurons, and participate in control of body water balance by regulating vasopressin secretion from the neurohypophysis.     

Segmental organization of rat lateral longissimus, a muscle involved in lordosis behavior: EMG and muscle nerve recordings

Following stimulation of lumbar cutaneous and high threshold muscle afferents in urethane-anesthetized ovariectomized rats, polysynaptic multi-unit responses were recorded from muscle nerves to lateral longissimus (LL) at a mean latency of 9.5 ms. These responses could be reciprocally evoked between segments with no apparent rostro-caudal asymmetry. Polysynaptic responses habituated to repeated stimulation within 0.5-1.5 min but could be evoked again after several minutes rest. In estradiol-treated rats, 60% of the 9.5 ms responses were followed by late discharges (greater than 40 ms) as compared to 24% in untreated control animals (P = 0.05). Following stimulation to the L3 and L4 motor nerves to lateral longissimus, EMG responses were recorded from overlapping fields of innervation. The conduction velocity of the fast muscle fibers was determined to be 7 m/s. The short length of most of the LL muscle fibers and the intersegmental recruitment of LL motoneurons by cutaneous afferents is compatible with the integrated action of axial muscles in the performance of lordosis behavior.     

Inputs to spinocerebellar tract neurones located in stilling's nucleus in the sacral segments of the rat spinal cord

Spinocerebellar neurones located in the sacral segments of the rat spinal cord have been identified electrophysiologically. The neurones studied were located 0.7-1.1 mm deep to the cord dorsum, lateral and dorsal to the central canal in the medial part of lamina VII. Neurones were identified as spinocerebellar by antidromic activation from the cerebellar surface, the lowest threshold stimulation sites being near the midline on the posterior lobe vermis (lobule VIII). Estimates of conduction velocities of the axons ranged from 15-32 m/s (mean 22.8 m/s) and are directly comparable to velocities of presumed ventral spinocerebellar tract neurones recorded in the same animals. In intact animals, activity was most strongly influenced by passive movement of the tail. Activation by proprioceptors was confirmed with nerve stimulation: all of the neurones studied were discharged by stimulation of nerves which innervate ipsilateral tail muscles. In many cases responses appeared close to the threshold of the nerve, indicating that the largest, fastest conducting afferents (group Ia muscle spindle primary afferents) were responsible for them. Latencies of EPSPs or spikes were brief and in many cases indicative of a monosynaptic connection. We conclude that this group of neurones is powerfully and monosynaptically excited by group I muscle afferents and thus resemble the cells of Clarke's column and cells of the central cervical nucleus, both of which occupy a similar location in the grey matter of more rostral segments.     

Projection of phrenic afferents to the external cuneate nucleus in the cat

The present study, performed on anesthetized, spontaneously breathing cats, deals with the projection of group I and II muscle afferents of the phrenic nerve (PN) to the external cuneate nucleus (ECN). Stimulation of the central end of the PN evoked a complex response in the ipsilateral ECN. Two principal components could be distinguished in this potential from the respective absolute refractory periods (ARP) and from the effect of antidromic stimulation in the ECN. Thus, the early group of waves may correspond to recordings of direct fibers and the later group to postsynaptic activations within the ECN. Similar to the forelimb nerves and intercostal nerves of the upper intercostal spaces, the larger muscle afferents of the PN project to the ECN.     

The effects of naloxone, morphine and methionine enkephalinamide on Ia afferent terminations in the cat spinal cord

Naloxone, morphine, Met5-enkephalinamide (MENKA) and procaine were administered microelectrophoretically near extracellularly stimulated extensor muscle group Ia afferent fibres and terminations in the lumbar spinal cord of cats anaesthetized with pentobarbitone sodium. Observations were made of effects on the electrical threshold, on the depolarizing action of GABA or piperidine-4-sulphonate (P4S), and on bicuculline-sensitive primary afferent depolarization (PAD) generated by tetanic stimulation of flexor muscle low threshold afferents. All 4 agents reversibly elevated the threshold of Ia fibres in the dorsal column and Ia terminations in the ventral horn. The depolarizations of terminations by GABA or P4S were also reduced, an effect, which for all except MENKA, probably accounted for a concomitant reduction in PAD. In the absence of a consistent effect on either threshold or depolarization by GABAmimetics, MENKA reversibly diminished PAD, an action blocked by naloxone. Intravenously administered naloxone, in doses known to enhance spinal monosynaptic excitation in the cat, had no effect on GABAergic PAD and little or no effect on Ia termination threshold. The results are discussed in relation to a naloxone-sensitive effect of MENKA which reduces transmitter release from GABAergic axo-axonic synapses on Ia terminals, but which does not account for the enhancement of spinal reflexes by naloxone.     

Activation of spinal cord interneurons which process inputs from the femoral-saphenous vein

Recordings were made from single spinal cord interneurons which could be activated by electrical stimulation of afferents terminating in the wall of the femoral-saphenous vein. Interneurons were either excited or both excited and inhibited by venous afferent stimulation. Most of the venous afferent-driven interneurons could also be driven by electrical activation of A-alpha beta muscle and cutaneous afferents. Stimulation of several different muscle nerves drove single interneurons.     

Fatigue-induced changes in group IV muscle afferent activity: differences between high- and low-frequency electrically induced fatigues

Recordings of group IV afferent activity of tibialis anterior muscle were performed in paralysed rabbits during runs of electrically induced fatigue produced by direct muscle stimulation at a high (100 Hz, high-frequency fatigue HFF) or a low rate (10 Hz, low-frequency fatigue LFF). In addition to analysis of afferent nerve action potentials, muscle force and compound muscle action potentials (M waves) elicited by direct muscle stimulation with single shocks were recorded. Changes in M wave configuration were used as an index of the altered propagation of membrane potentials and the associated efflux of potassium from muscle fibers. The data show that increased group IV afferent activity occurred during LFF as well as HFF trials and developed parallel with force failure. Enhanced afferent activity was significantly higher during LFF (maximal Δf_impulses=249±35%) than HFF (147±45%). No correlation was obtained between the responses of group IV afferents to LFF or to pressure exerted on tibialis anterior muscle. On the other hand, decreased M wave amplitude was minimal with LFF while it was pronounced with HFF. Close correlations were found between fatigue-induced activation of group IV afferents and decreases in force or M wave amplitude, but their strength was significantly higher with LFF compared to HFF. Thus, electrically induced fatigue activates group IV muscle afferents with a prominent effect of low-frequency stimulation. The mechanism of muscle afferent stimulation does not seem to be due to the sole increase in extracellular potassium concentration, but also by the efflux of muscle metabolites, present during fatiguing contractions at low rate of stimulation.     

Depression of the recurrent inhibition of extensor motoneurons by the action of group II afferents

The influence of varying the muscular afferent fiber input on both the normal firing rate (F_n) and the amount of recurrent inhibition (F_n−F_i) induced by a constant ventral root stimulation was investigated on tonic extensor motoneurons recorded from ventral root filaments in decerebrate cats. The afferent input was varied by graded electrical stimulation of the gastrocnemius nerves and by vibrating the triceps surae muscle (100 μm amplitude). When the input consisted solely of impulses in Ia afferents, as was the case during vibration, the mean recurrent inhibition F_n−F_i was 2.3 times greater than during nerve tetanization at 1.8 times threshold of group I (T_I). This strength generally excited all group I and some low-threshold group II afferents. Between 1.8 T_I and 8 T_I, F_n−F_i decreased by some 50%. The average F_n increased slightly and motoneurons with a phasic discharge pattern were recruited when the stimulus strength was raised so as to excite group II afferents; these cells were never recruited during vibration and nerve tetanization at 1.8 T_I. The results indicate the possibility of a disinhibitory action of secondary muscle spindle afferents on extensor motoneurons by reducing the recurrent inhibition.     

Demonstration of initial axon collaterals of cells of origin of the ventral spinocerebellar tract in the cat

Neurones of origin of the ventral spinocerebellar tract were stained with intracellularly applied horseradish peroxidase to investigate whether they give off any initial axon collaterals. The neurones were located in the fourth and fifth lumbar segments and were identified by their antidromic activation following stimulation in the contralateral superior cerebellar peduncle. Nine of the 23 neurones with well-stained axons were found to give off axon collaterals soon after the axons crossed the midline. The collaterals entered the contralateral ventral horn and branched within lamina VII and the dorsal part of lamina VIII. Collaterals were found arising only from neurones located in the middle of lamina VII and from axons which took a mediorostral direction. In all of these neurones excitatory postsynaptic potentials were evoked from group Ia afferents of at least some nerves, in addition to such potentials from Ib or unidentified group I afferents, and inhibitory postsynaptic potentials were evoked from group I and II afferents. The area of terminal branching of the collaterals suggests that they may supply contralateral ventral spinocerebellar neurones with information from muscles and/or mediate crossed reflexes from group I afferents.     

Convergence of muscle spindle afferents on single neurons of the cat dorsal spino-cerebellar tract and their synaptic efficacy

By means of tungsten microelectrodes, action potentials from axons within the dorsal spino-cerebellar tract (DSCT) and from muscle spindle afferents were recorded. A quantitative study was performed in monomuscular DSCT neurons which were excited predominantly by Ia fibers originating in the gastrocnemius muscles. In some experiments single Ia fibers were stimulated electrically while the impulse sequence of a DSCT neuron postsynaptic to the respective afferent fiber was recorded. The gastrocnemius DSCT neurons receive excitatory inputs from 10-18 Ia muscle spindle afferents. The efficacy of each of these inputs is very similar. Thus the neuronal activation decreased approximately linearly with the number of the excitatory afferents cut. Cross-correlograms between the impulse sequence of a Ia gastrocnemius muscle spindle afferent and a DSCT neuron postsynaptic to it exhibited an increased discharge probability of the DSCT neuron from 3-4 ms to 10 ms after the Ia action potential. With increasing impulse rates of the Ia afferent fibers, the excitatory efficacy of the single action potential decreased, but the overall excitation increased with the presynaptic discharge frequency, according to a hyperbolic function. This effect was tested by electrical stimulation of a single Ia axon exciting the DSCT neuron recorded. Interval histograms computed from DSCT neuron impulse trains at steady stretch conditions were predominantly monomodal. They can be well approximated by a Gaussian distribution. The coefficient of variation was independent of the mean activity. At impulse rates above 25 imp X s-1 a negative correlation between successive intervals was observed in first order joint interval diagrams. With an increasing mean discharge rate this correlation (expressed as the serial linear correlation coefficient of the first order r1,2) became stronger up to--0.62 at 90 imp X s-1. Only in a few neurons did the higher order linear correlation coefficients deviate significantly from zero. In 15% of the observed histograms double discharging (mean interval 3-5 ms) produced bimodal distributions. Under steady-state conditions the response of Ia-activated DSCT cells are linearly related to muscle stretch within a middle range of extensions. The differences between Ia impulse pattern and DSCT neuron impulse pattern at steady stretch are discussed. The number of large dendrites of the principal cells in the nucleus dorsalis (Clarke's column) corresponds to the number of excitatory afferent muscle fibers. It is assumed that each excitatory Ia axon sends one axon collateral to the DSCT neuron, forming a climbing type terminal mainly on one of the large dendrites of a DSCT cell.     

Effects of stimulus intensity, cervical cord tractotomies and cerebellectomy on somatosensory evoked potentials from skin and muscle afferents of cat hind limb

The somatosensory evoked potentials (SEPs) recorded from the sensory cortex were investigated by using graded stimulation of skin and muscle nerves from contralateral hind limb in the cat. Sections were made of the middle cervical cord to assess the pathways involved in mediating SEPs evoked by large and small diameter fibers. Dorsal column (DC) section caused a decrease of SEPs from skin group I afferents, and a small increase in those from group I muscle afferents. A subsequent section of dorso-lateral fasciculus (DLF) further decreased SEPs from skin and eliminated SEPs from muscle, evoked at low stimulus intensity. When the stimulus recruited group III fibres, SEPs were still present after DC and DLF section, both from skin and muscle nerves. Section of ALT in addition to DC confirmed a major role played by DLF (mainly spino-cervical tract of Morin) in transmitting impulses from muscle afferents; the role of DLF in mediating potentials evoked from skin is less remarkable than that of DC. Cerebellectomy did not change any SEP, however evoked. Previous results in the literature are discussed, taking into account the methodologies employed by various authors, and the possible interactions among pathways mediating SEPs.     

Synaptic effects from chemically activated fine muscle afferents upon α-motoneurones in decerebrate and spinal cats

In spinal and decerebrate cats fine muscle afferents (group III and IV) were selectively activated by intra-arterial injection of bradykinin and KCl into the gastrocnemius-soleus muscle. By this method the synaptic responses induced in lumbar α-motoneurones by fine muscle afferents could be examinedwithout interference of effect from large afferents. α-Motoneurones receiving EPSPs evoked by electrical stimulation of cutaneous and high threshold muscle afferents (mainly flexor motoneurones) responded to the activation of fine muscle afferents with a depolarization of their membrane and an increase in synaptic noise, while motoneurones in which IPSPs were evoked by electrical stimulation of cutaneous and high threshold muscle afferents (mainly extensor motoneurones), responded with hyperpolarization of their membrane. Cells with mixed excitatory-inhibitory electrically induced response characteristic responded with an increase in synaptic noise without substantial change in the level of their membrane potential to chemical stimulation of fine muscle afferents. The results indicate that one function of group IIIand IV muscle afferents is to participate in the complex reflex control performed by the flexor reflex system.     

The cortical distribution of muscle and cutaneous afferent projections from the human foot

Somatosensory evoked potentials to electrical stimulation of muscle and cutaneous afferents from the foot were recorded in normal human subjects using multiple channels centred on the vertex and referenced to the contralateral earlobe. Low-threshold muscle afferents were selectively activated by an insulated microelectrode inserted percutaneously at the motor point of abductor hallucis. Low-threshold cutaneous and joint afferents of the hallux or second toe were stimulated with ring electrodes. The posterior tibial and sural nerves were stimulated at the ankle through surface electrodes. The cerebral distribution of the initial cortical response (N33-P40) to stimulation of muscle afferents largely paralleled that to stimulation of its parent nerve, the posterior tibial nerve (which contains afferents of muscle, cutaneous and joint origin). They were maximal slightly posterior and ipsilateral to the vertex. The cutaneous-joint afferent projection from the hallux paralleled that from the sural nerve and both were less lateralized than the tibial and abductor hallucis projections.     

Contribution of cutaneous and muscle afferent fibres to cortical SEPs following median and radial nerve stimulation in man

Somatosensory evoked potentials were recorded after stimulation of motor and cutaneous nerves in the upper limb. Stimulation of the thenar motor branch of the median nerve and the deep motor branch of the radial nerve produced only broad, ill-defined and small-amplitude scalp-recorded responses. In contrast, stimulation of purely cutaneous nerves (digital and the superficial radial) gave responses of large amplitude. The cortical responses following combined deep and superficial radial nerve stimulation were of smaller amplitude than the two individual responses combined. These findings suggest that, contrary to an earlier report (Gandevia et al. 1984), muscle afferents do not make a major positive contribution to the scalp-recorded cortical responses produced by electrical stimulation of mixed nerves in the upper limb.     

Interfering cutaneous stimulation and the muscle afferent contribution to cortical potentials

Cerebral potentials were recorded in response to selective stimulation using microelectrodes of muscle afferents in motor fascicles innervating the intrinsic muscles of the foot or at the motor point of abductor hallucis. The early components of these potentials (P40, N50 and P60) were consistently attenuated by continuous tactile stimulation of related skin areas and by electrical stimulation of digital nerves, timed so that the digital volley reached cortex approximately 5 msec before the muscle afferent volley. The same conditioning cutaneous inputs also attenuated the cerebral potentials evoked by selective stimulation of cutaneous afferents. These findings confirm that there are intermodality and intramodality interactions between low-threshold cutaneous and muscle afferents and between cutaneous afferents, respectively. The findings indicate that 'interference phenomena' (Kakigi and Jones 1986) can occur between different afferent modalities, and within any one modality, and cannot be used to determine the afferent species responsible for the test evoked potential.     

Effect of superficial radial nerve stimulation on the activity of nigro-striatal dopaminergic neurons in the cat: Role of cutaneous sensory input

Summary The release of³H-dopamine (DA) continuously synthesized from³H-tyrosine was measured in the caudate nucleus (CN) and in the substantia nigra (SN) in both sides of the brain during electrical stimulation of the superficial radial nerve in cats lightly anaesthetized with halothane. Use of appropriate electrophysiologically controlled stimulation led to selective activation of low threshold afferent fibers whereas high stimulation activated all cutaneous afferents.Results showed that low threshold fiber activation induced a decreased dopaminergic activity in CN contralateral to nerve stimulation and a concomitant increase in dopaminergic activity on the ipsilateral side. Stimulation of group I and threshold stimulation of group II afferent fibers induced changes in the release of³H-DA mainly on the contralateral CN and SN and in the ipsilateral CN. High stimulation was followed by a general increase of the neurotransmitter release in the four structures.This shows that the nigro-striatal dopaminergic neurons are mainly-if not exclusively-controlled by cutaneous sensory inputs. This control, primarily inhibitory in the side contralateral to the stimulation, seems rather non-specific when high threshold cutaneous fibers are also activated. Such activations could contribute to restablish sufficient release of DA when the dopaminergic function is impaired as in Parkinson's disease.