iPSC-Cardiomyocyte Models of Brugada Syndrome—Achievements,Challenges and Future Perspectives

Brugada syndrome (BrS) is an inherited cardiac arrhythmia that predisposes to ventricular fibrillation and sudden cardiac death. It originates from oligogenic alterations that affect cardiac ion channels or their accessory proteins. The main hurdle for the study of the functional effects of those variants is the need for a specific model that mimics the complex environment of human cardiomyocytes. Traditionally, animal models or transient heterologous expression systems are applied for electrophysiological investigations, each of these models having their limitations. The ability to create induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), providing a source of human patient-specific cells, offers new opportunities in the field of cardiac disease modelling. Contemporary iPSC-CMs constitute the best possible in vitro model to study complex cardiac arrhythmia syndromes such as BrS. To date, thirteen reports on iPSC-CM models for BrS have been published and with this review we provide an overview of the current findings, with a focus on the electrophysiological parameters. We also discuss the methods that are used for cell derivation and data acquisition. In the end, we critically evaluate the knowledge gained by the use of these iPSC-CM models and discuss challenges and future perspectives for iPSC-CMs in the study of BrS and other arrhythmias.     

基于精细解剖结构的左心室心肌缺血仿真

为解释心肌缺血诱发心律失常的机理,基于精细的人体心脏解剖数据,构建了一个整合的人体心室组织模型,并模拟了心肌缺血对折返波的影响.在处理无通量边界问题上,采用相场法来自动处理复杂的边界条件.通过增加缺血区域内细胞外钾离子浓度值,模拟了心肌缺血环境下折返波的传播过程.实验结果表明,折返波会随着缺血程度的改变而变化.随着缺血...     

Responses of pigeon (Columba livia) Wulst neurons during acquisition and reversal of a visual discrimination task.

The avian visual "Wulst" is a target of the ascending thalamofugal visual pathway. In pigeons (Columba livia), lesion damage to the Wulst has little effect on simple visual discriminations, but impairs performance on tasks such as reversal learning. We recorded the responses of single Wulst neurons as pigeons were trained on the acquisition and subsequent reversal of a visual discrimination. Of the 64 units recorded, 54 (84%) displayed a significant difference in firing rate between some component of the task and the intertrial interval that separated trials. More important, 14 units (22%) displayed a significant change in firing rate exclusively to the S+ and/or S- as learning progressed either during acquisition or reversal. The responses of these 14 neurons indicate that learning during initial acquisition was as likely to correlate with a change in firing rate as during reversal, and some neuronal responses could be characterized as representing reward properties together with visual stimulus features. As such, responses of pigeon Wulst neurons indicate a role in representing aspects of learning as much as the physical/perceptual properties of visual stimuli. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

用于长期神经元放电记录的一体化可步进植入式神经电极

步进式电极是神经科学电生理记录的重要工具。传统电极支架的主要功能是电极丝支撑以及机械驱动电极丝的微推进。在慢性记录过程中,电极的位置可以推进到更深的脑组织中,从而记录更多的神经元放电活动。但传统电极支架制作和组装需要多个步骤和部件,组装过程烦琐困难、结构集成度低,且无法保证支撑板结构之间相互平行,增加了实验误差。该文提出一种可实现集成度高、结构稳定、组装容易的新型电极构架。与传统电极支架相比,新设计的电极支架具有更少的组件,且一体化的支架设计减少了不同支架之间的误差,有助于实验条件的统一。受力分析表明,该文提出的新电极具有优良的抗形变特性,且新电极比传统电极重量更小,可减轻实验小鼠头部负载压力。通过手术在小鼠大脑中植入电极,实验结果表明一体化记录电极可获得高质量的神经信号。因此,该文提出了一种新的电极设计思路,该思路有助于提高实验效率,并可应用于多种小动物在体电生理实验。     

Measurement of Electrophysiological Signals In Vitro Using High-Performance Organic Electrochemical Transistors

Biological environments use ions in charge transport for information transmission. The properties of mixed electronic and ionic conductivity in organic materials make them ideal candidates to transduce physiological information into electronically processable signals. A device proven to be highly successful in measuring such information is the organic electrochemical transistor (OECT). Previous electrophysiological measurements performed using OECTs show superior signal-to-noise ratios than electrodes at low frequencies. Subsequent development has significantly improved critical performance parameters such as transconductance and response time. Here, interdigitated-electrode OECTs are fabricated on flexible substrates, with one such state-of-the-art device achieving a peak transconductance of 139 mS with a 138 µs response time. The devices are implemented into an array with interconnects suitable for micro-electrocorticographic application and eight architecture variations are compared. The two best-performing arrays are subject to the full electrophysiological spectrum using prerecorded signals. With frequency filtering, kHz-scale frequencies with 10 µV-scale voltages are resolved. This is supported by a novel quantification of the noise, which compares the gate voltage input and drain current output. These results demonstrate that high-performance OECTs can resolve the full electrophysiological spectrum and suggest that superior signal-to-noise ratios could be achieved in high frequency measurements of multiunit activity.     

Self-Folding Graphene-Based Interface for Brain-Like Modular 3D Tissue (Adv. Funct. Mater. 37/2023)

Electrophysiology of 3D neuronal cultures is of rapidly growing importance for revealing cellular communications associated with neurodevelopment and neurological diseases in their brain-like 3D environment. Despite that the brain also exhibits an inherent modular architecture that is essential for cortical processing, it remains challenging to interface a modular network consisting of multiple 3D neuronal tissues. Here, a self-folding graphene-based electrode array is proposed that enables to reconstruct modular 3D neuronal tissue and investigate firing dynamics among moduli. A graphene-sandwiched parylene-C film self-folds into a cylindrical structure within which living cells can be encapsulated. Culture of encapsulated cells inside the folded graphene enables to spontaneously construct 3D cell aggregates and ensure firm contact between the graphene surface and encapsulated cells. As the inner graphene surface can be utilized as an electrode, the reliable cell–electrode contact allows for long-term electrical recording from multiple 3D aggregates. Additionally, the modular network consisting of multiple 3D aggregates exhibits richer firing patterns than a conventional homogenous 2D network, which demonstrates that the approach enables measurements of firing dynamics in complex 3D neuronal networks. The deformable graphene electrode will be a powerful platform for investigating complex cellular communications in brain-like 3D cultures.     

Recent advances in insect olfaction, specifically regarding the morphology and sensory physiology of antennal sensilla of the female sphinx moth Manduca sexta.

The antennal flagellum of female Manduca sexta bears eight sensillum types: two trichoid, two basiconic, one auriculate, two coeloconic, and one styliform complex sensilla. The first type of trichoid sensillum averages 34 microm in length and is innervated by two sensory cells. The second type averages 26 microm in length and is innervated by either one or three sensory cells. The first type of basiconic sensillum averages 22 microm in length, while the second type averages 15 microm in length. Both types are innervated by three bipolar sensory cells. The auriculate sensillum averages 4 microm in length and is innervated by two bipolar sensory cells. The coeloconic type-A and type-B both average 2 microm in length. The former type is innervated by five bipolar sensory cells, while the latter type, by three bipolar sensory cells. The styliform complex sensillum occurs singly on each annulus and averages 38-40 microm in length. It is formed by several contiguous sensilla. Each unit is innervated by three bipolar sensory cells. A total of 2,216 sensilla were found on a single annulus (annulus 21) of the flagellum. Electrophysiological responses from type-A trichoid sensilla to a large panel of volatile odorants revealed three different subsets of olfactory receptor cells (ORCs). Two subsets responded strongly to only a narrow range of odorants, while the third responded strongly to a broad range of odorants. Anterograde labeling of ORCs from type-A trichoid sensilla revealed that their axons projected mainly to two large female glomeruli of the antennal lobe.     

Amygdalar unit activity during three learning tasks: Eyeblink classical conditioning, Pavlovian fear conditioning, and signaled avoidance conditioning.

Neural activity in central and basolateral amygdala nuclei (CeA and BLA, respectively) was recorded during delay eyeblink conditioning, Pavlovian fear conditioning, and signaled barpress avoidance. During paired training, the CeA exhibited robust learning-related excitatory activity during all 3 tasks. By contrast, the BLA exhibited minimal activity during eyeblink conditioning, while demonstrating pronounced increases in learning-related excitatory responsiveness during fear conditioning and barpress avoidance. In addition, the relative amount of amygdalar activation observed appeared to be related to the relative intensity of the unconditioned stimulus and somatic requirements of the task. Results suggest the CeA mediates the Pavlovian association between sensory stimuli and the BLA mediates the modulation of instrumental responding through the assignment of motivational value to the unconditioned stimulus. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Reversible Photocontrol of Dopaminergic Transmission in Wild-Type Animals

Understanding the dopaminergic system is a priority in neurobiology and neuropharmacology. Dopamine receptors are involved in the modulation of fundamental physiological functions, and dysregulation of dopaminergic transmission is associated with major neurological disorders. However, the available tools to dissect the endogenous dopaminergic circuits have limited specificity, reversibility, resolution, or require genetic manipulation. Here, we introduce azodopa, a novel photoswitchable ligand that enables reversible spatiotemporal control of dopaminergic transmission. We demonstrate that azodopa activates D₁-like receptors in vitro in a light-dependent manner. Moreover, it enables reversibly photocontrolling zebrafish motility on a timescale of seconds and allows separating the retinal component of dopaminergic neurotransmission. Azodopa increases the overall neural activity in the cortex of anesthetized mice and displays illumination-dependent activity in individual cells. Azodopa is the first photoswitchable dopamine agonist with demonstrated efficacy in wild-type animals and opens the way to remotely controlling dopaminergic neurotransmission for fundamental and therapeutic purposes.     

Electroantennogram responses ofScolytus multistriatus (Coleoptera: Scolytidae) to its pheromone components and to associated compounds

Electroantennograms were recorded fromScolytus multistriatus in response to 4-methyl-3-heptanol, the four geometric isomers of multistriatin, and cubeb oil. Charateristic dose-response curves for response amplitude and the time required for the voltage trace to return to 1/2 baseline (recovery rate) were established. Recovery rates were significantly more rapid following stimulation with 4-methyl-3-heptanol or cubeb oil than with the multistriatin isomers. At most intensities, -multistriatin, the isomer that evokes behavioral response, gave significantly larger EAGs with significantly longer recovery rates than the other isomers. Results of differential adaptation experiments suggested that 4-methyl-3-heptanol interacted with the processes involving multistriatin and cubeb oil activity. However, cross-activity of acceptors for these compounds seems unlikely; single sensillum recordings are needed to ascertain the response spectra for individual receptor neurons.From a thesis submitted as partial fulfillment of the Masters of Science degree, Department of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, Syracuse, New York 13210.