Trains Of Electrical Stimuli Research Articles

Structure and contractile properties of the ostial muscle (musculus orbicularis ostii) in the heart of the American lobster

“Venous” blood enters the crustacean heart through bivalved ostia. Each ostium is a discrete anatomical unit that remains functional even when isolated from the heart. Muscle fibers produce overshooting action potentials that have a plateau of variable duration in response to nervous drive from the cardiac ganglion or during trains of electrical stimuli. Contractions show summation and facilitation when stimulated by trains of stimuli delivered at rates greater than 0.5 s−1 and 0.2 s−1, respectively. Contraction amplitude increases with stimulating impulse frequency and train duration. Maximum force occurs at 1.2 times the slack length. The morphology of ostial fibers resembles that of myocardial fibers. Interconnected bundles of myofilaments occur in both the ostial fibers and the myocardial fibers. In ostial and myocardial fibers, the myofilament bundles are invested by perforated sheets of sarcoplasmic reticulum, and these sheets interface with a network of sarcolemmal tubules to form dyadic interior couplings at the level of the sarcomeric H-bands. The contractile apparatus originates and terminates at intermediate junctions on the transverse cellular boundaries, and the lateral surfaces of the muscle fibers are linked by a modest number of communicating (gap) junctions.

Ca2+ waves during triggered propagated contractions in intact trabeculae. Determinants of the velocity of propagation.

During triggered propagated contractions, Ca2+ waves travel along cardiac trabeculae with a constant velocity (Vprop) ranging from 0. 34 to 5.47 mm/s. To explore the determinants of Vprop, we studied (1) the relationship between [Ca2+]i and Vprop and (2) the effect of low concentrations of caffeine on Vprop. Trabeculae were dissected from the right ventricle of rat hearts. [Ca2+]i was measured using electrophoretically injected fura-2 and an image-intensified CCD camera. Force was measured using a silicon strain gauge, and sarcomere length was measured using laser diffraction techniques. After induction of reproducible Ca2+ waves by trains of electrical stimuli (2.5 Hz) at 21.9+/-0.2 degrees C, the number of stimuli or [Ca2+]o was varied in 9 trabeculae. In 5 trabeculae, the effects of caffeine (0.1 to 1.0 mmol/L) at [Ca2+]o of 2.2+/-0.3 mmol/L were determined. All images were recorded under stable conditions of wave propagation. The increment in [Ca2+]i during the last electrically stimulated transient (DeltaCaT) and [Ca2+]i just before onset of the Ca2+ waves (CaD) were used to estimate the Ca2+ loading of the sarcoplasmic reticulum (SR) and the myoplasm, respectively. The ratio (DeltaCaW/DeltaCaT) of the [Ca2+]i increment during the waves (DeltaCaW) to DeltaCaT was used to estimate the probability of opening of the SR-Ca2+ release channel during wave propagation. As a result of an increase of the number of stimuli or [Ca2+]o, Vprop increased in proportion to (1) DeltaCaT (r=0.82); (2) CaD (r=0.88); (3) DeltaCaW (r=0.85); and (4) DeltaCaW/DeltaCaT (r=0.74). The addition of caffeine (</=0.3 mmol/L) increased Vprop for any DeltaCaT and any DeltaCaW, revealing an increased sensitivity of Vprop to DeltaCaT and DeltaCaW. In contrast, caffeine had little effect on the relationship between Vprop and CaD and no effect on that between Vprop and DeltaCaW/DeltaCaT. These results suggest that both the cellular Ca2+ loading and open probability of the SR-Ca2+ release channels determine the velocity of propagation of Ca2+ waves.

Windup leads to characteristics of central sensitization

Central sensitization refers to enhanced excitability of dorsal horn neurons and is characterized by increased spontaneous activity, enlarged receptive field (RF) areas, and an increase in responses evoked by large and small caliber primary afferent fibers. Sensitization of dorsal horn neurons often occurs following tissue injury and inflammation and is believed to contribute to hyperalgesia. Windup refers to the progressive increase in the magnitude of C-fiber evoked responses of dorsal horn neurons produced by repetitive activation of C-fibers. In the present study, we tested the hypothesis that windup leads to central sensitization. Recordings were made from rat nociceptive dorsal horn neurons classed as wide dynamic range. Windup was produced by conditioning stimuli in a train of 12 electrical pulses (0.5 ms duration) applied to the RF at intensities three times the threshold for excitation of C-fibers and at a frequency of 0.5 Hz. Single electrical stimuli applied outside the RF never evoked responses except when delivered following conditioning stimulation inside the RF, indicating an expansion of the RF following windup. C-Fiber conditioning stimuli applied outside the RF also increased the response evoked by a single stimulus and increased the total number of spikes evoked by a train of electrical stimuli delivered inside the RF. Although both A- and C-fibers were activated, conditioning stimuli did not alter subsequent responses evoked by stimulation of A-fibers. Enhanced responsivity to C-fiber input following windup produced by stimulation inside the RF at a frequency of 0.5 Hz could be maintained for approximately 100 s by stimuli delivered at 0.1 Hz, a frequency that itself cannot produce windup. It is concluded that neuronal events leading to windup also produce some of the classical characteristics of central sensitization including expansion of RFs and enhanced responses to C- but not A-fiber stimulation. Thus, windup may be a useful tool to study mechanisms underlying certain characteristics of central sensitization related to C-fiber activity.

A Quantitative Description of Short-Term Plasticity at Excitatory Synapses in Layer 2/3 of Rat Primary Visual Cortex

Cortical synapses exhibit several forms of short-term plasticity, but the contribution of this plasticity to visual response dynamics is unknown. In part, this is because the simple patterns of stimulation used to probe plasticity in vitro do not correspond to patterns of activity that occur in vivo. We have developed a method of quantitatively characterizing short-term plasticity at cortical synapses that permits prediction of responses to arbitrary patterns of stimulation. Synaptic responses were recorded intracellularly as EPSCs and extracellularly as local field potentials in layer 2/3 of rat primary visual cortical slices during stimulation of layer 4 with trains of electrical stimuli containing random mixtures of frequencies. Responses exhibited complex dynamics that were well described by a simple three-component model consisting of facilitation and two forms of depression, a stronger form that decayed exponentially with a time constant of several hundred milliseconds and a weaker, but more persistent, form that decayed with a time constant of several seconds. Parameters obtained from fits to one train were used to predict accurately responses to other random and constant frequency trains. Control experiments revealed that depression was not caused by a decrease in the effectiveness of extracellular stimulation or by a buildup of inhibition. Pharmacological manipulations of transmitter release and postsynaptic sensitivity suggested that both forms of depression are mediated presynaptically. These results indicate that firing evoked by visual stimuli is likely to cause significant depression at cortical synapses. Hence synaptic depression may be an important determinant of the temporal features of visual cortical responses.

Prejunctional modulation by nociceptin of nerve-mediated inotropic responses in guinea-pig left atrium

The superimposition of a train of electrical stimuli (15 Hz, 1 ms, 60 V for 2.5 s) to the electrically driven (3 Hz) isolated left atria from reserpine-pretreated guinea pigs in the presence of atropine (1 μM) produces a delayed positive inotropic response due to the antidromic activation of capsaicin-sensitive primary afferents and the release of the sensory neuropeptide, calcitonin gene-related peptide (CGRP). The novel opioid peptide nociceptin, inhibited ( E max 88% inhibition at 1 μM) in a concentration-dependent manner (10 nM–1 μM) (EC 50 33 nM) the delayed positive inotropic response induced by train electrical field stimulation, without affecting the positive inotropic response produced by exogenous CGRP (10 nM) or capsaicin (30 nM). The inhibitory effect of nociceptin on the delayed positive inotropic response induced by train electrical field stimulation was not antagonized by the opioid receptor antagonists naloxone, naltrindole and nor-binaltorphimine (1 μM each) nor was it modified by a cocktail of peptidase inhibitors (bestatin, captopril and thiorphan, 1 μM each). A significant inhibition by nociceptin (1 μM) was also observed toward the sympathetic positive inotropic response produced by EFS at 5 Hz in the presence of atropine (1 μM) and after in vitro capsaicin desensitization and toward the parasympathetic negative inotropic response produced by EFS at 10 Hz in atria from reserpine-pretreated guinea pigs and after in vitro capsaicin desensitization. We conclude that nociceptin exerts a prejunctional inhibitory effect on evoked release of CGRP from capsaicin-sensitive sensory nerve terminals in guinea-pig left atria. The effect of nociceptin occurs independently from the activation of μ-, δ- or κ-opioid receptors. Nociceptin, at appropriate frequency of stimulation, appears to exert a general inhibitory neuromodulation on transmitters release in guinea-pig left atria.

A slow excitatory postsynaptic current mediated by G-protein-coupled metabotropic glutamate receptors in rat ventral tegmental dopamine neurons.

Dopamine neurons in the substantia nigra and ventral tegmental area express metabotropic glutamate receptors, but activation of these receptors by synaptic release of neurotransmitter has not been demonstrated thus far. Patch pipettes were used to record membrane currents under voltage clamp from presumed dopamine-containing neurons in the whole-cell configuration in the rat brain slice. A short train of electrical stimuli delivered to bipolar electrodes placed in the slice evoked a slow excitatory postsynaptic current (EPSC; 50-300 pA at -70 mV) which peaked 560 ms after onset and lasted several seconds, with a decay time-constant of 630 ms. This slow EPSC was voltage-dependent, and was abolished by tetrodotoxin (0.5 microM) or by perfusate containing low calcium (0.5 mM) and high magnesium (10 mM). The metabotropic glutamate receptor antagonist (+/-)-alpha-methyl-4-carboxyphenylglycine (MCPG; 300 microM) blocked the slow EPSC, but L(+)-2-amino-3-phosphonopropionic acid (AP3; 300 microM) had no effect. The slow EPSC was largely occluded by inward current produced by the metabotropic receptor agonist trans-(+/-)-1-amino-1, 3-cyclopentanedicarboxylic acid (t-ACPD; 300 microM), and the EPSC was reduced > 90% during acute desensitization produced by prolonged perfusion with t-ACPD. (+/-)-2-Amino-4-phosphonobutyric acid (AP4; 300 microM), another metabotropic receptor agonist, reduced the slow EPSC but had no effect on currents evoked by t-ACPD applied by pressure-ejection from micropipettes. The slow EPSC was progressively reduced in amplitude when pipettes contained the G-protein inhibitor GDP-beta-S (0.5 mM). When pipettes contained GTP-gamma-S (0.5 mM), a non-hydrolysable analogue of GTP, onset of the slow EPSC was more rapid and its decay was significantly prolonged. These results demonstrate that a slow EPSC mediated by G-protein-coupled metabotropic glutamate receptors can be evoked in dopamine neurons.

Antifibrillatory effect of esmolol alone and in combination with lidocaine.

The primary objective of this study was to determine the effect of esmolol, administered alone and in combination with lidocaine, on ventricular fibrillation threshold (VFT) in pigs. A secondary objective was to determine the relationship between blood esmolol concentrations and VFT. We determined VFT using a train of electrical stimuli delivered to the right ventricle after eight paced beats at a basic cycle length of 285 ms. Current was increased in 2-mA increments until VF occurred. VFT determinations were performed during administration of esmolol 1,000 mu g/kg/min, during a continuous infusion of lidocaine, and during infusion of esmolol and lidocaine in combination. Mean increases in VFT from baseline during infusion of esmolol and lidocaine alone were 32.3 +/- 12.9 and 8.5 +/- 7.2 mA, respectively (p < 0.05, each drug compared with baseline; p < 0.05, esmolol vs. lidocaine). Mean increase in VFT from baseline during infusion of the combination was 52.0 +/- 22.0 mA (p < 0.05 as compared with baseline and with esmolol or lidocaine alone). The relationship between blood esmolol concentrations and VFT was described by a counterclockwise hysteresis curve, suggesting delay in equilibration of esmolol between blood and site of effect. The antifibrillatory efficacy of esmolol is significantly greater than that of lidocaine in this model. Administration of the two agents in combination resulted in significantly greater antifibrillatory efficacy than that associated with either drug administered alone.

Effect of reversible dorsal cold block on the persistence of inhibition generated by spinal reflexes.

The effects of bilateral focal cooling of dorsolateral thoracic spinal cord on segmental reflex pathways to the triceps surae muscles were assessed in decerebrate cats from the reflex forces produced by single shocks or trains of electrical stimuli applied to the ipsilateral caudal cutaneous sural and the contralateral tibial nerves. The validity of the dorsal cold block technique as a substitute for acute surgical dorsal hemisection was established by showing that focal cooling reliably reproduced the stretch-induced "clasp knife" inhibition of triceps surae reflexive force seen following dorsal hemisection. Under control (warm) conditions, the inhibitory components of electrically evoked ipsilateral sural and contralateral tibial reflexes faded rapidly during sustained trains, with a resultant production of large-amplitude reflex force as measured from either the entire triceps surae or from the medial gastrocnemius muscle alone. Dorsal cold block greatly reduced the amplitude of reflexive force evoked by sustained electrical stimulation of either nerve. Indeed, the cold block completely reversed the sign of train-evoked reflexes to a net inhibition of reflex force output in one-half of the sural and one-half of the contralateral tibial stimulation experiments. Peak transient forces evoked by single shocks to the sural or contralateral tibial nerves were also sometimes reduced, but this result was more variable than for prolonged nerve stimulation. The persistence of activity in segmental inhibitory pathways during dorsal cold block, as indicated by instances of reflex sign reversal, suggests that descending bulbospinal pathways traversing the dorsolateral funiculi may be responsible for "fading" of segmental inhibitory reflex components in decerebrate cats with intact spinal cords during sustained afferent input. The possibility that the enhanced magnitude and duration of segmental inhibition during cold block will increase the likelihood of disruption of the size principle for motoneuron recruitment is also discussed.

Depression of transmission from group II muscle afferents by electrical stimulation of the cuneiform nucleus in the cat.

The effects of short trains of electrical stimuli applied within the cuneiform nucleus and the subcuneiform region were examined on transmission from group I and group II muscle afferents to first-order spinal neurons. Variations in the effectiveness of transmission from these afferents were assessed from changes in the sizes of the monosynaptic component of extracellular field potentials evoked following stimulation of muscle nerves. Field potentials evoked from group II muscle afferents in the dorsal horn of the midlumbar and sacral segments and in the intermediate zone of the midlumbar segments were reduced when the test stimuli applied to peripheral nerves were preceded by conditioning stimulation of the cuneiform nucleus or the subcuneiform region. The depression occurred at conditioning-testing intervals of 20-400 ms, being maximal at intervals of 32-72 ms for dorsal horn potentials and 40-100 ms for intermediate zone potentials. At the shortest intervals, both group II and group I field potentials in the intermediate zone were depressed. Conditioning stimulation of the cuneiform nucleus depressed group II field potentials nearly as effectively as conditioning stimulation of the coerulear or raphe nuclei. We propose that the nonselective depression of transmission from group I and II afferents at short intervals is due to the activation of reticulospinal pathways by cells or fibers stimulated within the cuneiform area. We also propose that the selective depression of transmission from group II afferents at long intervals is mediated at least partly by monoaminergic pathways, in view of the similarity of the effects of conditioning stimulation of the cuneiform nucleus and of the brainstem monoaminergic nuclei and by directly applied monoamines (Bras et al. 1990). In addition, it might be caused by primary afferent depolarization mediated by non-monoaminergic fibers (Riddell et al. 1992).

Calcitonin Gene-Related Peptide (CGRP) Regulates Excitability and Refractory Period of the Guinea Pig Ureter

Previous studies have indicated that calcitonin gene-related peptide (CGRP), released from the peripheral endings of capsaicin-sensitive primary afferent neurons, may play a role as an inhibitory transmitter in the guinea pig ureter. The aim of this study was to compare the effect of capsaicin desensitization and administration of a CGRP receptor antagonist on the excitability and refractory period of the guinea pig ureter to electrical field stimulation. Electrical field stimulation using a long (5sec.) pulse width produced phasic contractions of the ureter which were unaffected by tetrodotoxin, that is, were produced through direct excitation of ureteral smooth muscle. Human αCGRP (1 to 10nM.) produced a concentration-dependent transient suppression of the evoked contractions, and its effect was prevented by the CGRP receptor antagonist human αCGRP(8-37) (1μM.). In vitro capsaicin pretreatment (10μM. for 15 minutes) to block neuropeptide release from peripheral endings of sensory nerves or administration of the CGRP receptor antagonist enhanced the responsiveness of the guinea pig ureter to electrical stimulation. In control ureters, the application of two trains of electrical stimuli failed to produce a second contraction at intertrain intervals greater than 20seconds. The intertrain interval required to obtain a second contraction averaging 50% of the amplitude of the first response (ITI50) of control ureters was about 50seconds. In vitro capsaicin pretreatment or administration of the CGRP receptor antagonist reduced the refractory period of the ureter to electrical field stimulation: ITI50 averaged 8.8 and 9.1seconds after capsaicin or CGRP antagonist pretreatment, respectively. These findings demonstrate that capsaicin pretreatment or blockade of CGRP receptors produced qualitatively and quantitatively similar excitatory effects on ureteral excitability and refractory period and are in general agreement with the idea that CGRP is a primary inhibitory transmitter in guinea pig ureter. Inhibition of motility by CGRP could be important for setting the frequency of ureteral peristalsis and suppression of latent pacemakers to prevent the occurrence of antiperistaltic waves.

Identification of reentry circuit sites during catheter mapping and radiofrequency ablation of ventricular tachycardia late after myocardial infarction.

Ventricular tachycardia reentry circuits in chronic infarct scars can contain slow conduction zones, which are difficult to distinguish from bystander areas adjacent to the circuit during catheter mapping. This study developed criteria for identifying reentry circuit sites using computer simulations. These criteria then were tested during catheter mapping in humans to predict sites at which radiofrequency current application terminated ventricular tachycardia. In computer simulations, effects of single stimuli and stimulus trains at sites in and adjacent to reentry circuits were analyzed. Entrainment with concealed fusion, defined as ventricular tachycardia entrainment with no change in QRS morphology, could occur during stimulation in reentry circuit common pathways and adjacent bystander sites. Pacing at reentry circuit common pathway sites, the stimulus to QRS (S-QRS) interval equals the electrogram to QRS interval (EG-QRS) during tachycardia. The postpacing interval from the last stimulus to the following electrogram equals the tachycardia cycle length. Pacing at bystander sites the S-QRS exceeds the EG-QRS interval when the conduction time from the bystander site to the circuit is short but may be less than or equal to the EG-QRS interval when the conduction time to the circuit is long. The postpacing interval, however, always exceeds the tachycardia cycle length. When conduction in the circuit slows during pacing, the S-QRS and postpacing intervals increase and the slowest stimulus train most closely reflects conduction times during tachycardia. Endocardial catheter mapping and radiofrequency ablation were performed during 31 monomorphic ventricular tachycardias in 15 patients with drug refractory ventricular tachycardia late after myocardial infarction. During ventricular tachycardia, trains of electrical stimuli or scanning single stimuli were evaluated before application of radiofrequency current at the same site. Radiofrequency current terminated ventricular tachycardia at 24 of 241 sites (10%) in 12 of 15 patients (80%). Ventricular tachycardia termination occurred more frequently at sites with entrainment with concealed fusion (odds ratio, 3.4; 95% confidence interval [CI], 1.4 to 8.3), a postpacing interval approximating the ventricular tachycardia cycle length (odds ratio, 4.6; 95% CI, 1.6 to 12.9) and an S-QRS interval during entrainment of more than 60 milliseconds and less than 70% of the ventricular tachycardia cycle length (odds ratio, 4.9; 95% CI, 1.4 to 17.1). Ventricular tachycardia termination was also predicted by the presence of isolated diastolic potentials or continuous electrical activity (odds ratio, 5.2; 95% CI, 1.8 to 15.5), but these electrograms were infrequent (8% of all sites). Combinations of entrainment with concealed fusion, postpacing interval, S-QRS intervals, and isolated diastolic potentials or continuous electrical activity predicted a more than 35% incidence of ventricular tachycardia termination during radiofrequency current application versus a 4% incidence when none suggested that the site was in the reentry circuit. Analysis of the postpacing interval and S-QRS interval suggested that 25% of the sites with entrainment with concealed fusion were in bystander areas not within the reentry circuit. At restudy 5 to 7 days later, 6 patients had no monomorphic ventricular tachycardia inducible, and inducible ventricular tachycardias were modified in 4 patients. None of these 10 patients have suffered arrhythmia recurrences during a follow-up of 316 +/- 199 days, although 4 continue to receive previously ineffective medications. Regions giving rise to reentry after myocardial infarction are complex and can include bystander areas, slow conduction zones, and isthmuses for impulse propagation at which radiofrequency current lesions can interrupt reentry.

Regular oscillations of synaptic activity in spinal networks in vitro

1. Spontaneous synaptic potentials were recorded in motoneurons grown in organotypic slice cultures of embryonic rat spinal cord. In 71 of 85 cells these potentials appeared without obvious temporal structure (random patterns); in the remaining 14 cells they appeared in bursts (rhythmic patterns). 2. Random activity patterns could be converted into rhythmic patterns by treating the cultures with strychnine, bicuculline, or both. The excitatory amino acid N-methyl-D-aspartate (NMDA) transiently increased the rate of spontaneous synaptic activity without inducing rhythmic patterns. The NMDA antagonist 7-chloro-kynurenate reduced the burst rate while leaving the burst length unchanged in rhythmic patterns. In random patterns it reduced the rate of spontaneous synaptic activity by 68%. 3. Histograms of interevent times of the random patterns were best fitted by the sum of two expontentials, suggesting that the random type of activity could not be described simply as a Poisson process but involved at least one additional mechanism. 4. Rhythmic patterns consisted of bursts of activity with a mean burst length of 2.2 s that were separated by interburst intervals with a mean length of 6.6 s. Within the bursts autocorrelograms revealed regular oscillations with a mean period of 226 ms in 6 of 11 experiments with rhythmic patterns. The period showed little variation between individual experiments (202-288 ms). In random patterns no oscillations were detected. 5. Within the spontaneous bursts the excitatory postsynaptic potentials (EPSPs) progressively declined in amplitude. A corresponding depression of EPSPs was observed when trains of electrical stimuli were applied at 5 Hz to the dorsal horns of the spinal cord slices.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemical switching device: biomimetic approach and application to neural network study

There are many examples of enzymes that share substrates or cofactors in a cyclic manner. Techniques have been developed that use cyclic enzyme systems to assay quantitatively small amounts of biochemical substances (cofactor, substrate), however, only a few studies of the control of these systems have been published. The author previously showed with computer simulations that cyclic enzyme systems have the reliability of on-off types of operation (McCulloch-Pitts' neuronic equation) capable of storing short-memory, and the applicability for a switching circuit in a biocomputer. This paper introduces a unique switching mechanism of cyclic enzyme system (basic switching element), and next, building the integrated biochemical switching system being composed of the basic switching element, shows the physiological phenomenon termed ‘selective elimination of synapses’ generally produced as a result of low-frequency train of electrical stimuli to the synapses (Kuroda, Y. (1989) Neurochem. Int. 14, 309–319).

Transmission from group II muscle afferents is depressed by stimulation of locus coeruleus/subcoeruleus, Kölliker-Fuse and raphe nuclei in the cat.

The effects of brief trains of electrical stimuli applied within the locus coeruleus and subcoeruleus, the Kölliker-Fuse nucleus and the raphe magnus, obscurus and pallidus nuclei were tested on transmission from group I and group II muscle afferent fibres in mid-lumbar spinal segments of chloralose anaesthetized cats. Changes in the effectiveness of transmission from these afferents were assessed from changes in the size of monosynaptic extracellular field potentials evoked by them. The depression of group II field potentials occurred at conditioning-testing intervals of 20-400 ms, and was maximal at intervals of 40-100 ms and 30-60 ms for potentials recorded in the intermediate zone and dorsal horn, respectively. At intervals up to about 30 ms it was combined with the depression of group I components of the intermediate zone field potentials. However, at longer intervals the conditioning stimuli depressed group II components of these potentials as selectively as monoamines applied ionophoretically at the recording site (Bras et al., 1989a, 1990). Thus, only the late depressive actions are considered as being possibly mediated by impulses in descending noradrenergic and/or serotonergic fibres. No major differences were found in the relative degree of depression of transmission from group II afferents by stimulation of the locus coeruleus/subcoeruleus, Kölliker-Fuse or raphe nuclei, either in the dorsal horn or in the intermediate zone. Since field potentials at these locations are preferentially depressed by ionophoretic application of serotonin and noradrenaline (Bras et al., 1990), and since the locus coeruleus/subcoeruleus, Kölliker-Fuse and raphe nuclei are interconnected, the study leads to the conclusion that both noradrenergic and serotonergic descending pathways can be activated by stimuli applied within either of them. Selective depression of field potentials of group II origin was also evoked by stimulation at other sites, e.g. the periaqueductal grey and medullary reticular formation, when conditioning-testing intervals were sufficiently long. Such a depression is considered to be secondary to activation of neurones of the locus coeruleus/subcoeruleus, Kölliker-Fuse or raphe nuclei and attributed to the spread of current or transsynaptic activation of these neurones, or to stimulation of their axon collaterals outside the nuclei rather than to other descending medullo-spinal systems. The non-selective depression of field potentials evoked by group I and group II afferents at shorter conditioning-testing intervals is proposed to be due to actions of reticulo-spinal pathways.

Halothane Alters Control of Intracellular Ca2+ Mobilization in Single Rat Ventricular Myocytes

In an attempt to understand the cellular mechanisms underlying volatile anesthetic-induced myocardial depression, halothane-induced negative inotropy was investigated in an animal model through continuous monitoring of intracellular Ca2+ concentration [( Ca2+]i) in rat ventricular myocytes loaded with fura-2. Single cells were stimulated with 15 mM caffeine or 15 mM extracellular K+ (K+O) or were paced by extracellular glass suction pipette electrode. With each stimulus modality, halothane (0.6-1.5%) caused a significant (P less than 0.05) and dose-dependent depression of the Ca2+ transient. Caffeine and electrically stimulated Ca2+ transients were reduced, in 1.5% halothane, to 35 +/- 14 and 42 +/- 8% of control, respectively. Resting or basal [Ca2+]i was unaffected by halothane. Halothane did not elicit spontaneous Ca2+ transients in these cells. Single cells stimulated by trains of electrical stimuli at 1.0, 1.5, and 2.0 Hz showed a change in [Ca2+]i from prestimulus levels to a stimulated baseline steady state that appeared to increase with stimulus frequency. Halothane at 0.7% increased the change in resting to stimulated baseline [Ca2+]i and depressed net transients (P less than 0.05) at 1.0 and 1.5 Hz. In contrast, 0.1 microM ryanodine depressed the Ca2+ transients in myocytes stimulated by trains of stimuli, but did not potentiate the change in stimulated baseline [Ca2+]i at any pacing rate. The results are consistent with the hypothesis that halothane reduces Ca2+i availability by causing a net loss of Ca2+ from the sarcoplasmic reticulum. The results from experiments using onset of pacing to induce a sudden increase in Ca2+i load in previously quiescent myocytes suggest that halothane may act to limit sarcoplasmic reticulum and/or sarcolemmal uptake/extrusion mechanisms, as compared to ryanodine, which depletes sarcoplasmic reticulum Ca2+ stores without affecting reuptake and extrusion.

Reflex activation of muscle spindles in human pretibial muscles during standing

1. Experiments were performed in standing subjects to determine whether low-threshold cutaneous and muscle afferents from mechanoreceptors in the human foot reflexly influence fusimotor neurons innervating pretibial flexor muscles. Recordings were made from 30 identified muscle-spindle afferents, four tendon-organ afferents, and one alpha-motor axon innervating the pretibial flexor muscles. The subjects stood without support or vision on a force platform while trains of electrical stimuli (5 stimuli, 300 Hz) were delivered at nonpainful intensities to the sural nerve or to the posterior tibial nerve at the ankle. 2. Seventeen of the 30 spindle endings had no background discharge, and none was activated by the sural or posterior tibial stimuli. Five silent afferents were given a background discharge by sustained pressure on the relevant tendon, but with two the discharge was dominated by a tremor rhythm obscuring any reflex response to the stimuli. Based on peristimulus time histograms (PSTHs), the sural stimuli then produced increases in discharge of two of the remaining three endings at latencies of 84 and 90 ms. These effects could not be explained by muscle stretch and are presumed to have been fusimotor mediated. 3. When the subjects stood freely without support or vision, 13 muscle-spindle endings had a background discharge, but with three endings tremor developed at the ankle and dominated the spindle discharge. Sural stimuli affected the discharge of five of nine endings unaffected by tremor. With three of these endings, there were changes in discharge that could be explained by muscle stretch.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothalamic effects on medullary reticular activation of deep back muscle EMG

Medullary reticular stimulation can activate deep back muscle EMG in urethane-anesthetized female rats. Midbrain central gray stimulation can facilitate brainstem reticular control over deep back muscles. Since these deep back muscles lateral longissimus (LL) and medial longissimus (ML) execute the vertebral dorsiflexion of lordosis behavior, and since the motor control hierarchy sketched above parallels lordosis behavior circuitry, we tested the hypothesis that medial hypothalamic lesions (which, in behavioral experiments, decrease lordosis) can also reduce medullary reticular activation of deep back muscle EMG. Urethane-anesthetized rats were tested systematically for amplitude of lateral longissimus (LL) and medial longissimus (ML) EMG responses to electrical stimulus trains applied to the nucleus gigantocellularis (NGC) of the medullary reticular formation, before and after electrolytic lesions of the ventromedial hypothalamus (n = 18) or control sites (n = 30). Bilateral ventromedial hypothalamic lesions were able to greatly reduce EMG responses in LL and ML, often with a time course similar to previous lordosis behavioral results. Surprisingly, lesions at the anterior ventromedial nucleus pole were particularly effective, and may reflect importance of intraventromedial local neurons. Although, on the average, various control lesions were less effective, the ventromedial hypothalamic effect was not unique. For example, it was possible to see an EMG decrease following lesions of the dorsomedial thalamus. Nevertheless, EMG loss was not well correlated with changes in the cortical EEG, and thus does not appear to be a simple consequence of changes in “arousal.” In conclusion, it appears that ventromedial hypothalamic neurons can affect medullary reticular control of back muscle EMG, but must share this role with other forebrain elements.

Fluctuations of membrane potential in isolated single ventricular myocytes of guinea-pig upon resumption of oxidative phosphorylation

Electrical and mechanical activities of guinea-pig single ventricular myocytes were investigated under conditions simulating hypoxia-reoxygenation. The localized movement of sarcomere was recorded simultaneously with membrane potential, and analyzed using microcomputer-based image processing. Exposure to 5 m m CN − caused progressive shortening of action potential duration and attenuation of twitch contraction. The myocytes became inexcitable about 30 to 70 min after the CN − treatment. On removal of CN −, the myocytes exhibited periodic miniature membrane depolarizations from the resting potential level (−95 mV). When depolarizations were smaller than 6 mV in amplitude and longer than 500 ms in duration, they were accompanied by localized sarcomere shortening like a propagating contractile wave (unifocal oscillation). Membrane depolarizations of larger amplitude and shorter duration were associated with a more uniform pattern of localized sarcomere shortening (multifocal oscillation). Trains of electrical stimuli applied during the washing out period caused transient augmentation of potential fluctuation and enhancement of synchronization of sarcomere shortening. These results suggest that non-uniform elevation of intracellular calcium concentration on the resumption of oxidative phosphorylation may initiate oscillatory fluctuations of membrane potential leading to abnormal spontaneous excitation.

Paraesthesiae and hypaesthesia following prolonged high-frequency stimulation of cutaneous afferents.

The activity of cutaneous afferents was recorded in human subjects using microelectrodes inserted into individual fascicles of the median nerve at the wrist before and after a 10 min train of electrical stimuli at 200 Hz delivered to the appropriate digital nerve (via ring electrodes) or to individual afferent axons (via the microelectrode). Changes in neural activity produced by the stimulation were correlated with the time course of paraesthesiae and with changes in the ability to detect cutaneous stimuli. From approximately 20 s after the end of the stimulus train, there was a progressive increase in neural activity, and individual afferents became spontaneously active and discharged in high-frequency bursts. At this time the subjects began to experience paraesthesiae. Repetitive stimulation proximal to a complete digital nerve block induced paraesthesiae that were felt distal to the block in the insensate digit, indicating that they did not arise from the unmyelinated terminal segment of the axon or from a stimulus-induced disorder of receptor function. Recordings of the compound action potential evoked by submaximal test stimuli were made after the 10 min stimulus train and revealed evidence of an early transient increase in excitability superimposed on a long-lasting decrease in excitability, reaching a nadir approximately 30-40 min after the end of the repetitive stimulation. In parallel recordings, there was no detectable change in the cutaneous afferent volley evoked by mechanical stimulation, paraesthesiae, can be attributed directly to a disturbance in peripheral afferent fibres, while the poststimulation negative symptoms such as hypaesthesia arise from stimulation-induced refractoriness at central synaptic relays.

Activation of human muscles at short muscle lengths during maximal static efforts.

1. Human muscle endurance is apparently enhanced during maximal voluntary contractions at short muscle lengths (McKenzie & Gandevia, 1987) but the ability of subjects to activate muscles fully at short lengths has not been established. Therefore this study examined the voluntary capacity to activate muscles fully at control (near resting) lengths and at decreased muscle lengths. Changes in mechanical properties of twitch responses to electrical stimulation of relaxed muscles at short muscle lengths were also documented. The abductor digiti minimi, elbow flexors and tibialis anterior were studied in five subjects. 2. For the three muscle groups, the mean reduction in twitch force between the control and short muscle lengths ranged from 46-51%. AT the short length there was a 9-13% reduction in the contraction time and a 21-27% reduction in the half-relaxation time. Maximal voluntary force declined by 21-49% at the short muscle length. A reduction in muscle length produced a shift to the right of the force-frequency curve as determined by brief trains of electrical stimuli. 3. During maximal efforts single or brief trains of two to four supramaximal stimuli, delivered to the parent nerve or motor point, failed to increase the force at a latency appropriate for onset of a muscle twitch in some but not all attempts. Each subject achieved 'maximal activation' of the muscle in a similar proportion of attempts at the control and short muscle lengths. 4. These results suggest that maximal voluntary activation of motoneurone pools is possible at short muscle lengths and that the central nervous system is able to maintain the discharge of motoneurones close to 'fusion' frequency despite a decrease in the temporal characteristics of the isometric twitch.