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P Legendre

Publications and source records attributed to P Legendre.

18 recordsLinked to original sources
PubMed1992

Intrinsic quantal variability due to stochastic properties of receptor-transmitter interactions.

Synaptic events at the neuromuscular junction are integer multiples of a quantum, the postsynaptic response to transmitter released from one presynaptic vesicle. At central synapses where quanta are small, it has been suggested they are invariant due to occupation of all postsynaptic receptors, a concept neglecting inherent fluctuations in channel behavior. If this did occur, the quantal release model would not apply there and could not be used to localize sites of synaptic modification. Monte Carlo simulations of quanta include transmitter diffusion and interactions with postsynaptic receptors that are treated probabilistically. These models suggest that when there are few postsynaptic channels available at a synapse, their stochastic behavior produces significant intrinsic variance in response amplitude and kinetics, and saturation does not occur. These results were confirmed by analysis of inhibitory quanta in embryonic and adult Mauthner cells involving a small and large number of channels, respectively. The findings apply to excitatory synapses as well.

Animals↗
PubMed1992

Analyzing multivariate flow cytometric data in aquatic sciences.

Flow cytometry has recently been introduced in aquatic ecology. Its unique feature is to measure several optical characteristics simultaneously on a large number of cells. Until now, these data have generally been analyzed in simple ways, e.g., frequency histograms and bivariate scatter diagrams, so that the multivariate potential of the data has not been fully exploited. This paper presents a way of answering ecologically meaningful questions, using the multivariate characteristics of the data. In order to do so, the multivariate data are reduced to a small number of classes by clustering, which reduces the data to a categorical variable. Multivariate pairwise comparisons can then be performed among samples using these new data vectors. The test case presented in the paper forms a time series of observations from which the new method enables us to study on the temporal evolution of cell types.

Data Interpretation, Statistical↗
PubMed1992

Expression of NMDA channels on cerebellar Purkinje cells acutely dissociated from newborn rats.

1. Conflicting evidence exists concerning the expression and properties of N-methyl-D-aspartate (NMDA) receptors on cerebellar Purkinje cells during development. We used whole-cell and single-channel recording to examine NMDA receptors on acutely dissociated Purkinje cells from newborn rats (postnatal day 0-4). 2. NMDA channels were present on > 80% of identified Purkinje cells and had pharmacological and single-channel properties that were indistinguishable from NMDA receptors on other neurons. In particular, responses were glycine-dependent and Mg2+ produced flickery open-channel block. 3. Our results demonstrate the transient expression of NMDA receptor/channels on Purkinje cells early in development. As NMDA receptors have been implicated in developmental plasticity in other regions of the CNS, a similar role is feasible during climbing fiber innervation of Purkinje cells.

Animals↗
PubMed1991

Maturation of a transient outward potassium current in mouse fetal hypothalamic neurons in culture.

The whole-cell voltage clamp technique was used to record potassium currents in mouse fetal hypothalamic neurons developing in culture medium from days 1 to 17. The neurons were derived from fetuses of IOPS/OF1 mice on the 14th day of gestation. The mature neurons (greater than six days in culture) showed both a transient potassium current and a non-inactivating delayed rectifier potassium current. These were identified pharmacologically by using the potassium channel blockers tetraethyl ammonium chloride and 4-aminopyridine, and on the basis of their kinetics and voltage sensitivities. The delayed rectifier potassium current had a threshold of-20 mV, a slow time-course of activation, and was sustained during the voltage pulse. The 4-aminopyridine-sensitive current was transient, and was activated from a holding potential more negative (-80 mV) than that required for evoking the delayed rectifier potassium current (-40 mV). The delayed rectifier potassium current was detectable from day 1 onwards, while the transient potassium current showed a distinct developmental trend. The time-constant of inactivation became faster with age in culture. The half steady-state inactivation potential showed a shift towards less negative membrane potentials with age, and the relationship was best described by a logarithmic regression equation. The developmental trend of the transient potassium current may relate functionally to the progressive morphological changes, and the appearance of synaptic connections during ontogenesis.

Animals↗
PubMed1991

Noncompetitive inhibition of gamma-aminobutyric acidA channels by Zn.

The action of zinc on chloride currents evoked by gamma-aminobutyric acid (GABA) was examined on cultured hippocampal neurons using whole cell voltage clamp and outside-out patch recording. Zn (5-30 microM) noncompetitively blocked responses evoked by GABA (0.5-100 microM), but did not affect either the time-to-peak or desensitization of the macroscopic current. In outside-out patches, Zn had no effect on the mean conductance or lifetime of the 19 or 30 pS openings of the GABA channel; however, the frequency of channel opening was markedly decreased in a voltage-independent manner. Zn inhibition of GABA responses appeared to be independent of the benzodiazepine binding site as Zn was effective in the presence of either diazepam or Ro15-1788, a competitive antagonist of benzodiazepine agonists and inverse agonists. In contrast to prior reports, Zn also inhibited GABA currents in a similar manner on cultured superior cervical ganglion neurons. These results suggest that Zn acts at an extracellular site on the GABAA receptor complex, which is distinct from either the GABA or benzodiazepine binding sites. The structural similarity of the Cys-Cys loop of the alpha and gamma GABAA receptor subunits to some Zn-binding proteins suggests one possible region for a Zn binding site.

Animals↗
PubMed1991

Ifenprodil blocks N-methyl-D-aspartate receptors by a two-component mechanism.

The inhibition of N-methyl-D-aspartate (NMDA) receptor channels by the vasodilatory and anti-ischemic agent ifenprodil was examined on cultured rat hippocampal neurons. Whole-cell and single-channel patch recordings were used. Ifenprodil inhibition of NMDA currents could be separated into two components, with IC50 values of 0.75 and 161 microM. The high and low affinity components were both voltage independent but could be separated by their kinetics and dependence on extracellular calcium and glycine. The maximal inhibition of inward current by ifenprodil (approximately 90%) was equally divided between the two components in 0.3 mM extracellular calcium and 500 nM glycine. The low affinity action of ifenprodil had rapid kinetics and appeared to result from allosteric inhibition of the glycine modulatory site on the NMDA receptor. The macroscopic kinetics of the high affinity component were slow. The rate of onset was concentration dependent, and complete recovery required 1-2 min. Unlike open-channel blockers, ifenprodil block was not use dependent, and pre-exposure to ifenprodil also reduced subsequent NMDA responses. Low concentrations of ifenprodil were less effective after calcium-dependent inactivation of whole-cell currents, but the IC50 was unaffected, suggesting that calcium and ifenprodil act on a common set of channels. On outside-out membrane patches, ifenprodil reduced the frequency of channel opening without altering the single-channel conductance. Open time histograms of the large conductance events revealed two mean open times of approximately 2 and 8 msec, but only the duration of the long openings was decreased by ifenprodil. This effect was concentration dependent and revealed a blocking rate constant of 6 x 10(7) M-1sec-1. However, the proportion of current blocked by low concentrations of ifenprodil was larger in outside-out patches than in whole-cell recordings, suggesting that intracellular factors may influence ifenprodil efficacy. These results indicate that high affinity ifenprodil binding is extracellular and does not require agonist binding or channel opening. Because low concentrations of ifenprodil only partially inhibited the current and affected only the long openings, ifenprodil may promote a modal shift in channel gating.

Animals↗
PubMed1990

The inhibition of single N-methyl-D-aspartate-activated channels by zinc ions on cultured rat neurones.

1. Single channels activated by N-methyl-D-aspartate (NMDA) were studied in outside-out patches of cultured hippocampal neurones in the presence of glycine and absence of magnesium. The effects of the transition metal ions zinc and cadmium on NMDA channels were tested by placing the membrane patch at the mouth of one of an array of large barrelled flow pipes. 2. Amplitude histograms revealed several conductance levels between 5 and 45 pS with the majority of NMDA-activated openings greater than 25 pS. Zinc (5-100 microM) and cadmium (30-100 microM) reduced the number of large conductance events in a voltage-independent manner. Zinc (30 microM) reduced the large conductance openings by approximately 70-80%. The small number of events under 20 pS precluded quantitative assessment of the effects of zinc and cadmium on these conductance levels. Zinc inhibition of the calculated macroscopic current due to NMDA-activated channels could be fitted with a single binding site isotherm with an IC50 of 12 microM. 3. Zinc and cadmium also reduced the mean open time of the two largest conductance events of 38 and 43 pS; this reduction was voltage independent. Open-time histograms were fitted with the sum of two exponentials. In the presence of 5 microM-NMDA at -60 mV, tau o2 = 10.49 ms and tau o1 = 1.47 ms; in 30 microM-zinc, tau o2 = 3.49 and tau o1 = 0.8 ms. The 'blocking' rate constant calculated at a membrane potential of +40 mV from the slope of 1/tau o2 vs. [zinc]o was 4 x 10(6) M-1 S-1. 4. Closed-time analysis revealed brief (tau c = 0.4-1.0 ms) zinc-insensitive gaps; longer closed-time intervals were not analysed since all patches contained more than one channel. Both burst duration and the number of bursts were reduced in the presence of zinc. 5. At holding potentials negative to -40 mV in magnesium-free solutions, zinc also induced high-frequency flickering of the open channel which included complete channel closures at 4 kHz filtering. No zinc-induced flickering was seen at positive membrane potentials. The flickering was dose dependent, becoming prominent at zinc concentrations above 30 microM. Cadmium did not induce flickering at concentrations up to 100 microM.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Effects of dopamine on voltage-dependent potassium currents in identified rat lactotroph cells.

The effects of dopamine (DA) on voltage-dependent potassium currents were investigated in rat lactotrophs maintained in primary culture. Lactotroph cells were identified using the reverse hemolytic plaque assay. Membrane currents and potentials of lactotroph cells were recorded using the patch-clamp recording technique in the 'whole-cell' configuration. In the presence of cobalt (2 mM), two types of voltage-dependent K+ currents were recorded, a voltage-activated delayed K+ current (IK) and a voltage-activated transient K+ current (IA). The current IK was activated at membrane potentials varying from -20 to +40 mV and did not inactivate during prolonged voltage steps (up to 25 s); it was blocked by tetraethylammonium (10 mM). The current IA was activated at membrane potentials higher than -45 mV and showed a voltage-dependent inactivation between -110 and -40 mV; it was slightly inhibited by 4-aminopyridine (5 mM). Under current-clamp conditions, the majority of the cells (60%) showed spontaneous Ca2(+)-dependent action potentials (APs) while silent cells (40%) were excitable by depolarizing current pulses. Bath application of 10 nM DA evoked a hyperpolarizing response, blocked spontaneous APs and decrease the amplitude of evoked APs. Only the hyperpolarizing response faded during the course of the whole cell recording experiments. Under voltage-clamp conditions, DA induced a reversible increase in both voltage-dependent outward K+ currents, without modifying their thresholds. Steady-state inactivation of IA was not affected by DA. These DA-induced responses were dose-dependent and they involved D2 receptor activation. They were mimicked by the specific D2 receptor agonist bromocriptine (10 nM) and blocked by the specific D2 receptor antagonist sulpiride (100 nM), the D1 antagonist SCH 23390 being ineffective. The ability of DA to increase voltage-dependent K+ currents cannot be observed without GTP in the recording pipette. It was pertussis-toxin-sensitive but was affected neither by bath application of 1 mM forskolin nor by the presence of 500 microM cyclic AMP with 500 microM 3-isobutyl-1-methylxanthine in the pipette solutions. We conclude that in lactotroph cells DA specifically increases two voltage-dependent K+ currents via a pertussis-toxin-sensitive guanine nucleotide regulatory protein and appears to be independent of intracellular cyclic AMP. This effect leads to a decrease in the excitability of the cell, explaining in part the inhibitory effect of DA on prolactin release.

4-Aminopyridine↗
PubMed1990

Dopamine inhibits two characterized voltage-dependent calcium currents in identified rat lactotroph cells.

The effects of dopamine (DA) on voltage-dependent Ca2+ currents were investigated in cultured rat lactotroph cells using the patch clamp recording technique. Each recorded cell was identified by the reverse hemolytic plaque assay. In the whole-cell configuration, two types of Ca2+ currents, L and T, were characterized on the basis of their kinetics, voltage sensitivity, and pharmacology. The L component had a threshold of -25 mV, showed little inactivation during a 150-msec voltage step, and was maximal at +10 mV. Cadmium ions (100 microM) significantly reduced its amplitude (75%). The T component was activated at a membrane potential close to -50 mV, was maximal at -10 mV, and showed a voltage-dependent inactivation between -90 and -30 mV. It was quickly inactivated during a maintained depolarization (time constant, 27 ms at -30 mV) and was strongly reduced (80%) by nickel ions (100 microM). Bath application of DA (10 nM) caused a markedly general depression of inward Ca2+ currents, acting differently on the T- and L-type currents. DA application shifted the voltage-dependence of the L-type current activation toward depolarization values (8 mV) without modifying its time- and voltage-dependent inactivation. In contrast, DA enhanced the inactivation of the T-type current by accelerating its time-dependent inactivation (25% decrease in the time constant of inactivation) and by shifting the voltage-dependence of the T-type current inactivation toward hyperpolarizing values (-63 mV in control vs. -77 mV in the presence of DA). These effects of DA were dose-dependent and involved the activation of a D2 receptor type. They were mimicked by bromocriptine application (10 nM), whereas sulpiride (100 nM) blocked the DA-evoked response. The D1 antagonist SCH 23390 was ineffective up to 100 microM. All of these DA-induced modifications in Ca2+ currents were abolished using a GTP-free pipette solution or after pretreatment of cells with pertussis toxin, suggesting that DA can regulate the function of Ca2+ channels through GTP-binding proteins (G-proteins). Our results show that DA acts simultaneously by reducing both voltage-dependent Ca2+ currents on lactotroph cells. Thus, DA reduces the entry of Ca2+ ions across the surface membrane and thereby influences electrical activity and the cytosolic free Ca2+ concentration involved in both basal and evoked PRL release.

Animals↗
PubMed1988

Excitatory effect of noradrenaline on pacemaker cells in spinal cord primary cultures.

Intracellular recordings were made from dissociated fetal mouse spinal cord neurons in primary culture. One particular type of neuron, with a large cell body (40-50 micron) and three to five thick neurites, exhibited rhythmic electrical activity of two different types, consisting of either spontaneous burst discharges or tonic action potential firing. Both types of activity appeared to be triggered by an endogenous membrane potential oscillation. Micropressure application of noradrenaline (10(-5) M in the delivery pipette) onto the surface of such cells evoked, in a dose-dependent manner, an increase in the input resistance with a depolarization of the membrane potential. The response to NA was potential-dependent. The maximum change in input resistance was observed at membrane potential values between -60 mV and -45 mV and the response was suppressed at membrane potentials lower than -80 mV. No modification of the response was observed in the presence of 50 mM of tetraethylammonium. The extrapolated reversal potential, close to -90 mV, was modified by increasing extracellular K+ concentration and unaltered by increasing the intracellular Cl- concentration. The decrease in K+ conductance induced by noradrenaline was Ca2+-dependent and reversibly suppressed by Ba2+ (6 mM) and Cd2+ (0.1 mM). This response to noradrenaline was suppressed in the presence of muscarine (10 microM) suggesting that noradrenaline decreases a K+ conductance related to M current. The noradrenaline evoked increase in input resistance was mediated by activation of an alpha 1 receptor site. Prazosin, an alpha 1 antagonist and phentolamine, an alpha 1 alpha 2 antagonist, reversibly suppressed the response in a competitive manner. Yohimbine, a competitive alpha 2 antagonist, also blocked the response, but in a noncompetitive manner. Clonidine, an alpha 2 agonist, isoprenaline, a beta agonist and L-alprenolol, a beta antagonist, had no effect.

Action Potentials↗
PubMed1987

[Development of electrical activity of hypothalamic cells in culture].

The development of the electrical activity of hypothalamic cells was studied using intracellular and patch clamp recording technics on cultured hypothalamic neurones from 14 days mouse embryos. After 24 h of incubation, 15% of recorded cells were spontaneously active. During the first five days of culture this ratio increased exponentially to reach 95% at day 5. Between the 5th and the 9th day the spontaneous activity progressively decreased although the majority of cells remained excitable. Spontaneous activity reappeared after the 9th day and was underlain by a synaptic potential activity. During the first five days of culture, only a TTX sensitive inward current was observed in all cell tested. A calcium inward current appeared after the first week of incubation. It was recorded on 40% of cells at day 11th and in 80% of cells tested after a month of incubation. Our results show that electrical activity of cultured hypothalamic neurones develop in three stages. A first stage characterized by a spontaneous electrical activity without post synaptic potential, an intermediate stage during which a Calcium inward current appeared corresponding to synaptic contact development and a third stage during which post synaptic potential activity was observed, corresponding to synaptic contact maturation.

Animals↗
PubMed1985

Evidence for bursting pacemaker neurones in cultured spinal cord cells.

Intracellular recordings were made from dissociated mouse spinal cord cells in primary culture. One type of spinal cord neurone, with a large cell body (40-50 micron), 3-5 short neurites, and a mean resting potential of -65 mV, was found to fire rhythmic bursts of action potentials with a phase duration of approximately 1s when the membrane potential was depolarized to -55 mV. These bursts did not arise from spontaneous synaptic input, but appeared to result from endogenous ionic conductance properties of the membrane resembling those observed in molluscan bursting pacemaker neurones. Ionic conductances underlying this bursting activity were studied pharmacologically by local application of ionic conductance blockers. Pacemaker potentials depended on Na+ conductance, since tetrodotoxin and Na-free medium were the most potent agents for blocking spontaneous rhythmic activity. However, a Ca2+ conductance was involved in the depolarizing phase of membrane potential oscillations, since Ba2+ application increased oscillation amplitude. Action potentials observed during the bursts were Na+- and Ca2+-dependent. They did not differ significantly from those observed in other spinal cord neurones in culture. Application of tetraethylammonium, CoCl2, BaCl2 and 4-aminopyridine revealed at least three different potassium conductances which controlled this bursting pacemaker activity. A delayed potassium conductance controlled spike duration, a Ca-dependent potassium conductance controlled the duration of the burst and underlay the hyperpolarizing phase terminating the burst, and finally, a transient potassium conductance appeared to be involved in the control of phase duration. The demonstration that spinal cord neurones growing in monolayer culture display typical bursting pacemaker activity raises the possibility that bursting pacemaker neurones in the mammalian spinal cord may be involved in a phasic pattern generator that could control such activities as walking and the respiratory rhythm.

Action Potentials↗
PubMed1977

[Treatment of bacterial endocarditis with oral amoxicillin and intra-muscular gentamicin].

Fourteen patients suffering from bacterial endocarditis due to a streptococcus or staphylococcus were treated using a combination of amoxicillin per os in a dose of 1 gram every 2 or 3 hours and gentamicin in a dose of 60 mg intramuscularly every 6 or 8 hours. Two patients failed to tolerate amoxicillin, which had to be replaced by penicillin G. Two others, after a period of improvement, relapsed and were cured by the substitution of penicillin G given intravenously, in place of amoxicillin. The ten remaining patients were cured after a normal period of time had elapsed. Two of them were even treated at home. Bactericidal powers of serum obtained by the combination were satisfactory at between 1/16 to 1/4096 one hour after the administration of the antibiotics. This therapeutic protocol is thus effective, and has the advantage of improving the patient's comfort. It should nevertheless be reserved for use against sensitive organisms in patients without digestive problems, the bactericidal power of the serum being verified.

Administration, Oral↗
PubMed1975

[Familial amyloid neuropathy of Corino Andrade. Ultrastructural study of the peripheral nerve in 2 patients].

The authors describe two typical cases of Portuguese amyloid neuropathy in immigrants. One of the patients had been ill for only a short time while the other's condition had been developing over more than ten years. The first patient's neuropathy was characterized by a perforating ulcer of the foot and loss of sensation. The neurological disorder as well as the effect on general health were much more serious in the second patient. Neuro-muscular biopsy was carried out on each of these patients and revealed excessive endoneural amyloid and very severe lesions of nerve parenchyma mainly secondary to Wallerian degeneration. The results were compared with the few existing ultrastructural studies on the peripheral nerve in amyloid neuropathies.

Adult↗