Principes du traitement pharmacologique de l'addiction

peer reviewe

A Novel Phase Portrait to Understand Neuronal Excitability

Fifty years ago, Fitzugh introduced a phase portrait that became famous for a twofold reason: it captured in a physiological way the qualitative behavior of Hodgkin-Huxley model and it revealed the power of simple dynamical models to unfold complex firing patterns. To date, in spite of the enormous progresses in qualitative and quantitative neural modeling, this phase portrait has remained the core picture of neuronal excitability. Yet, a major difference between the neurophysiology of 1961 and of 2011 is the recognition of the prominent role of calcium channels in firing mechanisms. We show that including this extra current in Hodgkin-Huxley dynamics leads to a revision of Fitzugh-Nagumo phase portrait that affects in a fundamental way the reduced modeling of neural excitability. The revisited model considerably enlarges the modeling power of the original one. In particular, it captures essential electrophysiological signatures that otherwise require non-physiological alteration or considerable complexication of the classical model. As a basic illustration, the new model is shown to highlight a core dynamical mechanism by which the calcium conductance controls the two distinct firing modes of thalamocortical neurons

How modeling can reconcile apparently discrepant experimental results: the case of pacemaking in dopaminergic neurons.

Midbrain dopaminergic neurons are endowed with endogenous slow pacemaking properties. In recent years, many different groups have studied the basis for this phenomenon, often with conflicting conclusions. In particular, the role of a slowly-inactivating L-type calcium channel in the depolarizing phase between spikes is controversial, and the analysis of slow oscillatory potential (SOP) recordings during the blockade of sodium channels has led to conflicting conclusions. Based on a minimal model of a dopaminergic neuron, our analysis suggests that the same experimental protocol may lead to drastically different observations in almost identical neurons. For example, complete L-type calcium channel blockade eliminates spontaneous firing or has almost no effect in two neurons differing by less than 1% in their maximal sodium conductance. The same prediction can be reproduced in a state of the art detailed model of a dopaminergic neuron. Some of these predictions are confirmed experimentally using single-cell recordings in brain slices. Our minimal model exhibits SOPs when sodium channels are blocked, these SOPs being uncorrelated with the spiking activity, as has been shown experimentally. We also show that block of a specific conductance (in this case, the SK conductance) can have a different effect on these two oscillatory behaviors (pacemaking and SOPs), despite the fact that they have the same initiating mechanism. These results highlight the fact that computational approaches, besides their well known confirmatory and predictive interests in neurophysiology, may also be useful to resolve apparent discrepancies between experimental results

Methyl-laudanosine: A new pharmacological tool to investigate the function of small-conductance Ca2+-activated K+ channels

ABSTRACT Small-conductance Ca 2ϩ -activated K ϩ channels (SK channels) underlie the prolonged postspike afterhyperpolarization (AHP) observed in many central neurons and play an important role in modulating neuronal activity. However, a lack of specific and reversible blockers of these channels hampers their study in various experimental conditions. Because previous work has shown that bicuculline salts block these channels, we examined whether related alkaloids, namely laudanosine quaternary derivatives, would produce similar effects. Intracellular recordings were performed on rat midbrain dopaminergic neurons and hippocampus CA1 pyramidal cells. Binding experiments were performed on rat cerebral cortex membranes. Laudanosine, methyl-laudanosine, and ethyl-laudanosine blocked the apamin-sensitive AHP of dopaminergic neurons with mean IC 50 values of 152, 15, and 47 M, respectively. The benzyl and butyl derivatives were less potent. Methyl-laudanosine had no effect on the I h current, action potential parameters, or membrane resistance of dopaminergic cells, or on the decrease in input resistance induced by muscimol, indicating a lack of antagonism at GABA A receptors. Interestingly, 100 M methyllaudanosine induced a significant increase in spiking frequency of dopaminergic neurons but not of CA1 pyramidal cells, suggesting the possibility of regional selectivity. Binding experiments on laudanosine derivatives were in good agreement with electrophysiological data. Moreover, methyl-laudanosine has no affinity for voltage-gated potassium channels, and its affinity for SK channels (IC 50 4 M) is superior to its affinity for muscarinic (IC 50 114 M) and neuronal nicotinic (IC 50 Ն367 M) receptors . Methyl-laudanosine may be a valuable pharmacological tool to investigate the role of SK channels in various experimental models. Other than neurotransmitter receptors and transporters, ion channels constitute an attractive target to develop new drugs that will be active on the central nervous system. Currently, the only ion channel that is well established as a central nervous system target is the voltage-gated Na ϩ channel, which is blocked by antiepileptic drugs such as phenytoin, carbamazepine, and lamotrigine Evidence suggests that SK-channel modulation may be interesting in a range of central nervous system disorders, This work was supported in part by Grant 3.4525.98 from the National Fund for Scientific Research (Brussels, Belgium). This work has been presented in meeting abstract form: Scuvée-Moreau J, Liégeois JF, and Seutin V (2002) Effect of laudanosine derivatives on the apamin-sensitive afterhyperpolarization of rat dopaminergic neurons-identification of methyl-laudanosine as a new specific blocker (Abstract). Fundam Clin Pharmacol 16:67

Profils d'expression différents des canaux Ca2+ somatiques dans les neurones dopaminergiques du mésencéphale

peer reviewedDopaminergic (DA) neurons located in the ventral midbrain continuously generate a slow endogenous pacemaker activity, the mechanism of which is still debated. It has been suggested that, in the substantia nigra pars compacta (SNc), the pacemaking relies more on Ca2+ channels and that the density of L type Ca2+ channels is higher in these DA neurons than in those located in the ventral tegmental area (VTA). This might lead to a higher Ca2+ load in SNc DA neurons, and explain their higher susceptibility to degeneration. However, direct evidence for this hypothesis is lacking. We found that the L-type current and channel density is indeed higher in the somata of rat SNc DA neurons, and that this current undergoes less inactivation in this region. Non stationary fluctuation analysis (NSFA) measurements showed a much higher number of L-type channels in the soma of SNc DA neurons, as well as a smaller single channel conductance, pointing to a possible different molecular identity of L-type channels in DA neurons from the two areas. A major consequence of this is that pacemaking and even more so bursting are associated with a larger Ca2+ entry through L-type channels in SN DA neurons than in their VTA counterparts. Our results establish a molecular and functional difference between two populations of midbrain DA neurons that may contribute to their differential sensitivity to neurodegeneration.Caractérisation des canaux calciques somatiques au sein des neurones dopaminergiques mésencéphalique

Gating kinetics and pharmacological properties of small conductance calcium-activated potassium channels.

peer reviewedSmall conductance calcium-activated potassium (SK) channels are a promising treatment target in atrial fibrillation. However, the functional properties that differentiate SK inhibitors remain poorly understood. The objective of this study was to determine how two unrelated SK channel inhibitors, apamin and AP14145, impact SK channel function in excised inside-out single channel recordings. Surprisingly, both apamin and AP14145 exert much of their inhibition by inducing a class of very long-lived channel closures (apamin: τ c,vl=11.8±7.1 s, and AP14145: τ c,vl=10.3±7.2 s), which were never observed under control conditions. Both inhibitors also induced changes to the three closed and two open durations typical of normal SK channel gating. AP14145 shifted the open duration distribution to favor longer open durations, whereas apamin did not alter open state kinetics. AP14145 also prolonged the two shortest channel closed durations (AP14145: τ c,s=3.50±0.81 ms, and τ c,i=32.0±6.76 ms vs. control: τ c,s=1.59±0.19 ms, and τ c,i=13.5±1.17 ms), thus slowing overall gating kinetics within bursts of channel activity. In contrast, apamin accelerated intra-burst gating kinetics by shortening the two shortest closed durations (τ c,s=0.75±0.10 ms and τ c,i=5.08±0.49 ms), and inducing periods of flickery activity. Finally, AP14145 introduced a unique form of inhibition by decreasing unitary current amplitude. SK channels exhibited two clearly distinguishable amplitudes (control: Ahigh=0.76±0.03 pA, and Alow=0.54±0.03 pA). AP14145 both reduced the fraction of patches exhibiting the higher amplitude (AP14145: 4/9 patches vs. control: 16/16 patches), and reduced the mean low amplitude (0.38±0.03 pA). Here we have demonstrated that both inhibitors introduce very long channel closures, but that each also exhibits unique effects on other components of SK gating kinetics and unitary current. The combination of these effects is likely to be critical for understanding the functional differences of each inhibitor in the context of cyclical Ca2+-dependent channel activation in vivo

Inhibition of KCa2.2 and KCa2.3 channel currents by protonation of outer pore histidine residues

Ion channels are often modulated by changes in extracellular pH, with most examples resulting from shifts in the ionization state of histidine residue(s) in the channel pore. The application of acidic extracellular solution inhibited expressed KCa2.2 (SK2) and KCa2.3 (SK3) channel currents, with KCa2.3 (pIC50 of ∼6.8) being approximately fourfold more sensitive than KCa2.2 (pIC50 of ∼6.2). Inhibition was found to be voltage dependent, resulting from a shift in the affinity for the rectifying intracellular divalent cation(s) at the inner mouth of the selectivity filter. The inhibition by extracellular protons resulted from a reduction in the single-channel conductance, without significant changes in open-state kinetics or open probability. KCa2.2 and KCa2.3 subunits both possess a histidine residue in their outer pore region between the transmembrane S5 segment and the pore helix, with KCa2.3 also exhibiting an additional histidine residue between the selectivity filter and S6. Mutagenesis revealed that the outer pore histidine common to both channels was critical for inhibition. The greater sensitivity of KCa2.3 currents to protons arose from the additional histidine residue in the pore, which was more proximal to the conduction pathway and in the electrostatic vicinity of the ion conduction pathway. The decrease of channel conductance by extracellular protons was mimicked by mutation of the outer pore histidine in KCa2.2 to an asparagine residue. These data suggest that local interactions involving the outer turret histidine residues are crucial to enable high conductance openings, with protonation inhibiting current by changing pore shape

Cent scientifiques répliquent à SEA (Suppression des Expériences sur l’Animal vivant) et dénoncent sa désinformation

La lutte contre la maltraitance animale est sans conteste une cause moralement juste. Mais elle ne justifie en rien la désinformation à laquelle certaines associations qui s’en réclament ont recours pour remettre en question l’usage de l’expérimentation animale en recherche