Showing papers in "The Journal of Physiology in 1990"

Properties of a hyperpolarization-activated cation current and its role in rhythmic oscillation in thalamic relay neurones.

Abstract: 1. The physiological and functional features of time-dependent anomalous rectification activated by hyperpolarization and the current which underlies it, Ih, were examined in guinea-pig and cat thalamocortical relay neurones using in vitro intracellular recording techniques in thalamic slices. 2. Hyperpolarization of the membrane from rest with a constant-current pulse resulted in time-dependent rectification, expressed as a depolarizing sag of the membrane potential back towards rest. Under voltage clamp conditions, hyperpolarizing steps to membrane potentials negative to approximately -60 mV were associated with the activation of a slow inward current, Ih, which showed no inactivation with time. 3. The activation curve of the conductance underlying Ih was obtained through analysis of tail currents and ranged from -60 to -90 mV, with half-activation occurring at -75 mV. The time course of activation of Ih was well fitted by a single-exponential function and was strongly voltage dependent, with time constants ranging from greater than 1-2 s at threshold to an average of 229 ms at -95 mV. The time course of de-activation was also described by a single-exponential function, was voltage dependent, and the time constant ranged from an average of 1000 ms at -80 mV to 347 ms at -55 mV. 4. Raising [K+]o from 2.5 to 7.5 mM enhanced, while decreasing [Na+]o from 153 to 26 mM reduced, the amplitude of Ih. In addition, reduction of [Na+]o slowed the rate of Ih activation. These results indicate that Ih is carried by both Na+ and K+ ions, which is consistent with the extrapolated reversal potential of -43 mV. Replacement of Cl- in the bathing medium with isethionate shifted the chloride equilibrium potential positive by approximately 30-70 mV, evoked an inward shift of the holding current at -50 mV, and resulted in a marked reduction of instantaneous currents as well as Ih, suggesting a non-specific blocking action of impermeable anions. 5. Local (2-10 mM in micropipette) or bath (1-2 mM) applications of Cs+ abolished Ih over the whole voltage range tested (-60 to -110 mV), with no consistent effects on instantaneous currents. Barium (1 mM, local; 0.3-0.5 mM, bath) evoked a steady inward current, reduced the amplitude of instantaneous currents, and had only weak suppressive effects on Ih. 6. Block of Ih with local application of Cs+ resulted in a hyperpolarization of the membrane from the resting level, a decrease in apparent membrane conductance, and a block of the slow after-hyperpolarization that appears upon termination of depolarizing membrane responses, indicating that Ih contributes substantially to the resting and active membrane properties of thalamocortical relay neurones.(ABSTRACT TRUNCATED AT 400 WORDS)

Electrical resistance across the blood-brain barrier in anaesthetized rats: a developmental study.

Abstract: 1. Ion permeability of the blood-brain barrier was studied by in situ measurement of transendothelial electrical resistance in anaesthetized rats aged between 17 days gestation and 33 days after birth, and by electron microscopic examination of lanthanum permeability in fetal and neonatal rats aged up to 10 days old. 2. The blood-brain barrier in 17- to 20-day fetuses had a resistance of 310 omega cm2 but was impermeable to lanthanum, and therefore had properties intermediate between leaky and tight epithelia. 3. From 21 days gestation, the resistance was 1128 omega cm2, indicating a tight blood-brain barrier and low ion permeability. There was little further change in barrier resistance after birth, and in 28- to 33-day rats, when the brain barrier systems are mature in other ways, vessels had a mean resistance of 1462 omega cm2. 4. In the tight blood-brain barrier, arterial vessels had a significantly higher resistance than venous vessels, 1490 and 918 omega cm2 respectively. In vessels less than 50 microns diameter and within the normal 60 min experimental period, there was no significant variation in vessel resistance. 5. Hyperosmotic shock caused a rapid decay in resistance (maximal within 5 min), and after disruption of the blood-brain barrier, vessel resistance was 100-300 omega cm2 in both arterial and venous vessels, and the effect was reversible. After the application of metabolic poisons (NaCN plus iodoacetate) and low temperature there was a similarly low electrical resistance. 6. It is concluded that the increase in electrical resistance at birth indicates a decrease in paracellular ion permeability at the blood-brain barrier and is required for effective brain interstitial fluid ion regulation.

Paired‐pulse depression of monosynaptic GABA‐mediated inhibitory postsynaptic responses in rat hippocampus.

Abstract: 1. Intracellular recording techniques were used to characterize monosynaptic inhibitory postsynaptic potentials (IPSPs) and currents (IPSCs) in rat hippocampal slices and to study the mechanism of paired-pulse depression of these synaptic responses. This was achieved by stimulation in stratum radiatum close (less than 0.5 mm) to an intracellularly recorded CA1 neurone after pharmacological blockade of all excitatory synaptic transmission. 2. Under these conditions, low-frequency stimulation (0.033 Hz) evoked a pure biphasic IPSP, which had a short and constant latency to onset. This IPSP was blocked by tetrodotoxin (1 microM) suggesting that it resulted from the electrical stimulation of the axons and/or cell bodies of a monosynaptic inhibitory pathway. 3. Picrotoxin (100 microM) abolished the early component of the biphasic IPSP/C. It left an intact, pure late IPSP/C (IPSP/CB) which had a latency to onset of 29 +/- 2 ms, latency to peak of 139 +/- 4 ms, a duration of 723 +/- 135 (range 390-1730) ms and a reversal potential of -93 +/- 2 mV. The duration was highly dependent on the stimulus intensity whereas the latency to onset was largely independent of the stimulus intensity. The IPSP/CB was reduced or abolished by 1 mM-phaclofen. 4. Phaclofen (1 mM) and 2-hydroxy-saclofen (0.1-1.0 mM) reversibly depressed (60-100%) the late component of the biphasic IPSP/C and, where maximally effective, left a pure, early IPSP/C (IPSP/CA). The IPSP/CA had a latency to onset of 3 ms or less, a latency to peak of 17 +/- 1 ms, a duration of 225 +/- 3 ms and a reversal potential of -75 +/- 2 mV. 5. Two shocks of identical strength were applied in close succession to characterize, and to study the mechanisms underlying, frequency-dependent depression of inhibitory synaptic responses. Paired-pulse depression was seen for both phases of the biphasic IPSP/C and of the pure IPSP/CB, recorded in the presence of picrotoxin. Paired-pulse depression was not accompanied by changes in the reversal potential of either component, indicating that it was caused by a reduction in the two synaptic conductances. Paired-pulse depression was greater when high stimulus intensities were employed. 6. Paired stimuli were applied at separation intervals of between 5 ms and 10 s to determine the temporal profile of frequency-dependent depression. Paired-pulse depression of both IPSCA and IPSCB was most pronounced at an interstimulus interval of 100-125 ms and ceased to occur at intervals greater than 5 10s.(ABSTRACT TRUNCATED AT 400 WORDS)

Activation of unmyelinated afferent fibres by mechanical stimuli and inflammation of the urinary bladder in the cat.

Abstract: 1. We examined the functional properties of unmyelinated primary afferent neurones innervating the pelvic viscera in twenty-five anaesthetized cats. The axons were isolated from the intact dorsal root and the intact or chronically de-efferented ventral root of the segment S2. All units were electrically identified with electrical stimulation of the pelvic nerve. 2. The responses of the neurones were studied with natural stimulation of the urinary bladder using innocuous and noxious increases of intravesical pressure and at the onset of an acute artificial inflammation induced by intraluminal injection of mustard or turpentine oil. 3. Out of 297 unmyelinated afferent units isolated from the dorsal root, seven were excited by an increase of the intravesical pressure during contractions and distension of the urinary bladder. These units were silent when the bladder was empty and had thresholds of 30-50 mmHg which are presumed to be noxious. Further increases of the intravesical pressure were accurately encoded by the discharge rate of the fibres. Out of sixty-eight unmyelinated afferent units isolated from the ventral root none was activated by these stimuli. 4. Intraluminal injection of mustard oil excited mechanosensitive units at short latency. The discharge was not closely related to changes of the intravesical pressure and the units displayed on-going activity after the irritant had been removed. This observation suggests that the units had also chemosensitive properties and that the receptive endings were located in the bladder wall. 5. In sixteen cats ninety-five afferent fibres that were not activated by noxious mechanical stimuli of the urinary bladder were systematically tested with intraluminal injections of mustard oil. This excited 7/67 dorsal root units and 4/28 ventral root units with short latency. Intraluminal application of turpentine oil, tested on twenty-six afferents in four animals, did not produce a rapid excitation. 6. Following the induction of an inflammation some previously non-mechanosensitive units started to respond to changes of intravesical pressure in the biologically relevant pressure range of the urinary bladder. 7. In conclusion, a small subpopulation (2.4%) of unmyelinated visceral afferents responds to high, presumably noxious, intravesical pressure and intraluminal application of chemical irritants. Acute inflammation excites a larger proportion of afferents (9.5%) that are not activated by acute noxious mechanical stimulation of the normal urinary bladder. In the inflamed bladder some previously non-mechanosensitive units started to respond to increases of intravesical pressure. These novel types of chemosensitive receptors may contribute considerably to the pathogenesis of visceral pain states.

Quantal analysis of inhibitory synaptic transmission in the dentate gyrus of rat hippocampal slices: a patch-clamp study

Abstract: 1. Synaptically connected neurones were identified in the granule cell layer of slices of 17- to 21-day-old rat hippocampus. Whole-cell current recording using the patch-clamp technique revealed synaptic currents ranging from less than 10 to 200 pA in symmetrical Cl- conditions, at a holding potential of -50 mV. These currents were blocked by 2 microM-bicuculline, indicating that they result from the activation of postsynaptic gamma-aminobutyric acid receptor (GABAA-receptor) channels. 2. Addition of tetrodotoxin (TTX, 1 microM) resulted in the loss of most currents of more than 40 pA in amplitude. Currents which disappeared after TTX treatment were assumed to be the result of spontaneous presynaptic action potentials. The currents seen in the absence of TTX are referred to as spontaneously occurring inhibitory postsynaptic currents (IPSCs); those remaining in the presence of TTX were defined as miniature IPSCs. 3. Similar currents were observed when recording in the whole-cell configuration while extracellular stimulation was applied to a nearby neurone. These currents were also completely blocked by 2 microM-bicuculline and by 0.5 microM-TTX. They were thus defined as stimulus-evoked IPSCs. 4. The half rise time of both miniature and stimulus-evoked IPSCs was fast (less than 1 ms). The time course of decay of both miniature IPSCs and stimulus-evoked IPSCs could be well fitted with the sum of two exponentials. At a membrane potential of -50 mV, the mean decay time constants of the two components were 2.0 +/- 0.38 and 54.4 +/- 18 ms (mean +/- S.D.) for miniature IPSCs (six cells) and 2.2 +/- 1.3 and 66 +/- 20 ms (three cells) for stimulus-evoked IPSCs. 5. Stimulus-evoked IPSCs varied in amplitude from less than ten to hundreds of picoamperes. In eight of eleven cells histograms of IPSC amplitudes showed several clear peaks which, when fitted with the sum of Gaussian curves, were found to be equidistant. This is consistent with the view that stimulus-evoked IPSC amplitudes vary in a quantal fashion. The quantal size varied between 7 and 20 pA, at a membrane potential of -50 mV. 6. Decreasing the Ca2+ and increasing the Mg2+ concentration in the extracellular solution decreased the number of peaks in the IPSC amplitude histogram but did not affect the size of the quantal event.(ABSTRACT TRUNCATED AT 400 WORDS)

Permeation of calcium through excitatory amino acid receptor channels in cultured rat hippocampal neurones.

Abstract: 1. N-methyl-D-aspartate (NMDA)-, quisqualate- and kainate-induced currents were recorded in cultured rat hippocampal neurones using the whole-cell voltage-clamp technique. To isolate the inward currents carried by Ca2+ and other divalent cations (Sr2+, Ba2+, Mn2+ and Mg2+), both Na+ and K+ in the control external solution were replaced with the impermeant cation N-methylglucamine (NMG). 2. Replacement of Na+, K+ and Ca2+ with NMG abolished NMDA-, quisqualate- and kinate-induced inward currents. In Na(+)-, K(+)-free (abbreviated simply as Na(+)-free) solution containing 10 mM-Ca2+ NMDA caused prominent inward currents at -60 mV. In this solution with the internal solution containing 165 mM-Cs+, the reversal potential of the NMDA-induced current was -5.0 +/- 0.7 mV (n = 36), indicating a value of PCa/PCs = 6.2 for the ratio of the permeability coefficients of Ca2+ and Cs+ according to the constant-field equation. 3. NMDA elicited inward current responses at -60 mV in Na(+)-, Ca2(+)-free solution containing 10 mM-Sr2+, Ba2+, or Mn2+, but not in Na(+)-free, 10 mM-Mg2+ solution. On the basis of reversal potential measurements, the permeability sequence of NMDA receptor channels among the divalent cations was determined to be Ba2+ (1.2) greater than Ca2+ (1.0) greater than Sr2+ (0.8) greater than Mn2+ (0.3) much greater than Mg2+ (less than 0.02). 4. The reversal potential of the quisqualate-induced current was more negative than -80 mV in Na(+)-free, 10 mM-Ca2+ solution, indicating a value of PCa/PCs less than 0.18. 5. Kainate-induced current responses were classified into two types. In the type I response the reversal potential of the kainate-induced current was more negative than -80 mV in Na(+)-free, 10 mM-Ca2+ solution, indicating that the Ca2+ permeability of this type of kainate channel is as low as that of the quisqualate channel. In the neurones which showed a type I response, there was a tendency of outward rectification in the current-voltage plots of the kainate response in control solution. 6. In the type II response kainate caused prominent inward currents at -60 mV in Na(+)-free, 10 mM-Ca2+ solution. The reversal potential was -23.3 +/- 5.6 mV (n = 17), indicating a permeability ratio PCa/PCs = 2.3. In the neurones which showed a type II response, a remarkable inward rectification was observed in the current-voltage plots of the kainate response in control solution. 7. Type II kainate channels showed relatively poor selectivity among divalent cations.(ABSTRACT TRUNCATED AT 400 WORDS)

Analysis of excitatory synaptic action in pyramidal cells using whole-cell recording from rat hippocampal slices.

Abstract: 1. The pharmacological and biophysical properties of excitatory synapses in the CA1 region of the hippocampus were studied using patch electrodes and whole-cell recording from thin slices. 2. Excitatory postsynaptic currents (EPSCs) had a fast component whose amplitude was voltage insensitive and a slow component whose amplitude was voltage dependent with a region of negative slope resistance in the range of -70 to -30 mV. 3. The voltage-dependent component was abolished by the N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5-phosphonovalerate (APV; 50 microM), which had no effect on the fast component. Conversely, the fast voltage-insensitive component was abolished by the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM) which had no effect on the slow component. 4. In Ringer solution with no added Mg2+ the current-voltage relation of the NMDA component was linear over a much larger voltage range than in the presence of 1.3 mM-Mg2+. 5. The NMDA component of the EPSC could be switched off with a hyperpolarizing voltage step at the soma. The kinetics of this switch-off was used to estimate the speed of clamp control of the subsynaptic membrane as well as the electrotonic distance from the soma. The kinetic analysis of the EPSC was restricted to synapses which were judged to be under adequate voltage control. 6. For those synapses that were close to the soma the time constant for decay for the non-NMDA component, which was voltage insensitive, ranged from 4-8 ms. 7. The rise time for the NMDA component was 8-20 ms and the time constant for decay ranged from 60-150 ms. 8. During increased transmitter release with post-tetanic potentiation or application or phorbol esters, both components of the EPSC increased to a similar extent. 9. These experiments provide a detailed description of the dual receptor mechanism operating at hippocampal excitatory synapses. In addition, the experiments provide an electrophysiological method for estimating the electrotonic distance of synaptic inputs.

Visual transduction in cones of the monkey Macaca fascicularis.

Abstract: 1. Visual transduction in macaque cones was studied by measuring the membrane current of single outer segments projecting from small pieces of retina. 2. The response to a brief flash of light was diphasic and resembled the output of a bandpass filter with a peak frequency near 5 Hz. After the initial reduction in dark current there was a rebound increase which resulted from an increase in the number of open light-sensitive channels. The response to a step of light consisted of a prominent initial peak followed by a steady phase of smaller amplitude. 3. Responses to dim light were linear and time-invariant, suggesting that responses to single photons were linearly additive. From the flash sensitivity and the effective collecting area the peak amplitude of the single photon response was estimated as about 30 fA. 4. With flashes of increasing strength the photocurrent amplitude usually saturated along a curve that was gentler than an exponential but steeper than a Michaelis relation. The response reached the half-saturating amplitude at roughly 650 photoisomerizations. 5. The response-intensity relation was flatter in the steady state than shortly after a light step was turned on, indicating that bright light desensitized the transduction with a delay. This desensitization was not due to a reduction in pigment content. In the steady state, a background of intensity I lowered the sensitivity to a weak incremental test flash by a factor 1/(1 + I/IO), where IO was about 2.6 x 10(4) photoisomerizations s-1, or about 3.3 log trolands for the red- and green-sensitive cones. 6. Bleaching exposures produced permanent reductions in flash sensitivity but had little effect on the kinetics or saturating amplitude of subsequent flash responses. The sensitivity reductions were consistent with the expected reductions in visual pigment content and gave photosensitivities of about 8 x 10(-9) microns2 (free solution value) for the red- and green-sensitive pigments. During a steady bleaching exposure the final exponential decline of the photocurrent had a rate constant given by the product of the light intensity and the photosensitivity. 7. In some cells it was possible to measure a light-induced increase in current noise. The power spectrum of the noise resembled the spectrum of the dim flash response and the magnitude of the noise was consistent with a single photon response roughly 20 fA in size. 8. The membrane current recorded in darkness was noisy, with a variance near 0.12 pA2 in the band 0-20 Hz.(ABSTRACT TRUNCATED AT 400 WORDS)

Localization and responses of neurones in the parieto-insular vestibular cortex of awake monkeys (Macaca fascicularis).

Abstract: 1. In four Java monkeys (Macaca fascicularis) 152 vestibular neurones were recorded in the parietal cortex located in the upper bank of the lateral sulcus near the posterior end of the insula. We called this region parieto-insular vestibular cortex (PIVC). PIVC extends about 6-8 mm in the anterior-posterior direction from the posterior part of the insula into the retroinsular region (stereotaxic co-ordinates: anterior 4-12 mm, lateral 16-19 mm and vertical 3-6 mm). 2. About two-thirds of the neurones recorded from this region responded to vestibular stimuli; the non-vestibular neurones responded predominantly to somatosensory stimulation of the neck and shoulder region. The PIVC area is a polysensory field, since almost all vestibular neurones were also activated by somatosensory and visual stimuli. Large-field optokinetic stimulation was the most effective visual stimulus. 3. With vestibular stimuli, responses to angular acceleration were dominant; steady tilt in darkness rarely led to any change in neuronal spontaneous activity. Of sixty-four neurones tested by rotation in more than one plane, fifty-four responded to excitation of semicircular canal receptors aroused by rotation in more than one of the three experimental planes (roll, yaw, pitch). Compared with vestibular brain stem units PIVC neurones discharged with a higher variability. 4. In the responses to horizontal rotation of the animal 38% type I, 53% type II and 9% type III units were recorded (classification according to Duensing & Schaefer, 1958). The gain measured with horizontal sinewave rotation was lower by a factor of about 4 in PIVC neurones as compared with the responses of vestibular neurones in the brain stem or thalamus (VPL). The phase response characteristics and the gain increase with increasing sinewave stimulus frequency, however, were in the same range as observed in neurones of the afferent vestibular system. 5. When the vestibular responses to sinusoidal rotation were tested in all three experimental planes (yaw, roll, pitch), the response strength as expressed by the amplitude of the peristimulus time histograms (PSTHs) differed for the three rotational planes. For different units the relative sensitivity to rotation in each of the three planes also differed. We concluded from this observation that different PIVC units had different optimum sensitivity planes for rotation with respect to the head co-ordinates, whereby all possible planes are represented.

Regulation of the intracellular free calcium concentration in single rat dorsal root ganglion neurones in vitro.

Abstract: 1. Simultaneous whole-cell patch-clamp and Fura-2 microfluorimetric recordings of calcium currents (ICa) and the intracellular free Ca2+ concentration ([Ca2+]i) were made from neurones grown in primary culture from the dorsal root ganglion of the rat. 2. Cells held at -80 mV and depolarized to 0 mV elicited a ICa that resulted in an [Ca2+]i transient which was not significantly buffered during the voltage step and lasted long after the cell had repolarized and the current ceased. The process by which the cell buffered [Ca2+]i back to basal levels could best be described with a single-exponential equation. 3. The membrane potential versus ICa and [Ca2+]i relationship revealed that the peak of the [Ca2+]i transient evoked at a given test potential closely paralleled the magnitude of the ICa suggesting that neither voltage-dependent nor Ca2(+)-induced Ca2+ release from intracellular stores made a significant contribution to the [Ca2+]i transient. 4. When the cell was challenged with Ca2+ loads of different magnitude by varying the duration or potential of the test pulse, [Ca2+]i buffering was more effective for larger Ca2+ loads. The relationship between the integrated ICa and the peak of the [Ca2+]i transient reached an asymptote at large Ca2+ loads indicating that Ca2(+)-dependent processes became more efficient or that low-affinity processes had been recruited. 5. Inhibition of Ca2+ influx with neuropeptide Y demonstrated that inhibition of a large ICa produced minor alterations in the peak of the [Ca2+]i transient, while inhibition of smaller currents produced corresponding decreases in the [Ca2+]i transient. Thus, inhibition of the ICa was reflected by a change in the peak [Ca2+]i only when submaximal Ca2+ loads were applied to the cell, implying that modulation of [Ca2+]i is dependent on the activation state of the cells. 6. Intracellular dialysis with the mitochondrial Ca2+ uptake blocker Ruthenium Red in whole-cell patch-clamp experiments removed the buffering component which was responsible for the more efficient removal of [Ca2+]i observed when large Ca2+ loads were applied to the cell. 7. When cells were superfused with 50 mM-K+, [Ca2+]i transients recorded from the cell soma returned to control levels very slowly. Pharmacological studies indicated that mitochondria were cycling Ca2+ during this sustained elevation in [Ca2+]i. In contrast, [Ca2+]i transients recorded from cell processes returned to basal levels relatively rapidly. 8. Extracellular Na(+)-dependent Ca2+ efflux did not significantly contribute to buffering [Ca2+]i transients in dorsal root ganglion neurone cell bodies.(ABSTRACT TRUNCATED AT 400 WORDS)

Noradrenergic and serotonergic modulation of a hyperpolarization-activated cation current in thalamic relay neurones.

Abstract: 1. Modulation of the hyperpolarization-activated cation current, Ih, by noradrenaline (NA) and serotonin (5-HT) was examined in guinea-pig and cat medial and lateral geniculate relay neurones using the in vitro slice technique. 2. In the absence of pharmacological antagonists, local application of NA resulted in a slow depolarization and decrease in apparent input conductance, a response which was blocked by local or bath application of the alpha 1-adrenoceptor antagonist prazosin. Application of NA after pharmacological block of alpha 1- and alpha 2-adrenoceptors, or application of 5-HT in all conditions, induced a 1-3 mV slow depolarization which was associated with a pronounced increase in apparent input conductance. This response to NA and 5-HT persisted during blocked synaptic transmission and was present in both the guinea-pig and cat medial and lateral geniculate nuclei. 3. The increase in membrane conductance elicited by NA was mimicked by the beta-specific agonist isoprenaline and blocked by the beta-antagonists propranolol and atenolol, indicating that it is mediated by beta-adrenoceptors. The response to 5-HT was blocked by the 5-HT1 and 5-HT2 antagonist methysergide, but not by the 5-HT2 antagonist ritanserin. Applications of either the 5-HT1A agonist ipsapirone or the partial agonist 8-hydroxy-dipropylaminotetralin (8-OHDPAT) were without effect. 4. Current versus voltage relationships obtained under voltage clamp revealed NA and 5-HT to cause a voltage-dependent inward shift at membrane potentials negative to approximately -60 mV. This response appeared to be shared by NA and 5-HT since maximal application of 5-HT greatly reduced or abolished the response to NA. 5. Application of NA and/or 5-HT during hyperpolarizing voltage steps in voltage clamp resulted in a marked increase in amplitude of the hyperpolarization-activated cation current, Ih. In addition, the rate of activation of Ih was strongly increased during activation of beta-adrenoceptors. 6. The activation curve of the conductance underlying Ih (Gh) was shifted by 4-6 mV on the voltage axis with NA and/or 5-HT. The positive shift of Gh activation in the voltage domain resulted in an increase in the amplitude of Gh which is active at resting, and more hyperpolarized, membrane potentials. The subsequent increase in resting membrane conductance decreased the responsiveness of thalamic neurones to hyperpolarizations of all durations. 7. Local or bath application of caesium blocked both Ih and the increase in membrane conductance in response to NA and 5-HT. By contrast, barium blocked neither Ih nor the responses to NA and 5-HT.(ABSTRACT TRUNCATED AT 400 WORDS)

Vestibular neurones in the parieto‐insular cortex of monkeys (Macaca fascicularis): visual and neck receptor responses.

Abstract: 1. One hundred and fifty-two vestibularly activated neurones were recorded in the parieto-insular vestibular cortex (PIVC) of four awake Java monkeys (Macaca fascicularis): sixty-two were tested systematically with visual stimulation and seventy-nine were tested with various somatosensory stimuli. With very few exceptions all vestibular neurones tested responded to visual and somatosensory stimulation, therefore being classified as polymodal vestibular units. 2. A most effective stimulus for all fifty-eight visually activated PIVC units was movement of a large structured visual pattern in an optimal direction. From forty-four units responsive to a horizontally moving optokinetic striped drum, twenty-nine were activated with optokinetic movement in the opposite direction to the activating vestibular stimulus ('synergistic' response), thirteen were activated optokinetically and vestibularly in the same direction ('antagonistic' responses) and two were biphasic. The gain of the optokinetic response to sinusoidal stimulation (average 0.28 (impulses s-1) (deg s-1)-1 at 0.2 Hz, 56 deg amplitude) was in a range similar to that of the vestibular gain at low frequencies. At 1 Hz some units only showed weak optokinetic responses or none at all, but the vestibular response was still strong. 3. With different 'conflicting' or 'enhancing' combinations of optokinetic and vestibular stimulation no generalized type of interaction was observed, but the responses varied from nearly 'algebraic' summation to no discernible changes in the vestibular responses by additional optokinetic stimuli. With all visual-vestibular stimulus combinations the responses to the vestibular stimulus remained dominant. 4. The optokinetic preferred direction was not related to gravitational coordinates since the optokinetic responses were related to the head co-ordinates and remained constant with respect to the head co-ordinates at different angles of steady tilt. 5. Almost all PIVC units were activated by somatosensory stimulation, whereby mainly pressure and/or movement of neck and shoulders (bilateral) and movement of the arm joints elicited vigorous responses. Fewer neurones were activated by lightly touching shoulders/arms or neck, by vibration and/or pressure to the vertebrae, pelvis and legs. 6. A most effective somatosensory stimulus was sinewave rotation of the body with head stationary. The gain of this directionally selective neck receptor response was in the range of vestibular stimulation. Interaction of vestibular and neck receptor stimulation was either of a cancellation or facilitation type.(ABSTRACT TRUNCATED AT 400 WORDS)

Anaerobic energy production and O2 deficit-debt relationship during exhaustive exercise in humans.

Abstract: 1. Eight subjects performed one-legged, dynamic, knee-extensor exercise, first at 10 W followed by 10 min rest, then at an intense, exhaustive exercise load (65 W) lasting 3.2 min. After 60 min recovery, exercise was performed for 8-10 min each at 20, 30, 40 and 50 W. Measurements of pulmonary oxygen uptake, heart rate, blood pressure, leg blood flow, and femoral arterial-venous differences of oxygen content and lactate were performed as well as determination of ATP, creatine phosphate (CP) inosine monophosphate (IMP) and lactate concentrations on biopsy material from the quadriceps muscle before and immediately after the intense exercise, and at 3, 10 and 60 min into recovery. 2. Individual linear relations (r = 0.95-1.00) between the power outputs for submaximal exercise and oxygen uptakes (leg and pulmonary) were used to estimate the energy demand during intense exercise. Pulmonary and leg oxygen deficits determined as the difference between energy demand and oxygen uptake were 0.46 and 0.48 l (kg active muscle)-1, respectively. Limb and pulmonary oxygen debts (oxygen uptake during 60 min of recovery - pre-exercise oxygen uptake) were 0.55 and 1.65 l (kg active muscle)-1, respectively. 3. During the intense exercise, muscle [ATP] decreased by 30% and [CP] by 60% from resting concentrations of 6.2 and 22.4 mmol (kg wet wt)-1, respectively, and [IMP] increased to 1.1 mmol (kg wet wt)-1. Muscle [lactate] increased from 2 to 28.1 mmol (kg wet wt)-1, and the concomitant net lactate release was 14.8 mmol (kg wet wt)-1 or about 1/3 of the total net lactate production. During recovery 70% of the accumulated lactate was released to the blood, and the nucleotides and CP returned to about 40 and 85% of pre-exercise values at 3 and 10 min of recovery, respectively. 4. Total reduction in ATP and CP (and elevation of IMP) during the intense exercise amounted to 16.4 mmol ATP (kg wet wt)-1, which together with the lactate production accounted for 83.1 mmol ATP (kg wet wt)-1. In addition 6-8 mmol ATP (kg wet wt)-1 are made available related to accumulation of glycolytic intermediates including pyruvate (and alanine). Estimated leg oxygen deficit corresponded to an ATP production of 94.7 mmol ATP kg-1; this value included 3.1 mmol kg-1 related to unloading of HbO2 and MbO2.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino acid-mediated EPSPs at primary afferent synapses with substantia gelatinosa neurones in the rat spinal cord.

Abstract: 1. Fast excitatory postsynaptic potentials (EPSPs) evoked by stimulation of A delta and C fibres were examined by intracellular recording from substantia gelatinosa (SG) neurones in a transverse slice preparation of adult rat spinal cord. 2. Single low-intensity stimuli applied to the dorsal root activated A delta fibres and evoked monosynaptic EPSPs in 70% of SG neurones. In 5% of SG neurones, increasing the intensity and duration of stimulation evoked solely C fibre-mediated EPSPs. About 20% of neurones received both A delta and C fibre input from primary afferents. 3. Low concentrations of tetrodotoxin (TTX, approximately 50 nM) blocked EPSPs evoked by stimulation of A delta fibres without affecting those evoked by C fibre stimulation. Higher concentrations of TTX (500 nM) also blocked C fibre-evoked responses. 4. EPSPs evoked by A delta and C fibre stimulation reversed in polarity at membrane potentials near 0 mV, similar to the reversal potential of spontaneous EPSPs and of the potential change evoked by exogenous glutamate. 5. A delta and C fibre-evoked EPSPs were depressed by kynurenate and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX); C fibre-evoked EPSPs appeared to be less sensitive. 6. In the presence of TTX, only 50% of SG neurones were depolarized by L-glutamate. However, neurones which exhibited no direct response to L-glutamate received afferent-evoked EPSPs which were sensitive to CNQX. In sensitive neurones, the depolarization evoked by L-glutamate was depressed by only approximately 15% in the presence of CNQX, whereas afferent-evoked EPSPs recorded from the same neurone were almost completely suppressed. Combined application of DL-2-amino-5-phosphonovaleric acid (APV) and CNQX depressed the response to L-glutamate by only approximately 25%. 7. These findings suggest that A delta and C fibres use L-glutamate or a related amino acid as a transmitter at synapses with substantia gelatinosa neurones. The postsynaptic actions of this transmitter are mediated predominantly by non N-methyl-D-aspartic acid (NMDA) receptors. The failure of CNQX and APV to completely block the L-glutamate-evoked depolarization of substantia gelatinosa neurones raises the possibility that exogenously applied L-glutamate activates a non-NMDA receptor distinct from that which mediates the actions of the synaptically released afferent transmitter.

Allosteric potentiation of quisqualate receptors by a nootropic drug aniracetam.

Abstract: 1. Allosteric potentiation of the ionotropic quisqualate (iQA) receptor by a nootropic drug aniracetam (1-p-anisoyl-2-pyrrolidinone) was investigated using Xenopus oocytes injected with rat brain mRNA and rat hippocampal slices. 2. Aniracetam potentiates the iQA responses induced in Xenopus oocytes by rat brain mRNA in a reversible manner. This effect was observed above the concentrations of 0.1 mM. Kainate. N-methyl-D-aspartate and gamma-aminobutyric acid responses induced in the same oocytes were not affected. 3. The specific potentiation of iQA responses was accompanied by an increase in the conductance change of iQA and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) responses, but the affinity of receptors for agonist and the ion-selectivity of the channels (reversal potentials) were not changed. 4. Aniracetam reversibly potentiated the iQA responses recorded intracellularly from the pyramidal cells in the CA1 region of rat hippocampal slices. The excitatory postsynaptic potentials (EPSPs) in Schaffer collateral-commissural-CA1 synapses were also potentiated by aniracetam. 5. Population EPSPs recorded in the mossy fibre-CA3 synapses as well as Schaffer-commissural synapses were also potentiated by aniracetam. The amplitudes of the potentiation were not changed by the formation of long-term potentiation.

Perceptual responses to microstimulation of single afferents innervating joints, muscles and skin of the human hand.

Abstract: 1. Microneurographic techniques were used to isolate single afferent axons within cutaneous and motor fascicles of the median and ulnar nerves at the wrist in thirteen subjects. Of the sixty-five identified afferents, eleven innervated the interphalangeal and metacarpophalangeal joints, sixteen innervated muscle spindles, three innervated Golgi tendon organs and thirty-five supplied the glabrous skin of the hand. 2. Intrafascicular stimulation through the recording microelectrode, using trains of constant-voltage positive pulses (0.3-0.8 V, 0.1-0.2 ms, 1-100 Hz) or constant-current biphasic pulses (0.4-13.0 microA, 0.2 ms, 1-100 Hz), evoked specific sensations from sites associated with some afferent species but not others. 3. Microstimulation of eight of the eleven joint afferent sites (73%) evoked specific sensations. With four, subjects reported innocuous deep sensations referred to the relevant joint. With the other four, the subjects reported a sensation of joint displacement that partially reflected the responsiveness of the afferents to joint rotation. 4. Microstimulation of fourteen of the sixteen muscle spindle afferent sites (88%) generated no perceptions when the stimuli did not produce overt movement. However, subjects could correctly detect the slight movements generated when the stimuli excited the motor axons to the parent muscle. 5. With seven of the nine rapidly adapting (type RA or FAI) cutaneous afferents (88%) microstimulation evoked sensations of 'flutter-vibration', and with two of eight slowly adapting (type SAI) afferents (25%) it evoked sensations of 'sustained pressure'. Of the eighteen SAII afferents, which were classified as such by their responses to planar skin stretch, the majority (83%) generated no perceptions, confirming previous work, but three evoked sensations of movements or pressure. 6. The present results suggest a relatively secure transmission of joint afferent traffic to perceptual levels, and it is concluded that the human brain may be able to synthesize meaningful information on joint displacement on the basis of impulses in a single joint afferent. This could partly compensate for the low responsiveness of individual joint afferents within the physiological range of joint displacements. Although single muscle spindle afferents can adequately encode joint position and movement, the results suggest that the brain needs the information from more than one muscle spindle afferent to perceive changes in joint angle.

GABAA receptor function is regulated by phosphorylation in acutely dissociated guinea-pig hippocampal neurones.

Abstract: 1. Current mediated by GABAA receptors was examined in pyramidal cells acutely dissociated from the hippocampus of mature guinea-pigs. Current responses were measured using whole-cell voltage-clamp recordings. An internal perfusion technique was used to change the intracellular contents during recording. 2. Application of GABA (100-300 microM) by short duration pressure pulses produced outward current responses at a holding potential of -10 mV. When recordings were made with intracellular solutions which did not contain Mg-ATP, GABA responses progressively decreased to less than 10% of their initial values after 10 min. This 'run-down' of the GABA response could not be accounted for by desensitization since the rate of run-down was not dependent upon agonist application. 3. The run-down of the GABAA response was reversed when Mg2+ (4 mM) and ATP (2 mM) were introduced into the intracellular perfusate. In addition to the presence of Mg-ATP, buffering of Ca2+ in the intracellular solution to low levels (approximately 10(-8) M) was also necessary to stabilize the GABAA response. 4. The role of a phosphorylation process in regulating the GABAA receptor was tested. After the GABA response stabilized, introduction of alkaline phosphatase (100 micrograms/ml) to the intracellular perfusate caused a complete run-down of the GABA response. 5. Stable GABA responses were obtained when ATP was replaced by ATP-gamma-S (adenosine 5'-O-(thiotriphosphate), an analogue of ATP that donates a thiophosphate group resulting in a product that is more resistant to hydrolysis. Following such treatment GABA responses declined more slowly after the introduction of intracellular alkaline phosphatase. 6. Run-down of GABA responses accelerated when intracellular Ca2+ concentration ([Ca2+]i) was elevated to about 5 x 10(-4) M. The run-down caused by elevated [Ca2+]i could be stopped and reversed by reducing [Ca2+]i to about 10(-8) M. 7. The introduction of ATP-gamma-S to the intracellular medium retarded the run-down of GABA responses caused by elevation of [Ca2+]i. 8. N-(6-Aminohexyl)-5-chloro-1-naphthalenesulphonamide (W-7), a calmodulin inhibitor, reduced the rate of run-down induced by elevated [Ca2+]i. 9. These results suggest that the function of the GABAA receptor is maintained by phosphorylation of the receptor or some closely associated regulatory molecule. Elevation of [Ca2+]i destabilizes the function of the GABAA receptor, probably by activating a Ca2+/calmodulin-dependent phosphatase.

The contractile response during steady lengthening of stimulated frog muscle fibres.

Abstract: 1. Steady lengthenings at different velocities (0.025-1.2 microns/s per half-sarcomere; temperature 2-5.5 degrees C) were imposed on isolated frog muscle fibres at the isometric tetanus plateau by means of a loudspeaker motor. The lengthening at the sarcomere level was measured by means of a striation follower either in fixed-end or in length-clamp mode. The force response was measured by a capacitance gauge transducer (resonance frequency 50 kHz). Preparations showing gross non-homogeneity during lengthening were excluded. 2. A steady tension was in all cases reached after about 20 nm per half-sarcomere of lengthening. Tension during this steady phase rose with speed of elongation up to 0.25-0.4 micron/s per half-sarcomere, when tension was 1.9-2 times isometric tetanic force (T0). Further increase in speed produced only very little increase in the steady tension. 3. During the transitory phase, before steady tension was reached, the tension rose monotonically if speed of lengthening was less than 0.25-0.3 micron/s per half-sarcomere; at higher speed the tension rose above the steady level, reaching a peak when extension was 10-14 nm per half-sarcomere, and then fell to the steady level. Tension at the peak continued to rise with speed of lengthening above 0.3 micron/s per half-sarcomere. 4. During the tension rise within the transitory phase of force response the segment elongated at a speed 15-20% lower than that imposed on the whole fibre, as a consequence of tendon compliance. 5. During the steady phase, non-homogeneity of lengthening speed began above a speed of lengthening which varied from fibre to fibre. At speeds below this value, segments elongated at the same speed as that imposed on the fibre. 6. Tension responses to large step stretches (up to 12 nm per half-sarcomere), applied at the plateau of isometric tetanus, showed that the instantaneous elasticity of contractile machinery is not responsible for the limit in force attained with high-speed lengthening. 7. Instantaneous stiffness was determined during the steady state of force response by superposing small steps (less than 1.5 nm per half-sarcomere) on steady lengthening at different velocities. Stiffness was 10-20% larger during lengthening than at the plateau of isometric tetanus and remained practically constant, independent of lengthening velocity, in the range of velocities used. 8. The results indicate that steady lengthening of a tetanized fibre induces a cross-bridge cycle characterized by fast detachment of the cross-bridge extended beyond a critical level.(ABSTRACT TRUNCATED AT 400 WORDS)

Excitation of the corticospinal tract by electromagnetic and electrical stimulation of the scalp in the macaque monkey

Abstract: 1. The responses evoked by non-invasive electromagnetic and surface anodal electrical stimulation of the scalp (scalp stimulation) have been studied in the monkey. Conventional recording and stimulating electrodes, placed in the corticospinal pathway in the hand area of the left motor cortex, left medullary pyramid and the right spinal dorsolateral funiculus (DLF), allowed comparison of the actions of non-invasive stimuli and conventional electrical stimulation. 2. Responses to electromagnetic stimulation (with the coil tangential to the skull) were studied in four anaesthetized monkeys. In each case short-latency descending volleys were recorded in the contralateral DLF at threshold. In two animals later responses were also seen at higher stimulus intensities. Both early and late responses were of corticospinal origin since they could be completely collided by appropriately timed stimulation of the pyramidal tract. The latency of the early response in the DLF indicated that it resulted from direct activation of corticospinal neurones: its latency was the same as the latency of the antidromic action potentials evoked in the motor cortex from the recording site in the DLF. 3. Scalp stimulation, which was also investigated in three of the monkeys, evoked short-latency volleys at threshold and at higher stimulus intensities these were followed by later waves. The short-latency volleys could be collided from the pyramid and, at threshold, had latencies compatible with direct activation of corticospinal neurones. The longer latency volleys were also identified as corticospinal in origin. 4. The latency of the early volley evoked by electromagnetic stimulation remained constant with increasing stimulus intensities. In contrast, with scalp stimulation above threshold the latency of the early volleys decreased considerably, indicating remote activation of the corticospinal pathway below the level of the motor cortex. In two monkeys both collision and latency data suggest activation of the corticospinal pathway as far caudal as the medulla. 5. The majority of fast corticospinal fibres could be excited by scalp stimulation with intensities of 20% of maximum stimulator output. Electromagnetic stimulation at maximum stimulator output elicited a volley of between 70 and 90% of the size of the maximal volley evoked from the pyramidal electrodes. 6. Electromagnetic stimulation was also investigated in one awake monkey during the performance of a precision grip task. Short-latency EMG responses were evoked in hand and forearm muscles. The onsets of these responses were approximately 0.8 ms longer than the responses evoked by electrical stimulation of the pyramid.(ABSTRACT TRUNCATED AT 400 WORDS)

Plasma potassium changes with high intensity exercise.

Abstract: 1. Exercise seems to change the extracellular potassium concentration far beyond the narrow limits seen in resting subjects. To examine alterations in plasma potassium concentration during exercise, twenty healthy, well-trained men ran on the treadmill at 6 deg inclination with catheters inserted in the femoral vein and artery. 2. During 1 min exhausting exercise plasma potassium concentration rose in parallel in the vein and artery, reaching peak post-exercise values of 8.34 +/- 0.23 mmol l-1 and 8.17 +/- 0.29 mmol l-1. After 3 min recovery the potassium concentration was 0.50 +/- 0.05 mmol l-1 below pre-exercise values. Both the rise of plasma potassium concentration during exercise and the decline during recovery followed exponential time courses with a half-time of 25 s. 3. Exercise at reduced intensity showed that the peak post-exercise potassium concentration was linearly related to the exercise intensity. Individual resting, peak and nadir values were proportionally related. 4. The increased potassium concentration during exercise can be explained in full by the electrical activity in the exercising muscles. Repeated 1 min exhausting exercise bouts revealed no relationship between potassium concentration and plasma pH nor glycogen break-down. 5. All of the observations fit a simple model of potassium efflux from active muscle and elimination from blood with the following characteristics: the efflux increases (decreases) stepwise at the onset (end) of exercise, and the efflux rate during exercise increases with exercise intensity. Potassium is eliminated from blood by a proportional regulator which may be the Na(+)-K+ pump of the exercising muscle. Extracellular potassium is indirectly linked to the pump stimulus, and the rate of reuptake is proportional to the extracellular accumulation. Thus no limited maximal power for potassium uptake was found. The post-exercise undershoot of 0.5 mmol l-1 can be explained by a higher gain of the pump after exercise. 6. The large, rapid changes in the plasma potassium concentration during and after exercise is due to the first order kinetics of the reuptake mechanism rather than to a limited power to take up potassium.

Mechanisms of caffeine activation of single calcium-release channels of sheep cardiac sarcoplasmic reticulum.

Abstract: 1. Calcium-release channels of sheep cardiac junctional sarcoplasmic reticulum were incorporated into planar phospholipid bilayers. Single-channel current fluctuations were recorded under voltage clamp conditions. 2. Channels incorporate into the bilayer with a fixed orientation and channel open probability is regulated by the calcium concentration at the cytosolic face of the membrane. 3. Addition of caffeine (0.5-2.0 mM) to the cytosolic side of the membrane increased the open probability of the calcium-activated calcium-release channel by increasing the frequency of opening without significant alteration to the durations of open events. This effect was observed at both 0.1 and 10 microM-activating cytosolic calcium. 4. Caffeine (0.5-2.0 mM) did not activate the channel at a subactivating cytosolic calcium concentration (80 pM). 5. At subactivating calcium concentrations, channels could be activated by higher concentrations of caffeine (greater than 5.0 mM) revealing a second, calcium-independent, mechanism for channel activation. Channel openings induced by these high concentrations of caffeine at subactivating calcium concentrations displayed different kinetics from those observed with calcium as the sole activating ligand or with combinations of calcium and low concentrations of caffeine. 6. Activation of channel opening by caffeine in the presence of calcium did not affect single-channel conductance. Channel openings produced by caffeine at subactivating cytosolic calcium concentrations had identical conductance and relative permeability to those seen on calcium activation. 7. Channels activated by caffeine at both activating and subactivating calcium concentrations were characteristically modified by ryanodine, Ruthenium Red, ATP and magnesium, implying that the same channel is involved under both conditions.

Synaptic excitation of inhibitory cells by single CA3 hippocampal pyramidal cells of the guinea-pig in vitro.

Abstract: 1. In simultaneous recordings from pairs of neurones in hippocampal slices from guinea-pigs, single action potentials fired by CA3 pyramidal cells could initiate inhibitory postsynaptic potentials (IPSPs) in nearby pyramidal cells. 2. The latencies of these IPSPs could be as short as 3 ms. However, they were mediated disynaptically via chemical, excitatory synapses, since inhibitory coupling was suppressed by an excitatory amino acid antagonist. 3. The properties of excitatory synapses made onto inhibitory cells were examined to assess the basis for this strong coupling. Inhibitory cells were identified either by showing that they inhibited another cell or by their characteristic firing pattern. 4. Excitatory postsynaptic potentials (EPSPs) elicited by single pyramidal cell action potentials had a mean amplitude of 1-4 mV and a time to peak of 1.5-4 ms. In most cases they decayed with a time constant similar to that of the inhibitory cell membrane. 5. EPSP amplitude increased with hyperpolarization of the postsynaptic membrane. Membrane polarization had little effect on EPSP shape. 6. EPSPs fluctuated in amplitude and transmission sometimes failed, suggesting transmission was quantal and that few quanta were released. 7. When presynaptic cells were made to fire bursts of action potentials, EPSPs in inhibitory cells were initially potentiated. 8. EPSPs could cause inhibitory cells to fire. The interval between pre- and postsynaptic spikes could be as short as 2.5 ms and the probability of spike transmission could be as high as 0.6. Some inhibitory cells which received feedback excitation were also excited in feedforward fashion by mossy fibre stimuli. 9. One pyramidal cell could activate several disynaptic inhibitory pathways terminating on another pyramidal cell. This suggests that excitatory synapses made by pyramidal cell axon collaterals onto inhibitory cells are divergent. 10. This strong, divergent excitation of inhibitory cells ensures recurrent inhibition is sufficiently widespread, rapid and potent to control the spread of activity by recurrent excitatory connections between CA3 pyramidal cells.

Tonic vibration reflexes elicited during fatigue from maximal voluntary contractions in man.

Abstract: 1. In the present study on human foot dorsiflexor muscles we have examined the effects of high-frequency (150 Hz) muscle vibration on weak or moderate voluntary contractions (maintained by constant effort) and on maximal voluntary contractions (MCVs) of (i) non-fatigued muscles, (ii) muscles fatigued by sustained MVCs and (iii) muscles deprived of gamma-fibre innervation by partial anaesthetic nerve block. The motor outcome of the voluntary dorsiflexion efforts was assessed by measuring the firing rates of single motor units in the anterior tibial (TA) muscle, the mean voltage EMG activity from the pretibial muscles and foot dorsiflexion force. 2. With the subject instructed to exert constant effort in maintaining a weak or moderate contraction, superimposed vibration caused an enhancement of EMG activity and contraction force. 3. Previous claims that muscle vibration has no facilitatory effect on motor output in MVCs were found to hold true for non-fatigued but not for fatigued muscles. Thus, the fatigue-induced decline in EMG activity and motor unit firing rates was counteracted by short periods (less than 10-20 s) of superimposed vibration. However, with longer vibration periods it seemed as if the initial facilitation converted into an opposite effect which accentuated the fatigue-induced decline in motor output and contraction force. 4. Like muscle fatigue, a partial anesthetic block of the deep peroneal nerve, supposedly interrupting transmission in gamma-motor fibres, caused a reduction of MVC motor unit firing rates which could be counteracted by muscle vibration. In prolonged MVCs performed during the block, motor unit firing rates did not show the normal progressive decline from an initially high level, but stayed at a relatively constant low level throughout the contraction period. 5. Even though alternative interpretations are possible, the results agree with the hypotheses (i) that in sustained MVCs, fatigue processes occur not only in extrafusal but also in intrafusal muscle fibres, (ii) that the intrafusal fatigue leads to a reduction of the voluntary drive conveyed to the alpha-motoneurones via the gamma-loop and (iii) that vibration-induced activity in group Ia afferents can act as a substitute for the diminished fusimotor drive.

Low‐threshold mechanoreceptive and nociceptive units with unmyelinated (C) fibres in the human supraorbital nerve.

Abstract: 1. In recordings from the human supraorbital nerve with tungsten microelectrodes, eleven afferent units with unmyelinated (C) axons were identified on the basis of their conduction velocities (0.6-1.4 m/s). 2. Eight units had low mechanical thresholds (less than or equal to 0.23 g) and could be activated up to their maximal firing rates of about 100 impulses/s by weak tactile stimuli, whereas three units had higher thresholds (5.5 g) and responded vigorously to noxious stimuli only. 3. During a skin indentation the low-threshold units adapted to an irregular low-frequency discharge, and release of the stimulus elicited a prominent off-response often ending with an after-discharge. Slow stroking was a particularly effective stimulus, even when done with cotton wool, whereas rapid stroking reduced the response. All types of stroking stimuli were occasionally followed by after-discharges. Repeated mechanical stimulation at short intervals resulted in a decline of the response, indicating receptor fatigue. For two units a response to skin cooling was observed. 4. The above low-threshold C units have all the main characteristics of the C mechanoreceptors known from the cat and primates but not previously proven to exist in man. The high-threshold C units are similar to the polymodal nociceptors found in other human skin areas.

Agonist-stimulated divalent cation entry into single cultured human umbilical vein endothelial cells

Abstract: 1. The free cytoplasmic Ca2+ concentration ([Ca2+]i) can be measured using Fura-2 in superfused single human umbilical vein endothelial cells. When an endothelial cell is stimulated by a maximal dose of histamine (100 microM), [Ca2+]i rises to a peak and then falls back to a maintained plateau which is due to a stimulated Ca2+ influx. 2. If extracellular Ca2+ is replaced by 50 microM-Mn2+ then 100 microM-histamine causes a rise in [Ca2+]i accompanied by a fluorescence quench that signals the stimulated entry of Mn2+ into the cytoplasm. 3. If in Ca2(+)-free solution a cell is stimulated by 100 microM-histamine for 120 s to discharge the internal Ca2+ store, and then exposed to 50 microM-Mn2+ after removal of the histamine, a similar stimulated Mn2+ entry is seen. This quench is unaffected by readdition of histamine and is not seen if the store is refilled by exposure to 1 mM-extracellular Ca2+ for 180 s before exposure to the Mn2+. 4. The refilling of the internal store by exposure to 1 mM-Ca2+ and the stimulated entry of Mn2+ are both blocked by 2 mM-Ni2+. 5. If [Ca2+]i is stimulated to produce repetitive spikes by a low dose of histamine (0.3-1 microM) in nominally Ca2(+)-free solution containing Mn2+, then the stimulated quench is uniform and is not modulated by the [Ca2+]i spiking. 6. If the internal store is discharged by exposure to histamine in Ca2(+)-free solution and then refilled for a short period then the cell is in a state where the internal store is partly full to an extent that depends on the duration of the refilling. In such an experiment, the rate of Mn2+ influx may be estimated by measuring the rate of quench during a short exposure to 50 microM-Mn2+. The rate of Mn2+ entry varies inversely with the degree of fullness of the internal Ca2+ store. 7. If a similar experiment is repeated but with the fullness of the internal store being varied by varying the period of the initial exposure to 100 microM-histamine, with no refilling, the same inverse relationship between Mn2+ influx and fullness of the internal store is obtained. 8. These experiments show that Mn2+ enters human umbilical vein endothelial cells following agonist stimulation by a pathway that is controlled by the degree of fullness of the internal store; it does not, however, enter the cytoplasm by exactly the same route as Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)

Prolonged muscle vibration reducing motor output in maximal voluntary contractions in man.

Abstract: 1. We observed in a previous study on the human foot dorsiflexor muscles that the fatigue-induced decline in motor output during sustained maximal voluntary contractions (MVCs) was temporarily counteracted during the initial phase of superimposed high-frequency (150 Hz) muscle vibration, whereas prolonged muscle vibration seemed to accentuate the fatigue-induced decline in gross EMG activity and motor unit firing rates. A more extensive investigation of this late effect of muscle vibration on MVCs was performed in the present study. 2. Prolonged periods of superimposed muscle vibration caused a reduction of EMG activity, motor unit firing rates and contraction force in both intermittent and sustained MVCs. This vibration-induced effect had the following main characteristics: (i) it developed slowly during the course of about 1 min of sustained vibration and subsided within 10-20 s after the end of vibration; (ii) it was much more pronounced in some subjects than in others (not age-dependent) and it was accentuated by preceding muscle exercise; (iii) it affected primarily the subject's ability to generate and/or maintain high firing rates in high-threshold motor units. 3. Since the effect developed while vibration at the same time exerted a tonic excitatory influence on the alpha-motoneurone pool (as evidenced by the presence of a tonic vibration reflex) it is argued that the vibration-induced suppression of motor output in MVCs probably does not depend on alpha-motoneurone inhibition, but on a reduced accessibility of these neurones to the voluntary commands. It is suggested that contributing mechanisms might be vibration-induced presynaptic inhibition and/or 'transmitter depletion' in the group Ia excitatory pathways which constitute the afferent link of the gamma-loop.

Presynaptic glutamate receptors depress excitatory monosynaptic transmission between mouse hippocampal neurones.

Abstract: 1. Whole-cell patch-clamp techniques were used to record the excitatory postsynaptic current (EPSC) in a cultured mouse hippocampal neurone that resulted from electrical stimulation of another neurone in the cell culture. 2. L-Glutamate (less than 1 microM) reversibly depressed the EPSC amplitude in 67% of the synapses tested. The average amplitude reduction was 40%. The depression by glutamate was not blocked by extracellular magnesium (0.8 mM) or 2-amino-5-phosphonovaleric acid (AP5, 100 microM), indicating that N-methyl-D-aspartate (NMDA) receptors were not involved. 3. The phosphonic derivative of glutamate, L-2-amino-4-phosphonobutyrate (L-AP4), also depressed the EPSC amplitude. Neither glutamate nor L-AP4 induced any detectable inward current at concentrations which produced a potent depression of the EPSC. Statistical analysis of the amplitude fluctuations of evoked synaptic currents showed that the depression induced by both glutamate and L-AP4 was due to a decrease in the probability of synaptic release, confirming a presynaptic site of action. 4. Kainate and quisqualate also depressed excitatory synaptic transmission, but this action was related to the postsynaptic inward current that they induced. Statistical analysis showed that this action was consistent with a purely postsynaptic site of action. 5. Paired EPSCs separated by 20 ms showed either depression or potentiation of the second synaptic response. There was a strong correlation between those EPSCs which exhibited paired pulse depression and those depressed by glutamate application. 6. gamma-Aminobutyric acid (GABA) and baclofen also depressed excitatory synaptic transmission. This depression was not blocked by picrotoxin (100 microM). GABA (10 microM) was effective in 85% of cell pairs tested, while baclofen (5 microM) depressed every EPSC tested. A presynaptic site of action for both substances was indicated by the statistical analysis. 7. The results indicate that both glutamate and GABA suppress excitatory synaptic transmission by an action at presynaptic sites. The glutamate-induced depression may result from activation of a distinct excitatory amino acid receptor for which L-AP4 is a specific agonist.

Regulation of transmitter release at the squid giant synapse by presynaptic delayed rectifier potassium current.

Abstract: 1 The three-microelectrode voltage clamp technique and pharmacological agents were used to examine the properties and functions of potassium currents in squid giant presynaptic terminals 2 Outward currents consisted of two components: a slow component which activated over hundreds of milliseconds and was blocked by extracellular application of tetraethylammonium (TEA) ions and a more rapidly activating component which was relatively insensitive to extracellular TEA 3 The more rapid component was studied in isolation by treating presynaptic terminals with extracellular TEA, as well as tetrodotoxin (to block sodium channel currents) and manganese (to block calcium channel currents) The magnitude of this current component was 1-2 mA cm-2 at 0 mV Rates of activation and deactivation were voltage dependent and little evidence of inactivation was seen for depolarizations less than several seconds in duration 4 The reversal potential of the current was -70 to -80 mV in normal saline and became more positive with elevated extracellular potassium concentrations, suggesting that potassium is the primary permeant ion Accumulation of extracellular potassium appeared to be marked during depolarizations that produced significant activation of the current 5 Extracellular application of 3,4-diaminopyridine (DAP) blocked the current with an apparent dissociation constant of 7 microM at 0 mV Intracellular applications of DAP and TEA also were effective in reducing this current These treatments, but not extracellular TEA application, broadened presynaptic action potentials and increased the magnitude and time-to-peak of postsynaptic currents elicited by the broadened presynaptic action potentials Postsynaptic currents were a sensitive and linear function of action potential duration; a 30% increase in action potential duration increased postsynaptic current amplitude by 190% 6 Estimation of the magnitude and time course of the presynaptic calcium current, based on previous measurements of calcium channel gating, indicated that action potential broadening produces a large increase in calcium current magnitude These calculations predict that a 30% increase in presynaptic action potential duration will increase the peak amplitude of the calcium current by approximately 170% and the total amount of calcium entry by approximately 230% This implies a linear relationship between transmitter release and calcium entry during an action potential and can be explained by assuming that calcium co-operatively triggers release within intracellular domains that do not overlap(ABSTRACT TRUNCATED AT 400 WORDS)

Reflex inhibition of human soleus muscle during fatigue

Abstract: 1. Human soleus muscles were fatigued under ischaemic conditions by intermittent stimulation at 15 Hz. When maximal voluntary plantarflexion was then attempted, the loss of torque was found to be associated with a reduction in voluntary EMG activity. 2. The decrease in EMG activity could not have been due to 'exhaustion' of descending motor drive in the central nervous system since fatigue had been induced by electrical stimulation of peripheral nerve fibres. Similarly, the decrease could not be explained by changes at the neuromuscular junction or muscle fibre membrane, since changes in the M wave (evoked muscle compound action potential) were relatively modest. 3. When the excitability of the soleus motoneurones was tested during fatigue, using the H (Hoffmann) reflex, it was found to be significantly reduced. Control experiments with ischaemia or electrical stimulation, but without fatigue, failed to demonstrate any significant effects on reflex excitability. 4. The findings in this study favour the concept of reflex inhibition of alpha-motoneurones during fatigue.

Opioid actions on single nucleus raphe magnus neurons from rat and guinea‐pig in vitro.

Abstract: 1. Intracellular recordings were made from neurons of the nucleus raphe magnus (NRM) from rat (n = 128) and guinea-pig (n = 115). Two types of cells were found in each, primary (103 in rat, 27 in guinea-pig) and secondary cells (25 in rat, 88 in guinea-pig). 2. Primary cells had input resistances of 186 +/- 9 M omega (n = 9) in rat and 255 +/- 50 M omega (n = 11) in guinea-pig. The action potential in each was about 1.5 ms in duration. Synaptic potentials were evoked by focal electrical stimulation and consisted of both gamma-aminobutyric acid (GABA) and excitatory amino acid components. 3. Morphine, [Met5]enkephalin (ME) and [D-Ala2,N-Me-Phe4, Gly5-ol]enkephalin (DAMGO) depressed the amplitude of the GABA-mediated synaptic potential by a maximum of 50-65% and had little effect on the excitatory amino acid-mediated synaptic potential. There was no effect of these opioids on the resting membrane potential or input resistance of primary cells in rat or guinea-pig. 4. Secondary cells had short duration action potentials (less than 1 ms) and an input resistance of 354 +/- 47 M omega in rat (n = 6) and 290 +/- 40 M omega in guinea-pig (n = 15). The synaptic potential observed in the cells of this group was mediated by activation of only excitatory amino acid receptors. 5. ME hyperpolarized and/or abolished the spontaneous firing in sixteen out of twenty-four neurons in the secondary group from rat and eight out of eighty-four neurons from guinea-pig. ME induced an outward current at -60 mV that reversed polarity at potentials more negative than -92 +/- 3 mV in rat (n = 6) and -98 +/- 2 mV in guinea-pig (n = 18). The reversal potential of the opioid current was shifted to less negative potentials when the external potassium concentration was increased, as predicted by the Nernst equation. 6. The morphology of the two types of cells were distinguishable in that primary cells were oval (29 x 18 microns in rat; 36 x 19 microns in guinea-pig) with two to four thick tapering dendrites that branched within 50 microns of the cell body. Secondary cells were generally round or oval (about 24 x 13 microns in rat; 27 x 17 microns in guinea-pig) with two to five thin non-tapering dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)