Chronic Spinal Animals Research Articles

The impact of training and neurotrophins on functional recovery after complete spinal cord transection: cellular and molecular mechanisms contributing to motor improvement.

Beneficial effects of locomotor training on the functional recovery after complete transection of the spinal cord indicate that in chronic spinal animals spontaneous recovery processes are enhanced and shaped by the training. The mechanisms of that use-dependent improvement are still not fully understood. This review tackles three aspects of this issue: (1) neurochemical attributes of functional improvement showing that concentrations of excitatory and inhibitory amino acids in the lumbar spinal segments, which were changed after transection, normalize after the training, or even raise beyond normal. As it does not translate to functional equilibrium between excitatory and inhibitory neurotransmission and may lead to hyperexcitability, the postsynaptic mechanisms which might be responsible for the hyperexcitability are discussed, including (i) dysfunction of K(+)-Cl(-) cotransporter KCC2, which controls the strength and robustness of inhibition, and (ii) altered function of 5-HT2 receptors, which may be targeted to restore KCC2 activity and intrinsic inhibition; (2) morphological changes of lumbar motoneurons and their inputs related to functional improvement of spinal animals, pointing to use-dependent diminution/reversal of the atrophy of the dendritic tree of the hindlimb motoneurons and of their synaptic impoverishment, which in paraplegic animals differs depending on the degree of disuse of the muscles; (3) the role of neurotrophins in motor improvement of spinal animals showing, that increases in neurotrophins due to training or due to efficient viral vector-based transgene expression, that might be responsible for the enrichment of the dendritic tree, elongation of processes and influence neurotransmitter systems in the areas subjected to plastic modifications after injury, correlate with improvement of locomotor functions.

The upright posture improves plantar stepping and alters responses to serotonergic drugs in spinal rats

Recent studies on the restoration of locomotion after spinal cord injury have employed robotic means of positioning rats above a treadmill such that the animals are held in an upright posture and engage in bipedal locomotor activity. However, the impact of the upright posture alone, which alters hindlimb loading, an important variable in locomotor control, has not been examined. Here we compared the locomotor capabilities of chronic spinal rats when placed in the horizontal and upright postures. Hindlimb locomotor movements induced by exteroceptive stimulation (tail pinching) were monitored with video and EMG recordings. We found that the upright posture alone significantly improved plantar stepping. Locomotor trials using anaesthesia of the paws and air stepping demonstrated that the cutaneous receptors of the paws are responsible for the improved plantar stepping observed when the animals are placed in the upright posture.We also tested the effectiveness of serotonergic drugs that facilitate locomotor activity in spinal rats in both the horizontal and upright postures. Quipazine and (±)-8-hydroxy-2-(dipropylamino)tetralin hydrobromide (8-OH-DPAT) improved locomotion in the horizontal posture but in the upright posture either interfered with or had no effect on plantar walking. Combined treatment with quipazine and 8-OH-DPAT at lower doses dramatically improved locomotor activity in both postures and mitigated the need to activate the locomotor CPG with exteroceptive stimulation. Our results suggest that afferent input from the paw facilitates the spinal CPG for locomotion. These potent effects of afferent input from the paw should be taken into account when interpreting the results obtained with rats in an upright posture and when designing interventions for restoration of locomotion after spinal cord injury.

Hindlimb Endpoint Forces Predict Movement Direction Evoked by Intraspinal Microstimulation in Cats

We measured the forces produced at the cat's hindpaw by microstimulation of the lumbar spinal cord and the movements resulting from those forces. We also measured the forces and movements produced by co- and sequential activation of two intraspinal sites. Isometric force responses were measured at nine limb configurations with the paw attached to a force transducer. The active forces elicited at different limb configurations were summarized as patterns representing the sagittal plane component of the forces produced at the paw throughout the workspace. The force patterns divided into the same distinct types found with the femur fixed. The responses during simultaneous activation of two spinal sites always resembled the response for activation of one of the two sites, i.e., winner-take-all, and we did not observe vectorial summation of the forces produced by activation of each site individually as reported in chronic spinal animals. The movements produced by activation of each of the sites were consistent with the force orientations, and different movements could be created by varying the sequence of activation of individual sites. Our results highlight the absence of a vectorial summation phenomenon during intraspinal microstimulation in decerebrate animals, and the preservation during movement of the orientation of isometric forces.

Functional consequences of motor unit recruitment order reversals following spinal cord transection in cat

Motoneuron recruitment order determinations were made for acute, 2-week chronic, and 3-month chronic spinal cats by comparing cutaneous nerve stimulation thresholds for evoking single unit tibialis anterior (TA) electromyogram (EMG) spikes of different sizes. Recruitment order was largely (∼80%) orderly (small spikes recruited at lower stimulus intensities than large spikes) in acute and 3-month chronic spinal animals. However, in 2-week chronic spinal animals recruitment order was reversed, with large units more often recruited at lower stimulus intensities than small units (∼65%). Morphological analyses of TA muscle fibers suggested that fiber size changes were unlikely to account for the dramatic alterations in recruitment order results of the 2-week chronic spinal animals. Additional studies suggested that the recruitment order reversal in the 2-week chronic animals coincided with an enhanced reflex neural output (increased recruitment or reflex gain) for the flexion reflex which compensated for disuse atrophy related decreases in flexor muscle force generation capability in these animals. The data from 2-week chronic spinal animals represent a functionally significant example of deviation from the normal size principle of motoneuron recruitment order as the corresponding reflex gain increases can enhance the rapidity of motor function recovery (standing, locomotion) following spinal injury.

Drastic Decrease in Isoflurane Minimum Alveolar Concentration and Limb Movement Forces after Thoracic Spinal Cooling and Chronic Spinal Transection in Rats

Individuals with spinal cord injury may undergo multiple surgical procedures; however, it is not clear how spinal cord injury affects anesthetic requirements and movement force under anesthesia during both acute and chronic stages of the injury. The authors determined the isoflurane minimum alveolar concentration (MAC) necessary to block movement in response to supramaximal noxious stimulation, as well as tail-flick and hind paw withdrawal latencies, before and up to 28 days after thoracic spinal transection. Tail-flick and hind paw withdrawal latencies were measured in the awake state to test for the presence of spinal shock or hyperreflexia. The authors measured limb forces elicited by noxious mechanical stimulation of a paw or the tail at 28 days after transection. Limb force experiments were also conducted in other animals that received a reversible spinal conduction block by cooling the spinal cord at the level of the eighth thoracic vertebra. A large decrease in MAC (to </= 40% of pretransection values) occurred after spinal transection, with partial recovery (to approximately 60% of control) at 14-28 days after transection. Awake tail-flick and hind paw withdrawal latencies were facilitated or unchanged, whereas reflex latencies under isoflurane were depressed or absent. However, at 80-90% of MAC, noxious stimulation of the hind paw elicited ipsilateral limb withdrawals in all animals. Hind limb forces were reduced (by >/= 90%) in both chronic and acute cold-block spinal animals. The immobilizing potency of isoflurane increases substantially after spinal transection, despite the absence of a baseline motor depression, or "spinal shock." Therefore, isoflurane MAC is determined by a spinal depressant action, possibly counteracted by a supraspinal facilitatory action. The partial recovery in MAC at later time points suggests that neuronal plasticity after spinal cord injury influences anesthetic requirements.

Serotonergic systems in the spinal cord of the amphibian urodelePleurodeles waltl

The role of the monoamine serotonin (5-HT) in modulating the neural networks underlying axial locomotor movements was studied in an adult amphibian urodele, Pleurodeles waltl. 5-HT was applied to an in vitro brainstem-spinal cord preparation of P. waltl, which displayed fictive axial locomotor patterns following bath application of N-methyl-D-aspartate (5 microM) with D-serine (10 microM). Our results showed that 5-HT (1-25 microM) produces a reversible increase in the cycle duration and the duration of rhythmic bursting activity recorded extracellularly from ventral roots innervating the axial musculature. When applied alone, 5-HT does not trigger axial locomotor activity. The distribution pattern of 5-HT immunoreactive (5-HT-ir) cells along the spinal cord was investigated both in intact and in chronic spinal animals. The number of 5-HT-ir cell bodies is higher at brachial levels and decreases through crural levels. Sparse oval or fusiform 5-HT-ir somata are present within the gray matter, just ventrolateral to the central canal. Longitudinal fibers were detected throughout the entire white matter, except in the medial part of the dorsal funiculi. Two columns of intensely labeled and profusely branching thick and thin fibers associated with numerous varicosities run continuously along the ventrolateral surface of the spinal cord. Three weeks following full spinal cord transection at the level of the second spinal root, all longitudinal processes had disappeared, indicating their supraspinal origin, whereas the ventrolateral plexes remained, suggesting that they originated from intraspinal 5-HT-ir cell bodies. Our data showing that spinal 5-HT is organized according to a rostrocaudal gradient suggest that the 5-HT systems of P. waltl are not related to the presence of limb motor pools but more likely are related to axial central pattern generators (CPGs) networks down the length of the spinal cord. The possible involvement of these two sources (descending vs. intraspinal) of 5-HT innervation in the modulation of the axial CPGs is discussed.

Vesicoanal, urethroanal, and urethrovesical reflexes initiated by lower urinary tract irritation in the rat.

Irritation of the urinary bladder causes activation of normally "silent" nociceptive primary afferent fibers. In the present study, it is reported that irritation of the urinary bladder or urethra with infusion of 0.5% acetic acid robustly activates motoneurons that innervate the striated muscle of the external anal sphincter via spinal reflex mechanisms. The activation of anal motoneurons following irritation of the bladder and urethra are termed vesicoanal and urethroanal reflexes, respectively. The reflexes can be mimicked by acute application of capsaicin to the bladder and urethra, and they show desensitization following prolonged topical application of capsaicin or following chronic systemic pretreatment with capsaicin. The reflexes can be demonstrated in chronic spinal cord-transected animals, indicating that the reflex pathways are organized within the spinal cord. The urethroanal reflex is also physiologically activated by urethral distension and/or increases in intraluminal pressure. In addition to activation of anal sphincter activity, slight distension, pressure increases, or instillation of 0.5% acetic acid into the urethra inhibited bladder contractions through activation of an inhibitory urethrovesical reflex. These reflexes are discussed in terms of clinical characteristics of urethritis and prostatitis. Anecdotally, it was discovered that the bladder can buffer acetic acid.

Morphine and Dextrorphan Lose Antinociceptive Activity but Exhibit an Antispastic Action in Chronic Spinal Rats

Within 3–4 weeks after spinal transection, morphine-induced antinociception, assessed with the tail flick reflex in rats, is profoundly reduced. The cause of this decrement is unknown. The present studies were conducted to determine whether this phenomenon reflects a general loss in opiate activity or a selective decline in opiate antinociception. This was accomplished by assessing the effect of morphine on two different responses, the tail flick reflex and the hindlimb spasticity that develops in chronic spinal rats. Because excitatory amino acid antagonists are also antinociceptive in acute spinal rats, the effect of one such drug, dextrorphan, on these two behaviors was also evaluated in chronic spinal animals. The antinociceptive and antispastic effect of subcutaneous (6 mg/kg) and intrathecal (5 μg) morphine injections were assessed in intact and chronic (21–28 days) spinal rats, whereas the effect of subcutaneous (25 and 40 mg/kg) and intrathecal (350 μg) dextrorphan was assessed in acute (1 day) and chronic spinal rats. The antinociceptive effect of both drugs was significantly reduced in chronic spinal animals, relative to saline controls. However, each drug treatment produced a significant antispastic effect in the same animals, indicating a selective decline in opiate action. This outcome also suggests that excitatory amino acid antagonists may be useful as adjunct antispastic agents.

Effect of capsaicin on micturition and associated reflexes in chronic spinal rats

The role of capsaicin-sensitive bladder afferents in micturition was studied in unanesthetized chronic spinal rats. Reflex voiding in response to tactile stimulation of the perigenital region appeared 5–9 days after spinal cord injury (SCI) whereas voiding induced by bladder distension occurred 2–3 weeks after SCI. The frequency and amplitude of reflex bladder contractions recorded under isovolumetric conditions were similar in chronic spinal and urethane-anesthetized CNS-intact rats. However, cystometrograms (CMGs) performed 6–8 weeks after SCI revealed that the chronic spinal rats had larger bladder capacities (1.86 ml) than CNS-intact rats (0.48 ml) and also exhibited multiple, small-amplitude, nonvoiding bladder contractions that were not detected in CNS-intact rats. Administration of capsaicin (50 mg/kg s.c.) acutely (onset 14–40 min) suppressed reflex bladder activity induced by bladder distension or by perigenital stimulation in chronic spinal animals. However, pretreatment of chronic spinal rats with capsaicin (125 mg/kg s.c.) 4 days before the experiment did not depress voiding reflexes or change bladder capacity but did eliminate the nonvoiding contractions. Inhibition of reflex bladder contractions by mechanical stimulation of rectoanal canal or the uterine cervix-vagina was not altered by pretreatment with capsaicin. These data indicate that capsaicin-sensitive bladder afferents are not essential for the initiation of reflex micturition in chronic spinal rats. However, these afferents do contribute to hyperactivity of the bladder during the filling phase of the CMG. Thus, capsaicin-sensitive bladder afferents should be evaluated as possible targets for the pharmacological treatment of bladder hyperreflexia in patients with SCI.

Effects of chronic spinalization on ankle extensor motoneurons. I. Composite monosynaptic Ia EPSPs in four motoneuron pools.

1. We examined the effects of 6-wk chronic spinalization at the L1-L2 level on composite monosynaptic Ia excitatory postsynaptic potentials (EPSPs) recorded in medial gastrocnemius (MG), lateral gastrocnemius (LG), soleus (SOL), and plantaris (PL) motoneurons. Amplitudes, rise times, and half-widths of composite monosynaptic Ia EPSPs evoked by low-strength electrical stimulation of peripheral nerves were measured in barbiturate-anesthetized cats and compared between unlesioned and chronic spinal preparations. 2. The mean amplitude of homonymous composite Ia EPSPs evoked by 1.2 times threshold (1.2T) stimulation and recorded in all four ankle extensor motoneuron pools increased 26% in chronic spinal animals compared with unlesioned controls. There was also an increased incidence of large-amplitude, short-rise time EPSPs. When the same data were separated according to individual motoneuron species, homonymous EPSP amplitudes in MG motoneurons were found to be unchanged. EPSPs recorded in LG motoneurons and evoked by stimulation of the combined LG and SOL nerve were increased by 46%. Mean EPSP amplitudes recorded in both SOL and PL motoneurons were larger after spinalization but statistical significance was only achieved when values from SOL and PL were combined to produce a larger sample size. 3. In LG motoneurons from chronic spinal animals, all EPSPs evoked by 1.2T stimulation of the LGS nerve were > or = 0.5 mV in amplitude. In unlesioned preparations, one fourth of the LG cells had EPSPs that were < or = 0.2 mV. 4. The mean amplitude of heteronymous EPSPs evoked by 2T stimulation of LGS and MG nerves and recorded in MG and LG motoneurons, respectively, doubled in size after chronic spinalization. Because homonymous EPSP amplitudes were unchanged in MG motoneurons, synaptic mechanisms and not passive membrane properties are likely responsible for increased heteronymous EPSP amplitudes in MG. 5. The mean 10-90% rise time of homonymous composite Ia EPSPs in pooled data from all motoneurons decreased 21% in 6-wk chronic spinal animals. Unlike EPSP amplitude, significant rise time decreases were found in all four motoneuron pools. Compared with the other motoneuron species, the mean homonymous rise time recorded in MG motoneurons was shortest and decreased the least in chronic spinal animals. Rise times of heteronymous Ia EPSPs in MG and LG motoneurons also decreased. The maximum rate of rise of homonymous EPSPs increased in all four motoneuron species. 6. The mean half-widths of Ia composite EPSPs decreased in 6-wk spinalized preparations in all motoneuron species.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of chronic spinalization on ankle extensor motoneurons. III. Composite Ia EPSPs in motoneurons separated into motor unit types.

1. In this paper we continue an examination of changes in composite Ia excitatory postsynaptic potentials (EPSPs) in ankle extensor motoneurons after 6-wk (L1-L2) spinal cordotomy. The ratio of rheobase to input resistance was used to divide motoneurons into three groups approximating fast-fatigable (FF), fast fatigue-resistant (FR), and slow (S) motor units in barbiturate-anesthetized cats. Homonymous monosynaptic Ia EPSPs evoked by low-strength [1.2 times threshold (T)] electrical stimulation and heteronymous EPSPs evoked by 2T stimulation were compared between groups of motoneurons in unlesioned and chronic spinal preparations. 2. The distribution of motor unit types of triceps surae and plantaris (PL) motoneurons according to the present classification scheme agrees well with that obtained elsewhere using mechanical typing. Chronic spinalization resulted in an increased proportion of type FF motoneurons in PL and type FR motoneurons in lateral gastrocnemius (LG) motoneurons. There was a numeric but insignificant increase in the proportion of fast medial gastrocnemius motor units. 3. Membrane time constant (tau m) and estimated total cell capacitance were significantly reduced in FF and S motoneurons in chronic spinal preparations. FF motoneurons from chronic spinal animals also had a reduced afterhyperpolarization duration. Mean values of membrane electrical properties in FR motoneurons were unaltered after spinalization. 4. Homonymous Ia EPSP changes after chronic spinalization occurred preferentially in type FR and S motor units. Amplitudes increased 69% in type FR and 38% type S motor units but were unchanged in type FF units. Furthermore, the amplitudes of heteronymous Ia EPSPs in type FF and S units in the chronic spinal preparation were almost double those in unlesioned preparations. 5. Homonymous EPSP 10-90% rise times decreased 25% in type FR motor units and 15% in type S motor units and were unchanged in type FF motor units. Homonymous EPSP half-width decreased in all three motoneuron groups. Normalization of EPSP rise time and half-width to tau m reduced the difference between EPSP shape indexes in unlesioned and chronic spinal preparations in type FF and S motoneurons but less so in type FR motoneurons. Normalized EPSP shape indexes in some type FR units were shorter after chronic spinalization than any in unlesioned preparations. 6. The increased amplitude and decreased rise time of Ia EPSPs in type FR motoneurons after spinalization occurred without changes in the electrical properties of type FR motor units.(ABSTRACT TRUNCATED AT 400 WORDS)

Effect of Mechanical Stimulation of the Skin on the External Urethral Sphincter Muscles in Anesthetized Cats.

Experiments were performed using anesthetized cats to examine the effects of cutaneous stimuli on the external urethral sphincter (EUS) activity and bladder motility, 1) when the bladder volume was changed from low to high, and in the latter case, in the presence or absence of micturition contractions, 2) in different neurological states, viz., in animals with an intact neuraxis and in acute or chronic spinal preparations. In animals with an intact neuraxis, when the volume of the bladder was low, the bladder was quiescent and noxious mechanical stimulation of the perineal skin produced abrupt and marked increases both in vesical pressure and EUS activity. Similar results were observed in acute spinal as well as chronic spinal animals. The effect of the stimulus was particularly potent in chronic spinal animals. In animals with an intact neuraxis, when the volume of the bladder was increased, rhythmic micturition contractions were produced and noxious stimulation of the perineal skin slowed the frequency of the large rhythmic micturition contractions of the bladder and increased the EUS activity. These effects in animals with an intact neuraxis were substantially similar to those in chronic spinal cats showing synergic states. The effects of these somatic stimuli on the integration and coordination of bladder motility (controlled mainly by the parasympathetic pelvic efferent nerves) and EUS activity (controlled by somatic pudendal efferent nerves) are discussed.

Electrophysiological study of micturition reflexes in rats.

Electrophysiological techniques were used to examine the asynchronous and evoked activity on postganglionic nerves to the urinary bladder in the urethananesthetized rat. Distension of the bladder (0.4-0.6 ml) evoked reflex contractions of the bladder (mean intravesical pressure 28 cmH2O) and efferent firing on postganglionic nerves. Electrical stimulation of afferent and efferent axons in the pelvic nerve elicited short-latency (0.3-11 ms) responses and long-latency (45-170 ms) reflexes on the nerves. The short-latency responses consisted of nonsynaptic axonal volleys with conduction velocities ranging from 0.5 to 11 m/s and synaptic responses with latencies of 6-11 ms. Stimulation of the pelvic nerve elicited late supraspinal reflexes (mean latency 122 +/- 28 ms) in 60% of normal rats and an early reflex (mean latency 56 +/- 5 ms) in 25% of those animals in which a late reflex was also identified. Early reflexes (mean latency 50 +/- 9 ms) were elicited in 100% of chronic spinal animals. The conduction time for the afferent and efferent limbs of the reflexes was calculated to be 7 and 58 ms, respectively, with a central delay of 57 ms for the late and less than 5 ms for the early reflex. It is concluded that sacral parasympathetic input to the urinary bladder of the rat is mediated by supraspinal and spinal reflex pathways. It is likely that in normal animals the late-occurring supraspinal reflex mediates micturition. The significance of the spinal reflex in the normal animals is uncertain; however, this reflex is essential for the generation of automatic micturition in chronic spinal preparations.

Characterization of postsynaptic potentials evoked by sural nerve stimulation in hindlimg motoneurons from acute and chronic spinal cats

The purpose of this study was to characterize the changes in postsynaptic potentials recorted in ankle extensor motoneurons resulting from activation of the sural nerve spinal cord transection in the adult cat. Eight acute and nine chronic animals were spinalized at T 12. Intracellular recordings from motoneurons innervating the triceps surae were performed. Sural nerve stimulations evoked complex synaptic potentials consisting of early and late components in all motoneurons. Early excitatory and inhibitory postsynaptic potentials (PSPs), as well as long latency excitatory postsnnaptic potentials were recorded and averaged for assessment of PSP amplitude and duration. Early PSPs, both excitatory and inhibitory, were significantly larger in the motoneurons of cats spinalized 4–6 months earlier. Central latency of excitatory potentials were similar in the two samples of motoneurons, but the central latency associated with the initial inhibitory PSP was significantly shorter in the recordings from motoneurons of chronic spinal cats. In most recordings, an additional inhibitory PSP followed the initial excitatory PSP in motoneurons, and this secoddary inhibitory PSP was similar in peak amplitude and duration in both samples of motoneurons. Also, a long latency excitatory PSP was recorded in a large percentage of motoneurons from both samples. This potential was typically of greater amplitude and longer duration in the motonuurons from chronic animals, when compared to recordings from acute animals. Although changes in amplitude and duration of PSP activity could be documented, there was no marked alteration in the frequency of occurrence of each PSP pattern recorded from the two preparations. This suggests that the synaptic pathways mediating the sural nerve reflexes have not qualitatively changed in the chronic spinal animals. The changes in amplitude and durations of the PSPs in the chronic spatial cat indicate, however, the quantitative changes have occurred. The quantitative changes have probably occurred in the interneuronal networks activated by cutaneous nerve (sural) stimulation, since it was shown that only minor changes in motoneuron membrane properties could be recorded in these same chronic spinal animals.

Characterization of hindlimb motoneuron membrane properties in acute and chronic spinal cats

The purpose of this study was to determine if changes in hindlimb motoneuron membrane electrical properties occur 4–6 months after spinal transection in the adult animal. Eight acute and nine chronic animals were spinalized at T 12. Intracellular recordings from motoneurons innervating the triceps surae were performed. Membrane electrical properties, including resting potential, action potential peak amplitude, afterhyperpolarization duration, rheobasic current, input resistance and axonal conduction velocity were measured. There were no statistical differences found between group means or frequency distributions in the membrane properties of motoneurons assessed from acute and chronic spinal animals. Thus, alteration of motoneuron membrane properties does not appear to be a major contributing factor to the hyperexcitable hindlimb reflex activity demonstrated by chronic spinal animals.

Development and characteristics of airstepping in chronic spinal cats

Airstepping, walking-like movements of the hindlimbs, is a commonly observed behavior in chronic spinal animals when they are held vertically. The purpose of this study was to: describe the development of airstepping after spinalization and compare it to the onset of segmental reflexes, characterize the EMG pattern of muscle activity during spontaneous airstepping, and examine the effects of sensory perturbation on the characteristic pattern. Airstepping was analyzed during three conditions of tonic sensory perturbation which included: tail pinching, tape applied to one hindpaw, and immobilization of the ankle and knee in a plaster cast. Seven adult cats were spinalized at T-12, and bipolar electrode wires were surgically implanted in selected hindlimb muscles at the hip, knee, and ankle. Testing began within 48 hr of transection. Segmental reflexes and paw-shake responses were present in the first week; however, the earliest observed airstepping occurred during tail pinching at 2 weeks after surgery, and the average onset of spontaneous airstepping (without exteroceptive stimuli) was at 33 days. The average cycle period of spontaneous airstepping (691 msec) was comparable to the shortest periods reported for fictive rhythms and to treadmill walking between 1 and 2 m/sec. Intralimb coordination was characterized by flexor and extensor synergies typical of locomotion, while interlimb coordination was characterized by alternating cycles similar to that reported for treadmill walking and fictive locomotion. Neither intralimb nor interlimb patterns of coordination were altered by conditions of sensory perturbation, although cycle period and EMG recruitment level were variable. Many characteristics of airstepping are similar to those of treadmill and fictive locomotion.(ABSTRACT TRUNCATED AT 250 WORDS)

Activity in sympathetic neurons supplying skin and skeletal muscle in spinal cats

In anesthetized cats with intact neuraxis, vasoconstrictor neurons supplying skeletal muscle (MVC) and hairy and hairless skin (CVC), and sudomotor neurons innervating sweat glands (SM), exhibit distinct reflex patterns. MVC and SM are largely under excitatory, CVC under inhibitory control of various afferent input systems from the body surface and from the viscera. In chronic spinal animals all 3 types of sympathetic neurons exhibit some resting activity without cardiac and respiratory modulation. Sixty to 150 days after isolation of the neural circuits within the spinal cord from their descending control systems by spinalization, these reflex patterns are very similar to those in animals with intact neuraxis. Important changes which do occur after spinalization are the following: CVC neurons are excited by stimulation of visceral afferents in spinal animals but inhibited in animals with intact neuraxis; noxious stimulation of skin leads to long-lasting after-effects in CVC and SM neurons in spinal animals. Comparison of reflexes among spinal animals and animals with intact neuraxis indicates that spinal circuits are probably important for the functioning of the sympathetic systems. It is possible that these circuits determine the typical reaction patterns seen in the sympathetic systems by integrating multisensory information from primary afferents and information from spinal descending fiber tracts.

Parasympathetic preganglionic neurons in the sacral spinal cord

Two types of preganglionic neurons have been identified in the sacral parasympathetic nucleus (SPN) of the cat. These neurons could be differentiated by various characteristics including axonal conduction velocities, morphology, location in the nucleus, organ of innervation and central reflex mechanisms controlling their activity. Neurons having myelinated axons (B-PGNs) with conduction velocities between 3.3 and 13 m/s were located in the lateral band of the SPN and innervated the urinary bladder. Neurons with unmyelinated axons (C-PGNs) with conduction velocities of 0.5–1.4 m/s were located in the dorsal band of the nucleus and innervated the large intestine. B-PGNs were excited by distension of the bladder and inhibited by distension or mechanical stimulation of the intestine, whereas C-PGNs exhibited the opposite responses to these stimuli. C-PGNs often exhibited a low level of spontaneous discharge in absence of stimulation but exhibited marked firing (3.5–10 spikes/s) during a defecation reflex elicited by mechanical stimulation of the rectum-anal canal. The excitatory responses were elicited by C-fiber afferents via a spinal reflex pathway. B-PGNs were inactive when intravesical pressure was below the threshold for inducing micturition (5 cm H 2O) but raising the pressure above the threshold induced firing consisting of repetitive bursts of action potentials occurring at relatively high frequencies (15–60 spikes/s). These bursts coincided with rhythmic bladder contractions. The frequency of bladder contractions and associated bursts of PGN-firing and the mean PGN-firing rate (2–8 spikes/s) increased as intravesical pressure was increased in steps between 5 and 30 cm H 2O. However, as indicated by interspike interval histograms, the frequency of firing within a burst of action potentials was unchanged. It is concluded that the micturition reflex pathway is organized as a simple on-off switching circuit and that B-PGNs receive a maximal synaptic input when intravesical pressure exceeds the micturition threshold. This circuit was triggered by vesical Aδ afferents via a spinobulbospinal pathway. Transection of the spinal cord interrupted the reflex pathway and blocked micturition. However, in chronic spinal animals a spinal reflex mechanism emerged which contributed to the recovery of bladder function. This mechanism, which was weak or non-existent in animals with an intact neuraxis, exhibited a number of important differences from the normal micturiton reflex, most notably being activated by a C-fiber afferent rather than a Aδ afferent limb. The mechanism underlying the emergence of C-fiber evoked bladder reflexes in spinal animals is uncertain.

Organization of the sympathetic innervation supplying the hairless skin of the cat's paw

The sympathetic outflow supplying the hairless skin of the cat's hind paw has been analyzed in brain-intact and chronic low spinal animals. For this purpose the activity of postganglionic axons in fascicles of the medial plantar nerve which innervate the central pad and the respective responses of the target organs (transient skin potentials, skin temperature) have been recorded. (1) Blood vessels and sweat glands in the hairless skin of the cat's hind paw are under efferent control of vasoconstrictor neurons, sudomotor neurons, and possibly vasodilatator neurons. (2) Vasoconstrictor neurons are largely under inhibitory control of various afferent input systems from the body surface and from the internal milieu, and sudomotor neurons under excitatory control in brain-intact as well as in chronic spinal cats. (3) The basic neuronal network or "machinery" for the this reciprocal organization is probably located in the spinal cord. (4) Effects of anesthetics on the sudomotor reflexes indicate that this spinal neuronal organization is controlled in a complex manner by descending spinal systems from the brain stem. (5) The existence of vasodilatator neurons, a third efferent control system supplying the hairless skin, which can only be activated by spinal cord warming in brain-intact as well as in chronic spinal cats is not very well established. Neurons with this property are rare and it is unclear whether they course through the sympathetic trunk or dorsal roots or through both. The axons of these neurons are unmyelinated. (6) The organization of the neuronal control of vessels and sweat glands in the hairless skin may be a paradigmatic model for studying the respective neuronal organization in the spinal cord and its descending control.

The sacral parasympathetic reflex pathway regulating colonic motility and defaecation in the cat.

1. The sacral parasympathetic outflow to the large intestine of the cat was studied by monitoring simultaneously intestinal motility and the efferent firing in postganglionic fibres on the serosal surface of the mid-distal colon. 2. Increases in efferent firing were noted during the occurrence of spontaneous propulsive activity (tonic pressure waves) or segmental contractions (slow rhythmic pressure waves). The neural discharge was not altered by transection of the lumbar sympathetic innervation to the colon but was blocked by interruption of the sacral parasympathetic outflow. 3. Electrical stimulation of pelvic nerve afferents arising in the colon or distension of the colon or rectum evoked reflex increases in efferent firing and sustained propulsive contractions that were associated with defaecation. Both responses were abolished by transection of the pelvic nerves or sacral dorsal roots. 4. Electrical stimulation of colonic afferent fibres also evoked synchronous reflex discharges in colonic efferents at latencies ranging from 180 to 300 msec. The discharges were enhanced during propulsive contractions, abolished by transection of the pelvic nerves but not altered by transection of the lumbar sympathetic nerves. 5. Sacral reflexes were present in cats with intact spinal cord and in chronic spinal animals (transection at T10-T12). The reflexes recovered within minutes to several hours after acute transection of the spinal cord. 6. Electrophysiological measurements indicated that the sacral reflexes to the large intestine were mediated by non-myelinated afferent and preganglionic efferent fibres. The central delay for the reflex was estimated to be 45-60 msec. 7. It is concluded that the sacral parasympathetic reflexes to the large intestine are mediated via a spinal pathway and have an essential role in the initiation of propulsive activity during defaecation.