Simultaneous conduction mapping and intracellular membrane potential recording in isolated atria

2016 ◽  
Vol 94 (5) ◽  
pp. 563-569 ◽  
Author(s):  
Melissa Neo ◽  
David G. Morris ◽  
Pawel Kuklik ◽  
Dennis H. Lau ◽  
Hany Dimitri ◽  
...  

We describe a novel approach for simultaneously determining regional differences in action potential (AP) morphology and tissue electrophysiological properties in isolated atria. The epicardial surface of rat atrial preparations was placed in contact with a multi-electrode array (9 × 10 silver chloride electrodes, 0.1 mm diameter and 0.1 mm pitch). A glass microelectrode (100 MΩ) was simultaneously inserted into the endocardial surface to record intracellular AP from either of 2 regions (A, B) during pacing from 2 opposite corners of the tissue. AP duration at 80% of repolarisation and its restitution curve was significantly different only in region A (p < 0.01) when AP was initiated at different stimulation sites. Alternans in AP duration and AP amplitude, and in conduction velocity were observed during 2 separate arrhythmic episodes. This approach of combining microelectrode array and intracellular membrane potential recording may provide new insights into arrhythmogenic mechanisms in animal models of cardiovascular disease.

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
S Glatstein ◽  
M Ghiringhelli ◽  
L Maizels ◽  
E Heller ◽  
E Maor ◽  
...  

Abstract Background One of the major barriers to an improved mechanistic understanding of atrial fibrillation (AF), and thus in the pipeline of drug development, has been a lack of appropriate tissue models, especially in small animals. Aim We propose an advanced anatomical ex-vivo model based on rat atria for acute assessment of AF susceptibility. This novel model could yield a better understanding of arrhythmia mechanisms as well as the development of potential therapeutic strategies for the prevention or termination of atrial arrhythmias. Methods Wistar rats atria (N=25) were isolated, flattened and pinned to a custom-made silicon plate. Atria were superfused with an oxygenized Tyrode's solution. Tissues were then loaded with a voltage-sensitive dye and mapped using a high-resolution optical mapping system. AF was induced with 1uM carbamylcholine (N=23) coupled with pacing maneuvers and treated with 30uM Vernakalant (N=10) or 10uM Flecainide (N=10). Finally, the feasibility of a new ablation technique (electroporation) was evaluated. Results Optical mapping results suggested that the superfusion procedure led to a fast atrial recovery. Sinus activity was conserved for all atria for a long period. All the anatomical landmarks were clearly visualized. The acquired optical signals were analyzed during sinus rhythm and pacing, which allowed the creation of detailed activation maps and measurements of action potential duration (APD) and conduction velocity (CV) at different pacing rates. The resulting APD restitution curves revealed electrical excitation at high pacing rates (cycle length between 50ms and 300ms) with a relatively flattened curve. AF was successfully induced and optically mapping confirmed the presence of reentrant activity. AF was successfully treated using Vernacalant and Flecainide. Finally, we demonstrated the feasibility of a new ablation approach (electroporation) for creation of a continuous linear lesion serving as a functional block. Conclusion The isolated superfused atria model, coupled with voltage-sensitive dyes, can be utilized for long-term high-resolution functional imaging of the atria during sinus rhythm, pacing and arrhythmogenic activity. This allows the study of the atrial electrophysiological properties, the mechanisms involved in AF initiation, perpetuation, and termination as well as the study of drug and new ablation modalities. FUNDunding Acknowledgement Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Research Council (ERC) Spontaneous activation of isolated atria


Sensors ◽  
2020 ◽  
Vol 20 (18) ◽  
pp. 5183
Author(s):  
Afraiz Tariq Satti ◽  
Jinsoo Park ◽  
Jangwoong Park ◽  
Hansang Kim ◽  
Sungbo Cho

Microneedle array electrodes (MNE) showed immense potential for the sensitive monitoring of the bioelectric signals by penetrating the stratum corneum with high electrical impedance. In this paper, we introduce a rigid parylene coated microneedle electrode array and portable electrocardiography (ECG) circuit for monitoring of ECG reducing the motion artifacts. The developed MNE showed stability and durability for dynamic and long-term ECG monitoring in comparison to the typical silver-silver chloride (Ag/AgCl) wet electrodes. The microneedles showed no mechanical failure under the compression force up-to 16 N, but successful penetration of skin tissue with a low insertion force of 5 N. The electrical characteristics of the fabricated MNE were characterized by impedance spectroscopy with equivalent circuit model. The designed wearable wireless ECG monitoring device with MNE proved feasibility of the ECG recording which reduces the noise of movement artifacts during dynamic behaviors.


1987 ◽  
Vol 58 (1) ◽  
pp. 180-194 ◽  
Author(s):  
F. R. Morales ◽  
P. A. Boxer ◽  
S. J. Fung ◽  
M. H. Chase

1. The electrophysiological properties of alpha-motoneurons in old cats (14–15 yr) were compared with those of adult cats (1–3 yr). These properties were measured utilizing intracellular recording and stimulating techniques. 2. Unaltered in the old cat motoneurons were the membrane potential, action potential amplitude, and slopes of the initial segment (IS) and soma dendritic (SD) spikes, as well as the duration and amplitude of the action potential's afterhyperpolarization. 3. In contrast, the following changes in the electrophysiological properties of lumbar motoneurons were found in the old cats: a decrease in axonal conduction velocity, a shortening of the IS-SD delay, an increase in input resistance, and a decrease in rheobase. 4. In spite of these considerable changes in motoneuron properties in the old cat, normal correlations between different electrophysiological properties were maintained. The following key relationships, among others, were the same in adult and old cat motoneurons: membrane potential polarization versus action potential amplitude, duration of the afterhyperpolarization versus motor axon conduction velocity, and rheobase versus input conductance. 5. A review of the existing literature reveals that neither chronic spinal cord section nor deafferentation (13, 21) in adult animals produce the changes observed in old cats. Thus we consider it unlikely that a loss of synaptic contacts was responsible for the modifications in electrophysiological properties observed in old cat motoneurons. 6. We conclude that during old age there are significant changes in the soma-dendritic portion of cat motoneurons, as indicated by the modifications found in input resistance, rheobase, and IS-SD delay, as well as significant changes in their axons, as indicated by a decrease in conduction velocity.


2003 ◽  
Vol 773 ◽  
Author(s):  
Shalini Prasad ◽  
Mo Yang ◽  
Xuan Zhang ◽  
Yingchun Ni ◽  
Vladimir Parpura ◽  
...  

AbstractCharacterization of electrical activity of individual neurons is the fundamental step in understanding the functioning of the nervous system. Single cell electrical activity at various stages of cell development is essential to accurately determine in in-vivo conditions the position of a cell based on the procured electrical activity. Understanding memory formation and development translates to changes in the electrical activity of individual neurons. Hence, there is an enormous need to develop novel ways for isolating and positioning individual neurons over single recording sites. To this end, we used a 3x3 multiple microelectrode array system to spatially arrange neurons by applying a gradient AC field. We characterized the electric field distribution inside our test platform by using two dimensiona l finite element modeling (FEM) and determined the location of neurons over the electrode array. Dielectrophoretic AC fields were utilized to separate the neurons from the glial cells and to position the neurons over the electrodes. The neurons were obtained from 0-2-day-old rat (Sprague-Dawley) pups. The technique of using electric fields to achieve single neuron patterning has implications in neural engineering, elucidating a new and simpler method to develop and study neuronal activity as compared to conventional microelectrode array techniques.


2019 ◽  
Vol 317 (6) ◽  
pp. C1256-C1267 ◽  
Author(s):  
Simon P. Wells ◽  
Helen M. Waddell ◽  
Choon Boon Sim ◽  
Shiang Y. Lim ◽  
Gabriel B. Bernasochi ◽  
...  

Cardiac arrhythmias of both atrial and ventricular origin are an important feature of cardiovascular disease. Novel antiarrhythmic therapies are required to overcome current drug limitations related to effectiveness and pro-arrhythmia risk in some contexts. Cardiomyocyte culture models provide a high-throughput platform for screening antiarrhythmic compounds, but comparative information about electrophysiological properties of commonly used types of cardiomyocyte preparations is lacking. Standardization of cultured cardiomyocyte microelectrode array (MEA) experimentation is required for its application as a high-throughput platform for antiarrhythmic drug development. The aim of this study was to directly compare the electrophysiological properties and responses to isoproterenol of three commonly used cardiac cultures. Neonatal rat ventricular myocytes (NRVMs), immortalized atrial HL-1 cells, and custom-generated human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were cultured on microelectrode arrays for 48–120 h. Extracellular field potentials were recorded, and conduction velocity was mapped in the presence/absence of the β-adrenoceptor agonist isoproterenol (1 µM). Field potential amplitude and conduction velocity were greatest in NRVMs and did not differ in cardiomyocytes isolated from male/female hearts. Both NRVMs and hiPSC-CMs exhibited longer field potential durations with rate dependence and were responsive to isoproterenol. In contrast, HL-1 cells exhibited slower conduction and shorter field potential durations and did not respond to 1 µM isoproterenol. This is the first study to compare the intrinsic electrophysiologic properties of cultured cardiomyocyte preparations commonly used for in vitro electrophysiology assessment. These findings offer important comparative data to inform methodological approaches in the use of MEA and other techniques relating to cardiomyocyte functional screening investigations of particular relevance to arrhythmogenesis.


1993 ◽  
Vol 70 (6) ◽  
pp. 2379-2390 ◽  
Author(s):  
F. Tell ◽  
A. Jean

1. Activation of N-methyl-D-aspartate (NMDA) receptors in caudal nucleus tractus solitarii (cNTS) neurons elicited endogenous rhythmic activities. We used an in vitro brain stem slice preparation to determine the ionic mechanisms underlying the generation of these activities. 2. Using intracellular recordings, we found several ionic conductances to be responsible for the electrophysiological properties of cNTS neurons. After addition of tetrodotoxin (TTX) to the perfusate, cNTS neurons were still able to generate action potentials (APs). Because these APs were suppressed by the addition of cobalt or by the reduction of calcium, they were likely due to calcium currents (ICa). In addition, the amplitude of the afterhyperpolarization (AHP) that followed a train of TTX-resistant APs was reduced in both low-calcium and cobalt-containing saline. It was therefore suggested that calcium-activated potassium (IKCa) currents were involved in the AHP. Accordingly, application of apamin, a blocker of slow IKCa, also decreased the AHP. cNTS neurons exhibited a delayed excitation phenomenon, characterized by a ramplike depolarization that delayed the onset of neuronal firing, when they were depolarized from hyperpolarizing potential. The underlying current was presumed to be an A-current (IKA), because this phenomenon was suppressed during application of 4-aminopyridine (4-AP). 3. Application of NMDA elicited different types of discharge patterns in cNTS neurons: a repetitive firing at depolarized levels of membrane potential (above -60 mV) and rhythmic patterns characterized by either rhythmic bursting or rhythmic single discharges at hyperpolarized levels (within membrane potential range of -60 to -85 mV). In all neurons, rhythmic patterns were superimposed on oscillations of membrane potential. They were characterized by a sudden shift of membrane potential, followed by a ramp-shaped phase of depolarization that preceded spike elicitation. Addition of TTX to the saline did not suppress NMDA-induced oscillations. Therefore rhythmic patterns were not driven by synaptic mechanisms but resulted from endogenous properties of cNTS neurons. 4. APs superimposed on NMDA-induced depolarizations presented the same characteristics as those elicited by positive current pulses. NMDA-elicited oscillations of membrane potential were eliminated by removing magnesium from the saline. Therefore oscillation generation was based primarily on the NMDA channel properties. 5. Intrinsic conductances of cNTS neurons interacted with NMDA-gated conductances to shape the depolarization waveform. Because removal of calcium from the saline suppressed endogenous oscillations, ICa currents were required for the expression of rhythmic activities. IKCa currents were involved in the repolarization phase of oscillations because apamin increased the duration of the oscillations.(ABSTRACT TRUNCATED AT 400 WORDS)


2016 ◽  
Vol 38 (3) ◽  
pp. 893-908 ◽  
Author(s):  
Ye Tao ◽  
Tao Chen ◽  
Zhongyu Liu ◽  
Weiwei Xu ◽  
Limin Qin ◽  
...  

Background/Aims: The pathphysiological properties of N-Methyl -N -nitrosourea (MNU) induced photoreceptor degeneration are similar to the hereditary retinitis pigmentosa (RP). The present study sought to explore the beneficial effects of the peonidin, a common aglycone form of anthocyanin, on the MNU induced photoreceptor degeneration via topographic measurements. Methods: The MNU administrated mouse received peonidin or vehicle injections, and then they were examined by electroretinography (ERG), multi electrode array (MEA), histological and immunohistochemistry studies. Results: The protective effects of peonidin on the MNU administrated retinas were systematically verified and quantified by topographic measures. The peonidin treatment could protect the photoreceptor against the MNU toxicity both functionally and morphologicaly. The most sensitive zone to peonidin therapy was sorted out, indicating that different rescuing kinetics existed between the retinal hemispheres and retinal quadrants. Moreover, the hyperactive spontaneous firing response and the debilitated light induced response in MNU administrated retinas could be partially reversed by peonidin treatment. To our knowledge, this was the first study to explore the pharmacological effects of peonidin on the electrophysiological properties of inner visual signal pathways. Conclusion: The peonidin could ameliorate the MNU induced photoreceptors degeneration and rectify the abnormities in the inner visual signal pathways. Future refinements of the knowledge cast insights into the discovery of a novel treatment for human RP.


2009 ◽  
Vol 69-70 ◽  
pp. 79-82 ◽  
Author(s):  
Yu Kui Wang ◽  
Zhen Long Wang ◽  
Mao Sheng Li ◽  
Wei Liang Zeng ◽  
M.H. Weng

In the paper, in order to overcome machining limits in throughput and precision because of positioning error and tool wear of a single tool electrode, a method for the microelectrode array fabrication by micro-WEDM is described and assessed. Characteristics of the microelectrode array fabrication by micro-WEDM, such as machining open voltage, pulse peak current, discharge duration and servo feed rate so on, are investigated through a series of experiments. A 10 10 squared electrode array is machined by micro-WEDM and the width of each squared electrode is about 40µm. The microelectrode array with good quality is obtained by applying decreased open voltage and peak current, increased discharge duration and optimized machining speed. Then micro hole-array is processed by applying obtained electrode array in micro-EDM method. The diameter of each squared hole in the array is about 50 µm due to appropriate control strategy that per micro pulse energy is decreased and periodic jump-down is applied during the machining process. Experiments have demonstrated that the combination process of microelectrode array fabricated by micro-WEDM and micro-hole array done by micro-EDM is a novel method of process which makes it more feasible and efficient to fabricate microelectrode array and high-density hole-array.


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