Serotonin type-4 receptors modulate the sensitivity of intramural mechanoreceptive afferents of the cat rectum.

It has been suggested that serotonin (5-hydroxytryptamine) type-4 (5-HT4) receptors modulate the sensitivity of intrinsic afferents of the intestinal mucosa. We studied the involvement of 5-HT4receptors in the modulation of extrinsic afferent sensitivity of the intestinal wall. During distension ramps, mechanoreceptive rectal afferents in sacral dorsal roots were examined in decerebrate anaesthesia-free cats using the selective 5-HT4receptor partial agonist, tegaserod (HTF 919), and the 5-HT4receptor antagonist, SB 203186. The static discharge rate of the afferents evoked by rectal distension decreased after intravenous (i.v.) administration of tegaserod at intraluminal pressures above 30 mmHg, with the most effective reduction occurring at 50 mmHg. The effect was dose-dependent, with maximal reduction occurring at 1.2 mg kg-1 bodyweight, and could be partly reversed by i.v. administration of SB 203186. Tegaserod did not alter the pressure-volume relationship (compliance) of the rectum. It is tentatively concluded that 5-HT4receptor activation has an inhibitory effect on intramural mechanoreceptors in the cat's rectum. Our results are in line with the observation that tegaserod relieves the sensory symptoms of patients suffering from irritable bowel syndrome.

Influences of pelvic floor structures and sacral innervation on the response to distension of the cat rectum.

The contributions to the rectal response to distension of the pelvic floor structures surrounding the rectum and of the sacral spinal innervation have never been studied. Using paralysed intercollicularly decerebrate, anaesthesia-free cats, we studied pressure-volume relationships during slow ramp distensions of the rectum. Results obtained from animals with intact pelvic cavities were compared with those following mobilization of the rectum from the pelvic floor musculature. To assess the influences of spinal outflow and afferent input, rectal pressure-volume relationships were measured in the mobilized rectum following bilateral sequential transection of the spinal roots S1 to S3, first dorsal, then ventral. Isolation of the rectum from the pelvic floor structures resulted in a decrease in balloon volume in the lower range of distension pressure but did not affect volumes at higher pressures. The only afferent effect was seen after sectioning of dorsal roots S1, which resulted in a decrease in balloon volume. The only efferent effect was seen after sectioning of ventral roots S3, which decreased balloon volume further. In conclusion, the rectal response to distension depends on the properties of the rectal wall. It may be influenced by somatic inputs, inputs from the myenteric nervous plexus, and from the parasympathetic and sympathetic nervous systems. Afferent inputs and spinal autonomic reflexes may decrease the tone of the rectal musculature during distension.

Discharge patterns of intramural mechanoreceptive afferents during selective distension of the cat's rectum.

The afferent input from the rectum to the central nervous system (CNS) has yet to be thoroughly characterized. The characteristics of mechanoreceptive rectal afferents have been studied in unanaesthetized decerebrate cats. Following lumbo-sacral laminectomy, single-unit activity (occasionally multi-unit activity) was recorded from centrally cut filaments of the sacral dorsal roots (predominantly S2), while a balloon was inflated in the rectum. Starting from their background activities (mean 15.1 imp sec-1, SD 7.6 imp sec-1), afferent discharge rate increased with increasing balloon pressure (mean threshold 6.3 mmHg, SD 3.6 mmHg). The dependence of firing rate on intrarectal pressure began to flatten out at 25 mmHg (mean; SD 10 mmHg). For 22 out of 29 units (76%) complete saturation occurred at 35 mmHg (mean; SD 15 mmHg) with a maximum discharge rate of 31 imp sec-1 (mean; SD 12.6 imp sec-1). In a number of recording sessions, cyclical rectal contractions were observed. In these cases, changes in firing of the units were closely related to changes in intrarectal pressure. Pressure-related afferent activity could be enhanced by parasympathomimetic drugs which augmented rectal contractions. We conclude that sacral dorsal roots contain afferents from low-threshold mechanoreceptors located in the rectal wall, and that these afferents monitor the filling state and contraction level of the rectum.

Learning "what" and "how" in a human motor task.

We studied the development of implicit and of verbally declared knowledge for normal human subjects who learned an unfamiliar motor task in one learning session. The exploratory nature of motor learning and a special period for optimizing skill were followed in real time. Subjects understood the goal for task success, but they had to learn a motor strategy of what pattern of serial movements to make and the tactics of how much to scale their amplitudes and timing. We compared the time course for acquiring tactical skill with that for acquiring knowledge of strategy and of tactics, and their necessary cues. Implicit and declarative knowledge were distinguished from one another by correlating subjects' verbal self-reports with movement kinematics and their results. Implicit generation of the correct strategy and of the tactics developed in an exploratory manner from the beginning of the learning session. Implicit strategy learning soon gave way to conscious efforts, but tactical learning remained implicit until its first unambiguous verbal declaration (with one exception). First strategy declarations were voiced before those for tactics, during trial-and-error learning that did not require task success, and referred to reversing the direction of hand movements (one exception). In contrast, first declarations of tactics almost always required actual or imminent success, referred to when direction was to be reversed, and it was achieved near the top of a sigmoid learning curve that rose to tactical skill (with one exception). During the sigmoid rise, movement amplitudes and timing were optimized in a distinct manner, although tactics usually adapted thereafter to movements of more moderate speed that could still be successful.

Evidence for D1 dopamine receptor-mediated modulation of the synaptic transmission from motor axon collaterals to Renshaw cells in the rat spinal cord.

The possible modulatory role of D1 dopamine receptors on the excitability of lumbar spinal Renshaw cells was studied in anesthetized rats spinalized at T4 level. Burst responses elicited by single electrical shocks to ipsilateral ventral roots L6 (frequency 0.5 Hz, stimulus width 0.1 ms) and spontaneous activity were recorded extracellularly using conventional 3 M KCl filled glass micropipettes. The specific D1 agonist SKF 38393 (0.5-1 mg/kg i.v.) enhanced Renshaw cell burst responses by 20-60% (n = 7) and increased their spontaneous discharge rate (n = 3). This effect was clearly antagonized by the specific D1 antagonist SCH 23390 (1 mg/kg i.v.) although SCH 23390 proved ineffective per se. We conclude that SKF 38393 induced facilitation was due to activation of the specific D1 receptors which could be the functional counterpart of the presynaptic D2 receptors described earlier by us in the same synapse.

Dopaminergic influence on the excitability of antidromically activated Renshaw cells in the lumbar spinal cord of the rat.

The interaction between dopaminergic and cholinergic systems in the mammalian central nervous system, which is thought to have important implications in the pathophysiology of major extrapyramidal disorders, has never been adequately demonstrated in vivo. Renshaw cell burst responses to single electrical shocks to lumbar ventral roots in spinalized and decerebrated rats were studied. In this monosynaptic cholinergic pathway, apomorphine, a dopaminergic receptor agonist, inhibited whereas the D2-antagonist sulpiride facilitated the burst responses. The mutual antagonism of the two drugs and the depression coupled with the faster decay of post-tetanic potentiation of Renshaw cells by apomorphine demonstrate the involvement of presynaptic D2-receptors through which dopamine can modulate acetylcholine-mediated central synaptic transmission in vivo. The study also provides further evidence for the involvement of the spinal cord in extrapyramidal disorders.

Presynaptic dopaminergic inhibition of the spinal reflex in rats.

Dopaminergic influence on spinal monosynaptic transmission was examined in rats. Monosynaptic mass reflex (MMR) was recorded from the ventral root L6 following supramaximal stimulation (0.2 Hz; 0.1 ms) to the ipsilateral dorsal root L6 in spinalized rat under pentobarbitone sodium (40 mg/kg, i.p.) anaesthesia. MMR was inhibited by intravenous administration of the dopaminergic agonist, apomorphine (50-200 ug/kg) in a dose-dependent manner. The attenuatory effect of apomorphine (200 ug/kg i.v.) on the reflex could be reversed by the dopaminergic antagonist haloperidol (0.5 mg/kg, i.v.). Under tetanic stimulation (200 Hz; 15s), the pretetanic relative inhibition induced by apomorphine (200 ug/kg, i.v.) was increased only for a short period immediately after the cessation of tetanic stimulation. The results indicate existence of presynaptic dopamine receptors on the afferent terminals converging on the motoneurone which may functionally modulate the spinal motor output.

Inhibitory connections of ipsilateral semicircular canal afferents onto Renshaw cells in the lumbar spinal cord of the cat.

1. In intercollicularly decerebrate cats, the excitability of lumbar spinal Renshaw cells (tested by single shocks to ventral roots and deafferented muscle nerves) decreased for 600-1000 ms after conditioning electrical stimulation of ipsilateral semicircular canal nerves. 2. Conditioning stimulation of posterior canal afferents and combined stimulation of anterior and lateral canal afferents were equally effective in causing inhibition of Renshaw cells. No significant differences were observed for Renshaw cells excitable from hind-limb flexor or extensor nerves. 3. Inhibition appeared when one to five stimuli were applied to the canal afferents and arrived at the spinal segmental level 11-15 ms after the onset of conditioning stimulation. 4. Evidence is adduced to suggest that the inhibitory effects on Renshaw cells following stimulation of semicircular canal afferents were mediated directly, i.e. they were not caused by alterations of motoneurone activity. 5. Excitation of Renshaw cells due to stimulation of the canal afferents was rarely observed; it could not be excluded that it was secondary to motoneurone discharges. 6. It is suggested that vestibular inhibition of Renshaw cells ensure a high gain of hind-limb alpha-motoneurones during postural adjustments following a massive disturbance of body equilibrium.

Chronic inflammatory polyneuropathy. Reduction of nerve conduction velocities in monkeys by systemic passive transfer of immunoglobulin G.

In chronic (relapsing) inflammatory polyneuropathy (CRIP), successful treatment with plasma exchange has led to the concept of pathogenic humoral factors. In 6 patients with CRIP, 5 of whom improved after plasma exchange, the potential pathogenic role of circulating immunoglobulin (Ig) fractions was tested by applying the systemic passive transfer model to marmoset monkeys. After continuous treatment with intramuscular injections for 2-8 weeks, monkeys injected with the crude immunoglobulin fractions or with purified IgG from 5 of the 6 patients showed a significant and partially reversible reduction of the motor nerve conduction velocity (mean 34%, P less than 0.001) when compared with pre-treatment values. In control animals the reduction was 4%. Morphological examination revealed only minor ultrastructural changes of the myelin sheath. Immunocytochemistry revealed that human IgG was able to cross the blood-nerve barrier. It is concluded that the circulating IgG-fraction of patients with CRIP contains a factor that may contribute to the disordered nerve function after crossing the blood-nerve barrier. It may be the removal of this particular factor which is responsible for the rapid recovery of nerve conduction in patients after plasma exchange.

Movement programming depends on understanding of behavioral requirements.

Use of programmed movements seems to depend on comprehension of the required behavior of which the movements are part. Once this understanding becomes evident by correct behavior, smooth movement take-offs and landings are programmed together. Monkeys were trained to perform a step-tracking task that required accurate self-paced elbow movements of moderate speed. Behavioral aspects of the task, such as correct successive movement directions and timing, were all learned at about the same rates. Use of programmed movements however, increased only after these behavioral requirements had been learned to near the 50% level, i.e., the beginning of behavioral sureness. Movements were programmed as a whole, accelerations together with decelerations, from earliest training onwards.

Dynamic properties of Renshaw cells: equivalence of responses to step changes in recruitment and discharge frequency of motor axons.

In decerebrate cats, the dynamic responses of Renshaw cells to step changes in input were determined separately both for changes in the number of alpha-axons excited and for changes in the frequency at which they were stimulated. Together, these two input variables to the Renshaw cells describe the level of activity in the motor output from the spinal cord. In either case, the dynamic responses of the interneurons depend only on their static activity before and after an input step occurs, but are otherwise indistinguishable. This favors the interpretation that the two input variables are equivalent under dynamic conditions, i.e., Renshaw cells respond to total motor output.

Participation of the principal olivary nucleus in neocerebellar motor control.

A new method for reversible cooling of the inferior olivary nucleus has been used in chronically prepared monkeys. Local olivary cooling depressed discharge of complex spikes of Purkinje cells in contralateral cerebellar cortex. Selective cooling of the principal olive (lateral and dorsal lamellae) produced movement oscillations at about 3-5 Hz of the contralateral arm during cooling in a monkey trained to make prescribed arm movements in the horizontal plane. The effects resemble those of dentate dysfunction. Selective cooling of the dorsal accessory olive and/or the overlying reticular formation, in 3 monkeys, produced during cooling a tendency for postural drift of the contralateral arm and for reduction of its movement amplitudes. These changes tended to vary together according to the degree of cooling. Arm oscillations did not occur. It is concluded that climbing fiber projections from the principal olivary nucleus are essential in the primate for optimal neocerebellar control of arm movements.

Static input-output relations in the spinal recurrent inhibitory pathway.

The static discharge rate of Renshaw cells (studied in deafferented, intercollicularly decerebrate cats) has a nonlinear dependence on the frequency of trains of stimulus impulses to alpha-motor axons in the ventral root. This dependence is well described by a rectangular hyperbola that approaches saturation with increasing stimulus frequency. The tendency to saturate is independent of the number of motor axons exciting a Renshaw cell. On average, the stimulus frequency at which the discharge rate reaches half its saturation value lies between 10 and 15 Hz. The effect of Renshaw cell activity -- measured as the antidromic inhibition of individual alpha-motoneurons -- reflects the forms of the static frequency characteristics. An electric circuit analog of the Renshaw cell membrane is presented which serves to explain the qualitative features of the static input-output relations; the nonlinearity is the result of synapses with linear properties acting together at the cell membrane.

X-ray controlled implantation of the brain stem.

Using X-rays to improve accuracy of stereotaxic implantation of the brain stem of Macaca irus monkeys overcomes the problems of moveable brain stem and inaccurate fixation in the stereotaxic frame. The method is independent of standard atlases and correct ear bar fixation. The key to the method is the establishment of a nomogram which relates the implantation target to bony landmarks. The nomogram is established by X-ray contrast studies which outline the brain stem, the implantation target in the brain stem being decided by superimposed histology. This target is nomographically related to bony landmarks seen both on the contrast X-ray and on a plain X-ray taken later at implantation with the monkey in the stereotaxic frame. Being able to locate the target on this plain X-ray, and having included on the X-ray a metal marker, the stereotaxic coordinates of which are known, the distance can be measured (allowing for X-ray magnification) between target and stereotaxic marker and thus one computes the target coordinates. Accuracy of implantation according to these coordinates is monitored with further plain X-rays.

Chronic implantation of gas cryoprobes in monkeys brain stem.

Cooling probe sheaths can accurately be implanted adjacent to brain stem nuclei of fascicularis monkeys making it possible to cool target nuclei repeatedly in long-term behavioral experiments. A description is given of the construction and use of flexible probe sheaths for chronic implantation. Appropriate therapy is detailed to counteract brain edema after implantation. A gas cooling system is described for effective cooling beyond the sheath tip. Effective operation of this system is proven by the effects of cooling the inferior olive on complex spike activity in contralateral cerebellar cortex.

Rat isolated phrenic nerve-diaphragm preparation for pharmacological study of muscle spindle afferent activity: effect of oxotremorine.

1. Muscle spindle afferent discharges exhibiting an approximately linear length-frequency relation could be recorded from the phrenic nerve in the isolated phrenic nerve-diaphragm preparation of the rat. 2. Muscle spindle afferent discharges could be identified by their characteristic "spindle pause" during muscle contraction and by their response to succinylcholine. 3. Cholinergic influence on spontaneous and stretch-induced afferent discharges was indicated by the augmentation produced by physostigmine and acetylcholine. (+)-Tubocurarine, but not atropine, prevented this augmentation indicating the presence of curariform cholinoceptors in muscle spindles. 4. Acetylcholine did not appear to be involved in the genesis of spindle afferent discharges as incubation with hemicholinium-3 and (+)-tubocurarine failed to affect the rate of spontaneous and stretch-induced spindle discharges. 5. Oxotremorine markedly increased the rate of spontaneous and stretch-induced spindle afferent discharges and this effect was prevented in the presence of hemicholinium-3 and (+)-tubocurarine. 6. These results with oxotremorine are of interest in connection with the observation that muscle spindle afferents and hyperactive in Parkinsonian patients.