Neuronal communication imposes a heavy metabolic burden in maintaining ionic gradients essential for action potential firing and synaptic signaling. Although cellular metabolism is known to regulate excitatory neurotransmission, it is still unclear whether the brain’s energy supply affects inhibitory signaling. Here we show that mitochondrial-derived reactive oxygen species (mROS) regulate the strength of postsynaptic GABAA receptors at inhibitory synapses of cerebellar stellate cells. Inhibition is strengthened through a mechanism that selectively recruits α3-containing GABAA receptors into synapses with no discernible effect on resident α1-containing receptors. Since mROS promotes the emergence of postsynaptic events with unique kinetic properties, we conclude that newly-recruited α3-containing GABAA receptors are activated by neurotransmitter released onto discrete postsynaptic sites. Although traditionally associated with oxidative stress in neurodegenerative disease, our data identifies mROS as a putative homeostatic signaling molecule coupling cellular metabolism to the strength of inhibitory transmission.

Adverse CNS effects account for a sizeable proportion of all drug attrition cases. These adverse CNS effects are mediated predominately by off-target drug activity on neuronal ion-channels, receptors, transporters and enzymes - altering neuronal function and network communication. In response to these concerns, there is growing support within the pharmaceutical industry for the requirement to perform more comprehensive CNS safety testing prior to first-in-human trials. Accordingly, CNS safety pharmacology commonly integrates several in vitro assay methods for screening neuronal targets in order to properly assess therapeutic safety. One essential assay method is the in vitro electrophysiological technique - the 'gold standard' ion channel assay. The in vitro electrophysiological method is a useful technique, amenable to a variety of different tissues and cell configurations, capable of assessing minute changes in ion channel activity from the level of a single receptor to a complex neuronal network. Recent advances in automated technology have further expanded the usefulness of in vitro electrophysiological methods into the realm of high-throughput, addressing the bottleneck imposed by the manual conduct of the technique. However, despite a large range of applications, manual and automated in vitro electrophysiological techniques have had a slow penetrance into the field of safety pharmacology. Nevertheless, developments in throughput capabilities and in vivo applicability have led to a renewed interest in in vitro electrophysiological techniques that, when complimented by more traditional safety pharmacology methods, often increase the preclinical predictability of potential CNS liabilities.

Activity-dependent strengthening of central synapses is a key factor driving neuronal circuit behavior in the vertebrate CNS. At fast inhibitory synapses, strengthening is thought to occur by increasing the number of GABA A receptors (GABARs) of the same subunit composition to preexisting synapses. Here, we show that strengthening of mouse cerebellar granule cell GABAergic synapses occurs by a different mechanism. Specifically, we show that the neuropeptide hormone, insulin, strengthens inhibitory synapses by recruiting ␣6-containing GABARs rather than accumulating more ␣1-containing receptors that are resident to the synapse. Because ␣6-receptors are targeted to functionally distinct postsynaptic sites from ␣1-receptors, we conclude that only a subset of all inhibitory synapses are strengthened. Together with our recent findings on stellate cells, we propose a general mechanism by which mature inhibitory synapses are strengthened. In this scenario, ␣1-GABARs resident to inhibitory synapses form the hardwiring of neuronal circuits with receptors of a different composition fulfilling a fundamental, but unappreciated, role in synapse strengthening.

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Voltage gated ion channels are central in defining the fundamental properties of the ventricular cardiac action potential (AP), and are also involved in the development of drug-induced arrhythmias. Many drugs can inhibit cardiac ion currents, including the Na current (I), L-type Ca current (Ica-L), and K currents (I, I, I, and I), and thereby affect AP properties in a manner that can trigger or sustain cardiac arrhythmias. Since publication of ICH E14 and S7B over a decade ago, there has been a focus on drug effects on QT prolongation clinically, and on the rapidly activating delayed rectifier current (I), nonclinically, for evaluation of proarrhythmic risk. This focus on QT interval prolongation and a single ionic current likely impacted negatively some drugs that lack proarrhythmic liability in humans. To rectify this issue, the Comprehensive in vitro proarrhythmia assay (CiPA) initiative has been proposed to integrate drug effects on multiple cardiac ionic currents with in silico modelling of human ventricular action potentials, and in vitro data obtained from human stem cell-derived ventricular cardiomyocytes to estimate proarrhythmic risk of new drugs with improved accuracy. In this review, we present the physiological functions and the molecular basis of major cardiac ion channels that contribute to the ventricle AP, and discuss the CiPA paradigm in drug development.