Changes In Membrane Excitability Research Articles

Decreased levels of s-nitrosylated Cx43 prevent arrhythmogenicity and myocardial infarction upon cardiac stress in duchenne muscular dystrophy.

Connexin43 (Cx43) plays a critical role in action potential propagation and cardiac contractility. In healthy cardiomyocytes, Cx43 is located at the intercalated disk, however, in cardiac diseases such as hypertrophic and dilated cardiomyopathy, it is remodelled to the lateral side of ventricular cardiomyocytes. In Duchenne muscular dystrophy (DMD) mice, lateralized cardiac Cx43 is found forming hemichannels at the plasma membrane that activate upon cardiac stress evoking triggered activity and arrhythmogenic behaviours. S-nitrosylated levels of Cx43 strongly correlate with changes in membrane excitability in cardiomyocytes from DMD mice. To unequivocally demonstrate a direct role of S-nitrosylated Cx43 in DMD cardiomyopathy, we developed a specific knockin mouse line, in which the Cx43 site for S-nitrosylation, cysteine 271, was specifically substituted for a serine (Cx43C271S). We then crossed the C271S mouse line with DMD mice to obtain DMD:Cx43C271S+/- mice with reduced levels of S-nitrosylated Cx43. Littermate mice (4-6-month-old) were used to assess isoproterenol-induced cardiac stress and, consequently, heart dysfunction in WT, DMD and DMD:Cx43C271S+/- mice. DMD mice displayed increased number of arrhythmogenic events, but without changes in RR and QRS complexes within 1-h post-isoproterenol treatment. However, elevated number of deadly arrhythmias, bradycardia, and increased QRS complex duration were detected in DMD mice for 24-h following isoproterenol stimulation. Strikingly, the number of arrhythmogenic events were significantly reduced in DMD:Cx43C271S+/- mice. β-adrenergic stimulation with isoproterenol induced cardiac hypertrophy, myocardial infarction, and 40% mortality in DMD mice, which was prevented in DMD:Cx43C271S+/- mice. Therefore, these findings strongly suggest that S-nitrosylation of Cx43 proteins at site cysteine 271 represents a fundamental nitric oxide-mediated mechanism involved in the induction of arrythmias and myocardial infarction in DMD mice after β-adrenergic stress.

Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X.

Dynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indicators that have been widely applied. Here, we demonstrate the performance of GCaMP-X in chronic Ca2+ imaging of cortical neurons, where GCaMP-X by design is to eliminate the unwanted interactions between the conventional GCaMP and endogenous (apo)CaM-binding proteins. By expressing in adult mice at high levels over an extended time frame, GCaMP-X showed less damage and improved performance in two-photon imaging of sensory (whisker-deflection) responses or spontaneous Ca2+ fluctuations, in comparison with GCaMP. Chronic Ca2+ imaging of one month or longer was conducted for cultured cortical neurons expressing GCaMP-X, unveiling that spontaneous/local Ca2+ transients progressively developed into autonomous/global Ca2+ oscillations. Along with the morphological indices of neurite length and soma size, the major metrics of oscillatory Ca2+, including rate, amplitude and synchrony were also examined. Dysregulations of both neuritogenesis and Ca2+ oscillations became discernible around 2-3 weeks after virus injection or drug induction to express GCaMP in newborn or mature neurons, which were exacerbated by stronger or prolonged expression of GCaMP. In contrast, neurons expressing GCaMP-X were significantly less damaged or perturbed, altogether highlighting the unique importance of oscillatory Ca2+ to neural development and neuronal health. In summary, GCaMP-X provides a viable solution for Ca2+ imaging applications involving long-time and/or high-level expression of Ca2+ probes.

Intrinsic threshold plasticity: cholinergic activation and role in the neuronal recognition of incomplete input patterns.

Activity-dependent changes in membrane excitability are observed in neurons across brain areas and represent a cell-autonomous form of plasticity (intrinsic plasticity; IP) that in itself does not involve alterations in synaptic strength (synaptic plasticity; SP). Non-homeostatic IP may play an essential role in learning, e.g. by changing the action potential threshold near the soma. A computational problem, however, arises from the implication that such amplification does not discriminate between synaptic inputs and therefore may reduce the resolution of input representation. Here, we investigate consequences of IP for the performance of an artificial neural network in (a) the discrimination of unknown input patterns and (b) the recognition of known/learned patterns. While negative changes in threshold potentials in the output layer indeed reduce its ability to discriminate patterns, they benefit the recognition of known but incompletely presented patterns. An analysis of thresholds and IP-induced threshold changes in published sets of physiological data obtained from whole-cell patch-clamp recordings from L2/3 pyramidal neurons in (a) the primary visual cortex (V1) of awake macaques and (b) the primary somatosensory cortex (S1) of mice in vitro, respectively, reveals a difference between resting and threshold potentials of ∼15mV for V1 and ∼25mV for S1, and a total plasticity range of ∼10mV (S1). The most efficient activity pattern to lower threshold is paired cholinergic and electric activation. Our findings show that threshold reduction promotes a shift in neural coding strategies from accurate faithful representation to interpretative assignment of input patterns to learned object categories. KEY POINTS: Intrinsic plasticity may change the action potential threshold near the soma of neurons (threshold plasticity), thus altering the input-output function for all synaptic inputs 'upstream' of the plasticity location. A potential problem arising from this shared amplification is that it may reduce the ability to discriminate between different input patterns. Here, we assess the performance of an artificial neural network in the discrimination of unknown input patterns as well as the recognition of known patterns subsequent to changes in the spike threshold. We observe that negative changes in threshold potentials do reduce discrimination performance, but at the same time improve performance in an object recognition task, in particular when patterns are incompletely presented. Analysis of whole-cell patch-clamp recordings from pyramidal neurons in the primary somatosensory cortex (S1) of mice reveals that negative threshold changes preferentially result from electric stimulation of neurons paired with the activation of muscarinic acetylcholine receptors.

Characterization of Inhibitory Capability on Hyperpolarization-Activated Cation Current Caused by Lutein (β,ε-Carotene-3,3'-Diol), a Dietary Xanthophyll Carotenoid.

Lutein (β,ε-carotene-3,3′-diol), a xanthophyll carotenoid, is found in high concentrations in the macula of the human retina. It has been recognized to exert potential effectiveness in antioxidative and anti-inflammatory properties. However, whether and how its modifications on varying types of plasmalemmal ionic currents occur in electrically excitable cells remain incompletely answered. The current hypothesis is that lutein produces any direct adjustments on ionic currents (e.g., hyperpolarization-activated cation current, Ih [or funny current, If]). In the present study, GH3-cell exposure to lutein resulted in a time-, state- and concentration-dependent reduction in Ih amplitude with an IC50 value of 4.1 μM. There was a hyperpolarizing shift along the voltage axis in the steady-state activation curve of Ih in the presence of this compound, despite being void of changes in the gating charge of the curve. Under continued exposure to lutein (3 μM), further addition of oxaliplatin (10 μM) or ivabradine (3 μM) could be effective at either reversing or further decreasing lutein-induced suppression of hyperpolarization-evoked Ih, respectively. The voltage-dependent anti-clockwise hysteresis of Ih responding to long-lasting inverted isosceles-triangular ramp concentration-dependently became diminished by adding this compound. However, the addition of 10 μM lutein caused a mild but significant suppression in the amplitude of erg-mediated or A-type K+ currents. Under current-clamp potential recordings, the sag potential evoked by long-lasting hyperpolarizing current stimulus was reduced under cell exposure to lutein. Altogether, findings from the current observations enabled us to reflect that during cell exposure to lutein used at pharmacologically achievable concentrations, lutein-perturbed inhibition of Ih would be an ionic mechanism underlying its changes in membrane excitability.

Sarcolemmal Excitability, M-Wave Changes, and Conduction Velocity During a Sustained Low-Force Contraction.

This study was undertaken to investigate whether sarcolemmal excitability is impaired during a sustained low-force contraction [10% maximal voluntary contraction (MVC)] by assessing muscle conduction velocity and also by analyzing separately the first and second phases of the muscle compound action potential (M wave). Twenty-one participants sustained an isometric knee extension of 10% MVC for 3min. M waves were evoked by supramaximal single shocks to the femoral nerve given at 10-s intervals. The amplitude, duration, and area of the first and second M-wave phases were computed. Muscle fiber conduction velocity, voluntary surface electromyographic (EMG), perceived effort, MVC force, peak twitch force, and temperature were also recorded. The main findings were: (1) During the sustained contraction, conduction velocity remained unchanged. (2) The amplitude of the M-wave first phase decreased for the first ~30s (−7%, p<0.05) and stabilized thereafter, whereas the second phase amplitude increased for the initial ~30s (+7%, p<0.05), before stabilizing. (3) Both duration and area decreased steeply during the first ~30s, and then more gradually for the rest of the contraction. (4) During the sustained contraction, perceived effort increased fivefold, whereas knee extension EMG increased by ~10%. (5) Maximal voluntary force and peak twitch force decreased (respectively, −9% and −10%, p<0.05) after the low-force contraction. Collectively, the present results indicate that sarcolemmal excitability is well preserved during a sustained 10% MVC task. A depression of the M-wave first phase during a low-force contraction can occur even in the absence of changes in membrane excitability. The development of fatigue during a low-force contraction can occur without alteration of membrane excitability.

Light/Clock Influences Membrane Potential Dynamics to Regulate Sleep States.

The circadian rhythm is a fundamental process that regulates the sleep–wake cycle. This rhythm is regulated by core clock genes that oscillate to create a physiological rhythm of circadian neuronal activity. However, we do not know much about the mechanism by which circadian inputs influence neurons involved in sleep–wake architecture. One possible mechanism involves the photoreceptor cryptochrome (CRY). In Drosophila, CRY is receptive to blue light and resets the circadian rhythm. CRY also influences membrane potential dynamics that regulate neural activity of circadian clock neurons in Drosophila, including the temporal structure in sequences of spikes, by interacting with subunits of the voltage-dependent potassium channel. Moreover, several core clock molecules interact with voltage-dependent/independent channels, channel-binding protein, and subunits of the electrogenic ion pump. These components cooperatively regulate mechanisms that translate circadian photoreception and the timing of clock genes into changes in membrane excitability, such as neural firing activity and polarization sensitivity. In clock neurons expressing CRY, these mechanisms also influence synaptic plasticity. In this review, we propose that membrane potential dynamics created by circadian photoreception and core clock molecules are critical for generating the set point of synaptic plasticity that depend on neural coding. In this way, membrane potential dynamics drive formation of baseline sleep architecture, light-driven arousal, and memory processing. We also discuss the machinery that coordinates membrane excitability in circadian networks found in Drosophila, and we compare this machinery to that found in mammalian systems. Based on this body of work, we propose future studies that can better delineate how neural codes impact molecular/cellular signaling and contribute to sleep, memory processing, and neurological disorders.

The Role of Parvalbumin Interneuron GIRK Signaling in the Regulation of Affect and Cognition in Male and Female Mice.

Pathological impairments in the regulation of affect (i.e., emotion) and flexible decision-making are commonly observed across numerous neuropsychiatric disorders and are thought to reflect dysfunction of cortical and subcortical circuits that arise in part from imbalances in excitation and inhibition within these structures. Disruptions in GABA transmission, in particular, that from parvalbumin-expressing interneurons (PVI), has been highlighted as a likely mechanism by which this imbalance arises, as they regulate excitation and synchronization of principle output neurons. G protein-gated inwardly rectifying potassium ion (GIRK/Kir3) channels are known to modulate excitability and output of pyramidal neurons in areas like the medial prefrontal cortex and hippocampus; however, the role GIRK plays in PVI excitability and behavior is unknown. Male and female mice lacking GIRK1 in PVI (Girk1flox/flox:PVcre) and expressing td-tomato in PVI (Girk1flox/flox:PVCre:PVtdtom) exhibited increased open arm time in the elevated plus-maze, while males showed an increase in immobile episodes during the forced swim test (FST). Loss of GIRK1 did not alter motivated behavior for an appetitive reward or impair overall performance in an operant-based attention set-shifting model of cognitive flexibility; however it did alter types of errors committed during the visual cue test. Unexpectedly, baseline sex differences were also identified in these tasks, with females exhibiting overall poorer performance compared to males and distinct types of errors, highlighting potential differences in task-related problem-solving. Interestingly, reductions in PVI GIRK signaling did not correspond to changes in membrane excitability but did increase action potential (AP) firing at higher current injections in PVI of males, but not females. This is the first investigation on the role that PVI GIRK-signaling has on membrane excitability, AP firing, and their role on affect and cognition together increasing the understanding of PVI cellular mechanisms and function.

Ion Channels Controlling Circadian Rhythms in Suprachiasmatic Nucleus Excitability.

Animals synchronize to the environmental day-night cycle by means of an internal circadian clock in the brain. In mammals, this timekeeping mechanism is housed in the suprachiasmatic nucleus (SCN) of the hypothalamus and is entrained by light input from the retina. One output of the SCN is a neural code for circadian time, which arises from the collective activity of neurons within the SCN circuit and comprises two fundamental components: 1) periodic alterations in the spontaneous excitability of individual neurons that result in higher firing rates during the day and lower firing rates at night, and 2) synchronization of these cellular oscillations throughout the SCN. In this review, we summarize current evidence for the identity of ion channels in SCN neurons and the mechanisms by which they set the rhythmic parameters of the time code. During the day, voltage-dependent and independent Na+ and Ca2+ currents, as well as several K+ currents, contribute to increased membrane excitability and therefore higher firing frequency. At night, an increase in different K+ currents, including Ca2+-activated BK currents, contribute to membrane hyperpolarization and decreased firing. Layered on top of these intrinsically regulated changes in membrane excitability, more than a dozen neuromodulators influence action potential activity and rhythmicity in SCN neurons, facilitating both synchronization and plasticity of the neural code.

Intrinsic memory of temporal intervals in cerebellar Purkinje cells.

The general consensus for learning and memory, including in the cerebellum, is that modification of synaptic strength via long-term potentiation (LTP) or long-term depression (LTD) are the primary mechanisms for the formation of memories. Recent findings suggest additional cellular mechanisms - referred to as 'intrinsic plasticity' - where a neuron's membrane excitability intrinsically changes. These mechanisms act like a dimmer and alter neuronal responsiveness by adjusting response amplitudes and spike thresholds. Here, I argue that classical conditioning of cerebellar Purkinje cell responses reveals yet another cell-intrinsic learning mechanism which significantly differs from both changes in synaptic strength and changes in membrane excitability. When the conditional (CS) and unconditional stimuli (US) are delivered directly to the Purkinje cell's immediate pre-synaptic afferents, the parallel fibres and the climbing fibre, the cell learns to respond to the CS with a pause in its spontaneous firing that reflects the interval between the two stimuli. The pause response has a delayed onset and adaptively timed maximum, offset and duration, determined by the previously experienced CS-US interval. The timing is not dependent on any network-generated time-varying input. This implies the existence of a timing mechanism and a memory substrate that encodes the duration of the CS-US interval inside the Purkinje cell. Such temporal interval learning is not simply a change that causes more or less firing in response to an input. Here, I review these findings in relation to the standard theory of synaptic strength changes and the network interactions believed to be necessary for generating time codes.

Semaphorin 3A induces acute changes in membrane excitability in spiral ganglion neurons in vitro.

The development and survival of spiral ganglion neurons (SGNs) are dependent on multiple trophic factors as well as membrane electrical activity. Semaphorins (Sema) constitute a family of membrane-associated and secreted proteins that have garnered significant attention as a potential SGN "navigator" during cochlea development. Previous studies using mutant mice demonstrated that Sema3A plays a role in the SGN pathfinding. The mechanisms, however, by which Sema3A shapes SGNs firing behavior are not known. In these studies, we found that Sema3A plays a novel role in regulating SGN resting membrane potential and excitability. Using dissociated SGN from pre-hearing (P3-P5) and post-hearing mice (P12-P15), we recorded membrane potentials using whole-cell patch clamp recording techniques in apical and basal SGN populations. Recombinant Sema3A was applied to examine the effects on intrinsic membrane properties and action potentials evoked by current injections. Apical and basal SGNs from newborn mice treated with recombinant Sema3A (100ng/ml) displayed a higher resting membrane potential, higher threshold, decreased amplitude, and prolonged latency and duration of spikes. Although a similar phenomenon was observed in SGNs from post-hearing mice, the resting membrane potential was essentially indistinguishable before and after Sema3A exposure. Sema3A-mediated changes in membrane excitability were associated with a significant decrease in K+ and Ca2+ currents. Sema3A acts through linopirdine-sensitive K+ channels in apical, but not in the basal SGNs. Therefore, Sema3A induces differential effects in SGN membrane excitability that are dependent on age and location, and constitutes an additional early and novel effect of Sema3A SGNs in vitro.

Sarcolemmal membrane excitability during repeated intermittent maximal voluntary contractions.

What is the central question of this study? Is impaired membrane excitability reflected by an increase or by a decrease in M-wave amplitude? What is the main finding and its importance? The magnitude of the M-wave first and second phases changed in completely different ways during intermittent maximal voluntary contractions, leading to the counterintuitive conclusion that it is an increase (and not a decrease) of the M-wave first phase that reflects impaired membrane excitability. The study was undertaken to investigate separately the changes in the first and second phases of the muscle compound action potential (M-wave) during 4min of intermittent maximal voluntary contractions (MVCs) of the quadriceps. M-waves were evoked by supramaximal single electrical stimulation to the femoral nerve delivered in the resting periods between 48 successive MVCs of 3s. The amplitude, duration and area of the M-wave first and second phases were measured separately, together with muscle conduction velocity and MVC force. During the intermittent MVCs, the amplitude of the M-wave first phase increased uninterruptedly for the first 3min (12-16%, P< 0.05) and stabilized thereafter, whereas the second phase initially increased for 55-75s (11-22%, P< 0.05), but decreased subsequently. The enlargement of the first phase occurred in parallel with an increase in its duration, and concomitantly with a decline in conduction velocity (maximal cross-correlations, 0.89-0.97; time lag, 0s). Also, a significant temporal association was found between the amplitude of the first phase and MVC force (time lag, 0s; maximal cross-correlations, 0.85-0.97). Conversely, there was no temporal association between the second phase amplitude and conduction velocity or MVC force (time lag, 73-117s; maximal cross-correlations, 0.65-0.77). It is concluded that the enlargement of the M-wave first phase is the electrical manifestation of impaired muscle membrane excitability. The results highlight the importance of independently analysing the first and second phases, as only the first phase can be used reliably to detect changes in membrane excitability, while the second might be affected by muscle architecture.

Muscle fiber conduction velocity of the vastus medilais and lateralis muscle after eccentric exercise induced-muscle damage

Change in muscle fiber conduction velocity (MFCV) has been reported after eccentric exercise induces muscle fiber damage, most likely due to a change in membrane permeability of the injured fiber. The extent of damage to the muscle fiber depends on the morphological and architectural characteristics of the muscle fibers. Morphological and architectural characteristics of the VMO muscle fibers are different from VL muscle. Thus, it is expected that eccentric exercise of quadriceps muscle results in a non-uniform fiber damage within the VMO and VL muscle and, as a consequence, non-uniform changes in membrane excitability and conduction velocity. The aim of the study was to investigate MFCV of the VMO and VL muscles before and 24 h after eccentric exercise. Multichannel surface EMG signals were concurrently recorded from the right VMO and VL muscles of 15 healthy men during sustained isometric contractions at 50% of the maximal force. Maximal voluntary force significantly reduced after eccentric exercise with respect to the pre-exercise condition (P < 0.0001). MFCV decreased over time during the sustained contractions at faster rates when assessed 24 h after exercise (VMO = −26.1; VL = −20.1) with respect to the pre-exercise condition (VMO = −9.1; VL = −13.7, P < 0.0001). Moreover, VMO showed a greater rate of reduction in MFCV over sustained contraction (26.1 ± 10.7%) in comparison with VL muscle (20.1 ± 8.5%, P < 0.025) 24 h after eccentric exercise. The result indicates that eccentric exercise contributes to a larger reduction in MFCV within the VMO muscle as compared to the VL muscle. This may abolish the ability of VMO to counteract the lateral pull of the VL muscle during knee extension, thereby leaving the knee complex more vulnerable to injury.

Circadian redox rhythms in the regulation of neuronal excitability.

Oxidation-reduction reactions are essential to life as the core mechanisms of energy transfer. A large body of evidence in recent years presents an extensive and complex network of interactions between the circadian and cellular redox systems. Recent advances show that cellular redox state undergoes a ~24-h (circadian) oscillation in most tissues and is conserved across the domains of life. In nucleated cells, the metabolic oscillation is dependent upon the circadian transcription-translation machinery and, vice versa, redox-active proteins and cofactors feed back into the molecular oscillator. In the suprachiasmatic nucleus (SCN), a hypothalamic region of the brain specialized for circadian timekeeping, redox oscillation was found to modulate neuronal membrane excitability. The SCN redox environment is relatively reduced in daytime when neuronal activity is highest and relatively oxidized in nighttime when activity is at its lowest. There is evidence that the redox environment directly modulates SCN K+ channels, tightly coupling metabolic rhythms to neuronal activity. Application of reducing or oxidizing agents produces rapid changes in membrane excitability in a time-of-day-dependent manner. We propose that this reciprocal interaction may not be unique to the SCN. In this review, we consider the evidence for circadian redox oscillation and its interdependencies with established circadian timekeeping mechanisms. Furthermore, we will investigate the effects of redox on ion-channel gating dynamics and membrane excitability. The susceptibility of many different ion channels to modulation by changes in the redox environment suggests that circadian redox rhythms may play a role in the regulation of all excitable cells.

Peripheral and central sensitization

BACKGROUND AND OBJECTIVES: Central sensitization is an important phenomenon for pain chronicity and is present in neuropathic pain. This study aimed at addressing some pathophysiological aspects of this phenomenon. CONTENTS: Some extra and intracellular aspects responsible for central sensitization genesis, especially phenotypic changes in plasticity of neurons involved in the process are described. CONCLUSION: Pain chronicity may result from changes in central nervous system neurons properties by the central sensitization phenomenon with constant changes in membrane excitability, inhibitory transmission reduction and increase in synaptic efficacy mediated by several convergent and divergent molecular mechanisms over a background of phenotypic and structural changes. Neuroplasticity deeply alters painful sensation, contributing for many clinical painful syndromes and may represent a major target for therapeutic intervention.

Increased excitability and excitatory synaptic transmission during in vitro ischemia in the neonatal mouse hippocampus

Objective: The present study tested the hypothesis that exposure to in vitro hypoxia–ischemia alters membrane properties and excitability as well as excitatory synaptic transmission of CA1 pyramidal neurons in the neonatal mouse. Methods: Experiments were conducted in hippocampal slices in P7–P9 C57Bl/6 mice using whole-cell patch clamp in current- and voltage-clamp mode. Passive membrane potential (Vm), input resistance (Rin) and active (action potential (AP) threshold and amplitude) membrane properties of CA1 pyramidal neurons were assessed at baseline, during 10min in vitro ischemia (oxygen-glucose deprivation (OGD)) and during reoxygenation. Spontaneous and miniature excitatory post-synaptic currents (s and mEPSCs) were studied under similar conditions. Results: OGD caused significant depolarization of CA1 pyramidal neurons as well as decrease in AP threshold and increase in AP amplitude. These changes were blocked by the application of tetrodotoxin (TTX), indicating Na+ channels’ involvement. Following 10min of reoxygenation, significant membrane hyperpolarization was noted and it was associated with a decrease in Rin. AP threshold and amplitude returned to baseline during that stage. sEPSC and mEPSC frequency increased during both OGD and reoxygenation but their amplitude remained unchanged. Additionally, we found that OGD decreases Ih (hyperpolarization activated current) in CA1 neurons from neonatal mice and this effect persists during reoxygenation. Significance: These results indicate that in vitro ischemia leads to changes in membrane excitability mediated by sodium and potassium channels. Further, it results in enhanced neurotransmitter release from presynaptic terminals. These changes are likely to represent one of the mechanisms of hypoxia/ischemia-mediated seizures in the neonatal period.

Neuronal Mitochondrial Trafficking Impairment: The Cause or a Consequence of Neuronal Dysfunction Caused by Amphetamine-Like Drugs

Drugs of abuse cause a variety of complex neuronal events at the cellular level, including changes in membrane excitability and neurotransmission, activation of complex signaling pathways, altered synaptic physiology and structural changes, and drug-evoked synaptic plasticity and neurotoxicity, which mediate both acuteand long-lasting effects and addiction. Neuronal mitochondria are highly dynamic organelles that, by undergoing fusion and fission events, are efficiently translocated along the neuronal processes, frequently changing direction, pausing or switching to persistent docking. The neuronal integrity and functionality are dependent upon the proper maintenance of a healthy mitochondrial population and their efficient distribution. There is a general consensus that mitochondrial-dependent pathways can provide a major understanding concerning pathological processes underlying neurotoxicity of drugs of abuse. As such, it is plausible to consider that alterations on mitochondrial trafficking may be key players on the neuronal effects mediated by these drugs. This work aims to provide a comprehensive and up-to-date review of the data linking mitochondrial trafficking impairment to amphetamine-like drugs, and, thus, contribute to a better understanding of their neuronal effects. Additionally, new research data describing alterations in neuronal mitochondrial trafficking for 3,4-methylenedioxymethamphetamine (MDMA; “ecstasy”) conjugated metabolites 5-(glutathion-S-yl)-N-methyl-α-methyldopamine [5-(GSH)-N-Me-α-MeDA] and 5-(N-acetylcystein-S-yl)-N-methyl-α- methyldopamine [5-(NAC)-N-Me-α-MeDA] are also provided.

Is Potassium a Ubiquitous Mediator of Vasodilation in the Central Nervous System?

Is Potassium a Ubiquitous Mediator of Vasodilation in the Central Nervous System?

P 176. Evidence for high-fidelity timing dependent synaptic plasticity of human motor cortex

A single transcranial magnetic stimulation (TMS) pulse typically evokes a short series of spikes in corticospinal neurons (known as indirect (I)-waves) which are thought to arise from transynaptic input. Delivering a second pulse at inter-pulse intervals (IPIs) corresponding to the timing of these I-waves leads to a facilitation of the response, and if stimulus pairs are delivered repeatedly, a persistent LTP-like increase in excitability can occur. This has been demonstrated at an IPI of 1.5 ms, which corresponds to the first I-wave interval, in an intervention referred to as ITMS (I-wave TMS), and it has been argued that this may have similarities with timing dependent plasticity models. Consequently we hypothesised that if the second stimulus is delivered so as not to coincide with I-wave timing it should lead to Ltd. We performed a cross-over study in ten subjects in which TMS doublets were timed to coincide (1.5 ms IPI, ITMS 1.5 ) or not coincide (2 ms IPI, ITMS 2 ) with I-wave firing. Single pulse motor evoked potential (MEP) amplitude, resting motor threshold (RMT) and short-interval cortical inhibition (SICI) were measured from the first dorsal interosseous (FDI) muscle. After ITMS 1.5 corticomotor excitability was increased by around ∼170% for 15 min ( p < 0.05) and returned to baseline by 20 min. Increasing the IPI by just 500 μs–2 ms reversed the after-effect and MEP amplitude was significantly reduced (−35%, p < 0.05) for 15 min before returning to baseline. This reduction was not associated with an increase in SICI, suggesting a reduction in excitatory transmission rather than an increase in inhibitory efficacy. RMT also remained unchanged suggesting that these changes were not due to changes in membrane excitability. Amplitude-matching ITMS 2 did not modulate excitability. The results are consistent with timing-dependent synaptic LTP/D-like effects, and suggest that there are plasticity mechanisms operating in the human motor cortex with a temporal resolution of the order of a few hundreds of microseconds.

Properties of spinal lamina III GABAergic neurons in naïve and in neuropathic mice

Nerve injury leads to Aβ-fibre-mediated mechanical allodynia that is in part due to an impaired GABAergic inhibition in the spinal cord dorsal horn. The properties and function of GABAergic neurons in spinal cord lamina III, an area where low-threshold mechanosensitive Aβ-fibres terminate are, however, largely unknown. We used transgenic mice, which express enhanced green fluorescent protein (EGFP) under control of the promoter GAD67. The morphology and neurochemical characteristics of GABAergic, EGFP-expressing neurons were characterized. We assessed active and passive membrane properties of spinal lamina III GABAergic neurons in naïve animals and animals with a chronic constriction injury (CCI) of the sciatic nerve. EGFP-expressing neurons in lamina III were predominantly islet cells (47%), whereas non-EGFP-expressing neurons were largely inverted stalked cells (40%). EGFP-expressing neurons accounted for about 25% of GABAergic neurons in lamina III. Forty-four percent co-expressed glycine, 10% neuronal nitric oxide synthase and 3% co-expressed parvalbumin. We found costaining with protein kinase CβII in 42% of EGFP-expressing neurons but no expression of protein kinase Cγ. Membrane properties and excitability of EGFP-and non-EGFP-expressing neurons from naïve and neuropathic animals were indistinguishable. The most frequent firing pattern was tonic firing (naïve: 35%, neuropathic: 37%) followed by gap firing (naïve: 33%, neuropathic: 25%). Delayed, initial burst and single-spike firing patterns made up the remainder in both groups. A change in membrane excitability or discharge pattern of this group of lamina III GABAergic neurons is unlikely the cause for mechanical allodynia in animals with CCI.

Evidence for high-fidelity timing-dependent synaptic plasticity of human motor cortex

A single transcranial magnetic stimulation (TMS) pulse typically evokes a short series of spikes in corticospinal neurons [known as indirect (I)-waves] which are thought to arise from transynaptic input. Delivering a second pulse at inter-pulse intervals (IPIs) corresponding to the timing of these I-waves leads to a facilitation of the response, and if stimulus pairs are delivered repeatedly, a persistent LTP-like increase in excitability can occur. This has been demonstrated at an IPI of 1.5 ms, which corresponds to the first I-wave interval, in an intervention referred to as ITMS (I-wave TMS), and it has been argued that this may have similarities with timing-dependent plasticity models. Consequently, we hypothesized that if the second stimulus is delivered so as not to coincide with I-wave timing, it should lead to LTD. We performed a crossover study in 10 subjects in which TMS doublets were timed to coincide (1.5-ms IPI, ITMS(1.5)) or not coincide (2-ms IPI, ITMS(2)) with I-wave firing. Single pulse motor-evoked potential (MEP) amplitude, resting motor threshold (RMT), and short-interval cortical inhibition (SICI) were measured from the first dorsal interosseous (FDI) muscle. After ITMS(1.5) corticomotor excitability was increased by ~60% for 15 min (P < 0.05) and returned to baseline by 20 min. Increasing the IPI by just 500 μs to 2 ms reversed the aftereffect, and MEP amplitude was significantly reduced (~35%, P < 0.05) for 15 min before returning to baseline. This reduction was not associated with an increase in SICI, suggesting a reduction in excitatory transmission rather than an increase in inhibitory efficacy. RMT also remained unchanged, suggesting that these changes were not due to changes in membrane excitability. Amplitude-matching ITMS(2) did not modulate excitability. The results are consistent with timing-dependent synaptic LTP/D-like effects and suggest that there are plasticity mechanisms operating in the human motor cortex with a temporal resolution of the order of a few hundreds of microseconds.