State-dependent Modulation Research Articles

A Novel High-Throughput Screening Assay for State-Dependent and Subunit-Dependent BK Channel Modulators

BK potassium channels are attractive drug targets for a wide variety of human disorders affecting almost every organ system. However, the therapeutic potential of many BK channel modulators is limited by lack of tissue specificity, reflecting that the pore forming a-subunit (Slo1) is encoded by a single gene. Properties of modulators that can potentially enhance tissue specificity include (a) sensitivity to regulatory subunits that are expressed in a tissue-dependent manner and (b) state-dependent action, conferring sensitivity to tissue-specific differences in membrane voltage and Ca2+ that activate BK channels. We have developed a novel 384-well fluorescent thallium-flux kinetic (FLIPR) assay to identify modulators with these properties. The screen includes assays on a panel of BK channel variants composed of hyperactive human Slo1 mutants (F380Y or R275C) in the presence or absence of different regulatory subunits (β1, β2, β2αFIW, β4, γ1), expressed in U2OS cells with Bacmam virus. Hyperactive Slo1 mutants allow both inhibitors and activators to be reliably detected under resting cellular conditions. Comparison of R275C and F380Y identifies state-dependent modulators since the voltage-sensors of R275C are constitutively activated while F380Y are not. A test screen of small molecules and arachnid venoms revealed a surprising degree of variant-selectivity and identified novel modulators with activities that depend on voltage-sensor activation and/or regulatory subunit expression. Thus, the screen detects modulators that would fail to be detected by conventional screens while also providing information about mechanism of action that should facilitate SAR analysis and development of potent tissue-selective modulators.

Brain state and polarity dependent modulation of brain networks by transcranial direct current stimulation.

Despite its widespread use in cognitive studies, there is still limited understanding of whether and how transcranial direct current stimulation (tDCS) modulates brain network function. To clarify its physiological effects, we assessed brain network function using functional magnetic resonance imaging (fMRI) simultaneously acquired during tDCS stimulation. Cognitive state was manipulated by having subjects perform a Choice Reaction Task or being at “rest.” A novel factorial design was used to assess the effects of brain state and polarity. Anodal and cathodal tDCS were applied to the right inferior frontal gyrus (rIFG), a region involved in controlling activity large‐scale intrinsic connectivity networks during switches of cognitive state. tDCS produced widespread modulation of brain activity in a polarity and brain state dependent manner. In the absence of task, the main effect of tDCS was to accentuate default mode network (DMN) activation and salience network (SN) deactivation. In contrast, during task performance, tDCS increased SN activation. In the absence of task, the main effect of anodal tDCS was more pronounced, whereas cathodal tDCS had a greater effect during task performance. Cathodal tDCS also accentuated the within‐DMN connectivity associated with task performance. There were minimal main effects of stimulation on network connectivity. These results demonstrate that rIFG tDCS can modulate the activity and functional connectivity of large‐scale brain networks involved in cognitive function, in a brain state and polarity dependent manner. This study provides an important insight into mechanisms by which tDCS may modulate cognitive function, and also has implications for the design of future stimulation studies.

The Interplay Between Cortical State and Perceptual Learning: A Focused Review.

Measurements of population activity in alert animals have demonstrated that the intrinsic response state of the cortex has profound effects on the neuronal representation of sensory inputs, raising the possibility that cortical state could influence the behavioral performance in perceptual learning (PL). PL is a process by which sensory experience leads to gradual and semi-permanent improvements in perceptual judgment, and it is generally agreed that these improvements are modulated by sensory cortical areas. Although the precise neural mechanisms underlying the improved perceptual judgment remain unclear, cortical state has been shown to impact the behavioral outcome of PL. We discuss several ways in which cortical state might influence PL based on the recent evidence for state-dependent modulation of sensory encoding. Conversely, training in a certain perceptual task feeds back to modulate cortical state, suggesting a bi-directional relationship between cortical state and behavioral outcomes of PL. We highlight the recent studies that shed light on the mechanism of the interplay between cortical state and PL.

EEG-triggered TMS reveals stronger brain state-dependent modulation of motor evoked potentials at weaker stimulation intensities

BackgroundCorticospinal excitability depends on the current brain state. The recent development of real-time EEG-triggered transcranial magnetic stimulation (EEG-TMS) allows studying this relationship in a causal fashion. Specifically, it has been shown that corticospinal excitability is higher during the scalp surface negative EEG peak compared to the positive peak of μ-oscillations in sensorimotor cortex, as indexed by larger motor evoked potentials (MEPs) for fixed stimulation intensity. ObjectiveWe further characterize the effect of μ-rhythm phase on the MEP input-output (IO) curve by measuring the degree of excitability modulation across a range of stimulation intensities. We furthermore seek to optimize stimulation parameters to enable discrimination of functionally relevant EEG-defined brain states. MethodsA real-time EEG-TMS system was used to trigger MEPs during instantaneous brain-states corresponding to μ-rhythm surface positive and negative peaks with five different stimulation intensities covering an individually calibrated MEP IO curve in 15 healthy participants. ResultsMEP amplitude is modulated by μ-phase across a wide range of stimulation intensities, with larger MEPs at the surface negative peak. The largest relative MEP-modulation was observed for weak intensities, the largest absolute MEP-modulation for intermediate intensities. These results indicate a leftward shift of the MEP IO curve during the μ-rhythm negative peak. ConclusionThe choice of stimulation intensity influences the observed degree of corticospinal excitability modulation by μ-phase. Lower stimulation intensities enable more efficient differentiation of EEG μ-phase-defined brain states.

State-Dependent Modulation of Visual Evoked Potentials in a Rodent Genetic Model of Electroencephalographic Instability

Despite normal sleep timing and duration, Egr3-deficient (Egr3−/−) mice exhibit electroencephalographic (EEG) characteristics of reduced arousal, including elevated slow wave (1–4 Hz) activity during wakefulness. Here we show that these mice exhibit state-dependent instability in the EEG. Intermittent surges in EEG power were found in Egr3−/− mice during wakefulness and rapid eye movement sleep, most prominently in the beta (15–35 Hz) range compared to wild type (Egr3+/+) mice. Such surges did not coincide with sleep onset, as the surges were not associated with cessation of electromyographic tone. Cortical processing of sensory information by visual evoked responses (VEP) were found to vary as a function of vigilance state, being of higher magnitude during slow wave sleep (SWS) than wakefulness and rapid eye movement sleep. VEP responses were significantly larger during quiet wakefulness than active wakefulness, in both Egr3−/− mice and Egr3+/+ mice. EEG synchronization in the beta range, previously linked to the accumulation of sleep need over time, predicted VEP magnitude. Egr3−/− mice not only displayed elevated beta activity, but in quiet wake, this elevated beta activity coincides with an elevated evoked response similar to that of animals in SWS. These data confirm that (a) VEPs vary as a function of vigilance state, and (b) beta activity in the EEG is a predictor of state-dependent modulation of visual information processing. The phenotype of Egr3−/− mice indicates that Egr3 is a genetic regulator of these phenomena.

State-Dependent Modification of Sensory Sensitivity via Modulation of Backpropagating Action Potentials.

Neuromodulators play a critical role in sensorimotor processing via various actions, including pre- and postsynaptic signal modulation and direct modulation of signal encoding in peripheral dendrites. Here, we present a new mechanism that allows state-dependent modulation of signal encoding in sensory dendrites by neuromodulatory projection neurons. We studied the impact of antidromic action potentials (APs) on stimulus encoding using the anterior gastric receptor (AGR) neuron in the heavily modulated crustacean stomatogastric ganglion (STG). We found that ectopic AP initiation in AGR's axon trunk is under direct neuromodulatory control by the inferior ventricular (IV) neurons, a pair of descending projection neurons. IV neuron activation elicited a long-lasting decrease in AGR ectopic activity. This modulation was specific to the site of AP initiation and could be mimicked by focal application of the IV neuron co-transmitter histamine. IV neuron actions were diminished after blocking H2 receptors in AGR's axon trunk, suggesting a direct axonal modulation. This local modulation did not affect the propagation dynamics of en passant APs. However, decreases in ectopic AP frequency prolonged sensory bursts elicited distantly near AGR's dendrites. This frequency-dependent effect was mediated via the reduction of antidromic APs, and the diminishment of backpropagation into the sensory dendrites. Computational models suggest that invading antidromic APs interact with local ionic conductances, the rate constants of which determine the sign and strength of the frequency-dependent change in sensory sensitivity. Antidromic APs therefore provide descending projection neurons with a means to influence sensory encoding without affecting AP propagation or stimulus transduction.

Circadian and Homeostatic Modulation of Multi-Unit Activity in Midbrain Dopaminergic Structures

Although the link between sleep disturbances and dopamine (DA)-related neurological and neuropsychiatric disorders is well established, the impact of sleep alterations on neuronal activity of midbrain DA-ergic structures is currently unknown. Here, using wildtype C57Bl mice, we investigated the circadian- and sleep-related modulation of electrical neuronal activity in midbrain ventral-tegmental-area (VTA) and substantia nigra (SN). We found no significant circadian modulation of activity in SN while VTA displayed a low amplitude but significant circadian modulation with increased firing rates during the active phase. Combining neural activity recordings with electroencephalogram (EEG) recordings revealed a strong vigilance state dependent modulation of neuronal activity with increased activity during wakefulness and rapid eye movement sleep relative to non-rapid eye movement sleep in both SN and VTA. Six-hours of sleep deprivation induced a significant depression of neuronal activity in both areas. Surprisingly, these alterations lasted for up to 48 hours and persisted even after the normalization of cortical EEG waves. Our results show that sleep and sleep disturbances significantly affect neuronal activity in midbrain DA structures. We propose that these changes in neuronal activity underlie the well-known relationship between sleep alterations and several disorders involving dysfunction of the DA circuitry such as addiction and depression.

Evidence of soft bound behaviour in analogue memristive devices for neuromorphic computing

The development of devices that can modulate their conductance under the application of electrical stimuli constitutes a fundamental step towards the realization of synaptic connectivity in neural networks. Optimization of synaptic functionality requires the understanding of the analogue conductance update under different programming conditions. Moreover, properties of physical devices such as bounded conductance values and state-dependent modulation should be considered as they affect storage capacity and performance of the network. This work provides a study of the conductance dynamics produced by identical pulses as a function of the programming parameters in an HfO2 memristive device. The application of a phenomenological model that considers a soft approach to the conductance boundaries allows the identification of different operation regimes and to quantify conductance modulation in the analogue region. Device non-linear switching kinetics is recognized as the physical origin of the transition between different dynamics and motivates the crucial trade-off between degree of analog modulation and memory window. Different kinetics for the processes of conductance increase and decrease account for device programming asymmetry. The identification of programming trade-off together with an evaluation of device variations provide a guideline for the optimization of the analogue programming in view of hardware implementation of neural networks.

Heterogeneity in the circadian and homeostatic modulation of multiunit activity in the lateral hypothalamus.

The lateral hypothalamus (LH) is a relatively large hypothalamic structure containing several neurochemically different, but spatially intermingled, neuronal populations. While the role of these neurons in the homeostatic regulation of diverse physiological and behavioral functions such as sleep/wake cycle has been studied extensively, the impact of sleep history on the electrophysiology of the LH and whether this effect is homogenous across LH is unknown. By combining multiunit activity (MUA) recordings in different regions of LH with electroencephalogram recordings in freely moving rats, we unravelled a heterogeneity of neural-activity patterns within different subregions of LH. This heterogeneity was evident in both the circadian and the vigilance state-dependent modulation of MUA. Interestingly, and consistent with this heterogeneity under baseline conditions, the magnitude of MUA suppression following 6 hr of sleep deprivation (SD) was also different within different locations of LH. Unlike the cortex and in contrast to the predictions of the synaptic homeostatic hypothesis, no correlation was found between the magnitude of activity increase during SD and the percentage of suppression of MUA during recovery sleep. These data provide in vivo evidence of a functional heterogeneity in the circadian and homeostatic modulation of neuronal activity in LH.

Precisely Timed Nicotinic Activation Drives SST Inhibition in Neocortical Circuits

Sleep, waking, locomotion, and attention are associated with cell-type-specific changes in neocortical activity. The effect of brain state on circuit output requires understanding of how neuromodulators influence specific neuronal classes and their synapses, with normal patterns of neuromodulator release from endogenous sources. We investigated the state-dependent modulation of a ubiquitous feedforward inhibitory motif in mouse sensory cortex, local pyramidal (Pyr) inputs onto somatostatin (SST)-expressinginterneurons. Paired whole-cell recordings in acute brain slices and invivo showed that Pyr-to-SST synapses are remarkably weak, with failure rates approaching 80%. Pharmacological screening revealed that cholinergic agonists uniquely enhance synaptic efficacy. Brief, optogenetically gated acetylcholine release dramatically enhanced Pyr-to-SST input, via nicotinic receptors and presynaptic PKA signaling. Importantly, endogenous acetylcholine release preferentially activated nicotinic, not muscarinic, receptors, thus differentiating drug effects from endogenous neurotransmission. Brain state- and synapse-specific unmasking of synapses may be a powerful way to functionally rewire cortical circuits dependent on behavioral demands.

Spatial Working Memory in Male Rats: Pre-Experience and Task Dependent Roles of Dopamine D1- and D2-Like Receptors.

The dopaminergic system is known to be involved in working memory processed by several brain regions like prefrontal cortex (PFC), hippocampus, striatum. In an earlier study we could show that Levodopa but not Modafinil enhanced working memory in a T-maze only during the early phase of training (day 3), whereas the later phase remained unaffected. Rats treated with a higher dose performed better than low dose treated rats. Here we could more specifically segregate the contributions of dopamine type 1- and 2- like receptors (D1R; D2R) to the training state dependent modulation of spatial working memory by intracerebroventricular (ICV) application of a D1R-like (SKF81297) and D2R-like agonist (Sumanirole) and antagonist (SCH23390, Remoxipride) at a low and high dose through 3 days of training. The D1R-like-agonist at both doses enhanced working memory at day 1 but only in the low dose treated rats enhancement persists over training compared to control rats. Rats treated with a high dose of a D1R-like-antagonist show persistent enhancement of working memory over training, whereas in low dose treated rats no statistical difference at any time point could be determined compared to controls. The D2R-like-agonist at both doses does not show an effect at any time point when compared to control animals, whereas the D2R-like antagonist at a low dose enhanced working memory at day 2. For the most effective D1R-like agonist, we repeated the experiments in a water maze working memory task, to test for task dependent differences in working memory modulations. Treated rats at both doses did not differ as compared to controls, but the temporal behavioral performance of all groups was different compared to T-maze trained rats. The results are in line with the view that spatial working memory is optimized within a limited range of dopaminergic transmission, however suggest that these ranges vary during spatial training.

Cholinergic Behavior State-Dependent Mechanisms of Neocortical Gain Control: a Neurocomputational Study.

The embodied mammalian brain evolved to adapt to an only partially known and knowable world. The adaptive labeling of the world is critically dependent on the neocortex which in turn is modulated by a range of subcortical systems such as the thalamus, ventral striatum, and the amygdala. A particular case in point is the learning paradigm of classical conditioning where acquired representations of states of the world such as sounds and visual features are associated with predefined discrete behavioral responses such as eye blinks and freezing. Learning progresses in a very specific order, where the animal first identifies the features of the task that are predictive of a motivational state and then forms the association of the current sensory state with a particular action and shapes this action to the specific contingency. This adaptive feature selection has both attentional and memory components, i.e., a behaviorally relevant state must be detected while its representation must be stabilized to allow its interfacing to output systems. Here, we present a computational model of the neocortical systems that underlie this feature detection process and its state-dependent modulation mediated by the amygdala and its downstream target the nucleus basalis of Meynert. In particular, we analyze the role of different populations of inhibitory interneurons in the regulation of cortical activity and their state-dependent gating of sensory signals. In our model, we show that the neuromodulator acetylcholine (ACh), which is in turn under control of the amygdala, plays a distinct role in the dynamics of each population and their associated gating function serving the detection of novel sensory features not captured in the state of the network, facilitating the adjustment of cortical sensory representations and regulating the switching between modes of attention and learning.

Attentional modulation of neuronal variability in circuit models of cortex.

The circuit mechanisms behind shared neural variability (noise correlation) and its dependence on neural state are poorly understood. Visual attention is well-suited to constrain cortical models of response variability because attention both increases firing rates and their stimulus sensitivity, as well as decreases noise correlations. We provide a novel analysis of population recordings in rhesus primate visual area V4 showing that a single biophysical mechanism may underlie these diverse neural correlates of attention. We explore model cortical networks where top-down mediated increases in excitability, distributed across excitatory and inhibitory targets, capture the key neuronal correlates of attention. Our models predict that top-down signals primarily affect inhibitory neurons, whereas excitatory neurons are more sensitive to stimulus specific bottom-up inputs. Accounting for trial variability in models of state dependent modulation of neuronal activity is a critical step in building a mechanistic theory of neuronal cognition.

State-dependent modulation of functional connectivity in early blind individuals

Resting-state functional connectivity (RSFC) studies have provided strong evidences that visual deprivation influences the brain's functional architecture. In particular, reduced RSFC coupling between occipital (visual) and temporal (auditory) regions has been reliably observed in early blind individuals (EB) at rest. In contrast, task-dependent activation studies have repeatedly demonstrated enhanced co-activation and connectivity of occipital and temporal regions during auditory processing in EB. To investigate this apparent discrepancy, the functional coupling between temporal and occipital networks at rest was directly compared to that of an auditory task in both EB and sighted controls (SC). Functional brain clusters shared across groups and cognitive states (rest and auditory task) were defined. In EBs, we observed higher occipito-temporal correlations in activity during the task than at rest. The reverse pattern was observed in SC. We also observed higher temporal variability of occipito-temporal RSFC in EB suggesting that occipital regions in this population may play the role of a multiple demand system. Our study reveals how the connectivity profile of sighted and early blind people is differentially influenced by their cognitive state, bridging the gap between previous task-dependent and RSFC studies. Our results also highlight how inferring group-differences in functional brain architecture solely based on resting-state acquisition has to be considered with caution.

State-Dependent Allosteric Inhibition of the Human SLC13A5 Citrate Transporter by Hydroxysuccinic Acids, PF-06649298 and PF-06761281

In the liver, citrate is a key metabolic intermediate involved in the regulation of glycolysis and lipid synthesis and reduced expression of the hepatic citrate SLC13A5 transporter has been shown to improve metabolic outcomes in various animal models. Although inhibition of hepatic extracellular citrate uptake through SLC13A5 has been suggested as a potential therapeutic approach for Type-2 diabetes and/or fatty liver disease, so far, only a few SLC13A5 inhibitors have been identified. Moreover, their mechanism of action still remains unclear, potentially limiting their utility for in vivo proof-of-concept studies. In this study, we characterized the pharmacology of the recently identified hydroxysuccinic acid SLC13A5 inhibitors, PF-06649298 and PF-06761281, using a combination of 14C-citrate uptake, a membrane potential assay and electrophysiology. In contrast to their previously proposed mechanism of action, our data suggest that both PF-06649298 and PF-06761281 are allosteric, state-dependent SLC13A5 inhibitors, with low-affinity substrate activity in the absence of citrate. As allosteric state-dependent modulators, the inhibitory potency of both compounds is highly dependent on the ambient citrate concentration and our detailed mechanism of action studies therefore, may be of value in interpreting the in vivo effects of these compounds.

Integrative properties and transfer function of cortical neurons initiating absence seizures in a rat genetic model.

Absence seizures are accompanied by spike-and-wave discharges in cortical electroencephalograms. These complex paroxysmal activities, affecting the thalamocortical networks, profoundly alter cognitive performances and preclude conscious perception. Here, using a well-recognized genetic model of absence epilepsy, we investigated in vivo how information processing was impaired in the ictogenic neurons, i.e. the population of cortical neurons responsible for seizure initiation. In between seizures, ictogenic neurons were more prone to generate bursting activity and their firing response to weak depolarizing events was considerably facilitated compared to control neurons. In the course of seizures, information processing became unstable in ictogenic cells, alternating between an increased and a decreased responsiveness to excitatory inputs, depending on the spike and wave patterns. The state-dependent modulation in the excitability of ictogenic neurons affects their inter-seizure transfer function and their time-to-time responsiveness to incoming inputs during absences. Epileptic seizures result from aberrant cellular and/or synaptic properties that can alter the capacity of neurons to integrate and relay information. During absence seizures, spike-and-wave discharges (SWDs) interfere with incoming sensory inputs and preclude conscious experience. The Genetic Absence Epilepsy Rats from Strasbourg (GAERS), a well-established animal model of absence epilepsy, allows exploration of the cellular basis of this impaired information processing. Here, by combining in vivo electrocorticographic and intracellular recordings from GAERS and control animals, we investigated how the pro-ictogenic properties of seizure-initiating cortical neurons modify their integrative properties and input-output operation during inter-ictal periods and during the spike (S-) and wave (W-) cortical patterns alternating during seizures. In addition to a sustained depolarization and an excessive firing rate in between seizures, ictogenic neurons exhibited a pronounced hyperpolarization-activated depolarization compared to homotypic control neurons. Firing frequency versus injected current relations indicated an increased sensitivity of GAERS cells to weak excitatory inputs, without modifications in the trial-to-trial variability of current-induced firing. During SWDs, the W-component resulted in paradoxical effects in ictogenic neurons, associating an increased membrane input resistance with a reduction in the current-evoked firing responses. Conversely, the collapse of cell membrane resistance during the S-component was accompanied by an elevated current-evoked firing relative to W-sequences, which remained, however, lower compared to inter-ictal periods. These findings show a dynamic modulation of ictogenic neurons' intrinsic properties that may alter inter-seizure cortical function and participate in compromising information processing in cortical networks during absences.

Phosphorylation state-dependent modulation of spinal glycine receptors alleviates inflammatory pain.

Diminished inhibitory neurotransmission in the superficial dorsal horn of the spinal cord is thought to contribute to chronic pain. In inflammatory pain, reductions in synaptic inhibition occur partially through prostaglandin E2- (PGE2-) and PKA-dependent phosphorylation of a specific subtype of glycine receptors (GlyRs) that contain α3 subunits. Here, we demonstrated that 2,6-di-tert-butylphenol (2,6-DTBP), a nonanesthetic propofol derivative, reverses inflammation-mediated disinhibition through a specific interaction with heteromeric αβGlyRs containing phosphorylated α3 subunits. We expressed mutant GlyRs in HEK293T cells, and electrophysiological analyses of these receptors showed that 2,6-DTBP interacted with a conserved phenylalanine residue in the membrane-associated stretch between transmembrane regions 3 and 4 of the GlyR α3 subunit. In native murine spinal cord tissue, 2,6-DTBP modulated synaptic, presumably αβ heteromeric, GlyRs only after priming with PGE2. This observation is consistent with results obtained from molecular modeling of the α-β subunit interface and suggests that in α3βGlyRs, the binding site is accessible to 2,6-DTBP only after PKA-dependent phosphorylation. In murine models of inflammatory pain, 2,6-DTBP reduced inflammatory hyperalgesia in an α3GlyR-dependent manner. Together, our data thus establish that selective potentiation of GlyR function is a promising strategy against chronic inflammatory pain and that, to our knowledge, 2,6-DTBP has a unique pharmacological profile that favors an interaction with GlyRs that have been primed by peripheral inflammation.

MGluR-mediated calcium signalling in the thalamic reticular nucleus

The thalamic reticular nucleus (TRN) plays a major role in modulating the transfer of information from the thalamus to the cortex. GABAergic inhibition via the TRN is differentially regulated by metabotropic glutamate receptors (mGluRs) and the effect of mGluRs on the membrane potential, on ion channels, and on the plasticity of electrical coupling of TRN neurons has been studied previously. Although mGluRs are generally known to trigger Ca2+ transients, mGluR-mediated Ca2+-transients in TRN neurons have not yet been investigated. In this study, we show that mGluRs can trigger Ca2+-transients in TRN neurons, that these transients depend on intracellular Ca2+-stores, and are mediated by IP3 receptors. Ca2+ transients caused by the group I mGluR agonist DHPG elicit a current that is sensitive to flufenamic acid and has a reversal potential around −40mV. Our results add mGluR-mediated Ca2+-signalling in the TRN to the state-dependent modulators of the thalamocortical system.

State-Dependent Modulation of Locomotion by GABAergic Spinal Sensory Neurons

The cerebrospinal fluid (CSF) constitutes an interface through which chemical cues can reach and modulate the activity of neurons located at the epithelial boundary within the entire nervous system. Here, we investigate the role and functional connectivity of a class of GABAergic sensory neurons contacting the CSF in the vertebrate spinal cord and referred to as CSF-cNs. The remote activation of CSF-cNs was shown to trigger delayed slow locomotion in the zebrafish larva, suggesting that these cells modulate components of locomotor central pattern generators (CPGs). Combining anatomy, electrophysiology, and optogenetics in vivo, we show that CSF-cNs form active GABAergic synapses onto V0-v glutamatergic interneurons, an essential component of locomotor CPGs. We confirmed that activating CSF-cNs at rest induced delayed slow locomotion in the fictive preparation. In contrast, the activation of CSF-cNs promptly inhibited ongoing slow locomotion. Moreover, selective activation of rostral CSF-cNs during ongoing activity disrupted rostrocaudal propagation of descending excitation along the spinal cord, indicating that CSF-cNs primarily act at the premotor level. Altogether, our results demonstrate how a spinal GABAergic sensory neuron can tune the excitability of locomotor CPGs in a state-dependent manner by projecting onto essential components of the excitatory premotor pool.

Octopaminergic modulation of temporal frequency tuning of a fly visual motion-sensitive neuron depends on adaptation level.

Several recent studies in invertebrates as well as vertebrates have demonstrated that neuronal response characteristics of sensory neurons can be profoundly affected by an animal’s locomotor activity. The functional consequences of such state-dependent modulation have been a matter of intense debate. In flies, a particularly interesting finding was that tethered walking or flying causes not only general response enhancement of visual motion-sensitive neurons, but also broadens their temporal frequency tuning towards higher values. However, in other studies such state-dependent alterations of neuronal tuning functions were not found. We hypothesize that these discrepancies were due to different adaptation levels of the motion-sensitive neurons, resulting from the use of different stimulation protocols. This is plausible, because the strength of adaptation during ongoing stimulation was shown to be affected by chlordimeform (CDM), an agonist of the insect neuromodulator octopamine, which mediates state-dependent modulation. Our results show that CDM causes broadening of the temporal frequency tuning of the blowfly’s visual motion-sensitive H1 neuron only in the adapted state, but not prior to the presentation of adapting motion. Thus, our study indicates that seemingly conflicting results on the locomotor state-dependence of neuronal tuning functions are consistent when considering the neurons’ adaptation level. Moreover, it demonstrates that stimulation history has to be considered when the significance of state-dependent modulation of sensory processing is interpreted.