Extracellular Action Potentials Research Articles

Electrochemical carbon fiber-based technique for simultaneous recordings of brain tissue PO2, pH, and extracellular field potentials

A method for simultaneous electrochemical detection of brain tissue PO2 (PtO2) and pH changes together with neuronal activity using a modified form of fast cyclic voltammetry with carbon fiber electrodes is described. This technique has been developed for in vivo applications and recordings from discrete brain nuclei in experimental animals. The small size of the carbon fiber electrode (⌀7 μm, length <100 μm) ensures minimal disruption of the brain tissue and allows recordings from small brain areas. Sample rate (up to 4 Hz) is sufficient to resolve rapid changes in PtO2 and pH that follow changes in neuronal activity and metabolism. Rapid switching between current and voltage recordings allows combined electrochemical detection and monitoring of extracellular action potentials. For simultaneous electrochemical detection of PtO2 and pH, two consecutive trapezoidal voltage ramps are applied with double differential-subtraction of the background current. This enables changes in current caused by protons and oxygen to be detected separately with minimal interference between the two. The profile of PtO2 changes evoked by increases in local neuronal activity recorded using the described technique was very similar to that of blood-oxygen-level-dependent responses recorded using fMRI. This voltammetric technique can be combined with fMRI and brain vessel imaging to study the metabolic mechanisms underlying neurovascular coupling response with much greater spatial and temporal resolution than is currently possible.

Ruthenium oxide based microelectrode arrays for in vitro and in vivo neural recording and stimulation

We have characterized the in vitro and in vivo extracellular neural recording and stimulation properties of ruthenium oxide (RuOx) based microelectrodes. Cytotoxicity and functional neurotoxicity assays were carried out to confirm the in vitro biocompatibility of RuOx. Material extract assays, in accordance to ISO protocol “10993-5: Biological evaluation of medical devices”, revealed no significant effect on neuronal cell viability or the functional activity of cortical networks. In vitro microelectrode arrays (MEAs), with indium tin oxide (ITO) sites modified with sputtered iridium oxide (IrOx) and RuOx in a single array, were developed for a direct comparison of electrochemical and recording performance of RuOx to ITO and IrOx deposited microelectrode sites. The impedance of the RuOx-coated electrodes measured by electrochemical impedance spectroscopy was notably lower than that of ITO electrodes, resulting in robust extracellular recordings from cortical networks in vitro. We found comparable signal-to-noise ratios (SNRs) for RuOx and IrOx, both significantly higher than the SNR for ITO. RuOx-based MEAs were also fabricated and implanted in the rat motor cortex to demonstrate manufacturability of the RuOx processing and acute recording capabilities in vivo. We observed single-unit extracellular action potentials with a SNR >22, representing a first step for neurophysiological recordings in vivo with RuOx based microelectrodes. Statement of significanceA critical challenge in neural interface technology is the development of microelectrodes that have recording and electrical stimulation capabilities suitable for bidirectional communication between the external electronic device and the nervous system. The present study explores the feasibility and functional capabilities of ruthenium oxide microelectrodes as a neural interface. Significant improvement in electrochemical properties and neuronal recordings are reported when compared to commercially available indium tin oxide and was similar to that of iridium oxide electrodes. The data demonstrate the potential for future development of chronic neural interfaces using ruthenium oxide based microelectrodes for recording and stimulation.

Reverse-filtering on extracellular action potential for waveform analysis

For further understanding the role of serotonergic neurons, unit recordings using behaving primates are increasingly needed. A widely used criterion to identify serotonergic neuron relies on the duration of extracellular action potential (EAP). However, the duration is inaccurate due to the passband limitation needed to carry out the spike sorting. To restore an original waveform, we conducted 1) averaging the EAPs collected from the unfiltered raw signal and 2) reverse-filtering the EAPs collected from the filtered raw signal. The reconstructed waveforms by these analyses well agreed with each other, suggesting that either analysis is applicable to restore the EAPs. Even using multivariate analyses, the reconstructed EAPs in dorsal raphe (DR) could not be divided into different clusters possibly related to neurochemicals, suggesting that the DR neurons in the previous studies that relied on waveform criterion might contain both serotonergic and non-serotonergic neurons. Between DR and orbitofrontal cortex (OFC), there were no differences in the duration of filtered EAPs. However, against the conventional criterion, the duration of the first crest in the unfiltered / restored EAPs in DR that might contain serotonergic neurons was shorter than that in OFC. This raises a possibility that the conventional waveform criterion needs further consideration.

A 10-bit reconfigurable ADC with SAR/SS mode for neural recording

A dual-mode, reconfigurable ADC for neural recording application is presented in this paper. In order to quantize the extracellular action potential (EAP or spike) of neural signal, the proposed ADC works as a 10 bits successive approximation register (SAR) ADC in the normal mode to process all the raw data. In other side, to reduce dynamic power, the ADC is designed to configure as a single-slope (SS) ADC in compression mode, and only essential parts of spike waveforms are processed. Fabricated in a 0.18 µm CMOS process, the proposed ADC consumes 2.83 μW in SAR mode and 24.71 μW in SS mode at supply voltage of 1.8 V, respectively. In SAR mode, the achieved SNDR and SFDR are 60.5 dB and 75.5 dB when a Nyquist frequency input signal is received @ the sampling rate of 25 kS/s. While in SS mode, the achieved SNDR and SFDR are 57.7 dB and 60.8 dB.

Anatomically accurate model of EMG during index finger flexion and abduction derived from diffusion tensor imaging.

This study presents a modelling framework in which information on muscle fiber direction and orientation during contraction is derived from diffusion tensor imaging (DTI) and incorporated in a computational model of the surface electromyographic (EMG) signal. The proposed model makes use of the principle of reciprocity to simultaneously calculate the electric potentials produced at the recording electrode by charges distributed along an arbitrary number of muscle fibers within the muscle, allowing for a computationally efficient evaluation of extracellular motor unit action potentials. The approach is applied to the complex architecture of the first dorsal interosseous (FDI) muscle of the hand to simulate EMG during index finger flexion and abduction. Using diffusion tensor imaging methods, the results show how muscle fiber orientation and curvature in this intrinsic hand muscle change during flexion and abduction. Incorporation of anatomically accurate muscle architecture and other hand tissue morphologies enables the model to capture variations in extracellular action potential waveform shape across the motor unit population and to predict experimentally observed differences in EMG signal features when switching from index finger abduction to flexion. The simulation results illustrate how structural and electrical properties of the tissues comprising the volume conductor, in combination with fiber direction and curvature, shape the detected action potentials. Using the model, the relative contribution of motor units of different sizes located throughout the muscle under both conditions is examined, yielding a prediction of the detection profile of the surface EMG electrode array over the muscle cross-section.

Novel method for action potential measurements from intact cardiac monolayers with multiwell microelectrode array technology

The cardiac action potential (AP) is vital for understanding healthy and diseased cardiac biology and drug safety testing. However, techniques for high throughput cardiac AP measurements have been limited. Here, we introduce a novel technique for reliably increasing the coupling of cardiomyocyte syncytium to planar multiwell microelectrode arrays, resulting in a stable, label-free local extracellular action potential (LEAP). We characterized the reliability and stability of LEAP, its relationship to the field potential, and its efficacy for quantifying AP morphology of human induced pluripotent stem cell derived and primary rodent cardiomyocytes. Rise time, action potential duration, beat period, and triangulation were used to quantify compound responses and AP morphology changes induced by genetic modification. LEAP is the first high throughput, non-invasive, label-free, stable method to capture AP morphology from an intact cardiomyocyte syncytium. LEAP can accelerate our understanding of stem cell models, while improving the automation and accuracy of drug testing.

A silicon-based spiky probe providing improved cell accessibility during in vitro slice recordings

AimsIn this study, we introduce an edge-type laminar silicon probe suitable for improved cell accessibility during in vitro brain slice recordings. With protruding contact sites, the spiky probe provides high signal yield and quality while approaching cells located deeper in the tissue. Methods. The spiky probe comprises an angled shank carrying 32 protruding contact sites with three possible spacing of 25 μm, 50 μm and 100 μm. The angled shank makes the probe compatible with large microscope objectives used typically in two-photon imaging, in vitro. Spiky probes were batch manufactured with high precision using the etching before grinding technology and bonded to a custom designed printed circuit board. A quantitative comparison of the performance was made against a commercially available surface probe. To investigate the long-term stability of contact sites, the spiky probe was repeatedly inserted in seven experiments (17 insertions, in total) using Wistar rat hippocampal slices, in vitro. Impedance magnitudes and phase angles were compared before and after the extensive usage. ResultsSingle unit activities were recorded with higher neuronal yield and higher amplitudes compared to the surface probe. Impedances did not increase significantly after reusing the probe in multiple experiments. Furthermore, we were able to detect extracellular action potentials from the same single unit on multiple, adjacent contact sites. Finally, we separated the recorded single units into putative cell types based on their extracellular waveforms. Conclusion and outlookThe spiky probes were proved to be optimal solutions to record extracellular spike waveforms, in vitro. The improved signal quality allowed us to distinguish between putative interneurons and principal cells. In future works, integrative experiments using these probes may provide multi-modal data for cellular electrophysiology resulting in a deeper understanding of single cell contributions to mass neural activity.

Modulation of Functional Connectivity Between Dopamine Neurons of the Rat Ventral Tegmental Area in vitro.

Micro Electrode Arrays were used to simultaneously record spontaneous extracellular action potentials from 10 to 30 dopamine neurons in acute brain slices from the lateral Ventral Tegmental Area (VTA) of the rat. The spike train of an individual neuron was used to characterize the firing pattern: firing rate, firing irregularity and oscillation frequency. Functional connectivity between a pair of neurons was quantified by the Paired Phase Consistency (PPC), taking the oscillation frequency as reference. Under baseline conditions the PPC was significantly different from zero and 42 of the 386 pairs of VTA neurons showed significant coupling. Fifty percent of the recorded dopamine neurons were part of the coupled VTA network. Raising extracellular potassium from 3.5 to 5 mM increased the mean firing rate of the dopamine neurons by 45%. The same increase could be induced by bath application of 300 μm glutamate. High potassium reduced the PPC, but it did not change during the glutamate application. Our findings imply that manipulating excitability has distinct and specific consequences for functional connectivity in the VTA network that cannot be directly predicted from the changes in neuronal firing rates. Functional connectivity reflects the spatial organization and synchronization of the VTA output and thus represents a unique element of the message that is sent to the mesolimbic projection area. It adds a dimension to pharmacological manipulation of the VTA micro circuit that might help to understand the pharmacological (side) effects of e.g., anti-psychotic drugs.

Optimal Electrode Size for Multi-Scale Extracellular-Potential Recording From Neuronal Assemblies.

Advances in microfabrication technology have enabled the production of devices containing arrays of thousands of closely spaced recording electrodes, which afford subcellular resolution of electrical signals in neurons and neuronal networks. Rationalizing the electrode size and configuration in such arrays demands consideration of application-specific requirements and inherent features of the electrodes. Tradeoffs among size, spatial density, sensitivity, noise, attenuation, and other factors are inevitable. Although recording extracellular signals from neurons with planar metal electrodes is fairly well established, the effects of the electrode characteristics on the quality and utility of recorded signals, especially for small, densely packed electrodes, have yet to be fully characterized. Here, we present a combined experimental and computational approach to elucidating how electrode size, and size-dependent parameters, such as impedance, baseline noise, and transmission characteristics, influence recorded neuronal signals. Using arrays containing platinum electrodes of different sizes, we experimentally evaluated the electrode performance in the recording of local field potentials (LFPs) and extracellular action potentials (EAPs) from the following cell preparations: acute brain slices, dissociated cell cultures, and organotypic slice cultures. Moreover, we simulated the potential spatial decay of point-current sources to investigate signal averaging using known signal sources. We demonstrated that the noise and signal attenuation depend more on the electrode impedance than on electrode size, per se, especially for electrodes <10 μm in width or diameter to achieve high-spatial-resolution readout. By minimizing electrode impedance of small electrodes (<10 μm) via surface modification, we could maximize the signal-to-noise ratio to electrically visualize the propagation of axonal EAPs and to isolate single-unit spikes. Due to the large amplitude of LFP signals, recording quality was high and nearly independent of electrode size. These findings should be of value in configuring in vitro and in vivo microelectrode arrays for extracellular recordings with high spatial resolution in various applications.

Towards pharmacological treatment screening of cardiomyocyte cells using Si nanowire FETs

Silicon nanowires (Si NWs) are the most promising candidates for recording biological signals due to improved interfacing properties with cells and the possibility of high-speed transduction of biochemical signals into detectable electrical responses. The recording of extracellular action potentials (APs) from cardiac cells is important for fundamental studies of AP propagation features reflecting cell activity and the influence of pharmacological substances on the signal. We applied a novel approach of using fabricated Si NW field-effect transistors (FETs) in combination with fluorescent marker techniques to evaluate the functional activity of cardiac cells. Extracellular AP signal recording from HL-1 cardiomyocytes was demonstrated. This method was supplemented by studies of the pharmacological effects of stimulations using noradrenaline (NorA) as a modulator of functional activity on a cellular and subcellular levels, which were also tested using fluorescent marker techniques. The role of calcium alteration and membrane potential were revealed using Fluo-4 AM and tetramethylrhodamine, methyl ester, perchlorate (TMRM) fluorescent dyes. In addition, chemical treatment with sodium dodecyl sulfate (SDS) solutions was tested. The results obtained demonstrate positive prospects for AP monitoring in different treatments for studies related to a wide range of myocardial diseases using lab-on-chip technology.

A Novel Sympathoexcitatory Pathway between The Deep Superior Colliculus and The Rostral Ventromedial Medulla

IntroductionThe superior colliculus plays key roles in the immediate processing of threatening sensory stimuli and generates rapid behavioural responses that are critical for survival. We have previously found that disinhibition of the deep superior colliculus (dSC) unmasks coordinated respiratory, sympathetic and somatomotor outputs that are independent of processing in higher centres. We postulate that these effects are a result of direct innervation of autonomic and respiratory medullary neurons based on the results of neuroanatomical tracing studies that identified dSC synaptic contacts on spinally projecting neurons within the rostral ventromedial medulla (RVMM). In the present study we investigate the physiological significance of this relay and compare effects of dSC stimulation to activation of dSC‐RVMM terminals.MethodsWe used an AAV vector to express Channelrhodopsin2 (ChR2) in the dSC and chronically implanted a fibre optic cannula into the dSC. After transgene expression electrophysiology experiments were conducted under urethane anesthesia with rats instrumented to record blood pressure, diaphragmatic EMG and splanchnic sympathetic nerve activity (SNA). One subset of experiments then positioned the optrode in the RVMM to deliver light to the terminal projections from the dSC. For the other subset of experiments, extracellular recordings were made of spinally projecting RVMM neurons.ResultsdSC photoactivation evoked an increase in respiratory frequency (35±8.7%, P=&lt;0.01, N=7) and heart rate (11±2.6 bpm, P=&lt;0.01, N=7), and a modest change in blood pressure when compared to controls. Stimulus‐triggered averaging of SNA revealed short‐latency excitatory potentials in most cases. Photoactivation of ChR2‐expressing dSC‐RVMM terminals evoked similar effects on cardiorespiratory outputs as dSC stimulation, with an increase in respiratory frequency (14 ±9.2%, P&lt;0.001, N=6) and blood pressure (13 ±2 mmHg P&lt;0.01, N=7), as well as qualitatively similar sympathoexcitation. In preliminary experiments that recorded extracellular action potentials in bulbospinal RVMM neurons we recorded excitatory effects of dSC stimulation in 6/8 spontaneously active neurons.ConclusionWe conclude that optogenetic activation of dSC neurons that project to the RVMM can drive cardiorespiratory effects that are consistent with our previous findings and that dSC stimulation evokes excitatory effects in some RVMM bulbospinal neurons. This confirms our hypothesis of the existence of a previously uncharacterized excitatory pathway between the dSC and RVMM. Ongoing experiments will functionally characterize RVMM neurons that receive dSC input.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Gold nanostructure microelectrode arrays for in vitro recording and stimulation from neuronal networks

An ideal microelectrode array (MEA) design should include materials and structures which exhibit biocompatibility, low electrode polarization, low impedance/noise, and structural durability. Here, the fabrication of MEAs with indium tin oxide (ITO) electrodes deposited with self-similar gold nanostructures (GNS) is described. We show that fern leaf fractal-like GNS deposited on ITO electrodes are conducive for neural cell attachment and viability while reducing the interfacial impedance more than two orders of magnitude at low frequencies (100–1000 Hz) versus bare ITO. GNS MEAs, with low interfacial impedance, allowed the detection of extracellular action potentials with excellent signal-to-noise ratios (SNR, 20.26 ± 2.14). Additionally, the modified electrodes demonstrated electrochemical and mechanical stability over 29 d in vitro.

How does the presence of neural probes affect extracellular potentials?

Objective. Mechanistic modeling of neurons is an essential component of computational neuroscience that enables scientists to simulate, explain, and explore neural activity. The conventional approach to simulation of extracellular neural recordings first computes transmembrane currents using the cable equation and then sums their contribution to model the extracellular potential. This two-step approach relies on the assumption that the extracellular space is an infinite and homogeneous conductive medium, while measurements are performed using neural probes. The main purpose of this paper is to assess to what extent the presence of the neural probes of varying shape and size impacts the extracellular field and how to correct for them. Approach. We apply a detailed modeling framework allowing explicit representation of the neuron and the probe to study the effect of the probes and thereby estimate the effect of ignoring it. We use meshes with simplified neurons and different types of probe and compare the extracellular action potentials with and without the probe in the extracellular space. We then compare various solutions to account for the probes’ presence and introduce an efficient probe correction method to include the probe effect in modeling of extracellular potentials. Main results. Our computations show that microwires hardly influence the extracellular electric field and their effect can therefore be ignored. In contrast, multi-electrode arrays (MEAs) significantly affect the extracellular field by magnifying the recorded potential. While MEAs behave similarly to infinite insulated planes, we find that their effect strongly depends on the neuron-probe alignment and probe orientation. Significance. Ignoring the probe effect might be deleterious in some applications, such as neural localization and parameterization of neural models from extracellular recordings. Moreover, the presence of the probe can improve the interpretation of extracellular recordings, by providing a more accurate estimation of the extracellular potential generated by neuronal models.

Capacity Analysis of a Peripheral Nerve Using Modulated Compound Action Potential Pulses

Artificial neural stimulation of a peripheral nerve can create an in-body data communications channel. We propose the stimulation of a peripheral nerve using energy-harvesting arrays of nanodevices, embedded in biocompatible tissue patches. The resulting extracellular compound action potential (CAP) pulse can provide a data bit-stream for communicating with an embedded receiver. Our objective is to determine the maximum achievable transmission range of a CAP along a nerve and the maximum sustainable bit rate. We model the generation of a CAP and then compute the reduction in amplitude and the spreading of the pulse with propagation distance. The channel capacity is calculated for ON–OFF keying (OOK) and digital pulse interval modulation. We show that the transmission range depends on the number and diameters of the activated neurons contributing to the CAP amplitude and width. Our modulation analysis demonstrates the effects of attenuation, background noise, the neural refractory period, and pulse broadening on the achievable bit rate. We show how a maximum OOK bit rate of 200 bit/s can be sustained over transmission distances greater than 100 mm. The proposed approach provides a low bit rate, unidirectional asynchronous transmission system that could, for example, deliver simple instructions to an embedded drug-delivery system.

The Axon Initial Segment is the Dominant Contributor to the Neuron's Extracellular Electrical Potential Landscape.

Extracellular voltage fields, produced by a neuron's action potentials, provide a widely used means for studying neuronal and neuronal-network function. The neuron's soma and dendrites are thought to drive the extracellular action potential (EAP) landscape, while the axon's contribution is usually considered less important. However, by recording voltages of single neurons in dissociated rat cortical cultures and Purkinje cells in acute mouse cerebellar slices through hundreds of densely packed electrodes, it is found, instead, that the axon initial segment dominates the measured EAP landscape, and, surprisingly, the soma only contributes to a minor extent. As expected, the recorded dominant signal has negative polarity (charge entering the cell) and initiates at the distal end. Interestingly, signals with positive polarity (charge exiting the cell) occur near some but not all dendritic branches and occur after a delay. Such basic knowledge about which neuronal compartments contribute to the extracellular voltage landscape is important for interpreting results from all electrical readout schemes. Finally, initiation of the electrical activity at the distal end of the axon initial segment (AIS) and subsequent spreading into the axon proper and backward through the proximal AIS toward the soma are confirmed. The corresponding extracellular waveforms across different neuronal compartments could be tracked.

Activity of swallowing-related neurons in the medulla in the perfused brainstem preparation in rats.

We aimed to investigate and validate the cellular activity patterns and the potential topographical organization of neurons of the medullary swallowing pattern generator (Sw-CPG). We used the perfused brainstem preparation as an innovative experimental model that allows for stable neuronal recording in the brainstem. Animal model. Experiments were conducted in 14 juvenile Wistar rats. The activities of the phrenic, vagus, and hypoglossal nerves were recorded at baseline, and fictive swallowing was elicited by stimulation of the superior laryngeal nerve. Extracellular action potentials of 72 swallowing-related neurons were recorded in the Sw-CPG of the dorsal medulla oblongata. Neurons could be classified into three types: sensory relay, and neurons that were excited or inhibited during fictive swallowing. Approximately one-third of the neurons likely received monosynaptic input from the laryngeal afferents. One-third of neurons recorded showed respiratory-related activity, most of which exhibited inspiratory modulation. The neurons were widely distributed in the nucleus tractus solitarius and reticular formation. The perfused brainstem preparation of rat fully preserves the Sw-CPG. The recorded cellular activities and general topographical organization of swallowing neurons are in accordance with previous in vivo studies. Thus, the perfused brainstem preparation is an ideal experimental model to advance the understanding of neuronal mechanisms underlying swallowing. NA Laryngoscope, 129:E72-E79, 2019.

Transparent Graphene/PEDOT:PSS Microelectrodes for Electro‐ and Optophysiology

AbstractConventional opaque electrodes in microelectrode array (MEA) technology obstruct the view of cells in their immediate vicinity (e.g., ≈50 µm) from which the strongest extracellular action potentials are recorded. This limitation has been overcome by transparent graphene electrodes which allow for optical access essential for novel applications such as optogenetics and calcium imaging. Downscaling, necessary for high resolution single‐unit electrophysiological recordings, has been a significant challenge due to inferior electrochemical impedance and correspondingly lower signal‐to‐noise ratio. Here, the combination of graphene with the conductive polymer poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) as a transparent microelectrode material for in vitro MEAs is presented and their application with optical imaging and electrophysiology is demonstrated. Optimal graphene/PEDOT:PSS microelectrodes display transparencies of 84% over the visible spectrum and impedance magnitude of (166 ± 13) kΩ at 1 kHz. The balance of transparency and 1 kHz impedance can be tuned from ≈90% and 700 kΩ to 50% and 42 kΩ.

Long-Term High-Density Extracellular Recordings Enable Studies of Muscle Cell Physiology.

Skeletal (voluntary) muscle is the most abundant tissue in the body, thus making it an important biomedical research subject. Studies of neuromuscular transmission, including disorders of ion channels or receptors in autoimmune or genetic neuromuscular disorders, require high-spatial-resolution measurement techniques and an ability to acquire repeated recordings over time in order to track pharmacological interventions. Preclinical techniques for studying diseases of neuromuscular transmission can be enhanced by physiologic ex vivo models of tissue-tissue and cell-cell interactions. Here, we present a method, which allows tracking the development of primary skeletal muscle cells from myoblasts into mature contracting myotubes over more than 2 months. In contrast to most previous studies, the myotubes did not detach from the surface but instead formed functional networks between the myotubes, whose electrical signals were observed over the entire culturing period. Primary cultures of mouse myoblasts differentiated into contracting myotubes on a chip that contained an array of 26,400 platinum electrodes at a density of 3,265 electrodes per mm2. Our ability to track extracellular action potentials at subcellular resolution enabled study of skeletal muscle development and kinetics, modes of spiking and spatio-temporal relationships between muscles. The developed system in turn enables creation of a novel electrophysiological platform for establishing ex vivo disease models.

Automatic spike sorting for high-density microelectrode arrays.

High-density microelectrode arrays can be used to record extracellular action potentials from hundreds to thousands of neurons simultaneously. Efficient spike sorters must be developed to cope with such large data volumes. Most existing spike sorting methods for single electrodes or small multielectrodes, however, suffer from the "curse of dimensionality" and cannot be directly applied to recordings with hundreds of electrodes. This holds particularly true for the standard reference spike sorting algorithm, principal component analysis-based feature extraction, followed by k-means or expectation maximization clustering, against which most spike sorters are evaluated. We present a spike sorting algorithm that circumvents the dimensionality problem by sorting local groups of electrodes independently with classical spike sorting approaches. It is scalable to any number of recording electrodes and well suited for parallel computing. The combination of data prewhitening before the principal component analysis-based extraction and a parameter-free clustering algorithm obviated the need for parameter adjustments. We evaluated its performance using surrogate data in which we systematically varied spike amplitudes and spike rates and that were generated by inserting template spikes into the voltage traces of real recordings. In a direct comparison, our algorithm could compete with existing state-of-the-art spike sorters in terms of sensitivity and precision, while parameter adjustment or manual cluster curation was not required. NEW & NOTEWORTHY We present an automatic spike sorting algorithm that combines three strategies to scale classical spike sorting techniques for high-density microelectrode arrays: 1) splitting the recording electrodes into small groups and sorting them independently; 2) clustering a subset of spikes and classifying the rest to limit computation time; and 3) prewhitening the spike waveforms to enable the use of parameter-free clustering. Finally, we combined these strategies into an automatic spike sorter that is competitive with state-of-the-art spike sorters.

Morphological and Biophysical Determinants of the Intracellular and Extracellular Waveforms in Nigral Dopaminergic Neurons: A Computational Study.

Action potentials (APs) in nigral dopaminergic neurons often exhibit two separate components: the first reflecting spike initiation in the dendritically located axon initial segment (AIS) and the second the subsequent dendro-somatic spike. These components are separated by a notch in the ascending phase of the somatic extracellular waveform and in the temporal derivative of the somatic intracellular waveform. Still, considerable variability exists in the presence and magnitude of the notch across neurons. To systematically address the contribution of AIS, dendritic and somatic compartments to shaping the two-component APs, we modeled APs of previously in vivo electrophysiologically characterized and 3D-reconstructed male mouse and rat dopaminergic neurons. A parsimonious two-domain model, with high (AIS) and lower (dendro-somatic) Na+ conductance, reproduced the notch in the temporal derivatives, but not in the extracellular APs, regardless of morphology. The notch was only revealed when somatic active currents were reduced, constraining the model to three domains. Thus, an initial AIS spike is followed by an actively generated spike by the axon-bearing dendrite (ABD), in turn followed mostly passively by the soma. The transition from being a source compartment for the AIS spike to a source compartment for the ABD spike satisfactorily explains the extracellular somatic notch. Larger AISs and thinner ABD (but not soma-to-AIS distance) accentuate the AIS component. We conclude that variability in AIS size and ABD caliber explains variability in AP extracellular waveform and separation of AIS and dendro-somatic components, given the presence of at least three functional domains with distinct excitability characteristics.SIGNIFICANCE STATEMENT Midbrain dopamine neurons make an important contribution to circuits mediating motivation and movement. Understanding the basic rules that govern the electrical activity of single dopaminergic neurons is therefore essential to reveal how they ultimately contribute to movement and motivation as well as what goes wrong in associated disorders. Our computational study focuses on the generation and propagation of action potentials and shows that different morphologies and excitability characteristics of the cell body, dendrites and proximal axon can explain the diversity of action potentials shapes in this population. These compartments likely make differential contributions both to normal dopaminergic signaling and could potentially underlie pathological dopaminergic signaling implicated in addiction, schizophrenia, Parkinson's disease, and other disorders.