Inhibitory Postsynaptic Potential Reversal Research Articles

Diversity of Neuron-specific K+-Cl−Cotransporter Expression and Inhibitory Postsynaptic Potential Depression in Rat Motoneurons

Motoneurons receive a robust recurrent synaptic inhibition by gamma-aminobutyric acid and glycine, which activate Cl(-) channels. Thus, Cl(-) homeostasis determines the efficacy of synaptic inhibition in the motoneurons. In situ hybridization reveals that the neuronal K(+)-Cl(-) cotransporter isoform 2 (KCC2), a major mechanism in maintaining a low Cl(-) concentration in neurons, is abundantly expressed in the facial, hypoglossal (XII), and spinal motoneurons innervating striated muscle, whereas the dorsal vagal motoneurons (DMVs) controlling smooth muscle exhibited little expression of KCC2. This raises a general interest in the correlation between KCC2 expression and inhibitory postsynaptic potential (IPSP) performance in the native circuits. Intracellular and whole-cell patch recordings revealed that an activity-dependent depression of IPSPs and positive shift of IPSP reversal potentials were more prominent in the DMV than in the XII. Cl(-) influx through Cl(-) channels was extruded more potently in the XII than in the DMV, suggesting that differences in Cl(-) extrusion account for these dynamic differences of IPSP. Cl(-) extrusion was inhibited by either furosemide or an increase in extracellular potassium concentrations. Thus, the rigid maintenance of IPSP and rapid Cl(-) extrusion in the XII reflects an intense expression of KCC2. KCC2 expression may strongly influence the IPSP depression and functional properties of the motoneurons innervating striated muscles.

Ionic mechanism of GABAA biphasic synaptic potentials in gustatory nucleus of the solitary tract.

Gamma-aminobutyric acid (GABA) is the principal neurotransmitter of synaptic inhibition in the gustatory nucleus of the solitary tract (rNST). High-frequency activation of GABA neurons in the rNST results in biphasic inhibitory postsynaptic potentials (IPSPs) that are initially hyperpolarizing but then became depolarizing. Our results indicate that high-frequency stimulation evokes redistribution of Cl- and K+ ions that shifts IPSP reversal potential in a more positive direction, which produces a biphasic or depolarizing IPSP.

Inhibitory control of somatodendritic interactions underlying action potentials in neocortical pyramidal neurons in vivo: An intracellular and computational study

The effect of synaptic inputs on somatodendritic interactions during action potentials was investigated, in the cat, using in vivo intracellular recording and computational models of neocortical pyramidal cells. An array of 10 microelectrodes, each ending at a different cortical depth, was used to preferentially evoke synaptic inputs to different somatodendritic regions. Relative to action potentials evoked by current injection, spikes elicited by cortical microstimuli were reduced in amplitude and duration, with stimuli delivered at proximal (somatic) and distal (dendritic) levels evoking the largest and smallest decrements, respectively. When the inhibitory postsynaptic potential reversal was shifted to around −50 mV by recording with KCl pipettes, synaptically-evoked spikes were significantly less reduced than with potassium acetate or cesium acetate pipettes, suggesting that spike decrements are not only due to a shunt, but also to voltage-dependent effects. Computational models of neocortical pyramidal cells were built based on available data on the distribution of active currents and synaptic inputs in the soma and dendrites. The distribution of synapses activated by extracellular stimulation was estimated by matching the model to experimental recordings of postsynaptic potentials evoked at different depths. The model successfully reproduced the progressive spike amplitude reduction as a function of stimulation depth, as well as the effects of chloride and cesium. The model revealed that somatic spikes contain an important contribution from proximal dendritic sodium currents up to ≈100 μm and ≈300 μm from the soma under control and cesium conditions, respectively. Proximal inhibitory postsynaptic potentials can prevent this dendritic participation thus reducing the spike amplitude at the soma. The model suggests that the somatic spike amplitude and shape can be used as a “window” to infer the electrical participation of proximal dendrites. Thus, our results suggest that inhibitory postsynaptic potentials can control the participation of proximal dendrites in somatic sodium spikes.

Effects of the Neuropeptide Thyrotropin-Releasing Hormone on GABAergic Synaptic Transmission of CA1 Neurons of the Rat Hippocampal Slice During Hypoxia

Barbieri, M. and A. Nistri. Effects of the neuropeptide thyrotropin-releasing hormone on GABAergic synaptic transmission of CA1 neurons of the rat hippocampal slice during hypoxia. Peptides 18(4) 585–591, 1997—Because thyrotropin-releasing hormone (TRH) has been suggested to improve recovery of brain neurons from hypoxia, which strongly impairs GABAergic synaptic transmission, the present electrophysiological study used intracellular recording from CA1 neurons of the rat hippocampal slice to examine the cellular mechanisms underlying this phenomenon. Hypoxia induced by superfusion with a medium devoid of oxygen evoked typical membrane hyperpolarization, fall in input resistance, and strong depression of monosynaptic, GABA A receptor-mediated fast inhibitory postsynaptic potentials (IPSPs). The depression of fast IPSPs during hypoxia was found to be due to a combination of factors such as shift in the IPSP reversal potential and membrane hyperpolarization. GABA B receptor-mediated slow IPSPs were comparatively less sensitive to hypoxia. TRH (10 μM), applied 1 min prior to hypoxia, selectively accelerated recovery of membrane potential and delayed return of fast IPSPs to control amplitude without changing the mechanisms responsible for depression of GABAergic transmission. In conclusion, despite a slower recovery of IPSPs, TRH facilitated earlier return of neuronal excitability after the hypoxic period.

Block of glutamate decarboxylase decreases GABAergic inhibition at the crayfish synapses: possible role of presynaptic metabotropic mechanisms.

1. The cytosolic concentration of a neurotransmitter is believed to be an important factor determining its release. The effects of 3-mercaptopropionic acid (MP) and aminooxyacetic acid (AOAA), glutamate decarboxylase (GAD) blockers, on GABAergic postsynaptic and presynaptic inhibitory neurotransmission were examined in the crayfish (Procambarus clarkii) opener neuromuscular synapses. 2. Intracellular recordings of evoked excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs) as well as loose macropatch clamp measurements of excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic currents (IPSCs) were used to evaluate the effects of the drugs, which were applied exclusively to the nerve bundle. 3. Under normal conditions, a stimulus train to the inhibitor preceding the excitor stimulation elicited a large reduction in EPSP amplitude in a time interval-dependent manner. This inhibition is effected by postsynaptic as well as presynaptic processes. 4. Treatment with MP or AOAA decreased the IPSP amplitude and its altered conductance but had no effect on the IPSP reversal potential or the resting potential of the cell. They did, however, slightly increase the Rin of the fiber. 5. Quantal analysis of single IPSCs revealed that GAD blockers increased the number of failures and thus reduced quantal content (m), diminished the probability of release (p), but did not affect the quantum current (q) or the statistical parameter (n), believed to be the number of available active zones. 6. Quantal analysis of EPSCs, released after interaction with the inhibitor, revealed a reduction in m without any effect on q. GAD blockers greatly reduced the efficacy of this inhibition without affecting the EPSC q. 7. GAD blockers increased the output of the excitor release sites by the following mechanisms: 1) increased EPSC, 2) increased EPSC facilitation, or 3) enhancement of spontaneous activity (miniature EPSCs). 8. Short time incubation with picrotoxin and CGP-35348 eliminated IPSCs and evoked inhibition. However, longer exposure (90 min) increased the excitor responses, similarly to the effects of GAD blockers. 9. Baclofen, a gamma-aminobutyric acid-B (GABAB) agonist, antagonized AOAA effects on evoked inhibition. 10. These results demonstrate that GAD blockers decrease postsynaptic and presynaptic inhibition by reducing both tonic and evoked release, most likely by diminishing p. 11. The reduction in GABA synthesis and release revealed a complex mechanism for GABAergic metabotropic regulation of inhibition efficacy and the release from the excitor glutamatergic terminals.

Inhibitory control of excitable dendrites in neocortex.

1. Many dendrites of pyramidal cells in mature neocortex express active Na+ and Ca2+ conductances. Dendrites are also the target of numerous inhibitory synapses. We examined the interactions between the intrinsic excitability of dendrites and synaptic inhibition using whole cell recordings from the apical dendrites of layer 5 pyramidal cells. Experiments were performed on slices of somatosensory cortex from mature rats. Slices were bathed in the glutamate receptor antagonists 2-amino-5-phosphonopentanoic acid and 6,7-dinitroquinoxaline-2,3-dione, and maintained at 32-36 degrees C. 2. In agreement with previous findings, intradendritic current injection evoked two distinct types of dendritic firing. Type I dendrites generated monophasic fast spikes, whereas type II dendrites showed more complex firing patterns, consisting of fast and slow spike components. 3. Stimulation of cortical layers 2/3 evoked fast inhibitory postsynaptic potentials (IPSPs) in all dendrites tested. IPSP reversal potentials were bimodally distributed, with means of about -53 and -85 mV when recorded with high-Cl(-)-concentration-filled electrodes. Interestingly, IPSP reversal potentials were correlated with the type of dendritic spiking pattern. 4. IPSPs were able to delay, completely block, or partially block spiking in dendrites, depending on the relative timing between inhibition and dendritic spiking. Slow, Ca(2+)-dependent spike components could be blocked selectively by IPSPs. Furthermore, inhibition could either phase advance or phase delay repetitive patterns of dendritic spiking, depending on the timing of the IPSP.

Activity‐induced decreased in early and late inhibitory synaptic conductances in hippocampus

The use dependence of inhibitory postsynaptic potentials (IPSPs) and their underlying conductances was studied in area CA1 of the hippocampal brain slice preparation, using a two-pulse paradigm in which paired activation of two separate synaptic inputs resulted in changes in the second, or "primed" response. In intracellular current-clamp recordings, the "primed" response, normally triphasic, exhibited a larger, wider excitatory PSP (EPSP) component and greatly reduced or absent IPSP components. Maximal widening occurred when the interval between synaptic stimuli was between 200 and 250 msec. Hyperpolarization of the postsynaptic cell reversed both the early IPSP and the direction of change of the width of the "primed" EPSP response, suggesting that the changes in the "primed" waveform were not due to the addition of an unidentified inward current(s). Furthermore, the reduction of the IPSPs during the "primed" response could not be accounted for by the fact that the membrane potential of the postsynaptic cell was hyperpolarized and therefore closer to IPSP reversal potential. Using single-electrode voltage-clamp techniques, we found that the early inhibitory conductance generally decreased by approximately 50%, with little if any change in reversal potential. The late inhibitory conductance also showed a priming-induced decrease of approximately 95%. Finally, "primed" four-pulse bursts of stimuli induced a larger depolarization in the postsynaptic cell than did unprimed bursts, also with an optimal interval of about 250 msec. We conclude that activation of certain synaptic pathways in the hippocampus results in a temporal window of 200-300 msec during which inhibitory synaptic activity is depressed and excitatory synaptic transmission is maximally effective, especially if the excitation occurs in short bursts. Such a mechanism would endow the inhibitory synaptic components of the hippocampus with a "gating" function to control long-term synaptic modification at excitatory synapses in the same region.

Serotonin‐mediated inhibitory postsynaptic potential in guinea‐pig prepositus hypoglossi and feedback inhibition by serotonin.

1. Intracellular recordings were made from neurones of the nucleus prepositus hypoglossi (PH) in slices of guinea-pig brain. Focal stimulation evoked an inhibitory postsynaptic potential (IPSP) that was typically 10-25 mV in amplitude and 1 s in duration. The IPSP reversal potential showed a Nernstian dependence on the external potassium concentration ([K+]o). 2. Spiperone blocked the IPSP with an IC50 of 40 nM, while ketanserin and (-)sulpiride had no effect. Cocaine (1 microM) prolonged the IPSP half-duration by 157%, and increased the amplitude by 28%. 3. 5-Hydroxytryptamine (5-HT, serotonin) hyperpolarized PH cells with an EC50 of 8.5 microM in control, and 135 nM in cocaine (10 microM). 8-Hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) also hyperpolarized PH cells with an EC50 of 16 nM, although the maximal effect was only 81% of the maximum 5-HT hyperpolarization. Spiperone produced a parallel, right shift of the 5-HT concentration-response curve; Schild analysis gave a Kd of 10 nM. Application of 5-HT to neurones voltage-clamped near their resting potential (about -55 mV) caused an outward current and an increase in membrane conductance. 4. The amplitude of the IPSP was reversibly decreased by non-hyperpolarizing concentrations of 5-HT and by the 5-HT1 receptor agonists 1-(m-trifluoromethylphenyl)piperazine (TFMPP) and 1-(3-chlorophenyl)piperazine (mCPP). The IC50 values for the latter two compounds were 50 nM and 1.5 microM, respectively; the maximal effect was a 90% inhibition. Neither compound affected the membrane potential nor changed the hyperpolarization induced by 5-HT. Quipizine competitively antagonized TFMPP with an estimated Kd of 165 nM. 5. When trains of stimuli were applied, an inhibition of the IPSP was observed following the first stimulus. At a frequency of 1 Hz, the inhibition was approximately 75%. This frequency-dependent 'run-down' of the IPSP was markedly attenuated by pre-treatment with TFMPP (1 microM). 6. It is concluded that the IPSP in PH cells is caused by 5-HT acting on 5-HT1A receptors to activate a potassium conductance. The release of 5-HT can be inhibited by activation of a presynaptic 5-HT1D receptor. This presynaptic receptor appears to be at least partly responsible for the run-down phenomenon, and may be involved in the physiological regulation of 5-HT synaptic transmission.

Relative contributions of passive equilibrium and active transport to the distribution of chloride in mammalian cortical neurons.

1. Active and passive factors affecting the chloride gradient of cortical neurons were assessed using intracellular recordings from neurons in slices of cingulate cortex maintained in vitro. The chloride equilibrium potential (ECl-) was estimated indirectly from the reversal potentials of responses to perisomatic gamma-aminobutyric acid (GABA) application and the Cl(-)-dependent inhibitory postsynaptic potential (IPSP). Under control conditions the mean resting potential (Vm; -69.7 mV) was not significantly different than the mean IPSP reversal potential (EIPSP; -70.1 mV). 2. Increasing the external potassium concentration ([K+]o) from 1 to 10 mM shifted the mean EIPSP from -80.4 to -61.8 mV. The mean EIPSP was approximately equal to the mean Vm at all [K+]oS. The conditions of Donnan equilibrium are not met in [K+]o less than 10 mM. 3. Polarization of Vm up to 20 mV away from EIPSP for 4 min with maintained current injection had no significant effect on EIPSP. 4. The GABA reversal potential was maintained 37-52 mV less negative than Vm after equilibration in saline in which the external chloride concentration had been reduced from 133 to 5 mM by substitution with isethionate. Vm and input resistance were not significantly different from control values in cells recorded under these conditions. 5. We conclude that Cl- is not passively distributed in cortical neurons, perhaps due to a low resting Cl- permeability. 6. Impalement with electrodes containing 2 M KCl resulted in a rapid 10 mV depolarizing shift in EIPSP that then remained relatively constant. Intracellular iontophoresis of Cl- resulted in a further depolarizing shift of EIPSP of 5-10 mV that returned to control in less than 1 min. The time course of recovery of IPSP amplitude could be fit with a single exponential having a mean time constant of 6.9 +/- 1.5 s and was independent of the amount of Cl- injected or stimulation frequency. 7. Reductions in temperature from 37 to 32 degrees C significantly increased the mean time constant of IPSP recovery from Cl- injection to 11.1 +/- 3.3 s, corresponding to Q10 = 2.6.(ABSTRACT TRUNCATED AT 400 WORDS)

GABAergic inhibition and the induction of spontaneous epileptiform activity by low chloride and high potassium in the hippocampal slice

Intracellular recordings from CA3b/c neurons in rat hippocampal slices showed that reduction of the extracellular Cl − concentration from 136 to 53 mM produced a positive (+ 10 mV) shift in the reversal potential of GABAergic inhibitory postsynaptic potentials (IPSPs). This shift was not significantly different from the shift produced by raising K + from 3.5 to 8.5 mM. Spontaneous interictal bursting occurred in both low Cl − and high K +. Extracellular recordings from the pyramidal cell layer in the CA3b/c region of hippocampal slices showed that bursts in 56 mM Cl − were of the same waveform and intensity as bursts produced by high K +. However the frequency of spontaneous bursting was much lower (6.6 ± 1.2/min, n = 10) in low Cl − compared to high K + (42.2 ± 3.0/min, n = 33). Burst frequency was a linear function of the shift in IPSP reversal potential produced by high K +, but not low Cl −. Replacing 60% of the Cl − with methylsulfate or isethionate was sufficient to produce spontaneous bursting, whereas it was necessary to replace 80% of the Cl − when propionate was used as a substitute. All 3 Cl − substitutes lowered the ionized Ca 2+ concentration, but raising the extracellular Ca 2+ concentration back to normal did not change the burst frequency. Since the amplitude of IPSPs is reduced to a similar extent in low Cl − and high K + solutions, whereas bursting is much faster in high K +, we suggest that impaired GABAergic inhibition is insufficient to fully account for spontaneous interictal bursting that is produced in hippocampal slices by raised extracellular K +.

Morphological and physiological properties of caudal medullary expiratory neurons of the cat

The activity of respiratory neurons in the caudal part of the nucleus retroambigualis (NRA) was recorded intracellularlyin decerebrated, spontaneously breathing cats. Spinal projections of these neurons were determined by antidromic stimulation at the C 3 segment. A high concentration of bulbospinal expiratory (BS-E) neurons was identified in the caudal NRA, whereas the inspiratory (I) neurons, not activated antidromically, were also found to be intermingled in the same region. The BS-E neurons had ramp-like depolarizing potentials during expiration, and repolarized rapidly at the onset of phrenic nerve discharge. The I neurons depolarized abruptly in the early I phase, and repolarized gradually thereafter, namely, they were early-I neurons. Intracellular current injections revealed postsynaptic inhibition of the BS-E neurons during inspiration, as evidenced by inhibitory postsynaptic potential reversal. Using the technique of intracellular labeling with horseradish peroxidase, seven well-stained expiratory cells located in the caudal NRA revealed detailed information about axonal morphology: the axon projected rostrally and dorsomedially for the first 2 mm after emerging from the soma, then turned caudally and ventrally along two different courses, and crossed the midline of the medulla almost at the same rostrocaudal level as the soma. No axon collaterals were observed along the length of the stained portion, indicating that the BS-E neurons cannot influence other respiratory neurons in the brainstem. It has been concluded that the NRA expiratory neurons are involved only in spinal action, and that they receive a postsynaptic inhibition during inspiration. The neighboring early-I neurons are likely to be candidates for the inhibitory inputs to the BS-E neurons.

Ammonia does not selectively block IPSPs in rat hippocampal pyramidal cells.

Intracellular recordings from CA1 pyramidal cells in the rat hippocampal slice preparation have been used to study the action of ammonia on inhibitory postsynaptic potentials (IPSPs). Concentrations of ammonia less than 2 mM had little effect on IPSPs or the action of iontophoretically applied gamma-aminobutyric acid (GABA). This concentration has been reported to be fully effective in blocking hyperpolarizing IPSPs in spinal cord and neocortex. Concentrations above 2 mM did cause a depolarizing shift in the IPSP and GABA reversal potentials, but this effect was accompanied by several generalized effects. The conductance increase during the IPSP but not during the GABA response was depressed, indicating that ammonia has a presynaptic depressant effect on the IPSP. Ammonia also depressed excitatory postsynaptic potentials (EPSPs), presynaptic fiber potentials, and pyramidal cell population spikes. In addition, the calcium-dependent potassium response elicited by depolarizing current pulses was depressed. This depression was due, in part, to a depolarizing shift in the reversal potential for this response. Responses recorded with potassium-sensitive microelectrodes indicate that ammonia releases potassium into the extracellular space. The possibility is discussed that the shifts in IPSP reversal potential seen with high concentrations of ammonia are a consequence of generalized nonspecific effects. We conclude that the relative insensitivity of hippocampal IPSPs to blockade by ammonia suggests that a mechanism fundamentally unlike an ammonia-sensitive chloride pump must maintain the hippocampal IPSP gradient.

Effects of phenytoin on pyramidal neurons of the rat hippocampus

The effectsof phenytoin (35 μg/ml) on membrane properties and inhibitory postsynaptic potentials (IPSPs) in CA1 and CA3 pyramidal neurons of the in vitro rat hippocampus were examined. No significant change was observed on input resistance or resting membrane potential. Action potential amplitude, overshoot, rate of rise and rate of decay were decreased. IPSP conductance increase and reversal potential, evoked in CA3 cells through mossy fiber stimulation and in CA1 cells through recurrent and Schaffer's collateral stimulation, were unaffected.

The effect of anion injection and changes in the external potassium and chloride concentration on the reversal potentials of the IPSP and acetylcholine

1.1. Certain neurones in the abdominal ganglion of the snail Helix aspersa show spontaneous inhibitory post-synaptic potentials (IPSP).2.2. Addition of acetylcholine normally causes a hyperpolarization of such neurones.3.3. Fifteen different anions, of increasing hydrated volume, were injected into the neurone. Those with a hydrated diameter greater than 1·24 times the hydrated potassium ion had no effect on the reversal potential of the IPSP and ACh. These anions were fluoride, bisulphite, bicarbonate, acetate, chromate, sulphate, phosphate and propionate.4.4. Injection of small anions, such as bromide, chloride, nitrate, thiocyante, chlorate, formate, altered the reversal potentials of the IPSP and ACh.5.5. Alteration of the external chloride concentration or the external potassium concentration affected the reversal potentials of the IPSP and ACh.6.6. Experiments in which the external concentrations of potassium and chloride were altered so that they either opposed or augmented each other indicate that the effect of the chloride ion was more marked and more rapid than that of the potassium ion on the IPSP and ACh reversal potentials.7.7. It is suggested that during the IPSP or when ACh is applied to the cell, pores open in the membranes so that approximately 90 per cent of the current is carried by the chloride ion and 10 per cent by the potassium ion.8.8. It is probable that acethylcholine is the transmitter at the snail inhibitory synapse.