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PubMed1996

FK506, a Ca2+/calmodulin-dependent phosphatase inhibitor, inhibits the induction of long-term potentiation in the rat hippocampus.

The effects of FK506, a Ca2+/calmodulin-dependent phosphatase 2B (calcineurin) inhibitor, on the synaptic potentials and long-term potentiation (LTP) were investigated, using extracellular recordings in the CA1 region of adult rat hippocampal slices. Bath application of FK506 (1-50 mu M) produced a dose-dependent and reversible inhibition on the field excitatory postsynaptic potentials. FK506 also showed an inhibitory effect on the induction of LTP evoked by theta-burst stimulation. Analyses of the fiber volley and paired-pulse facilitation revealed that FK506-induced effects were based on postsynaptic mechanisms. These results demonstrate that FK506 inhibits both the synaptic transmission and the LTP induction, and suggest that calcineurin plays a promoting role in the LTP processes in the hippocampus of adult rats.

Animals↗

PubMed1991

Inhibition of caudal medullary expiratory neurones by retrofacial inspiratory neurones in the cat.

1. Comparisons between the spike discharge of inspiratory neurons within the retrofacial area (RFN), and the membrane potential of expiratory neurones within the caudal medulla were made in pentobarbitone-anaesthetized, vagotomized, artificially ventilated cats. Spike-triggered averaging (STA) of synaptic potentials, triggered by the discharge of inspiratory RFN neurones, was utilized to test for synaptic connectivity. 2. Eighty-nine neurons with respiratory-phased discharge patterns were recorded in the vicinity of the RFN. Fifty-four neurones discharged at or slightly before the onset of the inspiratory burst activity of the phrenic nerve and continued firing throughout inspiration. Two continued to fire during post-inspiration. Forty-five of fifty-four inspiratory RFN neurones exhibited incrementing discharge patterns, six discharged with a plateau pattern, while only three neurones had a decrementing discharge pattern. 3. The membrane potential trajectories of caudal expiratory neurones revealed a typical wave of early inspiratory hyperpolarization. Occasionally, a second wave of hyperpolarization occurred during late inspiration, in conjunction with increased phrenic nerve activity. 4. Spike-triggered averaging revealed averaged inhibitory postsynaptic potentials (IPSPs), indicative of inhibitory synaptic connections, between eight and sixty-three pairs of RFN inspiratory and caudal expiratory neurones. 5. Inhibitory postsynaptic potentials detected by STA exhibited a relatively long latency and a slow time course. The IPSPs began, on average, 3.8 ms after an RFN action potential. The rise times, half-widths and durations of IPSPs were longer than expected for a monosynaptic somal input from myelinated axons of inspiratory RFN neurones. It is suggested that an inhibitory relay neurone in the immediate vicinity of the expiratory neurones is activated by a collateral of the RFN inspiratory neurone. 6. Retrofacial inspiratory neurones were antidromically activated only when high-intensity electrical stimulation was applied in the vicinity of caudal expiratory neurones. 7. The averaged IPSPs were preceded by diphasic and triphasic 'spike potentials'. The averaged spike potentials were highly entrained to the action potentials of RFN inspiratory neurones which triggered IPSPs. The spike potentials may be terminal potentials recorded from axons of RFN inspiratory neurones. 8. Evidence for convergence of synaptic inputs was obtained from STA tests in a caudal expiratory neurone receiving IPSPs from four RFN neurones. 9. The functional significance of this observation is discussed. We conclude that RFN inspiratory neurones exert a moderate inhibitory influence and act conjointly with other types of medullary inspiratory neurones.

Animals↗

PubMed1999

Electrical stimulation of the horizontal limb of the diagonal band of broca modulates population EPSPs in piriform cortex.

Electrical stimulation of the horizontal limb of the diagonal band of Broca (HDB) was coupled with recording of evoked potentials in the piriform cortex. Stimulation of the HDB caused an enhancement of the late, disynaptic component of the evoked potential elicited by stimulation of the lateral olfactory tract but caused a suppression of the synaptic potential elicited by stimulation of the posterior piriform cortex. The muscarinic antagonist scopolamine blocked both effects of HDB stimulation. The enhancement of disynaptic potentials could be due to cholinergic depolarization of pyramidal cells, whereas the suppression of potentials evoked by posterior piriform stimulation could be due to presynaptic inhibition of intrinsic fiber synaptic transmission by acetylcholine.

Animals↗

PubMed1973

On the mechanism of halothane anaesthesia.

1. The effects of halothane on the evoked potentials of in vitro preparations of guinea-pig olfactory cortex were studied.2. The evoked potentials recorded from the cortical surface comprised an initial diphasic wave - the lateral olfactory tract (l.o.t.) compound action potential - followed by a negative wave of 1-3 mV amplitude and about 10 msec duration. Superimposed on the negative wave was a number of positive peaks. The negative wave has been identified as an extracellularly recorded, monosynaptic, excitatory post-synaptic potential (e.p.s.p.) and the positive peaks have been shown to reflect the discharge of the cortical cell population in response to the evoked e.p.s.p. and are therefore termed ;population spikes'.3. When halothane (0.4-1.5%) was added to the gas stream that superfused the surface of the preparation the evoked e.p.s.p.s became smaller in amplitude and the size of the population spikes diminished. The l.o.t. compound action potential was unaffected by these levels of halothane. Higher levels of halothane (above 2%) further reduced the amplitude of the evoked e.p.s.p.s, abolished the population spikes, decreased the amplitude of the l.o.t. compound action potential and slowed its time course. The effects of halothane on the evoked potentials were dose-related and were independent (after the first 10 min of treatment) of the duration of the exposure to halothane.4. The decrease in the size of the population spike caused by the exposure to halothane implied that transmission through the cortical relay had been impaired. This was also shown by the decrease in the evoked activity of units in the prepiriform cortex. Of eleven units, eight were depressed by halothane (0.5-1.5%) two were unaffected and one showed a transient increase in the number of spikes generated in response to a l.o.t. volley.5. Halothane (up to 1.5%) had no effect on the threshold of the l.o.t. fibres to electrical stimulation or on that of the post-synaptic cells to synaptic excitation.6. Post-tetanic potentiation and frequency potentiation of the evoked e.p.s.p.s were enhanced in the presence of 1% halothane.7. It is concluded that halothane reduces excitatory synaptic transmission not by an increase in the electrical threshold of the post-synaptic cells to synaptic excitation but by interference with the process of chemical transmission either by reducing the output of transmitter from the pre-synaptic nerve terminal or by reducing the sensitivity of the post-synaptic membrane to the released transmitter substance.

Action Potentials↗

PubMed1981

Selective synapse formation during sprouting after partial denervation of the guinea-pig superior cervical ganglion.

1. The synaptic connexions established by sprouting of intact preganglionic sympathetic axons were examined by intracellular recording in vitro and by observing the sympathetic end organ responses to ventral root stimulation in vivo. 2. The superior cervical ganglion of the guinea-pig was partially denervated (70-85%) by crushing the cervical sympathetic trunk at the level of the subclavian artery, leaving the ansa subclavia intact. The intact nerve carried some preganglionic axons arising from each of the eight spinal cord segments (C8-T7) contributing innervation to the ganglion. 3. During the first 4 weeks after the operation, there was a two-to threefold increase in the number of steps in the synaptic response elicited in individual ganglion cells by graded stimulation of the ansa subclavia. There was also an increase in the amplitude of the synaptic potential elicited by each preganglionic axon. 4. This increase in the synaptic contribution of the intact nerve to neurones in the superior cervical ganglion after partial denervation was attributed to sprouting of residual preganglionic axons. A major contribution from collateral connexions between ganglion cells was ruled out by intracellular recording form neurones during antidromic stimulation of their axons in the inferior post-ganglionic nerve. 5. After sprouting, the specificity of the sympathetic end organ responses elicited by stimulation of the ventral roots of spinal segments T1 and T4 in vivo was indistinguishable from normal, although the strength of these responses increased from just perceptible acutely after partial denervation to near normal 3-6 weeks after the operation, when sprouting was largely complete. 6. These results show that intact preganglionic axons arising from different spinal levels established selective connexions with different classes of ganglion cells during sprouting.

Action Potentials↗

PubMed2005

Reluctant vesicles contribute to the total readily releasable pool in glutamatergic hippocampal neurons.

The size of the readily releasable pool (RRP) of vesicles is critically important for determining the size of postsynaptic currents generated in response to action potentials. However, discrepancies in RRP estimates exist among methods designed to measure RRP size. In glutamatergic hippocampal neurons, we found that hypertonic sucrose application yielded RRP size estimates approximately fivefold larger than values obtained with high-frequency action potential trains commonly assumed to deplete the RRP. This discrepancy was specific for glutamatergic neurons, because no difference was found between sucrose and train estimates of RRP size in GABAergic neurons. A small component of the difference in excitatory neurons was accounted for by postsynaptic receptor saturation. Train estimates of vesicle pool size obtained using more stimuli revealed that action potential-elicited EPSCs did not truly reach a steady state during shorter trains, and RRP estimates were closer to sucrose estimates made in the same neurons. This suggested that reluctant vesicles may contribute to the total available pool. Two additional lines of evidence supported this hypothesis. First, RRP estimates from strongly depolarizing hyperkalemic solutions closely matched those obtained with sucrose. Second, when Ca2+ influx was enhanced during trains, train estimates of pool size matched those obtained with sucrose. These data suggest that glutamatergic hippocampal neurons maintain a heterogeneous population of vesicles that can be differentially released with varying Ca2+ influx, thereby increasing the range of potential synaptic responses.

Action Potentials↗

PubMed1994

Improved stability of Drosophila larval neuromuscular preparations in haemolymph-like physiological solutions.

Neuromuscular preparations from third instar larvae of Drosophila are not well-maintained in commonly used physiological solutions: vacuoles form in the muscle fibers, and membrane potential declines. These problems may result from the Na:K ratio and total divalent cation content of these physiological solutions being quite different from those of haemolymph. Accordingly haemolymph-like solutions, based upon ion measurements of major cations, were developed and tested. Haemolymph-like solutions maintained the membrane potential at a relatively constant level, and prolonged the physiological life of the preparations. Synaptic transmission was well-maintained in haemolymph-like solutions, but the excitatory synaptic potentials had a slower time course and summated more effectively with repetitive stimulation, than in standard Drosophila solutions. Voltage-clamp experiments suggest that these effects are linked to more pronounced activation of muscle fiber membrane conductances in standard solutions, rather than to differences in passive muscle membrane properties or changes in postsynaptic receptor channel kinetics. Calcium dependence of transmitter release was steep in both standard and haemolymph-like solutions, but higher external calcium concentrations were required for a given level of release in haemolymph-like solutions. Thus, haemolymph-like solutions allow for prolonged, stable recording of synaptic transmission.

Animals↗

PubMed1978

Physiological and morphological identification of a nonpyramidal hippocampal cell type.

In intracellular recordings from the CA1 region of the hippocampal slice preparation, an infrequent cell type different from the predominant pyramidal cell has been characterized. The cells have very brief spikes, hyperpolarizing after-potentials, and tend to fire at a high spontaneous rate; unitary synaptic potentials are visible in the baseline. Stimulation in stratum radiatum evokes an EPSP which often triggers a short burst of spikes; an IPSP invariably follows the initial excitation. Intracellular injection of HRP into these cells shows them to be a non-pyramidal cell type with extensive local axon ramification; however, they do not resemble the classical basket cell. These cells are, nevertheless, candidates for an interneuron in hippocampus.

Animals↗

PubMed1989

Neurophysiological characterization of commissurally projecting dentate neurons in the rat.

Commissural neurons in the dentate hilus and in the deep dentate granule cell layer were recorded intracellularly in vivo, in conjunction with combined injection of the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) at sites of electrical stimulation. Two hilar neurons responded with short latency antidromic spikes to stimulation of the contralateral dentate infrapyramidal molecular layer, but did not show any synaptic potentials, suggesting that these neurons do not receive commissural hilar input, either directly or indirectly, from the stimulating sites. On the other hand, 3 dentate-hilar border neurons responded to the contralateral hilar stimulation with antidromic spikes, excitatory postsynaptic potentials (EPSPs), orthodromic spikes, and inhibitory PSPs (IPSPs), suggesting a rich synaptic interaction both commissurally and locally in this region. No direct commissural inhibition was observed in any of the cells. PHA-L injection at the stimulation site indicated that commissural hilar axon terminals project to a limited region of the contralateral molecular layer in a lamellar fashion, and have only a sparse distribution in the contralateral hilus. The results indicate that rapidly conducting commissural neurons in the dentate gyrus are themselves inhibited in an indirect manner by commissural fibers.

Animals↗

PubMed1982

Intracellular analysis of a postsynaptic action of adenosine in the rat hippocampus.

The effects of topical or iontophoretic application of adenosine (AD) were studied by means of intracellular recording techniques in rat hippocampal slices. AD applied in a microdrop containing up to 0.1 nmol of AD produced a 7 +/- 0.7 mV hyperpolarization associated with a 30% decrease in input resistance of the recorded neurons. The hyperpolarization was dependent on the resting membrane potential and had an apparent reversal potential of about -90mV. These effect were still present when synaptic activity was blocked. AD also caused a marked reduction in the magnitude of synaptic potentials in the recorded neurons. Iontophoretic application of AD in dendritic regions caused a suppression of EPSP without affecting resting membrane potential ionotophoresis of AD onto somata produced both hyperpolarization and decreased EPSP. It is suggested that AD produces both presynaptic depression of transmitter release and postsynaptic increase in K+ conductance.

4-Aminopyridine↗

PubMedDate not supplied

Activity following colonic distension in enteric sensory fibres projecting to the inferior mesenteric ganglion in the guinea pig.

In this investigation the characteristics of the response to colonic distension of afferent fibres projecting to the inferior mesenteric ganglion (IMG) of the guinea pig were studied in vitro. Intracellular membrane potential recordings were made from neurons in the IMG. Post-synaptic potentials arising from activity in afferent fibres were recorded in these cells following distensions of a small segment of colon placed in a separate and independently perfused organ bath. These afferent fibres showed both transient and sustained responses to distension. Application of a solution containing 0.25 mM Ca2+ and 10 mM2+ Mg2+ to the colon (but not to the IMG) reduced the overall response but did not abolish the activity in these fibres. It is concluded that some afferent fibres which respond to colonic distension project directly to the IMG.

Afferent Pathways↗

PubMed1995

Effects of a cAMP analogue simulate the distinct components of long-term potentiation in CA1 region of rat hippocampus.

Bath application of the cAMP analogue, dibutyryl cyclic adenosine 3',5'-monophosphate (dibutyryl cyclic AMP; dbcAMP) to rat hippocampal slices was found to potentiate both the CA1 population spike and population excitatory post-synaptic potential (EPSP) slope. dbcAMP (500-1000 microM) was applied to slices for 30 min; following washout the population EPSP slope was potentiated for at least 30 min to a mean value of 51% above the drug-free baseline value. The population spike was similarly potentiated to a mean value of 64% above baseline after dbcAMP washout. dbcAMP-induced population EPSP slope potentiation occluded long-term potentiation (LTP) induced by high frequency electrical stimulation, and LTP occluded dbcAMP-induced EPSP slope potentiation. Earlier investigations (Pockett et al., Neuroscience, 52 (1993) 229-236) using 200 microM dbcAMP reported similar potentiation of population spike but no potentiation of EPSP slope. These experiments support the hypothesis that the two components of LTP (Bliss and Lynch, In P.W. Landfield and S.A. Deadwyler (Eds.), Long-term Potentiation: from Biophysics to Behaviour, Alan R. Liss, New York, 1988, pp. 3-72) in the CA1 area of rat hippocampus both involve distinct cAMP-dependent mechanisms.

Animals↗

PubMed1990

GABAergic mechanisms in the CA3 hippocampal region during early postnatal life.

The developmental pattern of GABAergic neurons in the rat hippocampus during the first week of postnatal life shows several particularities both from a morphological and physiological point of view: (1) GABA immunoreactive neurons which are initially localized in a deep and superficial layer, progressively disappear from these two layers. From the end of the first postnatal week, GABAergic neuronal somata appear throughout the whole hippocampus, but GABA immunoreactive terminal structures are not frequent until the second postnatal week. (2) Intracellular observations in slices reveal the presence in CA3 pyramidal neurons between P0 and P6 (postnatal days) of spontaneous giant depolarizing potentials (GDPs); these are mediated by GABA acting on GABAA receptors and modulated presynaptically by NMDA receptors. During this period of development, GABA and GABAA analogues have a depolarizing action at resting membrane potential. Bicuculline at this developmental stage blocks completely spontaneous and evoked synaptic potentials. During the second postnatal week, when GABA responses shift from depolarizing to hyperpolarizing, bicuculline induces spontaneous interictal discharges. It is suggested that the positive feedback of the GABAergic interneuron on the pyramidal neuron during the first week of life may account for the generation of GDPS which may play an important role in synaptogenesis.

Action Potentials↗

PubMed2000

Long-term potentiation across rat cerebello-thalamic synapses in vitro.

We previously demonstrated that rat cerebello-thalamic synapses undergo an ultrastructural change, consistent with expression of long-term potentiation, in association with motor adaptation. The aim of these experiments was to determine if long-term potentiation can be expressed at these synapses physiologically. Excitatory post-synaptic potentials evoked by electrical stimulation of cut cerebello-thalamic afferents were recorded intracellularly from thalamic neurons in 17-24 day old rat brain slice preparations. The experimental protocol involved long periods of low frequency single-shock control stimuli interrupted by brief high frequency trains of conditioning stimuli. Conditioning at 100 Hz evoked long-term potentiation in 5/6 cells and long-term depression in 1/6 cells. Conditioning at 200 Hz was unsuccessful in evoking long-term potentiation but did evoke long-term depression in 1/3 cells and no change in 2/3 cells. We conclude that long-term potentiation can be evoked across rat cerebello-thalamic synapses in vitro.

Animals↗

PubMed1995

Arachidonic acid, but not sodium nitroprusside, stimulates presynaptic protein kinase C and phosphorylation of GAP-43 in rat hippocampal slices and synaptosomes.

Activation of protein kinase C (PKC) and phosphorylation of its presynaptic substrate, the 43-kDa growth-associated protein GAP-43, may contribute to the maintenance of hippocampal long-term potentiation (LTP) by enhancing the probability of neurotransmitter release and/or modifying synaptic morphology. Induction of LTP in rat hippocampal slices by high-frequency stimulation of Schaffer collateral-CA1 synapses significantly increased the PKC-dependent phosphorylation of GAP-43, as assessed by quantitative immunoblotting with a monoclonal antibody that recognizes an epitope that is specifically phosphorylated by PKC. The stimulatory effect of high-frequency stimulation on levels of immunoreactive phosphorylated GAP-43 was not observed when 4-amino-5-phosphonovalerate (50 microM), an N-methyl-D-aspartate (NMDA) receptor antagonist, was bath-applied during the high-frequency stimulus. This observation supports the hypothesis that a retrograde messenger is produced postsynaptically following NMDA receptor activation and diffuses to the presynaptic terminal to activate PKC. Two retrograde messenger candidates--arachidonic acid and nitric oxide (sodium nitroprusside was used to generate nitric oxide)--were examined for their effects in hippocampal slices on PKC redistribution from cytosol to membrane as an indirect measure of enzyme activation and PKC-specific GAP-43 phosphorylation. Bath application of arachidonic acid, but not sodium nitroprusside, at concentrations that produce synaptic potentiation (100 microM and 1 mM, respectively) significantly increased translocation of PKC immunoreactivity from cytosol to membrane as well as levels of immunoreactive, phosphorylated GAP-43. The stimulatory effect of arachidonic acid on GAP-43 phosphorylation was also observed in hippocampal synaptosomes. These results indicate that arachidonic acid may contribute to LTP maintenance by activation of presynaptic PKC and phosphorylation of GAP-43 substrate. The data also suggest that nitric oxide does not activate this signal transduction system and, by inference, activates a distinct biochemical pathway.

Animals↗

PubMed2004

Intracellular calcium store filling by an L-type calcium current in the basolateral amygdala at subthreshold membrane potentials.

The long-term changes that underlie learning and memory are activated by rises in intracellular Ca2+ that activate a number of signalling pathways and trigger changes in gene transcription. Ca2+ rises due to influx via L-type voltage-dependent Ca2+ channels (L-VDCCs) and release from intracellular Ca2+ stores have been consistently implicated in the biochemical cascades that underlie the final changes in memory formation. Here, we show that pyramidal neurones in the basolateral amygdala express an L-VDCC that is active at resting membrane potentials. Subthreshold depolarization of neurones either by current injection or summating synaptic potentials led to a sustained rise in cytosolic Ca2+ that was blocked by the dihydropyridine nicardipine. Activation of metabotropic receptors released Ca2+ from intracellular Ca2+ stores. At hyperpolarized potentials, metabotropic-evoked store release ran down with repeated stimulation. Depolarization of cells to -50 mV, or maintaining them at the resting membrane potential, restored release from intracellular Ca2+ stores, an effect that was blocked by nicardipine. These results show that Ca2+ influx via a low-voltage-activated L-type Ca2+ current refills inositol 1,4,5-trisphosphate (IP(3))-sensitive intracellular Ca2+ stores, and maintains Ca2+ release and wave generation by metabotropic receptor activation.

Algorithms↗

PubMed1992

Trimethyltin alters membrane properties of CA1 hippocampal neurons.

Trimethyltin produces pathological changes in the hippocampus, but the physiological mechanisms underlying its toxicity remain unclear. Intracellular recordings of CA1 neurons in rat hippocampal slices were conducted during bath application of 50 and 100 microM trimethyltin and 50 and 200 microM dimethyltin. Trimethyltin slowly depolarized the membrane potential without spontaneous spiking, and decreased input resistance and time constant. Trimethyltin decreased, in a dose-dependent manner, both the elicited action potential and the orthodromically-stimulated action potential amplitudes; increased the threshold current for both elicited and orthodromically-stimulated action potentials; and prolonged the duration of the orthodromic excitatory post-synaptic potential. These trimethyltin-induced effects were not readily reversed. On the other hand, dimethyltin at high concentrations only reduced the amplitude of the orthodromic action potential. Slice viability was compromised following exposure to trimethyltin, but not following dimethyltin. These data demonstrate that decreased membrane time constant and input resistance is an early and reliable indicator of the onset of trimethyltin-induced changes. The effects do not resemble those produced by excitotoxins, but rather share similarities with responses observed during slice hypoxia.

Action Potentials↗

PubMed2002

The life cycle of Ca(2+) ions in dendritic spines.

Spine Ca(2+) is critical for the induction of synaptic plasticity, but the factors that control Ca(2+) handling in dendritic spines under physiological conditions are largely unknown. We studied [Ca(2+)] signaling in dendritic spines of CA1 pyramidal neurons and find that spines are specialized structures with low endogenous Ca(2+) buffer capacity that allows large and extremely rapid [Ca(2+)] changes. Under physiological conditions, Ca(2+) diffusion across the spine neck is negligible, and the spine head functions as a separate compartment on long time scales, allowing localized Ca(2+) buildup during trains of synaptic stimuli. Furthermore, the kinetics of Ca(2+) sources governs the time course of [Ca(2+)] signals and may explain the selective activation of long-term synaptic potentiation (LTP) and long-term depression (LTD) by NMDA-R-mediated synaptic Ca(2+).

Action Potentials↗