Functional coupling between motor and sensory nerves through contraction of sphincters in the pudendal area of the female cat.

The question of whether skin receptors might help in the perception of muscle contraction and body movement has not been settled. The present study gives direct evidence of skin receptor firing in close coincidence with the contraction of the vaginal and anal sphincters. The distal stump of the sectioned motor pudendal nerve was stimulated. Single shocks induced a wavelike increase in the lumen pressure of the distal vagina and the anal canal, as well as constriction of the vaginal introitus and the anus. The constriction pulls on and moves the surrounding skin, which was initially detected visually. In the present experiments, a thin strain gauge that pressed on the skin surface detected its displacement. Single shocks to the motor nerve induced a wave of skin movement with maximal amplitude at 5 mm from the anus and propagated with decrement beyond 35 mm. The peripheral terminals of the sensory pudendal nerve and the posterior femoral nerve supply the skin that moves. Sensory axons from both nerves fired in response to both tactile stimulation and the skin movement produced by the constriction of the orifices (motor-sensory coupling). In cats with all nerves intact, a single shock to the sensory nerves induced reflex waves of skin movement and lumen pressure (sensory-motor coupling). Both couplings provide evidence for a feedforward action that might help to maintain the female posture during mating and to the perception of muscle contraction.

Structure, innervation, mechanical properties and reflex activation of a striated sphincter in the vestibule of the cat vagina.

Vaginal constriction might be important for reproduction in mammals, but existing information is both limited and controversial. This paper shows the structure, mechanical properties, innervation and reflex response of a striated sphincter in the vestibule of the cat vagina. A Foley catheter coupled to a pressure transducer detected in the lumen of the vestibule a pressure wave that was induced by stimulation of the external branch of the motor pudendal nerve. The peak pressure of the wave induced by bilateral stimulation (30.6 cm H(2)O) was about double of the peak pressure wave induced unilaterally. The tetanus/twitch amplitude ratio was 4.5. The sphincter that produces the increase in vaginal pressure fatigues slowly. Digital, point-to-point summation of unilateral waves was greater than the wave induced bilaterally. Summation of the pressure wave induced by the separate stimulation of the terminal motor branches was also greater than the wave induced by the entire motor nerve. This might reflect multiple innervation of muscle fibres. Single, controlled probing of the vaginal vestibule induced a reflex discharge in the motor nerve. Repetitive probing (10 Hz) induced a motor nerve post-discharge lasting >1 min. The vaginal sphincter is two-half rings of striated muscle fibres in the wall of the vaginal vestibule; the fibres end freely in the dorsal and ventral midlines. Penetration of the vestibule by the penis might trigger sustained contraction of the vaginal sphincter.

Pudendal nerve stimulation, interneurons post-discharge and delayed depolarization in hind limb motoneurons of the female cat.

The present experiments were done in the spinal female cat. In a prior work in the decerebrate female cat, stimulation of the sensory pudendal nerve (SPN) induced a depolarizing wave (LD) in hind limb motoneurons that outlasted the stimulus by up to 6 s. LD triggered self-sustained motoneuron firing (bistability). An intrinsic potential underlies bistable firing, which, in the cat, depends on two main factors; first, the integrity of pathways descending from the brain stem to the spinal cord and, second, the membrane potential of the motoneuron just before the stimulus; at high resting potential, excitatory short-lasting inputs induce transient but no sustained firing. Thus, no bistability occurs in the spinal cat or in hyperpolarized motoneurons of the decerebrate cat. LD might be an intrinsic potential that could also be absent in the spinal cat, or an extrinsic (synaptic) potential induced by spinal interneurons. In the latter case, the interneurons generating LD should show post-discharge as prolonged as LD. LD was produced in spinal cats and its amplitude did not change or increase slightly during hyperpolarizing pulses, which suggests that LD might be a synaptic response. Interneurons showing post-discharge to train of stimulation to SPN were located 100-200 microm above the pools of hind limb motoneurons. Some post-discharges were as prolonged as LD. We conclude that LD might be a synaptic response to local interneurons.

Gamma->alpha linkage and persistent firing of Ia fibers by pudendal nerve stimulation in the decerebrate cat.

The sensory pudendal nerve (SPN) was stimulated in decerebrate female cats. Spikes of single Ia muscle spindle afferents from the medial gastrocnemius (MG) muscle were recorded in dorsal root filaments. Electroneurography (ENG) was recorded in a cut nerve filament to the MG muscle; MG electromyography (EMG) was also recorded. Single shock to SPN induced discharges of small ENG spikes (SS) with similar amplitude to that of gamma spikes elicited by ventral root stimulation. Thus SS were identified as gamma spikes. The latency of the gamma discharge was approximately 15 ms. As expected, the onset of the gamma discharge preceded a discharge of Ia spikes; the time difference between both discharges was approximately 5 ms. After the initial bursts, the Ia and the gamma activities paused during 20-30 ms but later increased again to last approximately 1 s. After the shock, the EMG activity was depressed during approximately 50 ms; later, motor-unit spikes may show transient activation. Thus the onset of the gamma activation preceded the activation of motor units (gamma-->alpha link). Trains of shocks (1 or 100 Hz) to SPN induced a sustained increase in the frequency of gamma spikes, Ia spikes, and motor units that outlasted the train by 20-120 s. The sustained firing of Ia fibers might trigger or help to trigger and maintain the response of alpha-motoneurons.

Sustained firing of alpha and gamma hind limb motoneurons induced by stimulation of the pudendal nerve.

Axons from receptors in the cat vaginal wall run in the sensory pudendal nerve (SPN), and brief (<10 s) vaginal probing (VP) in the decerebrate cat produces a long-lasting (>1 min) contraction of the triceps surae (TS) muscles. The aim of the present project was to find out whether brief SPN stimulation also produces sustained TS response and, eventually, to study the mechanisms involved in it. Decerebrate female cats were used. In some cats, TS electromyography (EMG) and tension response were recorded; stimulation of left SPN with single or repetitive trains of shocks produced a bilateral TS response that outlasted the stimulus >1 min as VP did. In paralyzed cats (pancuronium; Panc), intracellular recordings were made from hind limb motoneurons (MNs). SPN stimulation produced a depolarization 1 min) electroneurographic (ENG) postdischarge in a small filament of the medial gastrocnemius (MG) nerve; the MG EMG postdischarge was also recorded. Large spikes (LS) and small spikes (SS) were distinguished in the ENG. During the postdischarge, LS frequency and the integrated EMG activity correlated well (r > 0.9); no correlation was found between SS and EMG. After Panc injection, LS postdischarge was absent but the SS postdischarge remained. LS followed by EMG potential were also evoked by brief TS stretch (reflex LS); single shocks to SPN only elicited SS that were not followed by EMG potential. It is concluded that alpha axons and gamma axons produced LS and SS, respectively, and that SPN activates gamma axons. It is proposed that, in the nonparalyzed cats, the stimulation of SPN with trains of shocks might cause an increase in the afferent inflow from muscle spindles to alpha MNs through the sustained firing of gamma MNs. The increased excitatory inflow would depolarize alpha MNs and allow bistable MN firing; Panc would decrease this inflow by blocking transmission to the spindle fibers.

Methods to find aponeurosis and tendon stiffness and the onset of muscle contraction.

A method to measure small movements of living tissues either large or small is presented. The method is based on the detection of changes in reflected infrared light. An optocoupler (coupled photodiode and photodetector) and a small (< 1 cm2) mirror were used. The optocoupler (OC) has a low cost and it can be calibrated easily. It can be also used as the transducer of a strain-gage. Three different uses are shown: (a) as a strain-gage transducer; (b) detection of tendon and aponeurosis movements in large muscles (cat soleus); (c) detection of the onset of muscle contraction. Movements of less than 1 microm can be detected with the aid of automatic averaging of the signals. Concerning the second use (b), it permits the estimation of tendon stretch. Concerning the third use, the onset of muscle movement precedes by at least 2 ms that of the force recorded at the tendon.

Effects of postganglionic nerve section on synaptic transmission in the superior cervical ganglion of the guinea-pig.

In sympathetic neurons, axotomy induces a marked depression in synaptic transmission. We asked whether the decrease in synaptic efficacy observed in a given axotomized cell is more severe if most of the postsynaptic neurons are also injured. Accordingly, we studied if the synaptic depression induced by axotomy in neurons with axons running in a postganglionic nerve is influenced by section of other postganglionic nerves. The excitatory postsynaptic potentials evoked by preganglionic stimulation were recorded intracellularly in an in vitro preparation of the superior cervical ganglion. Eight days after cutting the inferior postganglionic nerve, postsynaptic potentials recorded from neurons projecting through this nerve were smaller (median = 6 mV, n = 62) than the controls (median = 34 mV, n = 89), but were similar to those found after sectioning the inferior nerve plus most postganglionic nerves (median = 5 mV, n = 70). If the inferior nerve was left intact, but most postganglionic branches were cut, the synaptic potentials recorded from inferior nerve neurons were normal (median = 33 mV, n = 77). It is concluded that the synaptic depression induced by axotomy in a sympathetic neuron is not affected by axotomy of nearby ganglion cells, even if they share part of their presynaptic axons. This suggests that the effect of axotomy is restricted to the synaptic terminals on the injured neuron.

Inhibition of withdrawal responses by pelvic nerve electrical stimulation.

In urethane-anesthetized rats, the compound action potential of the pelvic nerve was found to consist of three different waves, two in the A delta fiber and one in the C-fiber range of conduction velocity. Electrical stimulation of the pelvic nerve produced a complete inhibition of the withdrawal response to noxious foot pinch or foot compression. The electromyographic (EMG) activity of the contralateral posterior biceps muscle was used to record the withdrawal response. The withdrawal response inhibition was related to the duration and the frequency of electrical stimulation. Low (5-10 Hz) and high (100-300 Hz) frequencies were ineffective in inhibiting the withdrawal response, whereas intermediate frequencies (20-80 Hz) produced a complete inhibition of the withdrawal response. Short (300 ms) trains of stimulation inhibited the withdrawal response only during the stimulation period. Longer trains of stimulation (500 ms-10 s) produced long-lasting inhibition of the response to noxious stimulation. The inhibition persisted for up to 20 s after the end of electrical stimulation of the pelvic nerve. A delta fiber stimulation was adequate to inhibit the withdrawal response in most (15 out of 17) of the animals. However, A delta plus C-fiber stimulation was needed to inhibit the response to noxious stimulation in two animals. In addition to inhibiting the response to noxious stimulation, pelvic nerve electrical stimulation reflexively activated abdominal muscles. On the basis of present findings using electrical stimulation, it can be suggested that, in the rat, A delta and C-visceral afferents of the pelvic nerve mediate the analgesic effect of vaginocervical probing pelvic and A delta afferents the contraction of abdominal muscles in the fetus-expulsion reflex.

Depression of fast axonal transport in axons demyelinated by intraneural injection of a neurotoxin from K. humboldtiana.

Tullidinol, a neurotoxin extracted from the Karwinskia humboldtiana fruit, dissolved in peanut oil was injected into the right sciatic nerve of adult cats. The contralateral sciatic nerve received an equivalent volume of peanut oil alone. The fast axonal transport of labeled ([3H]Leucine) protein was studied in sensory and motor axons of both sciatic nerves. The radioactive label was pressure injected either into the L7 dorsal root ganglion or the ventral region of the same spinal cord segment. Several days after the toxin injection, the cat limped and the Achilles tendon reflex was nearly absent in the right hind limb. The amount of transported label was decreased distal to the site of toxin injection. Proximal to this site, the transported material was damned. Sensory and motor axons showed similar changes. In addition, the toxin produced demyelination and axonal degeneration. Axonal transport and the structure of the axons were normal in the contralateral nerve. Both, Schwann cells and axons of the right sciatic nerve showed globular inclusions, presumably oil droplets containing the toxin. We conclude that Schwann cells and axons as well are tullidinol targets.

The cat pudendal nerve: afferent fibers responding to mechanical stimulation of the perineal skin, the vagina or the uterine cervix.

Some afferent fibers from the pudendal nerve of the female cat were stimulated by pressing on the perineal skin, the vagina or the uterine cervix. Three different types of skin mechanoreceptors were found: (1) with low threshold (< 20 mg) and slow-adapting discharges; (2) with high threshold (0.1-0.5 g) and slow-adapting discharges; and (3) with low threshold and fast-adapting discharges. Most of these receptors increased their firing frequency as the velocity of skin indentation was increased (velocity detectors). The average conduction velocity of the skin afferents was 29 +/- 9 m/s. The receptors located at the vagina showed a fast-adapting response to probing and were sensitive to the velocity of the probe movement. Most of these receptors, however, showed a slow adaptation when the vaginal wall was distended with a balloon. The conduction velocity in vaginal afferents was 37 +/- 16 m/s. Those receptors responding to pressure on the uterine cervix adapted slowly to constant pressure but were sensitive to the velocity of the pressure pulses. The conduction velocity in the afferents from the uterine cervix was 31 +/- 9 m/s.

Efficacy of a putative GABA analog on synaptic transmission in the cat spinal cord.

4-Hydroxy-4-phenylcaproamide (HPhCA), at high doses or rates of IV injection depressed the ventral root reflexes elicited by nerve or dorsal root stimulation. The D (direct) and I (synaptic) ventral root waves and the antidromic (A) dorsal root wave evoked by intraspinal stimulation were also depressed. Similar effects were produced when HPhCA was applied topically on the cord dorsum. At 80 mg/kg and 8 mg/kg/min, the spinal reflexes and the I wave were facilitated for 4 to 6 h, but the D and A waves were depressed. Intracellular recordings from motoneurons showed that HPhCA injection produced: hyperpolarization that lasted several hours, short lasting (< 20 min) facilitation of both EPSPs and IPSPs as well as spike-like potentials (SLPs) that were triggered by EPSPs even though the neuron was hyperpolarized. SLPs may reach the threshold for full spikes. Our results suggest that the spinal depression results from hyperpolarization of motoneurons and the initial facilitation appears to be presynaptic. The late facilitation may be produced by SLPs. HPhCA does not appear to mimic the actions of GABA in primary afferents fibers and motoneurons.

Prolonged inhibition of the flexor reflex by probing the cervix uteri in the cat.

In decerebrate or spinal cats, sustained mechanical stimulation of the cervix uteri inhibited the flexor reflex elicited by electrical stimulation of the foot pad during the probing period (160 s). After probing, 3-15 min were required for reflex recovery. No additional inhibition was produced if probing was repeated before recovery, but instead the reflex was facilitated. When probing was applied 5-10 min after reflex recovery the reflex was again abolished. The recovery, however, occurred earlier and was followed by facilitation. Probing the cervix with single mechanical pulses inhibited transiently (140-200 ms) the short latency reflex components, but the components with longer latencies are unaffected or facilitated. Distension of the vaginal wall with a balloon also inhibited the flexor reflex, but a transient, mild facilitation appeared several seconds after the distension. In general, whenever the inhibition decreases, the facilitation predominates. Our findings suggest that cervical probing or vaginal distension triggers both a long-lasting inhibition and a concomitant facilitation in different intraspinal flexor reflex pathways.

Sustained activation of the triceps surae muscles produced by mechanical stimulation of the genital tract of the female cat.

In decerebrate cats, controlled mechanical stimulation of the perivulvar skin, the vaginal wall or the cervix uteri induced visible hind limb extension. Pressing on the cervix uteri produced the greater response. To quantify these responses, the EMG activity and the tension developed by the normally inserted triceps surae muscles were recorded. The activity induced in these muscles by stimulation of the genital canal outlasted the stimulus by many seconds or a few minutes. These effects disappeared after spinalization at the T12 level. We propose that stimulation of the vaginal canal in the female cat may induce bistability of triceps surae motoneurones.

The influence of peripheral connections on wallerian degeneration.

Following electrophysiological techniques we investigated whether wallerian degeneration of the cat sural nerve may be influenced by (1) its peripheral connection and (2) the distance from the cell body. Distal stumps connected to their innervation sites peripherally showed less degeneration than proximal ones when isolated between two sections. However, when two isolated adjacent segments were produced by making 3 sections, the resulting degeneration was more pronounced in the distal segment. Thus, both the factors mentioned above appear to influence wallerian degeneration.

Serial dependency in the discharge pattern of dorsal spinocerebellar tract neurons: a computer simulation analysis.

We have developed a model in order to analyze the factors eventually responsible for the strong negative serial dependency between successive interspike intervals in the discharge of the Dorsal Spinocerebellar Tract (DSCT) neurons. This dependency is reflected, phenomenologically, by short intervals followed by long ones and, quantitatively, by the first order correlation coefficient (R1-2); which can be lower than -.6 (Jansen, Nicolaysen & Rudjord, 1966; Kröller and Grüsser, 1982). We have found that the lowest values of R1-2 are always related with model parameter values which were very similar to those obtained experimentally. It was observed that EPSP amplitude distribution plays an important role in the discharge patterns of the DSCT neurons. There is one fiber that elicits EPSPs greater than 6 mV, which is responsible for the genesis of the short intervals in the discharge. Long intervals are determined basically by a suprathreshold depolarization and the afterhyperpolarization processes.

Spectrophotometric and chromatographic detection of Karwinskia humboldtiana (tullidora) toxin in rat serum after tullidora ingestion.

The absorbance spectrum of tullidinol, a tullidora (Karwinskia humboldtiana) toxin, showed peaks at 260 and 410 nm when dissolved in ethanol; the second peak was linearly related to toxin concentration. Silica gel thin layer chromatograms of tullidinol solutions showed a single, yellow spot with an Rf of 0.76. A single dose of tullidora fruit homogenate was orally given to male Wistar rats; the absorbance spectrum of serum from tullidora-treated rats showed peaks at 300 nm and at 410 nm; the second peak was absent in serum from control rats. No spots were detected in chromatograms of normal serum, but a yellow spot with the same Rf as that of tullidinol was found in the serum of tullidora-treated rats. We conclude that tullidinol was detected in the serum of treated animals and that the methods we used for its detection may also help to diagnose accidental tullidora poisoning in cattle and human beings.

Depression of fast axonal transport produced by tullidora.

The fast axoplasmic transport of labeled proteins was studied in cats showing hindlimb paralysis 4-7 weeks after a single oral dose of tullidora (Karwinskia humboldtiana) toxins. The isotope (3H-leucine) was injected into the spinal ganglion and the contralateral spinal cord of the seventh lumbar segment in order to study transport in sensory and motor fibers. The axoplasmic transport in motor fibers of the sciatic nerve was clearly altered in tullidora-treated cats. The majority of these animals showed a gradual decline of radioactivity from the cord to the periphery instead of the clear-cut wave front always seen in normal cats. An apparent wave was seen in three treated cats but the wave peak was behind the normal position and the slope of the wave front was reduced. While the rate of transport indicated by the farthest extent of the foot of the slope was not in all cases significantly changed, the results all indicated a hindered transport by the reduced slope front in the distal segments of the motor axons. In contrast, the axoplasmic transport appeared normal in the sensory fibers of all but one tullidora-treated cat. Light and electron microscopy of medial gastrocnemius and sural (cutaneous) nerves revealed axonal constrictions and axolemal irregularities associated with organelle retention after tullidora treatment. Also, some mitochondria appeared swollen. These changes were more frequent and intense in the motor nerve fibers than in the cutaneous nerve fibers.

Axon-to-axon transmission in tullidora (buckthorn) neuropathy.

Oral administration of ether extracts of the tullidora ( Karwinskia humboldtiana ) fruit, which contains an identified neurotoxin, produced flaccid hind limb paralysis in cats after a latency of 4 to 7 weeks. Acute experiments were conducted after the paralysis was evident. Spinal roots of lumbar and sacral segments were transected as close as possible to the spinal cord and divided into several filaments. Stimulation of some filaments distal to the transection evoked action potentials in other filaments (axon-to-axon transmission or cross talk) after a latency of at least 8 ms. Cross-talk responses frequently consisted of multiple discharges. Axon-to-axon transmission was seen only between motor axons and disappeared when hind limb nerves were transected 10 to 15 cm from the spinal cord. Twin pulses were applied to a filament at various intervals; the pulse intensity was adjusted so that the conditioning pulse was subthreshold to elicit cross talk, but the test pulse frequently elicited it (temporal facilitation). In three fully studied fibers the facilitation was prolonged to 50 to 80 ms. In some cases, no cross talk was evoked in a given filament by individual stimulation of two other filaments, but simultaneous stimulation of the same filaments did evoke cross talk (spatial facilitation). Series of periodic bursts of activity spontaneously occurred in those axons responding with multiple discharges to single stimulation of other axons. At low temperatures (about 30 degrees C) the stimulus could trigger essentially similar series of bursts. Single motoneurons were intracellularly stimulated by brief depolarizing pulses. The action potential elicited by the stimulus was followed after several msec by a secondary train of discharges generated at the periphery ("back firing").

Tullidora (Karwinskia humboldtiana) toxin mainly affects fast conducting axons.

Cats were given a single oral dose of ether extracts from tullidora (Karwinskia humboldtiana) fruit which contains an identified neurotoxin. Acute experiments were performed 4-7 weeks after toxin administration when flaccid limb paralysis was evident. Normal cats were used as controls. The medial gastrocnemius, the soleus and the sural nerves were electrically stimulated and the unitary potentials evoked by the stimuli were extracellularly recorded from spinal root filaments to measure the conduction velocity of single fibres. In control cats, the average conduction velocity (CV) was greater in medial gastrocnemius motor fibres than in the afferent ones of the same nerve and the soleus motor axons, whereas in the sural nerve CV was less than in the aforementioned cases. The CV values and the proportion of fast conducting fibres (greater than 80 m/s) in each nerve were directly related (r = 0.99). In treated cats, CV diminished in all the nerves studied, but the conduction velocity was further reduced in the faster fibres. Consequently, the motor division of the medial gastrocnemius nerve, normally composed of a high proportion (57%) of fast fibres, was more affected by tullidora and the sural nerve, which has the lowest proportion (0.7%) of these type of fibres, was the less affected. Our findings suggest that the preferential involvement of motor nerves in the experimental tullidora (buckthorn) neuropathy, as well as the preservation of somatic sensation in quadriplegic children accidentally poisoned with tullidora, are related to the distribution of axonal diameters in peripheral nerves.