Probing Synaptic Signaling with Optogenetic Stimulation and Genetically Encoded Calcium Reporters.

Autor:	Borja GB; Q-State Biosciences Inc., Cambridge, MA, USA., Shroff H; Q-State Biosciences Inc., Cambridge, MA, USA., Upadhyay H; Q-State Biosciences Inc., Cambridge, MA, USA., Liu PW; Q-State Biosciences Inc., Cambridge, MA, USA., Baru V; Q-State Biosciences Inc., Cambridge, MA, USA., Cheng YC; Q-State Biosciences Inc., Cambridge, MA, USA., McManus OB; Q-State Biosciences Inc., Cambridge, MA, USA., Williams LA; Q-State Biosciences Inc., Cambridge, MA, USA., Dempsey GT; Q-State Biosciences Inc., Cambridge, MA, USA., Werley CA; Q-State Biosciences Inc., Cambridge, MA, USA. kit.werley@qstatebio.com.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2191, pp. 109-134.
DOI:	10.1007/978-1-0716-0830-2_8
Abstrakt:	Optogenetics provides a powerful approach for investigating neuronal electrophysiology at the scale required for drug discovery applications. Probing synaptic function with high throughput using optogenetics requires robust tools that enable both precise stimulation of and facile readout of synaptic activity. Here we describe two functional assays to achieve this end: (1) a pre-synaptic calcium assay that utilizes the channelrhodopsin, CheRiff, patterned optogenetic stimulus, and the pre-synaptically targeted calcium reporter jRGECO1a to monitor pre-synaptic changes in calcium influx and (2) a synaptic transmission assay in which CheRiff and cytosolic jRGECO1a are expressed in non-overlapping sets of neurons, enabling pre-synaptic stimulation and post-synaptic readout of activity. This chapter describes the methodology and practical considerations for implementation of these two assays.
Databáze:	MEDLINE
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