Serotype-based evaluation of an optogenetic construct in rat cortical astrocytes

Balachandar, Lakshmini; Borrego, Diana; Riera, Jorge J.

doi:10.1016/j.bbrc.2022.01.027

Cited by 4 publications

(2 citation statements)

References 36 publications

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“…In contrast, a recent study reported positive ChR2 expression in targeting astrocytes using mCherry promoter and adeno-associated viral (AAV) vectors. An optogenetic construct, AAV-GFAP-ChR2(H134R)-mCherry was characterized and resulted in high astrocytic transduction (approximately 88%) with AAV8, successfully establishing light-induced intervention of astrocyte activity in the rat brain cortex [ 33 ].…”

Section: Resultsmentioning

confidence: 99%

Optogenetically Engineered Neurons Differentiated from Human SH-SY5Y Cells Survived and Expressed ChR2 in 3D Hydrogel

et al. 2022

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The cases of brain degenerative disease will rise as the human population ages. Current treatments have a transient effect and lack an investigative system that is physiologically relevant for testing. There is evidence suggesting optogenetic stimulation is a potential strategy; however, an in vitro disease and optogenetic model requires a three-dimensional microenvironment. Alginate is a promising material for tissue and optogenetic engineering. Although it is bioinert, alginate hydrogel is transparent and therefore allows optical penetration for stimulation. In this study, alginate was functionalized with arginine-glycine-aspartate acid (RGD) to serve as a 3D platform for encapsulation of human SH-SY5Y cells, which were optogenetically modified and characterized. The RGD-alginate hydrogels were tested for swelling and degradation. Prior to encapsulation, the cells were assessed for neuronal expression and optical-stimulation response. The results showed that RGD-alginate possessed a consistent swelling ratio of 18% on day 7, and degradation remained between 3.7–5% throughout 14 days. Optogenetically modified SH-SY5Y cells were highly viable (>85%) after lentiviral transduction and neuronal differentiation. The cells demonstrated properties of functional neurons, developing beta III tubulin (TuJ1)-positive long neurites, forming neural networks, and expressing vGlut2. Action potentials were produced upon optical stimulation. The neurons derived from human SH-SY5Y cells were successfully genetically modified and encapsulated; they survived and expressed ChR2 in an RGD-alginate hydrogel system.

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Section: Resultsmentioning

confidence: 99%

Optogenetically Engineered Neurons Differentiated from Human SH-SY5Y Cells Survived and Expressed ChR2 in 3D Hydrogel

et al. 2022

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“…Optogenetics has also helped understand astrocytic roles in contextual fear memory 56 and in modulation of spike-wave discharges in absence epilepsy, thereby shedding light on possible therapeutic approaches 57 . Optogenetic intervention of astrocytes coupled with theoretical evaluations can help further the understanding of the intricacies in astrocytic responses to light and help establish robust control and applied in translational animal studies 58,59 .…”

Section: Use Of Optogenetics As the Stimulation Modalitymentioning

confidence: 99%

Characterization of optimal optogenetic stimulation paradigms to evoke calcium events in cortical astrocytes

Balachandar

Moncion

Riera

2023

Preprint

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Understanding the roles of astrocytic calcium signaling in multiple brain regulatory mechanisms including metabolism, blood flow, neuromodulation and neuroinflammation has remained one of the enduring challenges in glial biology. To delineate astrocytic contribution from concurrent neuronal activity, it is vital to establish robust control and manipulate astrocytes using a technique like optogenetics due to its high cellular specificity and temporal resolution. Lack of an experimental paradigm to induce controlled calcium signaling in astrocytes has hindered progress in the field. To address this, in this study, we systematically characterize and identify light stimulation paradigms for inducing regulated, on-demand increases in astrocytic calcium in cortical astrocytes in MlC1-ChR2(C128S)-EYFP mice. We identified paradigms 20%, 40% and 60% (of T = 100s) to elicit robust calcium responses upon multiple stimulations, while the 95% paradigm exhibited a response only during the first stimulation. We also quantified several parameters, including peak height, Full Width Half Maximum (FWHM), and latencies, and observe that the 20% paradigm has the highest peak ΔF/F0 among the paradigms across all stimulations and the lowest FWHM during the first stimulation. Overall, the 20% paradigm is a favorable choice for eliciting robust astrocytic calcium responses in astrocytes while performing multiple stimulations.

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Multichannel Wireless Neurosensing System for battery-free monitoring of neuronal activity

Moncion

Balachandar

Venkatakrishnan

et al. 2022

Biosensors and Bioelectronics

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Serotype-based evaluation of an optogenetic construct in rat cortical astrocytes

Cited by 4 publications

References 36 publications

Optogenetically Engineered Neurons Differentiated from Human SH-SY5Y Cells Survived and Expressed ChR2 in 3D Hydrogel

Optogenetically Engineered Neurons Differentiated from Human SH-SY5Y Cells Survived and Expressed ChR2 in 3D Hydrogel

Characterization of optimal optogenetic stimulation paradigms to evoke calcium events in cortical astrocytes

Multichannel Wireless Neurosensing System for battery-free monitoring of neuronal activity

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