Spiking activity in the visual thalamus is coupled to pupil dynamics across temporal scales.

The processing of sensory information, even at early stages, is influenced by the internal state of the animal. Internal states, such as arousal, are often characterized by relating neural activity to a single "level" of arousal, defined by a behavioral indicator such as pupil size. In this study, we expand the understanding of arousal-related modulations in sensory systems by uncovering multiple timescales of pupil dynamics and their relationship to neural activity. Specifically, we observed a robust coupling between spiking activity in the mouse dorsolateral geniculate nucleus (dLGN) of the thalamus and pupil dynamics across timescales spanning a few seconds to several minutes. Throughout all these timescales, 2 distinct spiking modes-individual tonic spikes and tightly clustered bursts of spikes-preferred opposite phases of pupil dynamics. This multi-scale coupling reveals modulations distinct from those captured by pupil size per se, locomotion, and eye movements. Furthermore, coupling persisted even during viewing of a naturalistic movie, where it contributed to differences in the encoding of visual information. We conclude that dLGN spiking activity is under the simultaneous influence of multiple arousal-related processes associated with pupil dynamics occurring over a broad range of timescales.

Spontaneous activity in cortical neurons is stereotyped and non-Poisson.

Neurons fire even in the absence of sensory stimulation or task demands. Numerous theoretical studies have modeled this spontaneous activity as a Poisson process with uncorrelated intervals between successive spikes and a variance in firing rate equal to the mean. Experimental tests of this hypothesis have yielded variable results, though most have concluded that firing is not Poisson. However, these tests say little about the ways firing might deviate from randomness. Nor are they definitive because many different distributions can have equal means and variances. Here, we characterized spontaneous spiking patterns in extracellular recordings from monkey, cat, and mouse cerebral cortex neurons using rate-normalized spike train autocorrelation functions (ACFs) and a logarithmic timescale. If activity was Poisson, this function should be flat. This was almost never the case. Instead, ACFs had diverse shapes, often with characteristic peaks in the 1-700 ms range. Shapes were stable over time, up to the longest recording periods used (51 min). They did not fall into obvious clusters. ACFs were often unaffected by visual stimulation, though some abruptly changed during brain state shifts. These behaviors may have their origin in the intrinsic biophysics and dendritic anatomy of the cells or in the inputs they receive.

In vivo extracellular recordings of thalamic and cortical visual responses reveal V1 connectivity rules.

The brain's connectome provides the scaffold for canonical neural computations. However, a comparison of connectivity studies in the mouse primary visual cortex (V1) reveals that the average number and strength of connections between specific neuron types can vary. Can variability in V1 connectivity measurements coexist with canonical neural computations? We developed a theory-driven approach to deduce V1 network connectivity from visual responses in mouse V1 and visual thalamus (dLGN). Our method revealed that the same recorded visual responses were captured by multiple connectivity configurations. Remarkably, the magnitude and selectivity of connectivity weights followed a specific order across most of the inferred connectivity configurations. We argue that this order stems from the specific shapes of the recorded contrast response functions and contrast invariance of orientation tuning. Remarkably, despite variability across connectivity studies, connectivity weights computed from individual published connectivity reports followed the order we identified with our method, suggesting that the relations between the weights, rather than their magnitudes, represent a connectivity motif supporting canonical V1 computations.

Robust effects of corticothalamic feedback and behavioral state on movie responses in mouse dLGN.

Neurons in the dorsolateral geniculate nucleus (dLGN) of the thalamus receive a substantial proportion of modulatory inputs from corticothalamic (CT) feedback and brain stem nuclei. Hypothesizing that these modulatory influences might be differentially engaged depending on the visual stimulus and behavioral state, we performed in vivo extracellular recordings from mouse dLGN while optogenetically suppressing CT feedback and monitoring behavioral state by locomotion and pupil dilation. For naturalistic movie clips, we found CT feedback to consistently increase dLGN response gain and promote tonic firing. In contrast, for gratings, CT feedback effects on firing rates were mixed. For both stimulus types, the neural signatures of CT feedback closely resembled those of behavioral state, yet effects of behavioral state on responses to movies persisted even when CT feedback was suppressed. We conclude that CT feedback modulates visual information on its way to cortex in a stimulus-dependent manner, but largely independently of behavioral state.

Corticothalamic feedback sculpts visual spatial integration in mouse thalamus.

En route from the retina to the cortex, visual information passes through the dorsolateral geniculate nucleus (dLGN) of the thalamus, where extensive corticothalamic (CT) feedback has been suggested to modulate spatial processing. How this modulation arises from direct excitatory and indirect inhibitory CT feedback pathways remains enigmatic. Here, we show that in awake mice, retinotopically organized cortical feedback sharpens receptive fields (RFs) and increases surround suppression in the dLGN. Guided by a network model indicating that widespread inhibitory CT feedback is necessary to reproduce these effects, we targeted the visual sector of the thalamic reticular nucleus (visTRN) for recordings. We found that visTRN neurons have large RFs, show little surround suppression and exhibit strong feedback-dependent responses to large stimuli. These features make them an ideal candidate for mediating feedback-enhanced surround suppression in the dLGN. We conclude that cortical feedback sculpts spatial integration in the dLGN, likely via recruitment of neurons in the visTRN.

Voltage distributions in extracellular brain recordings.

Extracellular recordings of brain voltage signals have many uses, including the identification of spikes and the characterization of brain states via analysis of local field potential (LFP) or EEG recordings. Though the factors underlying the generation of these signals are time varying and complex, their analysis may be facilitated by an understanding of their statistical properties. To this end, we analyzed the voltage distributions of high-pass extracellular recordings from a variety of structures, including cortex, thalamus, and hippocampus, in monkeys, cats, and rodents. We additionally investigated LFP signals in these recordings as well as human EEG signals obtained during different sleep stages. In all cases, the distributions were accurately described by a Gaussian within ±1.5 standard deviations from zero. Outside these limits, voltages tended to be distributed exponentially, that is, they fell off linearly on log-linear frequency plots, with variable heights and slopes. A possible explanation for this is that sporadically and independently occurring events with individual Gaussian size distributions can sum to produce approximately exponential distributions. For the high-pass recordings, a second explanation results from a model of the noisy behavior of ion channels that produce action potentials via Hodgkin-Huxley kinetics. The distributions produced by this model, relative to the averaged potential, were also Gaussian with approximately exponential flanks. The model also predicted time-varying noise distributions during action potentials, which were observed in the extracellular spike signals. These findings suggest a principled method for detecting spikes in high-pass recordings and transient events in LFP and EEG signals.NEW & NOTEWORTHY We show that the voltage distributions in brain recordings, including high-pass extracellular recordings, the LFP, and human EEG, are accurately described by a Gaussian within ±1.5 standard deviations from zero, with heavy, exponential tails outside these limits. This offers a principled way of setting event detection thresholds in high-pass recordings. It also offers a means for identifying event-like, transient signals in LFP and EEG recordings which may correlate with other neural phenomena.

LFP clustering in cortex reveals a taxonomy of Up states and near-millisecond, ordered phase-locking in cortical neurons.

During slow-wave sleep and anesthesia, mammalian cortex exhibits a synchronized state during which neurons shift from a largely nonfiring to a firing state, known as an Up-state transition. Up-state transitions may constitute the default activity pattern of the entire cortex (Neske GT. Front Neural Circuits 9: 88, 2016) and could be critical to understanding cortical function, yet the genesis of such transitions and their interaction with single neurons is not well understood. It was recently shown that neurons firing at rates >2 Hz fire spikes in a stereotyped order during Up-state transitions (Luczak A, McNaughton BL, Harris KD. Nat Rev Neurosci 16: 745-755, 2015), yet it is still unknown if Up states are homogeneous and whether spiking order is present in neurons with rates <2 Hz (the majority). Using extracellular recordings from anesthetized cats and mice and from naturally sleeping rats, we show for the first time that Up-state transitions can be classified into several types based on the shape of the local field potential (LFP) during each transition. Individual LFP events could be localized in time to within 1-4 ms, more than an order of magnitude less than in previous studies. The majority of recorded neurons synchronized their firing to within ±5-15 ms relative to each Up-state transition. Simultaneous electrophysiology and wide-field imaging in mouse confirmed that LFP event clusters are cortex-wide phenomena. Our findings show that Up states are of different types and point to the potential importance of temporal order and millisecond-scale signaling by cortical neurons.NEW & NOTEWORTHY During cortical Up-state transitions in sleep and anesthesia, neurons undergo brief periods of increased firing in an order similar to that occurring in awake states. We show that these transitions can be classified into distinct types based on the shape of the local field potential. Transition times can be defined to <5 ms. Most neurons synchronize their firing to within ±5-15 ms of the transitions and fire in a consistent order.

A Visual Guide to Sorting Electrophysiological Recordings Using 'SpikeSorter'.

Few stand-alone software applications are available for sorting spikes from recordings made with multi-electrode arrays. Ideally, an application should be user friendly with a graphical user interface, able to read data files in a variety of formats, and provide users with a flexible set of tools giving them the ability to detect and sort extracellular voltage waveforms from different units with some degree of reliability. Previously published spike sorting methods are now available in a software program, SpikeSorter, intended to provide electrophysiologists with a complete set of tools for sorting, starting from raw recorded data file and ending with the export of sorted spikes times. Procedures are automated to the extent this is currently possible. The article explains and illustrates the use of the program. A representative data file is opened, extracellular traces are filtered, events are detected and then clustered. A number of problems that commonly occur during sorting are illustrated, including the artefactual over-splitting of units due to the tendency of some units to fire spikes in pairs where the second spike is significantly smaller than the first, and over-splitting caused by slow variation in spike height over time encountered in some units. The accuracy of SpikeSorter's performance has been tested with surrogate ground truth data and found to be comparable to that of other algorithms in current development.

Verification of multichannel electrode array integrity by use of cross-channel correlations.

BACKGROUND: The use of multichannel electrode arrays (MEAs) presents a number of practical challenges to experimenters including correctly labelling different recording channel locations and identifying sites that may be non-functional or short-circuited. These challenges are likely to increase as the number of sites used in recording increases. NEW METHOD: This paper presents a simple method for assessing MEA integrity based on the observation that physiologically induced signal correlations between nearby channels fall off with distance. Channels that violate this relationship are flagged as being potentially problematic. RESULTS: The method is able to present to the user a list of potentially faulty channels for further inspection. Underlying problems include non-functional, shorted and mislocalised channels and channels carrying spurious noisy signals unrelated to those on other channels. COMPARISON WITH EXISTING METHODS: Computational methods which automatically screen MEAs for faulty electrode channels do not appear to exist in the literature. Currently a user would have to examine single channels, or channel pairs, individually, which would be very time-consuming. CONCLUSIONS: Shorted or mislocalised channels may be more prevalent in MEA recordings than users suspect. The paper presents a simple screening method for identifying such channels prior to carrying out spike-sorting.

Spike detection methods for polytrodes and high density microelectrode arrays.

This paper compares the ability of different methods to detect and resolve spikes recorded extracellularly with polytrode and high-density microelectrode arrays (MEAs). Detecting spikes on such arrays is more complex than with single electrodes or tetrodes since a single spike from a neuron may cause threshold crossings on several adjacent channels, giving rise to multiple events. These initial events have to be recognized as belonging to a single spike. Combining them is, in essence, a clustering problem. A conflicting need is to be able to resolve spike waveforms that occur close together in space and time. We first evaluated three different detection methods, using simulated data in which spike shape waveforms obtained from real recordings were added to noise with an amplitude and temporal structure similar to that found in real recordings. Performance was assessed by calculating the percentage of correctly identified spikes vs. the false positive rate. Using the best of these detection methods, two different methods for avoiding multiple detections per spike were tested: one based on windowing and the other based on clustering. Using parameters that avoided spatial and temporal duplication, the spatiotemporal resolution of the two methods was next evaluated. The method based on clustering gave slightly better results. Both methods could resolve spikes occurring 1 ms or more apart, regardless of their spatial separation. There was no restriction on the temporal resolution of spike pairs for units more than 200 µm apart.

Spike sorting for polytrodes: a divide and conquer approach.

In order to determine patterns of neural activity, spike signals recorded by extracellular electrodes have to be clustered (sorted) with the aim of ensuring that each cluster represents all the spikes generated by an individual neuron. Many methods for spike sorting have been proposed but few are easily applicable to recordings from polytrodes which may have 16 or more recording sites. As with tetrodes, these are spaced sufficiently closely that signals from single neurons will usually be recorded on several adjacent sites. Although this offers a better chance of distinguishing neurons with similarly shaped spikes, sorting is difficult in such cases because of the high dimensionality of the space in which the signals must be classified. This report details a method for spike sorting based on a divide and conquer approach. Clusters are initially formed by assigning each event to the channel on which it is largest. Each channel-based cluster is then sub-divided into as many distinct clusters as possible. These are then recombined on the basis of pairwise tests into a final set of clusters. Pairwise tests are also performed to establish how distinct each cluster is from the others. A modified gradient ascent clustering (GAC) algorithm is used to do the clustering. The method can sort spikes with minimal user input in times comparable to real time for recordings lasting up to 45 min. Our results illustrate some of the difficulties inherent in spike sorting, including changes in spike shape over time. We show that some physiologically distinct units may have very similar spike shapes. We show that RMS measures of spike shape similarity are not sensitive enough to discriminate clusters that can otherwise be separated by principal components analysis (PCA). Hence spike sorting based on least-squares matching to templates may be unreliable. Our methods should be applicable to tetrodes and scalable to larger multi-electrode arrays (MEAs).

Polytrodes: high-density silicon electrode arrays for large-scale multiunit recording.

We developed a variety of 54-channel high-density silicon electrode arrays (polytrodes) designed to record from large numbers of neurons spanning millimeters of brain. In cat visual cortex, it was possible to make simultaneous recordings from >100 well-isolated neurons. Using standard clustering methods, polytrodes provide a quality of single-unit isolation that surpasses that attainable with tetrodes. Guidelines for successful in vivo recording and precise electrode positioning are described. We also describe a high-bandwidth continuous data-acquisition system designed specifically for polytrodes and an automated impedance meter for testing polytrode site integrity. Despite having smaller interconnect pitches than earlier silicon-based electrodes of this type, these polytrodes have negligible channel crosstalk, comparable reliability, and low site impedances and are capable of making high-fidelity multiunit recordings with minimal tissue damage. The relatively benign nature of planar electrode arrays is evident both histologically and in experiments where the polytrode was repeatedly advanced and retracted hundreds of microns over periods of many hours. It was possible to maintain stable recordings from active neurons adjacent to the polytrode without change in their absolute positions, neurophysiological or receptive field properties.