strychnine and Spinal-Cord-Injuries

After spinal cord injury, central neuropathic pain develops in the majority of spinal cord injury patients. Spinal hemisection in rats, which has been developed as an animal model of spinal cord injury in humans, results in hyperexcitation of spinal dorsal horn neurons soon after the hemisection and thereafter. The hyperexcitation is likely caused by permanent elimination of the descending pain systems. We examined the change in synaptic transmission of substantia gelatinosa neurons following acute spinal hemisection by using an in vivo whole-cell patch-clamp technique.. An increased spontaneous action potential firings of substantia gelatinosa neurons was detected in hemisected rats compared with that in control animals. The frequencies and amplitudes of spontaneous excitatory postsynaptic currents and of evoked excitatory postsynaptic currentss in response to non-noxious and noxious stimuli were not different between hemisected and control animals. On the contrary, the amplitude and frequency of spontaneous inhibitory postsynaptic currents of substantia gelatinosa neurons in hemisected animals were significantly smaller and lower, respectively, than those in control animals (P < 0.01). Large amplitude and high-frequency spontaneous inhibitory postsynaptic currents, which could not be elicited by mechanical stimuli, were seen in 44% of substantia gelatinosa neurons in control animals but only in 17% of substantia gelatinosa neurons in hemisected animals. In control animals, such large amplitude spontaneous inhibitory postsynaptic currents were suppressed by spinal application of tetrodotoxin (1 µM). Cervical application of lidocaine (2%, 10 µl) also inhibited such large amplitude of inhibitory postsynaptic currents. The proportion of multi-receptive substantia gelatinosa neurons, which exhibit action potential firing in response to non-noxious and noxious stimuli, was much larger in hemisected animals than in control animals.. These suggest that substantia gelatinosa neurons receive tonic inhibition by spinal inhibitory interneurons which generate persistent action potentials. Spinal hemisection results in hyperexcitation of substantia gelatinosa neurons at least in part by eliminating the tonic descending control of spinal inhibitory interneurons from supraspinal levels.

Topics: Anesthetics, Intravenous; Animals; Bicuculline; Disease Models, Animal; Electric Stimulation; Functional Laterality; Hyperalgesia; Male; Neurons; Neurotransmitter Agents; Patch-Clamp Techniques; Physical Stimulation; Rats; Rats, Sprague-Dawley; Spinal Cord Injuries; Strychnine; Substantia Gelatinosa; Synaptic Transmission; Tetrodotoxin; Urethane

The spinal cord contains the circuitry to control posture and locomotion after complete paralysis, and this circuitry can be enabled with epidural stimulation [electrical enabling motor control (eEmc)] and/or administration of pharmacological agents [pharmacological enabling motor control (fEmc)] when combined with motor training. We hypothesized that the characteristics of the spinally evoked potentials after chronic administration of both strychnine and quipazine under the influence of eEmc during standing and stepping can be used as biomarkers to predict successful motor performance. To test this hypothesis we trained rats to step bipedally for 7 wk after paralysis and characterized the motor potentials evoked in the soleus and tibialis anterior (TA) muscles with the rats in a non-weight-bearing position, standing and stepping. The middle responses (MRs) to spinally evoked stimuli were suppressed with either or both drugs when the rat was suspended, whereas the addition of either or both drugs resulted in an overall activation of the extensor muscles during stepping and/or standing and reduced the drag duration and cocontraction between the TA and soleus muscles during stepping. The administration of quipazine and strychnine in concert with eEmc and step training after injury resulted in larger-amplitude evoked potentials [MRs and late responses (LRs)] in flexors and extensors, with the LRs consisting of a more normal bursting pattern, i.e., randomly generated action potentials within the bursts. This pattern was linked to more successful standing and stepping. Thus it appears that selected features of the patterns of potentials evoked in specific muscles with stimulation can serve as effective biomarkers and predictors of motor performance.

Topics: Animals; Biomechanical Phenomena; Disease Models, Animal; Electric Stimulation Therapy; Electromyography; Evoked Potentials, Motor; Female; Glycine Agents; Hindlimb; Muscle, Skeletal; Quipazine; Rats; Rats, Sprague-Dawley; Recovery of Function; Serotonin Receptor Agonists; Spinal Cord Injuries; Strychnine; Time Factors

Although spinal injury is a major cause of chronic disability, the mechanisms responsible for the lesion pathophysiology and their dynamic evolution remain poorly understood. Hence, current treatments aimed at blocking damage extension are unsatisfactory. To unravel the acute spinal injury processes, we have developed a model of the neonatal rat spinal cord in vitro subjected to kainate-evoked excitotoxicity or metabolic perturbation (hypoxia, aglycemia, and free oxygen radicals) or their combination. The study outcome is fictive locomotion one day after the lesion and its relation to histological damage. Excitotoxicity always suppresses locomotor network activity and produces large gray matter damage, while network bursting persists supported by average survival of nearly half premotoneurons and motoneurons. Conversely, metabolic perturbation simply depresses locomotor network activity as damage mainly concerns white rather than gray matter. Coapplication of kainate and metabolic perturbation completely eliminates locomotor network activity. These results indicate distinct cellular targets for excitotoxic versus dysmetabolic damage with differential consequences on locomotor pattern formation. Furthermore, these data enable to estimate the minimal network membership compatible with expression of locomotor activity.

Topics: Animals; Animals, Newborn; Chronic Disease; Convulsants; Excitatory Amino Acid Agonists; Glutamates; Interneurons; Kainic Acid; Locomotion; Lumbar Vertebrae; Nerve Net; Neurons; Neurotoxins; Rats; Spinal Cord Injuries; Strychnine; Wounds and Injuries

Previous work has shown that latent respiratory motor pathways known as crossed phrenic pathways are inhibited via a spinal inhibitory process; however, the underlying mechanisms remain unknown. The present study investigated whether spinal GABA-A and/or glycine receptors are involved in the inhibition of the crossed phrenic pathways after a C2 spinal cord hemisection injury. Under ketamine/xylazine anesthesia, adult, female, Sprague-Dawley rats were hemisected at the C2 spinal cord level. Following 1 week post injury, rats were anesthetized with urethane, vagotomized, paralyzed and ventilated. GABA-A receptor (bicuculline and Gabazine) and glycine receptor (strychnine) antagonists were applied directly to the cervical spinal cord (C3-C7), while bilateral phrenic nerve motor output was recorded. GABA-A receptor antagonists significantly increased peak phrenic amplitude bilaterally and induced crossed phrenic activity in spinal-injured rats. Muscimol, a specific GABA-A receptor agonist, blocked these effects. Glycine receptor antagonists applied directly to the spinal cord had no significant effect on phrenic motor output. These results indicate that phrenic motor neurons are inhibited via a GABA-A mediated receptor mechanism located within the spinal cord to inhibit the expression of crossed phrenic pathways.

Topics: Animals; Bicuculline; Body Weight; Efferent Pathways; Female; GABA Antagonists; gamma-Aminobutyric Acid; Glycine; Glycine Agents; Phrenic Nerve; Pyridazines; Rats; Rats, Sprague-Dawley; Receptors, Glycine; Respiratory Mechanics; Respiratory Muscles; Spinal Cord Injuries; Stereotaxic Techniques; Strychnine; Vagotomy

The spinal cord and spinal motoneurons are densely innervated by terminals of serotonin (5-HT) and norepinephrine (NE) neurons arising mostly from the brain stem, but also from intrinsic spinal neurons. Even after long-term spinal transection (chronic spinal), significant amounts (10%) of 5-HT and NE (monoamines) remain caudal to the injury. To determine the role of such endogenous monoamines, we blocked their action with monoamine receptor antagonists and measured changes in the sodium currents and firing in motoneurons. We focused on persistent sodium currents (Na PIC) and sodium spike properties because they are critical for enabling repetitive firing in motoneurons and are facilitated by monoamines. Intracellular recordings were made from motoneurons in the sacrocaudal spinal cord of normal and chronic spinal rats (2 mo postsacral transection) with the whole sacrocaudal cord acutely removed and maintained in vitro (cords from normal rats termed acute spinal). Acute and chronic spinal rats had TTX-sensitive Na PICs that were respectively 0.62 +/- 0.76 and 1.60 +/- 1.04 nA, with mean onset voltages of -63.0 +/- 5.6 and -64.1 +/- 5.4 mV, measured with slow voltage ramps. Application of 5-HT2A, 5-HT2C, and alpha1-NE receptor antagonists (ketanserin, RS 102221, and WB 4101, respectively) significantly reduced the Na PICs, and a combined application of these three monoamine antagonists completely eliminated the Na PIC, in both acute and chronic spinal rats. Likewise, reduction of presynaptic transmitter release (including 5-HT and NE) with long-term application of cadmium also eliminated the Na PIC. Associated with the elimination of the Na PIC in monoamine antagonists, the motoneurons lost their ability to fire during slow current ramps. At this point, the spike evoked by antidromic stimulation was not affected, suggesting that activation of the transient sodium current was not impaired. However, the spike evoked after a slow ramp depolarization was slightly reduced in height and rate-of-rise, suggesting decreased sodium channel availability as a result of increased channel inactivation. These results suggest that endogenous monoamine receptor activation is critical for enabling the Na PIC and decreasing sodium channel inactivation, ultimately enabling steady repetitive firing in both normal and chronic spinal rats.

Topics: Animals; Female; Motor Neurons; Norepinephrine; Patch-Clamp Techniques; Picrotoxin; Rats; Rats, Sprague-Dawley; Serotonin; Sodium Channels; Spinal Cord; Spinal Cord Injuries; Strychnine; Synaptic Transmission; Tetrodotoxin

We examined the influence of rectal distention on the spinal micturition reflex and the mechanism underlying its inhibition of bladder contraction.. Fourteen conscious female Sprague-Dawley rats were used in this study after transection of the lower thoracic cord. Isovolumetric cystometry was performed before and after distention of the rectum by inflation of a rectal balloon, followed by intrathecal injection of strychnine (a selective glycine receptor antagonist) or bicuculline (a GABA(A) receptor antagonist) into the lumbosacral cord.. Rectal distention (1.0-3.0 cm(3)) prolonged the interval, decreased the amplitude, and shortened the duration of bladder contraction, and eventually almost abolished bladder activity. After intrathecal injection of strychnine (0.001-10 microg) or bicuculline (0.001-1 microg) in animals with inhibition of bladder activity by rectal distention, the interval, amplitude, and duration of bladder contraction returned to baseline.. These results suggest that there is an inhibitory rectovesical reflex in the lumbosacral cord of rats with spinal cord injury, which modulates the spinal micturition reflex via glycinergic or GABAergic mechanisms.

Topics: Animals; Bicuculline; Female; GABA Antagonists; Glycine; Rats; Rats, Sprague-Dawley; Receptors, GABA; Receptors, Glycine; Rectum; Reflex; Spinal Cord; Spinal Cord Injuries; Strychnine; Urinary Bladder; Urination

Bath application of monoamines is a potent method for evoking locomotor activity in neonatal rats and mice. Monoamines also promote functional recovery in adult animals with spinal cord injuries by activating spinal cord networks. However, the mechanisms of their actions on spinal networks are largely unknown. In this study, we tested the hypothesis that monoamines establish rostrocaudal gradients of rhythmicity in the thoracolumbar spinal cord. Isolated neonatal mouse spinal cord preparations (P0-P2) were used. To assay excitability of networks by monoamines, we evoked a disinhibited rhythm by bath application of picrotoxin and strychnine and recorded neurograms from several thoracolumbar ventral roots. We first established that rostral and caudal segments of the thoracolumbar spinal cord had equal excitability by completely transecting preparations at the L3 segmental level and recording the frequency of the disinhibited rhythm from both segments. Next we established that a majority of ventral interneurons retrogradely labeled by calcium green dextran were active during network activity. We then bath applied combinations of monoaminergic agonists [5-HT and dopamine (DA)] known to elicit locomotor activity. Our results show that monoamines establish rostrocaudal gradients of rhythmicity in the thoracolumbar spinal cord. This may be one mechanism by which combinations of monoaminergic compounds normally stably activate locomotor networks.

Topics: Animals; Animals, Newborn; Benzazepines; Biogenic Monoamines; Dopamine; Dopamine Agonists; Dopamine Antagonists; Dose-Response Relationship, Drug; Drug Combinations; Evoked Potentials; Excitatory Amino Acid Agonists; Fluorescent Dyes; Functional Laterality; GABA Antagonists; Glycine Agents; In Vitro Techniques; Lumbosacral Region; Mice; N-Methylaspartate; Nerve Net; Neural Networks, Computer; Organic Chemicals; Periodicity; Picrotoxin; Quinpirole; Serotonin; Spinal Cord; Spinal Cord Injuries; Strychnine

The isolated thoracic cord of a neonatal rat in vitro generates tonic sympathetic activities in the splanchnic nerves. This tonic sympathetic nerve discharge (SND) has a prominent quasi-periodic oscillation at approximately 1-2 Hz. Bath application of bicuculline and strychnine, which removes endogenous GABA(A) and glycine receptor activities, transforms the quasi-periodic tonic SND into synchronized bursts (bSND). Picrotoxin, another GABA(A) receptor antagonist, also induces bSND. Serial transections of the thoracic cord (T1-12) were performed to identify the cord segments responsible for these tonic and bursting SNDs. Removal of T1-5 did not affect tonic SND. Nerve-cord preparation with either T6-8 or T10-12 segments could generate a substantial amount of tonic SND that retained comparable oscillating patterns. On the other hand, removal of T1-5 significantly reduced bSND amplitude without affecting its rhythmicity. Either T6-8 or T10-12 segments alone could generate bSND. Mid-point transection of T6-12 at T9 might split bSND rhythmogenesis, leading to the occurrence of bSND that could be attributed to two independent oscillators. Our results demonstrated that three segments within the T6-12 cord were sufficient to generate a rudimentary tonic and bursting SNDs. The thoracic cord segments, however, are dynamically interacting so that a full size bSND could only be produced with the intact thoracic cord.

Topics: Action Potentials; Animals; Animals, Newborn; Bicuculline; Dose-Response Relationship, Drug; Electrophysiology; GABA Antagonists; Glycine Agents; In Vitro Techniques; Magnesium; Oscillometry; Picrotoxin; Rats; Rats, Sprague-Dawley; Spinal Cord; Spinal Cord Injuries; Splanchnic Nerves; Strychnine; Sympathetic Nervous System; Thoracic Vertebrae

We examined the influence of lumbosacral glycinergic neurons on the spinobulbospinal and spinal micturition reflexes. Female rats were divided into intact rats, rats with acute injury to the lower thoracic spinal cord (SCI), and rats with chronic SCI. Under urethane anesthesia, isovolumetric cystometry was performed in each group before and after intrathecal (IT) injection of glycine or strychnine into the lumbosacral cord level. The glutamate and glycine levels of the lumbosacral cord were measured after injection of glycine or strychnine in intact and chronic SCI rats. Expression of strychnine-sensitive glycine receptor alpha-1 (GlyR alpha1) mRNA in the lumbosacral cord was also assessed in both rats. In chronic SCI rats, the interval and amplitude of bladder contractions were shorter and smaller when compared with intact rats. IT glycine (0.1-100 microg) prolonged the interval and decreased the amplitude of bladder contractions in both intact rats and chronic SCI rats. IT strychnine (0.01-10 microg) elevated the baseline pressure in intact rats and induced bladder contraction in acute SCI rats. On amino acid analysis, IT glycine (0.01-100 microg) decreased the glutamate level of the lumbosacral cord in intact rats, but not in chronic SCI rats. The glycine level of the lumbosacral cord was 54% lower in chronic SCI rats when compared with intact rats, while the GlyR alpha1 mRNA level did not change after SCI. These results suggest that glycinergic neurons may have an important inhibitory effect on the spinobulbospinal and spinal micturition reflexes at the level of the lumbosacral cord.

Topics: Acute Disease; Animals; Chronic Disease; Disease Models, Animal; Female; Glutamic Acid; Glycine; Glycine Agents; Injections, Spinal; Lumbosacral Region; Rats; Rats, Sprague-Dawley; Receptors, Glycine; Reflex; RNA, Messenger; Spinal Cord; Spinal Cord Injuries; Strychnine; Urinary Bladder; Urination

We have investigated the effects of ascending inhibitory pathways on two centrally generated rhythmic motor patterns in a simple vertebrate model, the young Xenopus tadpole. Tadpoles swim when touched, but when grasped respond with slower, stronger struggling movements during which the longitudinal pattern of motor activity is reversed. Surgical spinal cord transection to remove all ascending connections originating caudal to the transection (in tadpoles immobilized in alpha-bungarotoxin) did not affect "fictive" swimming generated more rostrally. In contrast, cycle period and burst duration both significantly increased during fictive struggling. Increases were progressively larger with more rostral transection. Blocking caudal activity with the anesthetic MS222 (pharmacological transection) produced equivalent but reversible effects. Reducing crossed-ascending inhibition selectively, either by midsagittal spinal cord division or rostral cord hemisection (1-sided transection) mimicked the effects of transection. Like transection, both operations increased cycle period and burst duration during struggling but did not affect swimming. The changes during struggling were larger with more rostral hemisection. Reducing crossed-ascending inhibition by spinal hemisection also increased the rostrocaudal longitudinal delay during swimming, and the caudorostral delay during struggling. Weakening inhibition globally with low concentrations of the glycine antagonist strychnine (10-100 nM) did not alter swimming cycle period, burst duration, or longitudinal delay. However, strychnine at 10-60 nM decreased cycle period during struggling. It also increased burst duration in some cases, although burst duration increased as a proportion of cycle period in all cases. Strychnine reduced longitudinal delay during struggling, making rostral and caudal activity more synchronous. At 100 nM, struggling was totally disrupted. By combining our results with a detailed knowledge of tadpole spinal cord anatomy, we conclude that inhibition mediated by the crossed-ascending axons of characterized, glycinergic, commissural interneurons has a major influence on the struggling motor pattern compared with swimming. We suggest that this difference is a consequence of the larger, reversed longitudinal delay and the extended burst duration during struggling compared with swimming.

Topics: Afferent Pathways; Aminobenzoates; Anesthetics; Animals; Bungarotoxins; Cordotomy; Escape Reaction; Glycine; Glycine Agents; Larva; Motor Activity; Neurotoxins; Periodicity; Spinal Cord; Spinal Cord Injuries; Strychnine; Swimming; Xenopus laevis

Topics: Animals; Bicuculline; Cats; Electric Stimulation; Electromyography; Female; GABA Antagonists; Glycine Agents; Locomotion; Neuronal Plasticity; Posture; Spinal Cord; Spinal Cord Injuries; Strychnine

Effect of intravenous administration of GABA antagonists on gastric acid secretion in perfused stomachs was studied in rats anesthetized with urethane. Bethanechol (BeCh)-stimulated acid secretion was definitely inhibited by bicuculline, a GABA antagonist, and strychnine, a glycine antagonist, but not by picrotoxin and pentylenetetrazol, GABA antagonists. The inhibitory effect of bicuculline and strychnine was accompanied by vigorous convulsions. Only the bicuculline-induced inhibition was still seen in d-tubocurarine paralyzed rats, and it was abolished in spinal rats. 2-Deoxy-D-glucose (2-DG)-stimulated acid secretion was apparently depressed by all the GABA antagonists of bicuculline, picrotoxin and pentylenetetrazol. The inhibitory effect of picrotoxin, but not bicuculline, on the 2-DG stimulation was still elicited in spinal rats. Inhibition of acid secretion stimulated by pentobarbital, a centrally acting secretagogue, was produced by picrotoxin and pentylenetetrazol in spinal rats. These findings suggest that bicuculline acts centrally to inhibit acid secretion stimulated both peripherally by BeCh and centrally by 2-DG, besides nonspecific mechanisms due to convulsions, and the action would be directed to centers which are implicated in stimulation of the sympatho-adrenomedullary system; picrotoxin and pentylenetetrazol also act centrally to inhibit 2-DG- or pentobarbital-stimulated acid secretion through depression of the central vagal tone which leads to inhibition of gastric acid secretion.

Topics: Animals; Bethanechol Compounds; Bicuculline; Deoxyglucose; GABA Antagonists; Gastric Acid; Injections, Intravenous; Male; Paralysis; Pentylenetetrazole; Picrotoxin; Rats; Rats, Inbred Strains; Spinal Cord Injuries; Stomach; Strychnine; Tubocurarine

Topics: Action Potentials; Aminobutyrates; Animals; Cats; Colon; Electric Stimulation; Female; Ganglia, Autonomic; Glycine; Homocysteine; Male; Muscle Contraction; Muscle, Smooth; Neurons; Parasympathetic Nervous System; Perineum; Reaction Time; Rectum; Reflex; Sacrococcygeal Region; Skin Physiological Phenomena; Spinal Cord; Spinal Cord Injuries; Stimulation, Chemical; Strychnine; Touch; Urinary Bladder; Urinary Bladder Diseases; Urination; Urination Disorders

Topics: Animals; Fractures, Bone; Paralysis; Rabbits; Seizures; Spinal Cord Injuries; Strychnine