Scalable Measurements of Intrinsic Excitability in Human iPS Cell-Derived Excitatory Neurons Using All-Optical Electrophysiology

Williams, Luis A.; Joshi, Vaibhav; Murphy, Michael P.; Ferrante, John; Werley, Christopher A.; Brookings, Theodore; McManus, Owen B.; Grosse, Johannes; Davies, Ceri H.; Dempsey, Graham T.

doi:10.1007/s11064-018-2694-5

“…The ability of OSCA to robustly detect changes in network activity profiles, coupled with the flexibility of utility in neuronal culture preparations from any host species or preparation, especially those where genetic manipulation is impossible, cumbersome, or expensive, such as patient-derived human induced pluripotent stem cell-derived neurons ( Williams et al, 2019 ; Puppo et al, 2021 ), make this technique directly amenable for high-throughput screening. In recent years, there has been a resurgence in phenotypic screening for discovering new drug candidates, drug targets and for neurotoxicological testing, with a focus on in vitro models that translate to in vivo and ultimately the clinic ( Moffat et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Optical Spike Detection and Connectivity Analysis With a Far-Red Voltage-Sensitive Fluorophore Reveals Changes to Network Connectivity in Development and Disease

Walker

¹

,

Raliski

²

,

Karbasi

³

et al. 2021

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The ability to optically record dynamics of neuronal membrane potential promises to revolutionize our understanding of neurobiology. In this study, we show that the far-red voltage sensitive fluorophore, Berkeley Red Sensor of Transmembrane potential-1, or BeRST 1, can be used to monitor neuronal membrane potential changes across dozens of neurons at a sampling rate of 500 Hz. Notably, voltage imaging with BeRST 1 can be implemented with affordable, commercially available illumination sources, optics, and detectors. BeRST 1 is well-tolerated in cultures of rat hippocampal neurons and provides exceptional optical recording fidelity, as judged by dual fluorescence imaging and patch-clamp electrophysiology. We developed a semi-automated spike-picking program to reduce user bias when calling action potentials and used this in conjunction with BeRST 1 to develop an optical spike and connectivity analysis (OSCA) for high-throughput dissection of neuronal activity dynamics. The high temporal resolution of BeRST 1 enables dissection of firing rate changes in response to acute, pharmacological interventions with commonly used inhibitors like gabazine and picrotoxin. Over longer periods of time, BeRST 1 also tracks chronic perturbations to neurons exposed to amyloid beta 1–42 (Aβ 1–42), revealing modest changes to spiking frequency but profound changes to overall network connectivity. Finally, we use OSCA to track changes in neuronal connectivity during maturation in culture, providing a functional readout of network assembly. We envision that use of BeRST 1 and OSCA described here will be of use to the broad neuroscience community.

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“…Application of optogenetic tools greatly exceeds neurons and cardiac cells, as demonstrated through reports on stem cells [ 33 ], connective tissue cells [ 49 ], plant cells [ 50 ], yeasts [ 51 , 52 ] and bacteria [ 53 ] that are starting to amass supported by both transient and transgenic formats.…”

Section: Generation Of Optogenetically Modified Cell Lines and The Complementary Tools For Their Characterizationmentioning

confidence: 99%

“…Other challenges and breakthroughs in the development of analytical tools for cell-based biosensing for specific applications can be found in recent review works [ 3 , 4 , 42 , 110 , 111 ], and thus, we limit the presentation. Instead, a special note is devoted to multimodal functional imaging approaches potentially suitable for ‘all-optical’ electrophysiological cell-based assays under current scrutiny [ 33 , 79 , 112 ]. ‘All-optical’ electrophysiology concerns the derivation of cellular electrical properties (for instance, membrane and action potentials, ion channels characterization) using only optical means.…”

Section: (Label-free) Analytical Tools Relevant For Optogenetic Cell Platformsmentioning

confidence: 99%

Advanced Optogenetic-Based Biosensing and Related Biomaterials

Gheorghiu

¹

,

Polonschii

²

,

Popescu

³

et al. 2021

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The ability to stimulate mammalian cells with light, brought along by optogenetic control, has significantly broadened our understanding of electrically excitable tissues. Backed by advanced (bio)materials, it has recently paved the way towards novel biosensing concepts supporting bio-analytics applications transversal to the main biomedical stream. The advancements concerning enabling biomaterials and related novel biosensing concepts involving optogenetics are reviewed with particular focus on the use of engineered cells for cell-based sensing platforms and the available toolbox (from mere actuators and reporters to novel multifunctional opto-chemogenetic tools) for optogenetic-enabled real-time cellular diagnostics and biosensor development. The key advantages of these modified cell-based biosensors concern both significantly faster (minutes instead of hours) and higher sensitivity detection of low concentrations of bioactive/toxic analytes (below the threshold concentrations in classical cellular sensors) as well as improved standardization as warranted by unified analytic platforms. These novel multimodal functional electro-optical label-free assays are reviewed among the key elements for optogenetic-based biosensing standardization. This focused review is a potential guide for materials researchers interested in biosensing based on light-responsive biomaterials and related analytic tools.

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“…The ability of OSCA to robustly detect changes in network activity profiles, coupled with the flexibility of utility in neuronal culture preparations from any host species or preparation, especially those where genetic manipulation is impossible, cumbersome, or expensive, such as patient-derived human induced pluripotent stem cell-derived neurons (Williams et al, 2019), make this technique directly amenable for high-throughput screening. In recent years there has been a resurgence in phenotypic screening for discovering new drug candidates, drug targets and for neurotoxicological testing, with a focus on in vitro models that translate to in vivo and ultimately the clinic (Moffat et al, 2017).…”

Section: Looking To the Future: A Role For Osca As A Complementary Scmentioning

confidence: 99%

Optical spike detection and connectivity analysis with a far-red voltage-sensitive fluorophore reveals changes to network connectivity in development and disease

Walker

¹

,

Raliski

²

,

Karbasi

³

et al. 2020

Preprint

View full text Add to dashboard Cite

The ability to optically record dynamics of neuronal membrane potential promises to revolutionize our understanding of neurobiology. In this study, we show that the far-red voltage sensitive fluorophore, Berkeley Red Sensor of Transmembrane potential −1, or BeRST 1, can be used to monitor neuronal membrane potential changes across dozens of neurons at a sampling rate of 500 Hz. Notably, voltage imaging with BeRST 1 can be implemented with affordable, commercially available illumination sources, optics, and detectors. BeRST 1 is well-tolerated in cultures of rat hippocampal neurons and provides exceptional optical recording fidelity, as judged by dual fluorescence imaging and patch-clamp electrophysiology. We developed a semi-automated spike-picking program to reduce user bias when calling action potentials and used this in conjunction with BeRST 1 to develop an optical spike and connectivity analysis workflow (OSCA) for high-throughput dissection of neuronal activity dynamics in development and disease. The high temporal resolution of BeRST 1 enables dissection of firing rate changes in response to acute, pharmacological interventions with commonly used inhibitors like gabazine and picrotoxin. Over longer periods of time, BeRST 1 also tracks chronic perturbations to neurons exposed to amyloid beta (Aβ1-42), revealing modest changes to spiking frequency but profound changes to overall network connectivity. Finally, we use OSCA to track changes in neuronal connectivity during development, providing a functional readout of network assembly. We envision that use of BeRST 1 and OSCA described here will be of use to the broad neuroscience community.Significance StatementOptical methods to visualize membrane potential dynamics provide a powerful complement to Ca2+ imaging, patch clamp electrophysiology, and multi-electrode array recordings. However, modern voltage imaging strategies often require complicated optics, custom-built microscopes, or genetic manipulations that are impractical outside of a subset of model organisms. Here, we describe the use of Berkeley Red Sensor of Transmembrane potential, or BeRST 1, a far-red voltage-sensitive fluorophore that can directly visualize membrane potential changes with millisecond resolution across dozens of neurons. Using only commercially available components, voltage imaging with BeRST 1 reveals profound changes in neuronal connectivity during development, exposes changes to firing rate during acute pharmacological perturbation, and illuminates substantial increases in network connectivity in response to chronic exposure to amyloid beta.

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Scalable Measurements of Intrinsic Excitability in Human iPS Cell-Derived Excitatory Neurons Using All-Optical Electrophysiology

Cited by 17 publications

References 42 publications

Optical Spike Detection and Connectivity Analysis With a Far-Red Voltage-Sensitive Fluorophore Reveals Changes to Network Connectivity in Development and Disease

Optical Spike Detection and Connectivity Analysis With a Far-Red Voltage-Sensitive Fluorophore Reveals Changes to Network Connectivity in Development and Disease

Advanced Optogenetic-Based Biosensing and Related Biomaterials

Optical spike detection and connectivity analysis with a far-red voltage-sensitive fluorophore reveals changes to network connectivity in development and disease

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