Mechanisms of intracortical I-wave facilitation elicited with paired-pulse magnetic stimulation in humans

In order to elucidate the mechanisms underlying intracortical I-wave facilitation elicited by paired-pulse magnetic stimulation, we compared intracortical facilitation of I1-waves with that of I3-waves using single motor unit and surface electromyographic (EMG) recordings from the first dorsal interosseous muscle (FDI). We used a suprathreshold first stimulus (S1) and a subthreshold second stimulus (S2). In most experiments, both stimuli induced currents in the same direction. In others, S1 induced posteriorly directed currents and S2 induced anteriorly directed currents. When both stimuli induced anteriorly directed currents (I1-wave effects), an interstimulus interval (ISI) of 1.5 ms resulted in extra facilitation of the responses to S1 alone. The latency of this effect was equivalent to that of the I2-wave from S1. When S1 evoked posteriorly directed currents (I3-wave recruitment), facilitation occurred at a latency corresponding to the I3-wave from S1. This facilitation occurred at an ISI of 1.5 ms when both S1 and S2 flowed posteriorly, and at an ISI of approximately 3.5 ms when S1 was posteriorly and S2 was anteriorly directed. Based on these findings, we propose the following mechanisms for intracortical I-wave facilitation. When S1 and S2 induce currents in the same direction, facilitation is produced by summation between excitatory postsynaptic potentials (EPSPs) elicited by S1 and subliminal depolarization of interneurones elicited by S2 directly. When S1 and S2 induce currents in the opposite direction, facilitation is produced by the same mechanism as above or by temporal and spatial summation of EPSPs elicited by two successive stimuli at interneurones or corticospinal neurones of the motor cortex.     

Characterisation of paired-pulse transcranial magnetic stimulation conditions yielding intracortical inhibition or I-wave facilitation using a threshold-hunting paradigm

Short-interval, paired-pulse transcranial magnetic stimulation (TMS) is usually used to demonstrate intracortical inhibition. It was shown recently that with short-interval, paired-pulse TMS a facilitation – called intracortical I-wave facilitation – can also be demonstrated. It was the aim of this study to investigate which stimulus conditions lead to intracortical inhibition and what conditions yield an intracortical I-wave facilitation in a hand muscle of normal subjects. Paired-pulse TMS responses with an interstimulus interval of 1.2 ms were obtained from the abductor digiti minimi muscle of four normal subjects. A threshold-hunting paradigm with hunting through first or second stimulus variation was used to obtain a curve of threshold-pair strengths. All subjects showed two branches of stimulus interaction on this diagram. If the first stimulus of a threshold pair was below approximately 65% of resting motor threshold it modified the response primarily due to the second stimulus through intracortical inhibition. However, if the first stimulus of a threshold pair exceeded approximately 65% of resting motor threshold it became responsible for the spinal action-potential initiation. The subsequent second stimulus served as a ”booster” for the ongoing intracortical I-wave activity, making it impossible to observe the intracortical inhibition evoked by the first stimulus. Received: 25 March 1999 / Accepted: 8 June 1999     

Short-interval paired-pulse inhibition and facilitation of human motor cortex: the dimension of stimulus intensity

Paired transcranial magnetic stimulation has greatly advanced our understanding of the mechanisms which control excitability in human motor cortex. While it is clear that paired-pulse excitability depends on the exact interstimulus interval (ISI) between the first (S1) and second stimulus (S2), relatively little is known about the effects of the intensities of S1 and S2, and the effects of manipulating neurotransmission through the GABA_A receptor. When recording the motor evoked potential (MEP) from the resting abductor digiti minimi (ADM) muscle, using a fixed ISI of 1.5 ms, and expressing the interaction between S1 and S2 as MEP_S1+S2/(MEP_S1+ MEP_S2), then a systematic variation of the intensities of S1 and S2 revealed short-interval intracortical facilitation (SICF) if S1 and S2 were approximately equal to MEP threshold (RMT), or if S1 > RMT and S2 < RMT. In contrast, short-interval intracortical inhibition (SICI) occurred if S1 < RMT and S2 > RMT. Contraction of the ADM left SICI unchanged but reduced SICF. The GABA_A receptor agonist diazepam increased SICI and reduced SICF in the resting ADM while diazepam had no effect during ADM contraction. Surface EMG and single motor unit recordings revealed that during ADM contraction SICI onset was at the I3-wave latency of S2, whereas SICF typically 'jumped up' by one I-wave and started with the I2-wave latency of S2. Findings suggest that SICI is mediated through a low-threshold GABA_A receptor-dependent inhibitory pathway and summation of IPSP from S1 and EPSP from S2 at the corticospinal neurone. In contrast, SICF originates through non-synaptic facilitation at the initial axon segment of interneurones along a high-threshold excitatory pathway.     

Long-term potentiation in the hippocampus in conditions of inhibition of caspase-3: Analysis of facilitation in paired-pulse stimulation

Treatment of hippocampal slices with the caspase-3 inhibitor Z-DEVD-FMK led to a decrease in the magnitude of long-term potentiation (LTP), which developed over time. Testing with paired stimuli separated by an interval of 70 msec showed that after caspase-3 inhibition, as compared with control slices, the second response in the pair showed no increase in amplitude in conditions of LTP. In these conditions, the magnitude of LTP depended on differences in the amplitudes of the first and second responses before induction of LTP. LTP was absent in slices with initially highly efficient afferent stimulation and correspondingly low levels of facilitation in paired-pulse stimulation. It is suggested that inhibition of caspase-3 prevents the structural rearrangements in LTP associated with the involvement of new synapses and neurons in the response. __________ Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 91, No. 8, pp. 915–926 August, 2005.     

Frequency-dependent shift from paired-pulse facilitation to paired-pulse depression at unitary CA3-CA3 synapses in the rat hippocampus

Paired recordings between CA3 interconnected pyramidal neurons were used to study the properties of short-term depression occurring in these synapses under different frequencies of presynaptic firing (   n = 22  ). In stationary conditions (0.05-0.067 Hz) pairs of presynaptic action potentials (50 ms apart) evoked EPSCs whose amplitude fluctuated from trial to trial with occasional response failures. In 15/20 cells, paired-pulse ratio (PPR) was characterized by facilitation (PPF) while in the remaining five by depression (PPD). Increasing stimulation frequency from 0.05-0.067 Hz to 0.1-1 Hz induced low frequency depression (LFD) of EPSC amplitude with a gradual increase in the failure rate. Overall, 9/12 cells at 1 Hz became almost 'silent'. In six cells in which the firing rate was sequentially shifted from 0.05 to 0.1 and 1 Hz, changes in synaptic efficacy were so strong that PPR shifted from PPF to PPD. The time course of depression of EPSC1 could be fitted with single exponentials with time constants of 98 and 36 s at 0.1 and 1 Hz, respectively. In line with the inversion of PPR at 1 Hz, the time course of depression of EPSC2 was faster than EPSC1 (7 s). Recovery from depression could be obtained by lowering the frequency of stimulation to 0.025 Hz. These results could be explained by a model that takes into account two distinct release processes, one dependent on the residual calcium and the other on the size of the readily releasable pool of vesicles.     

Lithium enhances neuronal muscarinic excitation by presynaptic facilitation

The mechanisms underlying the psychotropic actions of lithium are not established, but modulation of endogenous brain neurotransmitter systems is likely to be important. Several interactions of lithium with muscarinic responses have been reported, including a marked potentiation of seizures produced by muscarinic agonists. Because the mechanism by which lithium augments muscarinic seizures may be related to the mechanism by which it produces its psychotropic effects, we have studied the interaction of lithium and muscarinic agonists in vitro. Using rat hippocampal slices, we found that a muscarinic agonist, pilocarpine, increased postsynaptic neuronal excitability, but simultaneously decreased synaptic transmission because of presynaptic inhibition. Lithium did not alter pilocarpine's postsynaptic excitatory actions, but reversed its presynaptic inhibitory action, leading to markedly increased action potential firing. These presynaptic effects are not caused by alterations in presynaptic action potential shape or reliability of conduction, and do not involve pertussis toxin-sensitive G proteins. Activation of protein kinase C with phorbol-12,13-dibutyrate, or inhibition with H-7 and sphingosine, did not affect muscarinic presynaptic inhibition, but abolished lithium's ability to enhance synaptic transmission, suggesting that this effect of lithium involves protein kinase C. We propose that presynaptic facilitation accounts for lithium's potentiation of muscarinic seizures. Since these effects occur with concentrations of lithium used clinically, similar presynaptic effects in endogenous brain neurotransmitter systems may be important for lithium's psychotropic actions.     

The mechanisms of multi-component paired-pulse facilitation of neurotransmitter release at the frog neuromuscular junction

We have studied the mechanisms of paired-pulse facilitation (PPF) of neurotransmitter release in isolated nerve-muscle preparations of the frog cutaneous pectoris muscle. In normal extracellular Ca²⁺ concentration ([Ca²⁺]_o, 1.8 mM), as the interpulse interval was increased from 5 to 500 ms, PPF decayed as a sum of two exponential components: a larger but shorter first component (F1) and a smaller but more prolonged second component (F2). In low [Ca²⁺]_o (0.5 mM), both F1 and F2 increased, and a third “early” component (Fe) appeared whose amplitude was larger and whose duration was shorter than F1 or F2. In the presence of the “fast” Ca²⁺ buffer BAPTA-AM, Fe disappeared, whereas F1 and F2 decreased in amplitude and duration. In contrast, the “slow” Ca²⁺ buffer EGTA-AM caused a decrease of Fe and reduction or complete blockade of F2, without any changes of F1. In solutions containing Sr²⁺ (1 mM), the magnitude of Fe was decreased, F1 was significantly reduced and shortened, but F2 was unaffected. Application of the calmodulin inhibitor W-7 (10 μM) at normal [Ca²⁺]_o produced a marked decrease of F2, and at low [Ca²⁺]_o, a complete blockade of Fe. These results suggest that PPF at frog motor nerve terminals is mediated by several specific for different PPF components intraterminal Ca²⁺ binding sites, which trigger neurotransmitter release. These sites have a higher affinity for Ca²⁺ ions and are located farther from the release-controlling Ca²⁺ channels than the Ca ²⁺ sensor that mediates phasic release.     

Opioid receptors mediate a postsynaptic facilitation and a presynaptic inhibition at the afferent synapse of axolotl vestibular hair cells

This study was designed to determine the effects of opiate drugs on the electrical activity of afferent neurons and on the ionic currents of hair cells from semicircular canals. Experiments were done on larval axolotls (Ambystoma tigrinum). The multiunit spike activity of afferent neurons was recorded in the isolated inner ear under both resting conditions and mechanical stimulation. Ionic currents were recorded using voltage clamp of hair cells isolated from the semicircular canal. In the isolated inner-ear preparation, microperfusion of either non-specific opioid receptor antagonist naloxone (10 nM to 1 mM), mu receptor agonist [D-Ala(2), N-Me-Phe(4),Gly(5)-ol]-enkephalin (1 pM to 10 microM), or kappa receptor antagonist nor-binaltorphimine (10 nM to 100 microM) elicited a dose-dependent long-lasting (>5 min) increase of the electrical discharge of afferent neurons. The mu receptor agonist funaltrexamine (1 nM to 100 microM) and the kappa receptor agonist U-50488 (1 nM to 10 microM) diminished the basal spike discharge of vestibular afferents. The delta receptor agonist D-Pen(2)-D-Pen(5)-enkephalin (1 nM to 10 mM) and the antagonist naltrindole (1 nM to 10 mM) were without a significant effect. The only drug that displayed a significant action on hair-cell ionic currents was trans-(+/-)-3,4-dichloro-N-methyl-N-(2-[1-pyrrolidinyl]-cyclohexyl) benzeneacetamide methanesulfonate (U-50488) that reduced the Ca(2+) current in a dose-dependent fashion. On its own, mu receptor agonist [D-Ala(2), N-Me-Phe(4),Gly(5)-ol]-enkephalin (0.01 and 10 microM) significantly potentiated the response of afferent neurons to the excitatory amino acid agonist (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (0.1 microM), while synaptic transmission was blocked by the use of high-Mg(2+), low-Ca(2+) solutions. Our data indicate that the activity of vestibular afferent neurons may be regulated in a complex fashion by opioid receptors: mu opioid receptors mediating an excitatory, postsynaptic modulatory input to afferent neurons, and kappa receptors mediating an inhibitory, presynaptic input to hair cells.     

Contribution of presynaptic Na(+) channel inactivation to paired-pulse synaptic depression in cultured hippocampal neurons

Paired-pulse depression (PPD) of synaptic transmission is important for neuronal information processing. Historically, depletion of the readily releasable pool of synaptic vesicles has been proposed as the major component of PPD. Recent results suggest, however, that other mechanisms may be involved in PPD, including inactivation of presynaptic voltage-dependent sodium channels (NaChs), which may influence coupling of action potentials to transmitter release. In hippocampal cultures, we have examined the potential role and relative contribution of presynaptic NaCh inactivation in excitatory postsynaptic current (EPSC) PPD. Based on current- and voltage-clamp recordings from somas, our data suggest that NaCh inactivation could potentially participate in PPD. Paired stimulation of somatic action potentials (20- to 100-ms interval) results in subtle changes in action potential shape that are mimicked by low concentrations of tetrodotoxin (TTX) and that appear to be generated by a combination of fast and slow recovery from NaCh inactivation. Dilute concentrations of TTX dramatically depress glutamate release. However, we find evidence for only minimal contribution of NaCh inactivation to EPSC PPD under basal conditions. Hyperpolarization of presynaptic elements to speed recovery from inactivation or increasing the driving force on Na(+) ions through active NaChs had minimal effects on PPD while more robustly reversing the effects of pharmacological NaCh blockade. On the other hand, slight depolarization of the presynaptic membrane potential, by elevating extracellular [K(+)](o), significantly increased PPD and frequency-dependent depression of EPSCs during short trains of action potentials. The results suggest that NaCh inactivation is poised to modulate EPSC amplitude with small tonic depolarizations that likely occur with physiological or pathophysiological activity.     

In search of lost presynaptic inhibition

This chapter presents an historical review on the development of some of the main findings on presynaptic inhibition. Particular attention is given to recent studies pertaining the differential GABAa control of the synaptic effectiveness of muscle, cutaneous and articular afferents, to some of the problems arising with the identification of the interneurons mediating the GABAergic depolarization of primary afferents (PAD) of muscle afferents, on the influence of the spontaneous activity of discrete sets of dorsal horn neurons on the pathways mediating PAD of muscle and cutaneous afferents, and to the unmasking of the cutaneous-evoked responses in the lumbosacral spinal cord and associated changes in tonic PAD that follow acute and chronic section of cutaneous nerves. The concluding remarks are addressed to several issues that need to be considered to have a better understanding of the functional role of presynaptic inhibition and PAD on motor performance and sensory processing and on their possible contribution to the shaping of a higher coherence between the cortically programmed and the executed movements.     

A comparison of paired-pulse facilitation of AMPA and NMDA receptor-mediated excitatory postsynaptic currents in the hippocampus

Paired-pulse facilitation of excitatory synaptic transmission was investigated in the CA1 region of rat hippocampal slices using whole-cell patch-clamp recording. To optimise the measurement of excitatory synaptic transmission, -amino-butyric acid (GABA)-mediated synaptic inhibition was eliminated using both GABA_A and GABA_B antagonists. Pure -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) or N-methyl-d-aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs) were then isolated pharmacologically. Paired-pulse facilitation of either AMPA or NMDA receptor-mediated EPSCs (EPSC_A and EPSC_N, respectively) was investigated using two stimuli of identical strength delivered at intervals of between 25 and 1000 ms. The paired-pulse facilitation profiles of both EPSC_A and EPSC_N were similar. Pairedpulse facilitation of EPSC_A was independent of holding potential. In contrast paired-pulse facilitation of EPSC_N was markedly voltage-dependent; maximum facilitation was recorded at hyperpolarised membrane potentials. At positive membrane potentials there was little or no paired-pulse facilitation and, in most neurones, pairedpulse depression was observed. Voltage-dependence of paired-pulse facilitation of EPSC_N was similar in the presence or nominal absence of Mg²⁺ in the bathing medium, and was unaffected by extensive dialysis of neurones with 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid (BAPTA). These data are consistent with a presynaptic locus for paired-pulse facilitation of EPSC_A. However, paired-pulse facilitation of EPSC_N involves postsynaptic factors.     

A parametric assessment of GABA antagonist effects on paired-pulse facilitation in the rat anterior cingulate cortex

Paired-pulse facilitation (PPF) is a form of short-term plasticity that can be used qualitatively to characterize the synaptic effects of neuroactive compounds. As we have shown previously, CNQX has a marked effect on PPF which can be measured quantitatively. The aim of the present study was to examine quantitatively possible differences in the effects of the post- and pre-synaptic GABA antagonists on PPF in vitro. Experiments were performed on slices taken from the coronal anterior cingulate cortex (ACC) of Sprague-Dawley rats. The stimuli consisted of a pair of biphasic pulses with an inter-pulse interval of 40ms. Evoked extracellular field potentials in layers 2/3 of the ACC were recorded. Quantitative assessment of PPF was achieved by calculating two parameters, the PPFmax (theoretical maximal PPF) and the Stmax (stimulus intensity that produces the PPFmax). Picrotoxin treatment produced increases in both the PPFmax and Stmax, by increasing the stimulus producing the half-maximal effect. In contrast, CGP-55845 treatment produced an increase in only the PPFmax, which was due to an alteration in the asymptotic values of the response amplitudes. Our findings show that the effect of different GABA receptor antagonists on short-term synaptic facilitation in the ACC may be assessed and specified quantitatively.     

Invertebrate presynaptic inhibition and motor control

Conclusion Presynaptic inhibition is a widespread mechanism among invertebrates and its study has developed quite independently from that which has occurred in vertebrates. However, it is striking that recent studies on presynaptic inhibition in sensory afferents of Arthropod have shown great similarities with presynaptic control exerted in the mammalian spinal cord primary afferents (Gossard et al. 1989; Rudomin et al. 1991): in both cases, presynaptic inhibition of sensory terminals involves inhibitory PADs associated with GABA, operating via Cl^- ion channels. Sensory modulation exerted in this way may affect the reflex gain. Paired recordings on invertebrate preparations have made it possible to explain some of these mechanisms in greater detail, as was the case in lamprey (Alford et al. 1991).Presynaptic mechanisms and particularly presynaptic inhibition have been studied mainly in neurons where the spike-initiating zone (initial segment of cell body in classic vertebrate neuron or the equivalent in the neurite of invertebrate neuron) is far from the output synapse. In such neurons spikes are actively conveyed in axons before they reach the output synapse. This is the case for primary afferents and for descending INs. In such cases presynaptic inhibition exerts a clear-cut effect onto spike-triggered synaptic transmission. However, a given axon generally divides into several branches, and therefore presynaptic inhibition activity onto some branches may not affect others. This possibility is likely to exist in local INs and is important to consider, in view of our understanding of how neuronal networks achieve their observed behavior. In the past, such networks were considered as input-output reflex chains; more recently, active properties that are essential for pattern generation (i.e., voltage-dependent and calcium-dependent conductances responsible for nonlinear properties of the membrane potential such as pacemaker potential and plateau properties), have been shown to exist in the component neurons; even more recently, connectivity as well as active properties on neurons have been demonstrated to be controlled by INs that could de novo rebuild new networks with specific functional role (Meyrand et al. 1994); the time has now come to consider that each neuron is not behaving as a single unit, but is rather made up of different compartments subjected to different local controls and local active properties (Coleman and Nusbaum 1994). This idea has already been supported by the demonstration that some neurons had several spike-initiating zones; it is reinforced by the finding that target cells are capable of presynaptically modulating their input fibers (Nusbaum et al. 1992). These findings seriously complicate the functional schema we had drawn of neuronal networks. However, this is certainly one of the most important challenges for the next decade, in view of understanding how the brain works.     

Mechanisms of serotonergic facilitation of a command neuron

The lateral giant (LG) command neuron of crayfish responds to an attack directed at the abdomen by triggering a single highly stereotyped escape tail flip. Experimentally applied serotonin (5-hydroxytrptamine, 5-HT) can increase or decrease LG's excitability, depending on the concentration, rate, and duration of 5-HT application. Here we describe three physiological mechanisms that mediate serotonergic facilitation of LG. Two processes strengthen electrical coupling between the primary mechanosensory afferent neurons and LG: first, an early increase in the conductance of electrical synapses between primary afferent neurons and LG dendrites and second, an early increase in the membrane resistance of LG dendrites. The increased coupling facilitates LG's synaptic response and it promotes recruitment of weakly excited afferent neurons to contribute to the response. Third, a delayed increase in the membrane resistance of proximal regions of LG increases the cell's input resistance near the initial segment. Together these mechanisms contribute to serotonergic facilitation of LG's response.     

Feed-forward facilitation of glutamate release by presynaptic GABA(A) receptors

Disynaptic GABAergic inputs from Schaffer collateral (SC) afferents on to the soma of glutamatergic CA1 pyramidal neurons are involved in feed-forward inhibition in the hippocampal neural circuits. Here we report the functional roles of presynaptic GABA(A) receptors on SC afferents projecting to CA1 pyramidal neurons. Muscimol (0.5 microM), a selective GABA(A) receptor agonist, increased SC-evoked EPSC amplitude and decreased paired-pulse ratio in the slice preparation, in addition, it facilitated spontaneous glutamate release on to mechanically dissociated CA1 pyramidal neurons in an external Ca2+-dependent manner. In field recordings, muscimol at low concentrations (< or = 0.5 microM) increased not only the excitability of SC afferents but glutamate release, however, it at high concentrations (> or = 1 microM) changed bidirectionally. These results suggest that the moderate activation of presynaptic GABA(A) receptors depolarizes SC afferents and enhances SC-mediated glutamatergic transmission. When endogenous GABA was disynaptically released by brief trains of stimulation of SC afferents, the axonal excitability in addition to glutamate release was increased. The effects of endogenous GABA on the excitability of SC afferents were blocked by either SR95531 or AMPA receptor blockers, which would be expected to block disynaptic feed-forward neural circuits. The present results provide a novel form of presynaptic modulation (feed-forward facilitation) of glutamatergic transmission by presynaptic GABA(A) receptors within the intrinsic hippocampal neural circuits.     

Depression of the fast IPSP underlies paired-pulse facilitation in area CA1 of the rat hippocampus.

1. Intracellular recordings from CA1 pyramidal neurons in the rat hippocampal slice have been used to study synaptic transmission after maximal orthodromic stimulation of the Schaffer collateral-commissural fibers. Paired-pulse stimulation was used to investigate how the first (conditioning) stimulation influenced the response to the second (test) stimulation. 2. When the test stimulation was delivered up to approximately 4 s after the conditioning stimulation, the late phase of the excitatory postsynaptic synaptic potential (EPSP) was increased ("late-phase facilitation") whereas the fast (f-) and the slow (s-) inhibitory postsynaptic potentials (IPSPs) were depressed. 3. In terms of appearance and time course, facilitation of the intracellularly recorded EPSP was similar to that of the extracellularly recorded field EPSP in stratum radiatum. 4. The s-IPSP is not involved in facilitation of the EPSP. To show this, we counteracted the s-IPSP either by repolarizing the membrane potential to the resting level or by intracellularly injecting the quaternary lignocaine derivative QX 314. Facilitation of the late phase of the EPSP was unaffected by either procedure. 5. The conditioned response was modified in two ways when the stimulation was delivered at the equilibrium potential for the f-IPSP (Ef-IPSP) and the s-IPSP had been blocked by intracellular injection of QX 314. The amplitude of the EPSP was increased, and the repolarizing phase was delayed with an apparent depolarizing shift of Ef-IPSP. This effect was present at pulse intervals greater than 20 ms and was maximal after 150 ms. Facilitation could be detected at interpulse intervals of up to 4 s. 6. The gamma-aminobutyric acid-B (GABAB) agonist baclofen (1 microM) reduced late-phase facilitation by preferentially increasing the unconditioned response, such that this came to resemble a conditioned response in control medium. 7. The f-IPSP was isolated pharmacologically to investigate its role in the facilitation of the EPSP. This was done by blocking the s-IPSP with QX314 and the EPSP with a mixture of the N-methyl-D-aspartate (NMDA) receptor blocker, 2-amino-5-phosphonovaleric acid (APV, 50 microM), and the non-NMDA receptor blocker 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM). An f-IPSP was then evoked by stimulating the interneurons directly. This potential could be blocked by the GABAA receptor antagonist bicuculline (20 microM), thereby confirming the successful isolation of GABAAergic transmission. 8. With paired-pulse stimulation, the amplitude of the conditioned f-IPSP was depressed.(ABSTRACT TRUNCATED AT 400 WORDS)     

Crayfish neuromuscular facilitation activated by constant presynaptic action potentials and depolarizing pulses

1. Experiments were conducted to test the hypothesis that facilitation of transmitter release in response to repetitive stimulation of the exciter motor axon to the crayfish claw opener muscle is due to an increase in the amplitude or duration of the action potential in presynaptic terminals. No consistent changes were found in the nerve terminal potential (n.t.p.) recorded extracellularly at synaptic sites on the surface of muscle fibres.2. Apparent changes in n.t.p. are attributed to three causes.(i) Some recordings are shown to be contaminated by non-specific muscle responses which grow during facilitation.(ii) Some averaged n.t.p.s exhibit opposite changes in amplitude and duration which suggest a change in the synchrony of presynaptic nerve impulses at different frequencies.(iii) Some changes in n.t.p. are blocked by gamma-methyl glutamate, an antagonist of the post-synaptic receptor, which suggests that these changes are caused by small muscle movements.3. The only change in n.t.p. believed to represent an actual change in the intracellular signal is a reduction in n.t.p. amplitude to the second of two stimuli separated by a brief interval.4. Tetra-ethyl ammonium ions increase synaptic transmission about 20% and prolong the n.t.p. about 15%. This result suggests that an increase in n.t.p. large enough to increase transmission by the several hundred per cent occurring during facilitation would be detected.5. The nerve terminals are electrically excitable, and most synaptic sites have a diphasic or triphasic n.t.p., which suggests that the motor neurone terminals are actively invaded by nerve impulses.6. When nerve impulses are blocked in tetrodotoxin, depolarization of nerve terminals increases the frequency of miniature excitatory junctional potentials (e.j.p.s), and a phasic e.j.p. can be evoked by large, brief depolarizing pulses. Responses to repetitive or paired depolarizations of constant amplitude and duration exhibit a facilitation similar to that of e.j.p.s evoked by nerve impulses.7. It is concluded that facilitation in the crayfish claw opener is not due to a change in the presynaptic action potential, but is due to some change at a later step in the depolarization-secretion process.