Variance analysis of excitatory postsynaptic potentials in cat spinal motoneurons during posttetanic potentiation

1. Fluctuations in the peak amplitudes of composite excitatory postsynaptic potentials (EPSPs) in cat spinal motoneurons were analyzed during posttetanic potentiation (PTP). Each of a series of identical tetanic stimulus trains delivered to a muscle nerve was followed by 45 test stimuli applied at 2-s intervals. The mean peak amplitude and mean peak variance were calculated for EPSPs evoked by all those stimuli following a tetanus with the same time interval. It was assumed that the variance arises primarily from the probabilistic all-or-none behavior of single synaptic boutons and background noise due to spontaneous synaptic activity and thermal noise in the recording system. The variance was corrected for the contribution from additive Gaussian background noise. 2. If it is assumed that individual synaptic boutons behave independently, corrected mean peak variance and mean peak amplitude are related by a parabolic function. The expected parabolic relationship was seen in 9 of 31 cases studied, and the parameters of the best parabolic fit to the data allowed estimation of some synaptic properties. From these parameters, the mean amplitude of the unit EPSP (v) was estimated to be 102.1 +/- 57.4 (SD) microV. An average of 3.7 boutons comprised each Ia-motoneuron contact system. 3. On average, only 27% of all synaptic boutons given off by the stimulated Ia fibers to one motoneuron were active and releasing transmitter during unpotentiated reflex transmission. The remaining 73% of the synapse population was intermittently silent. The population of boutons which took part in synaptic transmission could be divided into two subpopulations, one with a release probability P = 1 and a second with a mean release probability P = 0.13 +/- 0.086. 4. We conclude that synaptic boutons connecting Ia afferents to motoneurons exist in two populations, one having a high and one a low probability of transmitter release. Transmitter release is quantal, resulting in a unit EPSP of approximately 100 microV measured at the motoneuron soma.     

5-Hydroxytryptamine depresses reticulospinal excitatory postsynaptic potentials in motoneurons of the lamprey.

Application of 5-hydroxytryptamine (5-HT) to the lamprey spinal cord in vitro reversibly depressed the chemical component of excitatory post-synaptic potentials recorded intracellularly in motoneurons and evoked by stimulation of single reticulospinal Müller cells. The depression could be produced either by local application of small volumes of 10 mM 5-HT to the surface of the spinal cord or by bath-application of 1 or 10 microM 5-HT. No effect on the input resistance of the postsynaptic cells or their sensitivity to glutamate, the suspected transmitter at this synapse, could be detected, suggesting the possibility of a presynaptic action of 5-HT at this synapse in the lamprey.     

Dual effect of GABA on descending monosynaptic excitatory postsynaptic potential in frog lumbar motoneurons

Monosynaptic excitatory postsynaptic potentials (EPSPs) evoked by stimulating ipsilateral ventrolateral column (VLC) in the thoracic section were recorded in lumbar motoneurons within the isolated spinal cord of the frog Rana ridibunda. Bath application of the selective GABAB receptor agonist (-)-baclofen (0.05 mM) caused a reduction in the peak amplitude of VLC EPSP. Baclofen did not cause any consistent change in the membrane potential or in the EPSP waveform within frog motoneurones. The selective GABA(B) receptor antagonist saclofen (0.1 mM) completely blocked the effect of (-)-baclofen on VLC EPSP. A decrease in VLC EPSP peak amplitude was also observed during GABA (0.5 mM) application. Unlike (-)-baclofen, inhibition of VLC EPSP induced by GABA was accompanied by a shortening of the EPSP time course and a reduction in membrane input resistance within lumbar motoneurons. The decrease in VLC EPSP peak amplitude induced by (-)-baclofen and GABA was accompanied by an increase in the paired-pulse facilitation. These data provide evidence for a dual pre- and postsynaptic GABAergic inhibition of the VLC monosynaptic EPSP in lumbar motoneurons within the frog spinal cord.     

Excitatory postsynaptic currents in response to different synaptic inputs of frog spinal motoneurons

Excitatory postsynaptic currents (EPSCs) evoked by the primary afferents (dorsal root; DR) and the descending lateral column (LC) fibers were studied in frog spinal motoneurons under voltage clamp with two separate electrodes. The average rise time and half-width of the EPSCs were shorter for LC-EPSCs than for DR-EPSCs, though the values of the parameters for LC- and DR-EPSCs were distributed within a similar range. The relation between the amplitudes of the EPSP and EPSC was almost linear. The amount of current required to generate a 1 mV increment in the EPSP was 5.0 +/- 2.3 nA for the DR-EPSC and 3.8 +/- 1.2 nA for the LC-EPSC. The decay time was shortened by hyperpolarization and prolonged by depolarization in DR- and LC-EPSCs and spontaneous EPSCs. The reversal potential ranged from -30 to -5 mV and was almost identical for DR- and LC-EPSCs and spontaneous EPSCs in individual motoneurons. The current-voltage relation was linear from -100 to +50 mV for these EPSCs. Spontaneous EPSCs became more prominent and frequent during a large hyperpolarization or a large depolarization. These results suggest that the ionic mechanisms underlying EPSC are similar for the functionally different excitatory synapses located on motoneurons.     

Miniature excitatory postsynaptic potentials in embryonic motoneurons grown in slice cultures of spinal cord,dorsal root ganglia and skeletal muscle

Summary Miniature excitatory postsynaptic potentials (mEPSPs) were recorded in motoneurons grown in organotypic cocultures of embryonic rat spinal cord, dorsal root ganglia and muscle in the presence of TTX. The motoneurons were electrically compact with a mean electrotonic length of 0.6. Spontaneous EPSPs were found in most of these motoneurons. With TTX the large EPSPs disappeared, whereas in more than half of the experiments mEPSPs persisted with a range in size of 1 to 4 mV (mean: 2.1 mV), probabely originating from the spontaneous release of single vesicles. The net inward charge transfer at the soma ranged from 0.12 to 0.34 pC. The mEPSPs were heterogeneous in size even within pools of potentials that were homogeneous in shape. They had similar shapes and amplitudes as the smallest spontaneous unitary EPSPs mediated by presynaptic impulses, suggesting that for the smallest afferents not more than one vesicle was released per afferent impulse. Both the miniature and the TTX-sensitive EPSPs were readily blocked by the glutamate antagonist DNQX.     

Separation of temperature sensitive and temperature insensitive components of the postsynaptic potentials in the frog motoneurons

Intracellular measurements were made in the in situ spinal cord of the frog at temperatures below 5 degrees C. Responses to volleys in the sciatic nerve, in the descending fibres and in the motor axons were studied. About 30% of the motoneurons responded to sciatic volleys with 1-3 ms segmental latency, which was short enough to assume electrotonic mediation of these responses. Another group of motoneurons responded with 6-8 ms latency, i.e. with the expected delay at chemical synapses at low temperature. Latency distribution of the sciatic-evoked postsynaptic potentials was clearly bimodal in contrast with that found at higher temperatures. Postsynaptic discharges occurred with rather long latency and they were attributed to chemically-mediated excitation. Some of the postsynaptic potentials to descending volleys also occurred with quite short latency, indicating possible electrotonic transmission from supraspinal centres to motoneurons. Latency distribution of the action potentials evoked from the motor axons was bimodal, corresponding to the different, i.e. antidromic and recurrent facilitatory, mechanism of these spikes. Calculated Q10 ratios for the sciatic-evoked reflex discharges and the afferent fibre volleys were about 2.3 and 1.8, respectively. We concluded that cooling helps to separate postsynaptic potentials according to their electrotonic and chemical mediation and that electrotonic excitation does not seem to have a primary role in the generation of postsynaptic discharges initiated by dorsal root volleys in the frog.     

Summation of excitatory postsynaptic potentials in electrically-coupled neurones

Dendritic electrical coupling increases the number of effective synaptic inputs onto neurones by allowing the direct spread of synaptic potentials from one neurone to another. Here we studied the summation of excitatory postsynaptic potentials (EPSPs) produced locally and arriving from the coupled neurone (transjunctional) in pairs of electrically-coupled Retzius neurones of the leech. We combined paired recordings of EPSPs, the production of artificial excitatory postsynaptic potentials (APSPs) in neurone pairs with different coupling coefficients and simulations of EPSPs produced in the coupled dendrites. Summation of the EPSPs produced in the dendrites was always linear, suggesting that synchronous EPSPs are produced at two or more different pairs of coupled dendrites and not in both sides of any one gap junction. The different spatio-temporal relationships explored between pairs of EPSPs or APSPs produced three main effects. (1) Synchronous pairs of EPSPs or APSPs exhibited an elongation of their decay phase compared to singe EPSPs. (2) Asymmetries in the amplitudes between the pair of EPSPs added a “hump” to the smallest EPSP. (3) Modelling the inputs near the electrical synapse or anticipating the production of the transjunctional APSP increased the amplitude of the compound EPSP. The magnitude of all these changes depended on the coupling coefficient of the neurones. We also show that the hump improves the passive conduction of EPSPs by adding low frequency components. The diverse effects of summation of local and alien EPSPs shown here endow electrically-coupled neurones with a wider repertoire of adjustable integrative possibilities.     

Cortically induced postsynaptic potentials in hypoglossal motoneurons after axotomy

Cortically induced postsynaptic potentials were studied in normal and axotomized cat hypoglossal motoneurons. In normal protruder motoneurons innervating tongue protruder muscles, we have demonstrated that stimulation of the orbital gyrus, at the point optimum for inducing lapping movements of the tongue by repetitive stimuli, produced inhibitory postsynaptic potentials or excitatory postsynaptic potentials followed by predominant inhibitory postsynaptic potentials. The cortically induced excitatory postsynaptic potential in normal protruder motoneurons was composed of only the short-latency component. In protruder motoneurons 30, 40, 60 and 80 days after axotomy, we have demonstrated that the number of protruder motoneurons responding with two components of excitatory postsynaptic potentials (the short- and the long-latency component) to cortical stimulation increased in correspondence with the lapse of days after axotomy and that the amplitude of cortically induced inhibitory postsynaptic potentials in axotomized protruder motoneurons was reduced in size as compared with normal protruder motoneurons.     

Nicotinic cholinoceptor-mediated excitatory postsynaptic potentials in rat nucleus ambiguus

In rat brainstem slice preparations, intracellular recording from neurons (n = 39) in the compact formation of the nucleus ambiguus (AMB_c) revealed spontaneous and miniature excitatory postsynaptic potentials (EPSPs; n = 11) that, along with acetylcholine-induced depolarization, were enhanced by physostigmine (10 μM; n = 2) and blocked by dihydro-β-erythroidine 1–5 pmol (n = 4). Retrograde neuronal tracing combined with choline acetyltransferase immunocytochemistry demonstrated that the AMB_c receives a projection from a subpopulation of cholinergic neurons in the zona intermedialis reticularis parvicellularis. Electrical stimulation of this region in slices evoked fast EPSPs in AMB_c neurons (n = 23) that were inhibited by dihydro-β-erythroidine 2–5 pmol (n = 8), but not by methscopolamine 1 pmol (n = 2). The present findings strongly support the existence of a cholinergic nicotinic synapse mediating fast transmission in brainstem vagal motoneurons.     

Inhibitory postsynaptic potentials in lumbar motoneurons remain depolarizing after neonatal spinal cord transection in the rat

GABA and glycine are excitatory in the immature spinal cord and become inhibitory during development. The shift from depolarizing to hyperpolarizing inhibitory postsynaptic potentials (IPSPs) occurs during the perinatal period in the rat, a time window during which the projections from the brain stem reach the lumbar enlargement. In this study, we investigated the effects of suppressing influences of the brain on lumbar motoneurons during this critical period for the negative shift of the reversal potential of IPSPs (E(IPSP)). The spinal cord was transected at the thoracic level on the day of birth [postnatal day 0 (P0)]. E(IPSP), at P4-P7, was significantly more depolarized in cord-transected than in cord-intact animals (E(IPSP) above and below resting potential, respectively). E(IPSP) at P4-P7 in cord-transected animals was close to E(IPSP) at P0-P2. K-Cl cotransporter KCC2 immunohistochemistry revealed a developmental increase of staining in the area of lumbar motoneurons between P0 and P7 in cord-intact animals; this increase was not observed after spinal cord transection. The motoneurons recorded from cord-transected animals were less sensitive to the experimental manipulations aimed at testing the functionality of the KCC2 system, which is sensitive to [K(+)](o) and blocked by bumetanide. Although bumetanide significantly depolarized E(IPSP), the shift was less pronounced than in cord-intact animals. In addition, a reduction of [K(+)](o) affected E(IPSP) significantly only in cord-intact animals. Therefore influences from the brain stem may play an essential role in the maturation of inhibitory synaptic transmission, possibly by upregulating KCC2 and its functionality.     

Depression of postsynaptic potentials by high-frequency stimulation in embryonic motoneurons grown in spinal cord slice cultures.

1. In embryonic cocultures of spinal cord, dorsal root ganglia, and muscle, excitatory postsynaptic potentials (EPSPs) were recorded in motoneurons during focal electrical stimulation of the dorsal root ganglia or the spinal cord. 2. EPSPs were depressed in amplitude at high-frequency stimulation relative to a control frequency of 0.5 Hz by 47 and 75% at 5 and 10 Hz, respectively. This was true for composite EPSPs and unitary EPSPs. 3. The depression showed a wide range of variability between individual experiments. The degree of depression at 5 Hz was negatively correlated to the rate of spontaneous excitatory input the motoneurons received. There was no correlation to the soma size, the average amplitude of the EPSPs, the rheobase, or the input resistance of the motoneurons. 4. An increase in latency of EPSPs was observed concomitant with or preceding the synaptic depression in most experiments. Total transmission failures, which were absent at low-frequency stimulation, appeared during depression. 5. Large incremental steps in amplitude could be seen during depression, suggesting that several release sites were switched off and on together. 6. Decreasing the extracellular calcium concentration from 5 to 1 mM led to a decrease in the frequency sensitivity of the synaptic efficacy and to a decrease of the EPSP amplitude and latency. 7. Measurements of the antidromic conduction of action potentials evoked in the axons and recorded in the somata of dorsal root ganglion cells revealed an increase in latency and the appearance of conduction failures at stimulation frequencies of 1-10 Hz. The frequency modulation of conduction was decreased in 1 mM compared with 5 mM external calcium. 8. Together these findings suggest that conduction failures in the presynaptic axons contribute to the synaptic depression of EPSPs in embryonic motoneurons.     

Frequency-dependent amplification of stretch-evoked excitatory input in spinal motoneurons

Voltage-dependent calcium and sodium channels mediating persistent inward currents (PICs) amplify the effects of synaptic inputs on the membrane potential and firing rate of motoneurons. CaPIC channels are thought to be relatively slow, whereas the NaPIC channels have fast kinetics. These different characteristics influence how synaptic inputs with different frequency content are amplified; the slow kinetics of Ca channels suggest that they can only contribute to amplification of low frequency inputs (<5 Hz). To characterize frequency-dependent amplification of excitatory postsynaptic potentials (EPSPs), we measured the averaged stretch-evoked EPSPs in cat medial gastrocnemius motoneurons in decerebrate cats at different subthreshold levels of membrane potential. EPSPs were produced by muscle spindle afferents activated by stretching the homonymous and synergist muscles at frequencies of 5-50 Hz. We adjusted the stretch amplitudes at different frequencies to produce approximately the same peak-to-peak EPSP amplitude and quantified the amount of amplification by expressing the EPSP integral at different levels of depolarization as a percentage of that measured with the membrane hyperpolarized. Amplification was observed at all stretch frequencies but generally decreased with increasing stretch frequency. However, in many cells the amount of amplification was greater at 10 Hz than at 5 Hz. Fast amplification was generally reduced or absent when the lidocaine derivative QX-314 was included in the electrode solution, supporting a strong contribution from Na channels. These results suggest that NaPICs can combine with CaPICs to enhance motoneuron responses to modulations of synaptic drive over a physiologically significant range of frequencies.     

Quantal analysis of excitatory postsynaptic potentials induced in hippocampal neurons by activation of granule cells

Summary The values of quantal content (m) and quantal amplitude (q) of excitatory postsynaptic potentials (EPSPs) elicited in CA3 neurons by activation of granule cells were estimated in thin hippocampal sections maintained in vitro. For this purpose, DL-homocysteate was administered to granule cells, and trains of EPSPs that were typical for single granule cell activation were recorded from individual CA3 neurons. The amplitudes of the first and second EPSPs in each train were measured. From the mean and variance of the amplitudes of the EPSPs, the values of q and m were calculated. The values of m and q for the first EPSPs were estimated at 8.3 and 0.28 mV, respectively, on the average. Potentiation of the second EPSPs was accompanied by a two-fold increase in the values of m without changes in the values of q. Therefore, frequency potentiation in synapses between mossy fibers and CA3 neurons may be explained by an increase in number of released quanta. Amplitudes of EPSPs were found to fluctuate in a manner described by Poisson's law.Supported by a grant from the Ministry of Education of Japan     

Voltage dependence of excitatory postsynaptic potentials of rat neocortical neurons

1. The properties of excitatory postsynaptic potentials (EPSPs) of rat neocortical neurons were investigated with a fast single-electrode current-voltage clamp in vitro. Typically, apparently pure EPSPs were obtained by selection of electric stimuli of low intensity. 2. The amplitude and time integral of the EPSP increased when the neuron was depolarized. At threshold for generation of action potentials, the amplitude of EPSPs was increased by approximately 30% [from 5.0 +/- 2.1 to 6.3 +/- 1.0 (SD) mV, n = 12]. The integral of EPSPs was maximally about fourfold (3.7 +/- 1.5, n = 16) larger than at resting membrane potential (Em). The mechanisms involved in this augmentation of EPSPs were further investigated. 3. The amplitude and the time integral of excitatory postsynaptic currents (EPSCs) decreased linearly with shifts in command potential from -100 to -60 mV. The decrease of the EPSC integral with depolarization indicates that the enhancement of the EPSP may be brought about by recruitment of a voltage-dependent inward current. 4. Evoking EPSPs at various delays after the onset of small depolarizing current pulses (0.3-0.6 nA, 600 ms) revealed that augmentation decays with time. The integral of EPSPs evoked approximately 80 ms after the onset of the current pulse was 3.7 (+/- 1.5, n = 16) times larger than at Em. The integral of EPSPs evoked at 480 ms. however, were only twofold (+/- 0.7, n = 16) larger. Hence EPSPs evoked after a delay of 80 ms were 1.7-fold (+/- 0.4, n = 24) larger than EPSPs evoked after 480 ms. EPSCs were independent of the delay of stimulation at all potentials. 5. Intracellular application of the lidocaine derivative N-(2,6-dimethyl-phenylcarbamoylmethyl) triethylammonium bromide (QX 314) at 100 mM from pipettes rapidly abolished fast action potentials and inward rectification. During comparable depolarizations the increase in EPSP integrals was much smaller in QX 314-treated neurons than in controls. On average, the integral of EPSPs evoked at 70-90 ms was 1.7 times (+/- 1.0) larger than at Em, and the integral of EPSPs evoked with larger delays was close to the value obtained at resting Em (0.9 +/- 0.3, n = 8). The ratio of EPSP integrals early versus late (1.8 +/- 0.5) is comparable to controls, suggesting that QX 314-sensitive currents are unlikely to be involved in the time-dependent enhancement. 6. Mimicking EPSPs by brief depolarizations atop long depolarizations revealed a time- and voltage-dependent enhancement comparable to that of EPSPs.(ABSTRACT TRUNCATED AT 400 WORDS)     

Influences of morphology and topography of motoneurons and muscle spindle afferents on amplitude of single fiber excitatory postsynaptic potentials in cat

Summary Excitatory postsynaptic potentials (e.p.s. p.s) elicited by impulses in single muscle spindle afferent fibers from the medial gastrocnemius (m.g.) muscle were recorded intracellularly from homonymous and heteronymous motoneurons in order to study factors that influence the amplitudes of such responses. Impulses in large afferent fibers elicited larger single-fiber e.p.s.p.s than those in smaller afferents. Mean e.p.s.p. amplitudes were related exponentially to afferent conduction velocities of both Ia and spindle group II fibers. The closer a motoneuron was to the spinal entry point of an afferent fiber, the larger was the mean e.p.s.p. amplitude evoked in it. Impulses in the same afferent fiber elicited larger e.p.s.p.s in small than large motoneurons when the two cells were located at the same craniocaudal levels. Other factors being equal, the single-fiber e.p.s.p.s evoked in homonymous and heteronymous motoneurons were approximately equal in amplitude. Relatively simple morphological and topographical explanations for the findings were advanced and their functional significance for orderly recruitment in partitioned and unpartitioned reflexes was described.