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The functions of the diverse retinal ganglion cell types in primates and the parallel visual pathways they initiate remain poorly understood. Here, unusual physiological and computational properties of the ON and OFF smooth monostratified ganglion cells are explored. Large-scale multi-electrode recordings from 48 macaque retinas revealed that these cells exhibit irregular receptive field structure composed of spatially segregated hotspots, quite different from the classic center-surround model of retinal receptive fields. Surprisingly, visual stimulation of different hotspots in the same cell produced spikes with subtly different spatiotemporal voltage signatures, consistent with a dendritic contribution to hotspot structure. Targeted visual stimulation and computational inference demonstrated strong nonlinear subunit properties associated with each hotspot, supporting a model in which the hotspots apply nonlinearities at a larger spatial scale than bipolar cells. These findings reveal a previously unreported nonlinear mechanism in the output of the primate retina that contributes to signaling spatial information., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The potential role of cardiac MR imaging in the diagnosis, risk-stratification, and treatment of patients with cardiac arrhythmias is rapidly evolving. Beyond substrate identification and characterization, it is now feasible to perform MR imaging-based noninvasive virtual programmed stimulation to identify potential ablation targets. The next phase of MR imaging-guided electrophysiology may involve intraprocedural guidance of electrophysiologic interventions using real-time MR imaging. This review discusses the spectrum of MR imaging applications in electrophysiology ranging from those that are established in clinical practice to those that are in the experimental phase of development., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The detection of bursts and also of response onsets is often of relevance in understanding neurophysiological data, but the detection of these events is not a trivial task. We build on a method that was originally designed for burst detection using the so-called burst surprise as a measure. We extend this method and provide a proper significance measure. Our method consists of two stages. In the first stage we model the neuron's interspike interval (ISI) distribution and make an i.i.d. assumption to formulate our null hypothesis. In addition we define a set of 'surprising' events that signify deviations from the null hypothesis in the direction of 'burstiness'. Here the so-called (strict) burst novelty is used to measure the size of this deviation. In the second stage we determine the significance of this deviation. The (strict) burst surprise is used to measure the significance, since it is the negative logarithm of the significance probability. After showing the consequences of a non-proper null hypothesis on burst detection performance, we apply the method to experimental data. For this application the data are divided into a period for parameter estimation to express a proper null hypothesis (model of the ISI distribution), and the rest of the data is analyzed by using that null hypothesis. We find that assuming a Poisson process for experimental spike data from motor cortex is rarely a proper null hypothesis, because these data tend to fire more regularly and thus a gamma process is more appropriate. We show that our burst detection method can be used for rate change onset detection, because a deviation from the null hypothesis detected by (strict) burst novelty also covers an increase of firing rate.

Genetically encoded fluorescent indicators typically consist of the sensitive and reporter protein domains connected with the amino acid linkers. The final performance of a particular indicator may depend on the linker length and composition as strong as it depends on the both domains nature. Here we aimed to optimize interdomain linkers in VSD-FR189-188-a recently described red fluorescent protein-based voltage indicator. We have tested 13 shortened linker versions and monitored the dynamic range, response speed and polarity of the corresponding voltage indicator variants. While the new indicators didn't show a contrast enhancement, some of them carrying very short interdomain linkers responded 25-fold faster than the parental VSD-FR189-188. Also we found the critical linker length at which fluorescence response to voltage shift changes its polarity from negative to positive slope. Our observations thus make an important contribution to the designing principles of the fluorescent protein-derived voltage indicators.

Membrane potential (Vm) of vascular smooth muscle cells determines vessel tone and thus blood flow to an organ. Changes in the expression and function of ion channels and electrogenic pumps that regulate Vm in disease conditions could potentially alter Vm, vascular tone, and blood flow. Thus, a basic understanding of electrophysiology and the methods necessary to accurately record Vm in healthy and diseased states are essential. This method will allow modulating Vm using different pharmacological agents to restore Vm. Although there are several methods, each with its advantages and disadvantages, this article provides protocols to record Vm from cannulated resistance vessels such as the middle cerebral artery using the microelectrode impalement method. Middle cerebral arteries are allowed to gain myogenic tone in a myograph chamber, and the vessel wall is impaled using high resistance microelectrodes. The Vm signal is collected through an electrometer, digitized, and analyzed. This method provides an accurate reading of the Vm of a vessel wall without damaging the cells and without changing the membrane resistance.

Various methods are used for analyzing the status of the human body. In digestive system, myoelectrical activity is recorded from the abdominal surface. Frequency and power of Gastrointestinal (GI) activity is usually assessed using power spectral analysis. However, spectral analysis of GI electrical activity is a nonspecific test, failing to distinguish diseases. Other dimensions of GI electrical waves may help identify diseases more precisely. Studying complex systems is one of the approaches that can be taken for research on body systems. The dynamic variability of a complex system is estimated by nonlinear methods. Previous studies have identified how functional dynamic variability of organs differ in normal and disease conditions. Variation of electroencephalogram, heart rate, and respiratory rate are known examples of system dynamic fluctuation. If we consider the digestive tract as a complex system, the dynamic variation of the waves in the GI system could be analyzed by nonlinear methods to possibly help clinicians assess various conditions of the GI system in healthy subjects and patients., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Drugs targeting different calcium channel subtypes have strong therapeutic potential for future drug development for cardiovascular disorders, neuropsychiatric diseases and cancer. This study aims to design and synthesize a new series of C2 substituted dihydropyrimidines to mimic the structure features of third generation long acting dihydropyridine calcium channel blockers and dihydropyrimidines analogues. The target compounds have been evaluated as blockers for Ca V 1.2 and Ca V 3.2 utilizing the whole-cell patch clamp technique. Among the tested compounds, compound 7a showed moderate calcium channel blockade activity against Ca V 3.2. Moreover, the predicted physicochemical properties and pharmacokinetic profiles of the target compounds recommend that they can be considered as drug-like candidates. The results highlight some significant information for the future design of lead compounds as calcium channel blockers., (Copyright © 2019. Published by Elsevier Inc.)

In this work, single-channel current recordings were used to selectively detect individual ssDNA strands in the vestibule of the α-hemolysin (α-HL) protein nanopore. The sensing mechanism was based on the detection of the intrinsic topological change of target ssDNA molecules after the hybridization with complementary PNA fragments. The readily distinguishable current signatures of PNA-DNA duplexes reversible association with the α-HL's vestibule, in terms of blockade amplitudes and kinetic features, allows specific detection of nucleic acid hybridization.

Background: The long-term predicted value of microvolt T-wave alternans (MTWA) for ventricular tachyarrhythmia in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC) remains unclear. Our study explored the characteristics of MTWA and its prognostic value when combined with an electrophysiologic study (EPS) in patients with ARVC., Methods: All patients underwent non-invasive MTWA examination with modified moving average (MMA) analysis and an EPS. A positive event was defined as the first occurrence of sudden cardiac death, documented sustained ventricular tachycardia (VT), ventricular fibrillation, or the administration of appropriate implantable cardioverter defibrillator therapy including shock or anti-tachycardia pacing., Results: Thirty-five patients with ARVC (age 38.6 ± 11.0 years; 28 males) with preserved left ventricular (LV) function were recruited. The maximal TWA value (MaxValt) was 17.0 (11.0-27.0) μV. Sustained VT was induced in 22 patients by the EPS. During a median follow-up of 99.9 ± 7.7 months, 15 patients had positive clinical events. When inducible VT was combined with the MaxValt, the area under the curve improved from 0.739 to 0.797. The receiver operating characteristic curve showed that a MaxValt of 23.5 μV was the optimal cutoff value to identify positive events. The multivariate Cox regression model for survival showed that MTWA (MaxValt, hazard ratio [HR], 1.06; 95% confidence interval [CI], 1.01-1.11; P = 0.01) and inducible VT (HR, 5.98; 95% CI, 1.33-26.8; P = 0.01) independently predicted positive events in patients with ARVC., Conclusions: MTWA assessment with MMA analysis complemented by an EPS might provide improved prognostic ability in patients with ARVC with preserved LV function during long-term follow-up.

In the heart, Ca 2+ influx through L-type Ca 2+ channels triggers Ca 2+ release from the sarcoplasmic reticulum. In most mammals, this influx occurs during the ventricular action potential (AP) plateau phase 2. However, in murine models, the influx through L-type Ca 2+ channels happens in early repolarizing phase 1. The aim of this work is to assess if changes in the open probability of 4-aminopyridine (4-AP)-sensitive Kv channels defining the outward K + current during phase 1 can modulate Ca 2+ currents, Ca 2+ transients, and systolic pressure during the cardiac cycle in intact perfused beating hearts. Pulsed local-field fluorescence microscopy and loose-patch photolysis were used to test the hypothesis that a decrease in a transient K + current (I to ) will enhance Ca 2+ influx and promote a larger Ca 2+ transient. Simultaneous recordings of Ca 2+ transients and APs by pulsed local-field fluorescence microscopy and loose-patch photolysis showed that a reduction in the phase 1 repolarization rate increases the amplitude of Ca 2+ transients due to an increase in Ca 2+ influx through L-type Ca 2+ channels. Moreover, 4-AP induced an increase in the time required for AP to reach 30% repolarization, and the amplitude of Ca 2+ transients was larger in epicardium than endocardium. On the other hand, the activation of I to with NS5806 resulted in a reduction of Ca 2+ current amplitude that led to a reduction of the amplitude of Ca 2+ transients. Finally, the 4-AP effect on AP phase 1 was significantly smaller when the L-type Ca 2+ current was partially blocked with nifedipine, indicating that the phase 1 rate of repolarization is defined by the competition between an outward K + current and an inward Ca 2+ current., (© 2019 López-Alarcón et al.)

Electrophysiological examination of penile nerves involves detection of somatic and autonomic nerves, including dorsal nerve somatosensory evoked potential, cavernous muscle reflex, skin sympathetic response, cavernous electromyography, and others, which plays an important role in the diagnosis of male sexual dysfunction, such as ED and premature ejaculation, as well as in the selection of treatment methods and evaluation of therapeutic effects. Due to the complex mechanisms of the brain, spinal cord and peripheral nerve pathways relating to erectile and ejaculatory centers in men and many unknown areas in penile electrophysiological techniques, penile electrophysiological examination has not been widely accepted by doctors and patients. Further studies are needed to provide more scientific and convincing evidence in this field.

In contrast to the long-standing notion that the role of individual neurons in population activity is vanishingly small, recent studies have shown that electrical activation of only a single cortical neuron can have measurable effects on global brain state, movement, and perception. Although highly important for understanding how neuronal activity in cortex is orchestrated, the cellular and network mechanisms underlying this phenomenon are unresolved. Here, we first briefly review the current state of knowledge regarding the phenomenon of single-cell induced network modulation and discuss possible underpinnings. Secondly, we show proof of principle for an experimental approach to elucidate the mechanisms of single-cell induced changes in cortical activity. The setup allows simultaneous recordings of the spiking activity of multiple neurons across all layers of the cortex using a multi-electrode array, while manipulating the activity of one individual neuron in close proximity to the array. We demonstrate that single cells can be recorded and stimulated reliably for hundreds of trials, conferring high statistical power even for expectedly small effects of single-neuron spiking on network activity. Preliminary results suggest that single-cell stimulation on average decreases the firing rate of local network units. We expect that characterization of the spatiotemporal spread of single-cell evoked activity across layers and columns will yield novel insights into intracortical processing.

Background: Physiological studies of sensorial systems often require the acquisition and processing of data extracted from their multiple components to evaluate how the neural information changes in relation to the environment changes. In this work, a comparative study about methodological aspects of two electrophysiological approaches is described., New Method: Extracellular recordings from deep vibrissal nerves were obtained by using a customized microelectrode Utah array during passive mechanical stimulation of rat´s whiskers. These recordings were compared with those obtained with bipolar electrodes. We also propose here a simplified empirical model of the electrophysiological activity obtained from a bundle of myelinated nerve fibers., Results: The peripheral activity of the vibrissal system was characterized through the temporal and spectral features obtained with both recording methods. The empirical model not only allows the correlation between anatomical structures and functional features, but also allows to predict changes in the CAPs morphology when the arrangement and the geometry of the electrodes changes., Comparison With Existing Method(s): This study compares two extracellular recording methods based on analysis techniques, empirical modeling and data processing of vibrissal sensory information., Conclusions: This comparative study reveals a close relationship between the electrophysiological techniques and the processing methods necessary to extract sensory information. This relationship is the result of maximizing the extraction of information from recordings of sensory activity., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Different neuron types serve distinct roles in neural processing. Extracellular electrical recordings are extensively used to study brain function but are typically blind to cell identity. Morphoelectrical properties of neurons measured on spatially dense electrode arrays have the potential to distinguish neuron types. We used high-density silicon probes to record from cortical and subcortical regions of the mouse brain. Extracellular waveforms of each neuron were detected across many channels and showed distinct spatiotemporal profiles among brain regions. Classification of neurons by brain region was improved with multichannel compared with single-channel waveforms. In visual cortex, unsupervised clustering identified the canonical regular-spiking (RS) and fast-spiking (FS) classes but also indicated a subclass of RS units with unidirectional backpropagating action potentials (BAPs). Moreover, BAPs were observed in many hippocampal RS cells. Overall, waveform analysis of spikes from high-density probes aids neuron identification and can reveal dendritic backpropagation. NEW & NOTEWORTHY It is challenging to identify neuron types with extracellular electrophysiology in vivo. We show that spatiotemporal action potentials measured on high-density electrode arrays can capture cell type-specific morphoelectrical properties, allowing classification of neurons across brain structures and within the cortex. Moreover, backpropagating action potentials are reliably detected in vivo from subpopulations of cortical and hippocampal neurons. Together, these results enhance the utility of dense extracellular electrophysiology for cell-type interrogation of brain network function.

Insect vector behavior and biology can be affected by pathogen-induced changes in the physiology and morphology of the host plant. Herein, we examined the temporal effects of Squash vein yellowing virus (family Potyviridae, genus Ipomovirus) infection on the settling, oviposition preference, and feeding behavior of its whitefly vector, Bemisia tabaci (Gennadius) Middle East-Asia Minor 1 (MEAM1), formerly known as B. tabaci biotype B. Settling and oviposition behavioral choice assays were conducted on pairs of infected and mock-inoculated watermelon (Citrullus lanatus (Thunb) Matsum and Nakai) (Cucurbitales: Cucurbitaceae) at 5-6 days post inoculation (DPI) and 10-12 DPI. Electropenetrography, or electrical penetration graph (both abbreviated EPG), was used to assess differences in feeding behaviors of whitefly on mock-inoculated, 5-6 and 10-12 DPI infected watermelon plants. Whiteflies showed no preference in settling or oviposition on the infected and mock-inoculated plants at 5-6 DPI. However, at 10-12 DPI, whiteflies initially settled on infected plants but then preference of settling shifted to mock-inoculated plants after 8 h. Only at 10-12 DPI, females laid significantly more eggs on mock-inoculated plants than infected plants. EPG revealed no differences in whitefly feeding behaviors among mock-inoculated, 5-6 DPI infected and 10-12 DPI infected plants. The results highlighted the need to examine plant disease progression and its effect on vector behavior and performance, which could play a crucial role in Squash vein yellowing virus spread., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America.)

Objective: Hippocampal slice cultures spontaneously develop chronic epilepsy several days after slicing and are used as an in vitro model of post-traumatic epilepsy. Here, we describe a hybrid microfluidic-microelectrode array (μflow-MEA) technology that incorporates a microfluidic perfusion network and electrodes into a miniaturized device for hippocampal slice culture based antiepileptic drug discovery., Methods: Field potential simulation was conducted to help optimize the electrode design to detect a seizure-like population activity. Epilepsy-on-a-chip model was validated by chronic electrical recording, neuronal survival quantification, and anticonvulsant test. To demonstrate the application of μflow-MEA in drug discovery, we utilized a two-stage screening platform to identify potential targets for antiepileptic drugs. In Stage I, lactate and lactate dehydrogenase biomarker assays were performed to identify potential drug candidates. In Stage II, candidate compounds were retested with μflow-MEA-based chronic electrical assay to provide electrophysiological confirmation of biomarker results., Results and Conclusion: We screened 12 receptor tyrosine kinases inhibitors, and EGFR/ErbB-2 and cFMS inhibitors were identified as novel antiepileptic compounds., Significance: This epilepsy-on-a-chip system provides the means for rapid dissection of complex signaling pathways in epileptogenesis, paving the way for high-throughput antiepileptic drug discovery.

The dorsal raphe nucleus (DRN), located in the brainstem, is involved in several functions such as sleep, temperature regulation, stress responses, and anxiety behaviors. This nucleus contains the largest population of serotonin expressing neurons in the brain. Serotonergic DRN neurons receive tonic γ-aminobutyric acid (GABA)inhibitory inputs from several brain areas, as well as from interneurons within the same nucleus. Serotonergic and GABAergic neurons in the DRN can be distinguished by their size, location, pharmacological responses, and electrophysiological properties. GABAergic neurons regulate the excitability of DRN serotonergic neurons and the serotonin release in different brain areas. Also, it has been shown that GABAergic neurons can synchronize the activity of serotonergic neurons across functions such as sleep or alertness. Moreover, dysregulation of GABA signaling in the DRN has been linked to psychiatric disorders such as anxiety and depression. This review focuses on GABAergic transmission in the DRN. The interaction between GABAergic and serotonergic neurons is discussed considering some physiological implications. Also, the main electrophysiological and morphological characteristics of serotonergic and GABAergic neurons are described.

Recordings of neural activity can be used to aid communication, control prosthetic devices or alleviate disease symptoms. Chronic recordings require a high signal-to-noise ratio that is stable for years. Current cortical devices generally fail within months to years after implantation. Development of novel devices to increase lifetime requires valid testing protocols and a knowledge of the critical parameters controlling electrophysiological performance. Here we present electrochemical and electrophysiological protocols for assessing implantable electrodes. Biological noise from neural recording has significant impact on signal-to-noise ratio. A recently developed surgical approach was utilised to reduce biological noise. This allowed correlation of electrochemical and electrophysiological behaviour. The impedance versus frequency of modified electrodes was non-linear. It was found that impedance at low frequencies was a stronger predictor of electrophysiological performance than the typically reported impedance at 1 kHz. Low frequency impedance is a function of electrode area, and a strong correlation of electrode area with electrophysiological response was also seen. Use of these standardised testing protocols will allow future devices to be compared before transfer to preclinical and clinical trials.

Imagine how flicking through your photo album and seeing a picture of a beach sunset brings back fond memories of a tasty cocktail you had that night. Computational models suggest that upon receiving a partial memory cue ('beach'), neurons in the hippocampus coordinate reinstatement of associated memories ('cocktail') in cortical target sites. Here, using human single neuron recordings, we show that hippocampal firing rates are elevated from ~ 500-1500 ms after cue onset during successful associative retrieval. Concurrently, the retrieved target object can be decoded from population spike patterns in adjacent entorhinal cortex (EC), with hippocampal firing preceding EC spikes and predicting the fidelity of EC object reinstatement. Prior to orchestrating reinstatement, a separate population of hippocampal neurons distinguishes different scene cues (buildings vs. landscapes). These results elucidate the hippocampal-entorhinal circuit dynamics for memory recall and reconcile disparate views on the role of the hippocampus in scene processing vs. associative memory.

The technique of microneurography has advanced the field of neuroscience for the past 50 years. While there have been a number of reviews on microneurography, this paper takes an objective approach to exploring the impact of microneurography studies. Briefly, Web of Science (Thomson Reuters) was used to identify the highest citation articles over the past 50 years, and key findings are presented in a decade-by-decade highlight. This includes the establishment of microneurography in the 1960s, the acceleration of the technique by Gunnar Wallin in the 1970s, the international collaborations of the 1980s and 1990s, and finally the highest impact studies from 2000 to present. This journey through 50 years of microneurographic research related to peripheral sympathetic nerve activity includes a historical context for several of the laboratory interventions commonly used today (e.g., cold pressor test, mental stress, lower body negative pressure, isometric handgrip, etc.) and how these interventions and experimental approaches have advanced our knowledge of cardiovascular, cardiometabolic, and other human diseases and conditions.

Stomatal movement, which plays an essential role in plant transpiration and photosynthesis, is controlled by ion channels that mediate K+ and anion fluxes across the plasma membrane (PM) of guard cells. These channels in dicots are accurately regulated by various physiological factors, such as pH, abscisic acid (ABA) and Ca2+; however, the data in monocots are limited. Here the whole-cell patch-clamping technique was applied to analyze the properties and regulations of PM K+ channels in maize guard cells. The results indicated that the hyperpolarization-activated inward-rectifying channels were highly K+-selective. These inward K+ (Kin) channels were sensitive to extracellular K+. Their slope factor (S) decreased when the apoplastic K+ concentration decline, causing a positive shift of the half-activation potential (V1/2). Their activities were promoted by apoplastic acidification but inhibited by apoplastic and cytosolic alkalization. Nevertheless, the outward K+ (Kout) channel activities were uniquely promoted by cytosolic alkalization. Both apoplastic and cytosolic ABA inhibited Kin channels independent of cytosolic Ca2+ ([Ca2+]cyt). And two Ca2+-dependent mechanisms with different Ca2+ affinities may mediate resting- and high-[Ca2+]cyt-induced inhibition on Kin channels, respectively. However, resting [Ca2+]cyt impaired the inhibition of Kin channels induced by apoplastic ABA, not cytosolic ABA. Furthermore, the result that high [Ca2+]cyt attenuated ABA-induced inhibition highlighted the importance of [Ca2+]cyt for Kin channel regulation. There may exist a Ca2+-dependent regulation of the Ca2+-independent ABA signaling pathways for Kin channel inhibition. These results provided an electrophysiological view of the multiple level regulations of PM K+ channel activities and kinetics in maize guard cells., (� The Author(s) 2018. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The noninvasive estimation of neuronal receptive field (RF) properties in vivo allows a detailed understanding of brain organization as well as its plasticity by longitudinal following of potential changes. Visual RFs measured invasively by electrophysiology in animal models have traditionally provided a great extent of our current knowledge about the visual brain and its disorders. Voxel-based estimates of population RF (pRF) by functional magnetic resonance imaging (fMRI) in humans revolutionized the field and have been used extensively in numerous studies. However, current methods cannot estimate single-neuron RF sizes as they reflect large populations of neurons with individual RF scatter. Here, we introduce an approach to estimate RF size using spatial frequency selectivity to checkerboard patterns. This method allowed us to obtain noninvasive, average single-neuron RF estimates over a large portion of human early visual cortex. These estimates were significantly smaller compared with prior pRF methods. Furthermore, fMRI and electrophysiology experiments in nonhuman primates demonstrated an exceptionally good match, validating the approach., Competing Interests: The authors declare no conflict of interest., (Copyright © 2019 the Author(s). Published by PNAS.)

Mammalian inspiratory rhythm is generated from a neuronal network in a region of the medulla called the preBötzinger complex (pBC), which produces a signal driving the rhythmic contraction of inspiratory muscles. Rhythmic neural activity generated in the pBC and carried to other neuronal pools to drive the musculature of breathing may be studied using various approaches, including en bloc nerve recordings and transverse slice recordings. However, previously published methods have not extensively described the brainstem-spinal cord dissection process in a transparent and reproducible manner for future studies. Here, we present a comprehensive overview of a method used to reproducibly cut rhythmically-active brainstem slices containing the necessary and sufficient neuronal circuitry for generating and transmitting inspiratory drive. This work builds upon previous brainstem-spinal cord electrophysiology protocols to enhance the likelihood of reliably obtaining viable and rhythmically-active slices for recording neuronal output from the pBC, hypoglossal premotor neurons (XII pMN), and hypoglossal motor neurons (XII MN). The work presented expands upon previous published methods by providing detailed, step-by-step illustrations of the dissection, from whole rat pup, to in vitro slice containing the XII rootlets.

The waveform of presynaptic action potentials (APs) regulates the magnitude of Ca 2+ currents (I Ca ) and neurotransmitter release. However, how APs control the timing of synaptic transmission remains unclear. Using the calyx of Held synapse, we find that Na + and K + channels affect the timing by changing the AP waveform. Specifically, the onset of I Ca depends on the repolarization but not depolarization rate of APs, being near the end of repolarization phase for narrow APs and advancing to the early repolarization phase for wide APs. Increasing AP amplitude has little effect on the activation but delays the peak time of I Ca . Raising extracellular Ca 2+ concentration increases the amplitude of I Ca yet does not alter their onset timing. Developmental shortening of APs ensures I Ca as a tail current and faithful synaptic delay, which is particularly important at the physiological temperature (35 °C) as I Ca evoked by broad pseudo-APs can occur in the depolarization phase. The early onset of I Ca is more prominent at 35 °C than at 22 °C, likely resulting from a temperature-dependent shift in the activation threshold and accelerated gating kinetics of Ca 2+ channels. These results suggest that the timing of Ca 2+ influx depends on the AP waveform dictated by voltage-gated channels and temperature.

2D electrophysiology is often used to determine the electrical properties of neurons. In the brain however, neurons form extensive 3D networks. Thus, performing electrophysiology in a 3D environment provides a closer situation to the physiological condition and serves as a useful tool for various applications in the field of neuroscience. In this study, 3D electrophysiology is established within a fiber-reinforced matrix to enable fast readouts from transfected cells, which are often used as model systems for 2D electrophysiology. Using melt electrowriting (MEW) of scaffolds to reinforce Matrigel, 3D electrophysiology is performed on a glycine receptor-transfected Ltk-11 mouse fibroblast cell line. The glycine receptor is an inhibitory ion channel associated when mutated with impaired neuromotor behavior. The average thickness of the MEW scaffold is 141.4 ± 5.7 µm, using 9.7 ± 0.2 µm diameter fibers, and square pore spacings of 100, 200, and 400 µm. For the first time, the electrophysiological characterization of glycine receptor-transfected cells is demonstrated with respect to agonist efficacy and potency in a 3D matrix. With the MEW scaffold reinforcement not interfering with the electrophysiological measurement, this approach can now be further adapted and developed for different kinds of neuronal cultures to study and understand pathological mechanisms under disease conditions., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The triple stimulation technique (TST) was rarely used in multiple sclerosis (MS). This study aimed to compare TST and motor evoked potentials (MEP) for the quantification of motor dysfunction. Central motor conduction based on MEP (four limbs) and TST (upper limbs) was assessed in 28 MS patients with a median Expanded Disability Status Scale (EDSS) of 4. EDSS, timed 25-foot walk (T25FW), grasping strength and motor components of the MS functional composite were evaluated. Regression analysis was used to assess the relationship between MEP, TST and clinical findings. TST was negatively correlated with EDSS (r = - 0.74, p < 0.0001) and to a lesser extent with T25FW (r = - 0.47, p < 0.05), and grasping strength (r = - 0.43, p < 0.05). A multiple regression analysis underlined the better correlation between clinical data and TST (R 2  = 0.56, p < 0.0005) than with MEP (0.03 < R 2  < 0.22, p > 0.05). This study evidenced the value of TST as a quantification tool of motor dysfunction. TST appeared to reflect a global disability since it was correlated not only to hand function but also to walking capacity.

Extracellular recording studies have revealed diverse and selective neural responses in the primary auditory cortex (A1) of awake animals. However, we have limited knowledge on subthreshold events that give rise to these responses, especially in non-human primates, as intracellular recordings in awake animals pose substantial technical challenges. We developed a novel intracellular recording technique in awake marmosets to systematically study subthreshold activity of A1 neurons that underlies their diverse and selective spiking responses. Our findings showed that in contrast to predominantly transient depolarization observed in A1 of anesthetized animals, both transient and sustained depolarization (during or beyond the stimulus period) were observed. Comparing with spiking responses, subthreshold responses were often longer lasting in duration and more broadly tuned in frequency, and showed narrower intensity tuning in non-monotonic neurons and lower response threshold in monotonic neurons. These observations demonstrated the enhancement of stimulus selectivity from subthreshold to spiking responses in individual A1 neurons. Furthermore, A1 neurons classified as regular- or fast-spiking subpopulation based on their spike shapes exhibited distinct response properties in frequency and intensity domains. These findings provide valuable insights into cortical integration and transformation of auditory information at the cellular level in auditory cortex of awake non-human primates.

Junctional and AV nodal reentrant tachycardia share common electrocardiographic features, but they differ in their management and outcomes after catheter ablation. This case concerns a 60-year-old female who presented with recurrent episodes of a relatively slow, regular supraventricular arrhythmia. Electrocardiographic features of the arrhythmia were discordant regarding its underlying mechanism. However, careful analysis of 12-lead electrocardiograms, with focus on the effect of spontaneous premature beats, pointed out the arrhythmia etiology. Electrophysiological study and pacing maneuvers defined the arrhythmic substrate that was successfully treated by catheter ablation., (© 2018 Wiley Periodicals, Inc.)

Neurons are classified into several morphological types according to the locations of their somata and the branching patterns of their axons and dendrites. Recent studies suggest that these morphological features are related to their physiological properties, including firing characteristics, responses to neuromodulators, and wiring patterns. Therefore, rapid morphological identification of electrophysiologically recorded neurons promises to advance our understanding of neuronal circuits. One of the most common anatomical cell identification methods is neuronal reconstruction with biocytin delivered through whole-cell patch-clamp pipettes. However, conventional reconstruction methods usually take longer than 24 h and limit the throughput of electrophysiological experiments. Here, we developed a quick, simple cell reconstruction method by optimizing the tissue clearing protocol ScaleSQ. We found that adding 200 mM NaCl almost entirely prevented tissue swelling without compromising optical clearing ability. This solution, termed IsoScaleSQ, allowed us to increase the transparency of the gray matter of 500-µm-thick slices within 30 min, meaning that the total time required to reconstruct whole-cell recorded neurons was reduced to 1 h. This novel method will improve the efficacy and effectiveness of electrophysiological experiments linked to cell morphology.

The dorsal root ganglia (DRG) contain cell bodies of primary afferent neurons, which are frequently studied by recording extracellularly with penetrating microelectrodes inserted into the DRG. We aimed to isolate single- and multi-unit activity from primary afferents in the lumbar DRG using non-penetrating electrode arrays and to characterize the relationship of that activity with limb position and movement. The left sixth and seventh lumbar DRG (L6-L7) were instrumented with penetrating and non-penetrating electrode arrays to record neural activity during passive hindlimb movement in 7 anesthetized cats. We found that the non-penetrating arrays could record both multi-unit and well-isolated single-unit activity from the surface of the DRG, although with smaller signal to noise ratios (SNRs) compared to penetrating electrodes. Across all recorded units, the median SNR was 1.1 for non-penetrating electrodes and 1.6 for penetrating electrodes. Although the non-penetrating arrays were not anchored to the DRG or surrounding tissues, the spike amplitudes did not change (<1% change from baseline spike amplitude) when the limb was moved passively over a limited range of motion (~20 degrees at the hip). Units of various sensory fiber types were recorded, with 20% of units identified as primary muscle spindles, 37% as secondary muscle spindles, and 24% as cutaneous afferents. Our study suggests that non-penetrating electrode arrays can record modulated single- and multi-unit neural activity of various sensory fiber types from the DRG surface.

Actin and microtubules form cellular cytoskeletal network, which mediates cell shape, motility and proliferation and are key targets for cancer therapy. Changes in cytoskeletal organization dramatically affect mechanical properties of the cells and correlate with proliferative capacity and invasiveness of cancer cells. Changes in the cytoskeletal network expectedly lead to altered nonmechanical material properties including electrical conductivity as well. Here we applied, for the first time, microtubule and actin based electrical measurement to monitor changes in the electrical properties of breast cancer cells upon administration of anti-tubulin and anti-actin drugs, respectively. Semiconductive behavior of microtubules and conductive behavior of actins presented different bioelectrical responses (in similar frequencies) of the cells treated by anti-tubulin with respect to anti-actin drugs. Doped silicon nanowires were applied as the electrodes due to their enhanced interactive surface and compatibility with electronic fabrication process. We found that treatment with Mebendazole (MBZ), a microtubule destabilizing agent, decreases electrical resistance while treatment with Paclitaxel (PTX), a microtubule stabilizing agent, leads to an increase in electrical resistance. In contrast, actin destabilizing agents, Cytochalasin D (CytD), and actin stabilizing agent, Phalloidin, lead to an increased and decreased electrical resistance, respectively. Our study thus provides proof-of-principle of the usage of determining the electrical function of cytoskeletal compartments in grading of cancer as well as drug resistance assays.

Objective: The prokinetic action of erythromycin is clinically useful under conditions associated with gastrointestinal hypomotility. Although erythromycin is known to affect the electrogastrogram, no studies have examined the effects that erythromycin has on gastric slow wave magnetic fields., Methods: In this study, gastric slow wave activity was assessed simultaneously using noninvasive magnetogastrogram (MGG), electrogastrogram, and mucosal electromyogram recordings. Recordings were obtained for 30 min prior to and 60 min after intravenous administration of erythromycin at dosages of 3 and 6 mg/kg., Results: MGG recordings showed significant changes in the percentage power distribution of gastric signal after infusion of both 3 and 6 mg/kg erythromycin at t = 1-5 min that persisted for t = 30-40 min after infusion. These changes agree with the changes observed in the electromyogram. We did not observe any statistically significant difference in MGG amplitude before or after injection of either 3 or 6 mg/kg erythromycin. Both 3 and 6 mg/kg erythromycin infusion showed retrograde propagation with a statistically significant decrease in slow wave propagation velocity 11-20 min after infusion. Propagation velocity started returning toward baseline values after approximately 21-30 min for the 3 mg/kg dosage and after 31-40 min for a dosage of 6 mg/kg., Conclusion: Our results showed that the magnetic signatures were sensitive to disruptions in normal slow wave activity induced by pharmacological and prokinetic agents such as erythromycin., Significance: This study shows that repeatable noninvasive bio-electro-magnetic techniques can objectively characterize gastric dysrhythmias and may quantify treatment efficacy in patients with functional gastric disorders.

Background: The precise detection of cortical sleep spindles is critical to basic research on memory consolidation in rodents. Previous research using automatic spindle detection algorithms often lacks systematic parameter variations and validations., New Method: We present a method to systematically tune and validate algorithm parameters in automatic spindle detection algorithms using a moderate number of human raters., Results: Comparing a Hilbert transform-based algorithm to a ground truth constructed by six human raters, this method produced a parameter set yielding an F1 score of 0.82 at 10 ms resolution. The algorithm performance fell within the range of human agreement with the ground truth. Both human and algorithm failures arose largely from disagreement in spindle boundaries rather than spindle occurrence. With no additional tuning, the algorithm performed similarly in recordings from different days or rats., Comparison With Existing Methods: Most spindle detection algorithms do not perform systematic parameter variations and validation using a ground truth. To our knowledge, our study is the first in which rodent spindle data is scored by humans, and in which an automatic spindle detection algorithm is evaluated with respect to this ground truth. The rodent data from this study make it possible to compare our algorithm with others previously tested on human data., Conclusions: We present a general ground truth based approach for the tuning and validation of spindle extraction algorithms and suggest that algorithms aimed at extracting precise spindle timing in rats should use a systematic approach for parameter tuning., (Copyright © 2018. Published by Elsevier B.V.)

Electrophysiology is a direct measure of neuronal processes, and it is uniquely sensitive to canonical neural operations that underlie emergent psychological operations. These qualities make it well suited for discovery of aberrant neural mechanisms that underlie complicated disease states. This technique is routinely utilized in vitro, in vivo, and in outpatient neurological clinics, offering a translatable bridge between animal models and human patients. The bench-to-bedside potential of this approach is unparalleled, yet it also remains undeveloped due to the slow inertia of legacy techniques and interpretations. In this review, I discuss these strengths of the method, and I detail compelling reasons why future advancements can have a direct and tangible influence over clinical practice. I hope to motivate a blurring of traditional boundaries between preclinical, computational, imaging, and clinical fields by advancing electrophysiology as a common hub for methodological integration and theoretical advancement., (© 2018 Society for Psychophysiological Research.)

Objective: The use of microelectrode array (MEA) recordings is a very effective neurophysiological method because it is able to continuously and noninvasively obtain the spatiotemporal information of electrical activity from many neurons constituting a neural network. Very recently, studies have been published that used MEAs for the measurement of a low-frequency component of electrical activity as an indicator of diverse activity of cultured neurons. The occurrence of low-frequency activities has electrophysiological information that does not include the information from fast spikes. However, there is no in vitro experimental model suitable for measuring the low-frequency activities (slow-waves) for further study., Methods: Neural clusters consisting of dozens of neurons were placed directly onto each electrode of an MEA from which fast spikes and slow-waves were measured., Results: We obtained sufficient data on the early development patterns of the slow-waves and the spikes measured from many independent neural clusters confirming that the slow-waves occurred first before the emergence of the spikes in the neural clusters. We also showed that changes in the occurrence frequency of the slow-waves for synaptic blockers were measured from a large number of independent cultures., Conclusion: Microsized neural cluster arrays, which can be combined with conventional MEAs, are suitable for multiple simultaneous recordings of slow-waves., Significance: Our technology provides a simple but useful method to study the generation of a low-frequency component of the electrical activity in cultured neural networks that are not yet well known as well as to expand the use of conventional MEAs.

Extracellular recordings by means of high-density microelectrode arrays (HD-MEAs) have become a powerful tool to resolve subcellular details of single neurons in active networks grown from dissociated cells. To extend the application of this technology to slice preparations, we developed models describing how extracellular signals, produced by neuronal cells in slices, are detected by microelectrode arrays. The models help to analyze and understand the electrical-potential landscape in an in vitro HD-MEA-recording scenario based on point-current sources. We employed two modeling schemes, (i) a simple analytical approach, based on the method of images (MoI), and (ii) an approach, based on finite-element methods (FEM). We compared and validated the models with large-scale, high-spatiotemporal-resolution recordings of slice preparations by means of HD-MEAs. We then developed a model-based localization algorithm and compared the performance of MoI and FEM models. Both models provided accurate localization results and a comparable and negligible systematic error, when the point source was in saline, a condition similar to cell-culture experiments. Moreover, the relative random error in the x-y-z-localization amounted only up to 4.3% for z-distances up to 200 μm from the HD-MEA surface. In tissue, the systematic errors of both, MoI and FEM models were significantly higher, and a pre-calibration was required. Nevertheless, the FEM values proved to be closer to the tissue experimental results, yielding 5.2 μm systematic mean error, compared to 22.0 μm obtained with MoI. These results suggest that the medium volume or "saline height", the brain slice thickness and anisotropy, and the location of the reference electrode, which were included in the FEM model, considerably affect the extracellular signal and localization performance, when the signal source is at larger distance to the array. After pre-calibration, the relative random error of the z-localization in tissue was only 3% for z-distances up to 200 μm. We then applied the model and related detailed understanding of extracellular recordings to achieve an electrically-guided navigation of a stimulating micropipette, solely based on the measured HD-MEA signals, and managed to target spontaneously active neurons in an acute brain slice for electroporation.

A multielectrode system that can address widely separated targets at multiple sites across multiple brain regions with independent implant angling is needed to investigate neural function and signaling in systems and circuits of small animals. Here, we present the systemDrive, a novel multisite, multiregion microdrive that is capable of moving microwire electrode bundles into targets along independent and nonparallel drive trajectories. Our design decouples the stereotaxic surgical placement of individual guide cannulas for each trajectory from the placement of a flexible drive structure. This separation enables placement of many microwire multitrodes along widely spaced and independent drive axes with user-set electrode trajectories and depths from a single microdrive body, and achieves stereotaxic precision with each. The system leverages tight tube-cannula tolerances and geometric constraints on flexible drive axes to ensure concentric alignment of electrode bundles within guide cannulas. Additionally, the headmount and microdrive both have an open-center design to allow for the placement of additional sensing modalities. This design is the first, in the context of small rodent chronic research, to provide the capability to finely position microwires through multiple widely distributed cell groups, each with stereotaxic precision, along arbitrary and nonparallel trajectories that are not restricted to emanate from a single source. We demonstrate the use of the systemDrive in male Long-Evans rats to observe simultaneous single-unit and multiunit activity from multiple widely separated sleep-wake regulatory brainstem cell groups, along with cortical and hippocampal activity, during free behavior over multiple many-day continuous recording periods.

Intracellular recordings in vivo remains the best technique to link single-neuron electrical properties to network function. Yet existing methods are limited in accuracy, throughput, and duration, primarily via washout, membrane damage, and movement-induced failure. Here, we introduce flexible quartz nanopipettes (inner diameters of 10-25 nm and spring constant of ∼0.08 N/m) as nanoscale analogs of traditional glass microelectrodes. Nanopipettes enable stable intracellular recordings (seal resistances of 500 to ∼800 MΩ, 5 to ∼10 cells/nanopipette, and duration of ∼1 hr) in anaesthetized and awake head-restrained mice, exhibit minimal diffusional flux, and facilitate precise recording and stimulation. When combined with quantum-dot labels and microprisms, nanopipettes enable two-photon targeted electrophysiology from both somata and dendrites, and even paired recordings from neighboring neurons, while permitting simultaneous population imaging across cortical layers. We demonstrate the versatility of this method by recording from parvalbumin-positive (Pv) interneurons while imaging seizure propagation, and we find that Pv depolarization block coincides with epileptic spread. Flexible nanopipettes present a simple method to procure stable intracellular recordings in vivo., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

A complete characterization of temperature -and voltage-activated TRP channel gating requires a precise determination of the absolute probability of opening in a wide range of voltages, temperatures, and agonist concentrations. We have achieved this in the case of the TRPM8 channel expressed in Xenopus laevis oocytes. Measurements covered an extensive range of probabilities and unprecedented applied voltages up to 500 mV. In this chapter, we describe animal care protocols of patch-clamp pipette preparation, temperature control methods, and analysis of ionic currents to obtain reliable absolute open channel probabilities.

Background: Quantal exocytosis of oxidizable neurotransmitters can be detected as spikes of amperometric current using electrochemical microelectrodes. Measurements of spike parameters indicate the maximal transmitter flux, flux duration, and amount of transmitter released from individual vesicles. Automated analysis algorithms need to reject spikes that overlap in time. In addition, many spikes are preceded by small amplitude "foot" signals, attributed to slow release of transmitter through a fusion pore. Accurate pre-spike baseline determination is essential for estimating fusion-pore duration and the amount of transmitter released through the fusion pore., New Method: We developed an estimation approach that is based on fitting a multi-exponential function to the data. Our previously described matched-filter algorithm is used to identify the sections of data to fit and provides seed values to facilitate convergence of the iterative fit. The new estimation algorithm includes overlap rejection, a two-step fitting procedure and a novel baseline estimation procedure., Results: Histograms of spike parameters demonstrate excellent agreement of the new approach with manually computed parameters., Comparison With Existing Methods: Parameter estimates generated using the new approach are closer to blind manual estimates than commonly used existing methods. The improved performance is due to better detection of valid spikes and rejection of overlapping spikes. Moreover, since the complete time course of the spike is fit to a function, more complete information about the spike time course is captured., Conclusions: The matched-filter seeded algorithm reliably rejects overlaps and estimates spike and foot signal parameters in a fully automated manner., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Metabotropic glutamate receptors (mGluRs) are one of the major types of glutamatergic receptors contributing to synaptic plasticity mechanisms such as long-term potentiation (LTP) and long-term depression. Interestingly, activation of mGluRs alone can engage metaplastic mechanisms that create a new neuronal state, facilitating the induction and maintenance of future LTP. Here we describe typical methods used to investigate mGluR-induced metaplasticity in acute hippocampal slices. While this chapter focuses on in vitro field electrophysiological investigations, many of the principles can be applied to single-cell recordings as well as in vivo electrophysiology and indeed many types of metaplasticity phenomena.

Development of the whole-cell patch-clamp electrophysiology technique has allowed for enhanced visualization and experimentation of ionic currents in neurons of mammalian tissue with high spatial and temporal resolution. Electrophysiology has become an exceptional tool for identifying single cellular mechanisms underlying behavior. Specifically, the role of glutamatergic signaling through α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors underlying behavior has been extensively studied. Here we will discuss commonly used protocols and techniques for performing whole-cell patch-clamp recordings and exploring AMPA and NMDA receptor-mediated glutamatergic responses and alterations in the context of substance abuse.

The recent years have seen unprecedented growth in the manufacturing of neurotechnological tools. The latest technological advancements presented the neuroscientific community with neuronal probes containing thousands of recording sites. These next-generation probes are capable of simultaneously recording neuronal signals from a large number of channels. Numerically, a simple 128-channel neuronal data acquisition system equipped with a 16 bits A/D converter digitizing the acquired analog waveforms at a sampling frequency of 20 kHz will generate approximately 17 GB uncompressed data per hour. Today's biggest challenge is to mine this staggering amount of data and find useful information which can later be used in decoding brain functions, diagnosing diseases, and devising treatments. To this goal, many automated processing and analysis tools have been developed and reported in the literature. A good amount of them are also available as open source for others to adapt them to individual needs. Focusing on extracellularly recorded neuronal signals in vitro, this chapter provides an overview of the popular open-source tools applicable on these signals for spike trains and local field potentials analysis, and spike sorting. Towards the end, several future research directions have also been outlined.

Since their introduction in the early 1970s, microelectrode arrays (MEAs) have been dominating the electrophysiology market thanks to their reliability, extreme robustness, and usability. Over the past 40 years, silicon technology has also played a role in the advancement of the field, and CMOS-based in vitro and in vivo systems are now able to achieve unprecedented spatial resolutions, giving the possibility to unveil hidden behavior of cellular aggregates down to the subcellular level. However, both the MEAs and silicon-based electronic devices present unavoidable problems such as their expensiveness, the usual rigidity of the employed materials, and the need of an (usually bulky) external reference electrode. Possible interesting alternatives to these incredibly useful devices unexpectedly lie in the field of organic electronics, thanks to the fast-growing pace of improvement that this discipline has undergone in the last 10-15 years. In this chapter, a particular organic transistor called organic charge-modulated field-effect transistor (OCMFET) will be presented as a promising bio-electronic interface, and a complete description of its employment as a detector of cellular electrical activity and as an ultrasensitive pH sensor will be provided, together with the discussion about the possibility of using such a device as an innovative multisensing tool for both electrophysiology and (neuro)pharmacology.

Probing of glutamate receptors at sub-millisecond time scale is a key element needed for understanding of their response kinetics, neural signal transduction, and, in a wider context, intercellular cross talk. One of the classical techniques used to obtain this type of data in electrophysiology is placing a recording pipette in front of a double-barrel solution application pipette, which provides a rapid switch between two solutions. Here we describe a modification of this classical technique, which utilizes a solution application pipette with several loading capillaries. Such a system is capable of replacing multiple application solutions within 7-12 s time intervals. This modified protocol enables ultrafast application of several solutions at the same set of receptors, thus allowing powerful paired-sample statistical approaches. In addition, the same experimental equipment fabricated for the ultra-resolution probing of receptor kinetics can also be applied in several other types of electrophysiological experiments.

High-density microelectrode arrays (HD-MEAs) are increasingly being used for the observation and manipulation of neurons and networks in vitro. Large-scale electrode arrays allow for long-term extracellular recording of the electrical activity from thousands of neurons simultaneously. Beyond population activity, it has also become possible to extract information of single neurons at subcellular level (e.g., the propagation of action potentials along axons). In effect, HD-MEAs have become an electrical imaging platform for label-free extraction of the structure and activation of cells in cultures and tissues. The quality of HD-MEA data depends on the resolution of the electrode array and the signal-to-noise ratio. In this chapter, we begin with an introduction to HD-MEA signals. We provide an overview of the developments on complementary metal-oxide-semiconductor or CMOS-based HD-MEA technology. We also discuss the factors affecting the performance of HD-MEAs and the trending application requirements that drive the efforts for future devices. We conclude with an outlook on the potential of HD-MEAs for advancing basic neuroscience and drug discovery.

Neurotoxicity can be defined by the ability of a drug or chemical to alter the physiology, biochemistry, or structure of the nervous system in a manner that may negatively impact the health or function of the individual. Electrophysiological approaches have been utilized to study the mechanisms underlying neurotoxic actions of drugs and chemicals for over 50 years, and in more recent decades, high-throughput patch-clamp approaches have been utilized by the pharmaceutical industry for drug development. The use of microelectrode array recordings to study neural network electrophysiology is a relatively newer approach, with commercially available systems becoming available only in the early 2000s. However, MEAs have been rapidly adopted as a useful approach for neurotoxicity testing. In this chapter, I will review the use of MEA approaches as they have been applied to the field of neurotoxicity testing, especially as they have been applied to the need to screen large numbers of chemicals for neurotoxicity and developmental neurotoxicity. In addition, I will also identify challenges for the field that when addressed will improve the utility of MEA approaches for toxicity testing.

The aim of this review was to provide an overview of the most important stages in the development of cellular electrophysiology. The period covered starts with Bernstein's formulation of the membrane hypothesis and the measurement of the nerve and muscle action potential. Technical innovations make discoveries possible. This was the case with the use of the squid giant axon, allowing the insertion of "large" intracellular electrodes and derivation of transmembrane potentials. Application of the newly developed voltage clamp method for measuring ionic currents, resulted in the formulation of the ionic theory. At the same time transmembrane measurements were made possible in smaller cells by the introduction of the microelectrode. An improvement of this electrode was the next major (r)evolution. The patch electrode made it possible to descend to the molecular level and record single ionic channel activity. The patch technique has been proven to be exceptionally versatile. In its whole-cell configuration it was the solution to measure voltage clamp currents in small cells. See also: https://doi.org/10.14814/phy2.13860 & https://doi.org/10.14814/phy2.13862., (© 2019 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)