Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice

Yasmine Kamen; Ragnhildur Þóra Káradóttir

doi:10.1016/j.xpro.2021.100439

Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice

Yasmine Kamen
, Ragnhildur Þóra Káradóttir

Læknadeild

Háskóli Íslands

Rannsóknarafurð: Framlag til fræðitímarits › Grein › ritrýni

Útdráttur

Single-cell electrophysiological recordings combined with dye loading and immunohistochemistry provide unparalleled single-cell resolution of cell physiology, morphology, location, and protein expression. When correlated with bulk RNA sequencing, these data can define cell identity and function. Here, we describe a protocol to prepare acute brain slices from embryonic and postnatal mice for whole-cell patch clamp, dye loading and post-hoc immunohistochemistry, and cell isolation for bulk RNA sequencing. While we focus on oligodendrocyte precursor cells, this protocol is applicable to other brain cells. For complete details on the use and execution of this protocol, please refer to Spitzer et al. (2019).

Upprunalegt tungumál	Enska
Númer greinar	100439
Síður (frá-til)	100439
Fræðitímarit	STAR Protocols
Bindi	2
Númer tölublaðs	2
DOI	https://doi.org/10.1016/j.xpro.2021.100439
Útgáfustaða	Útgefið - 18 jún. 2021

Athugasemd

Funding Information: We would like to thank Dr. Maike Paramor for comments on the protocol, Kimberley Evans for comments on the protocol and assistance with the graphics, Dr Omar de Faria Jr for assistance with the figures, and our following funders: the European Research Council (ERC: the European Union’s Horizon 2020 research and innovation programme grant agreement No 771411 ; R.T.K.); the Wellcome (Studentship award 102160/Z/13/Z; Y.K.); the Fonds de recherche du Québec-Santé (a scholarship, Y.K.); The Cambridge Commonwealth European & International Trust (a scholarship, Y.K.); and the Lister Institute of Preventive Medicine (a research prize, R.T.K.). The funders had no role in decision to publish or preparation of the manuscript. Funding Information: We would like to thank Dr. Maike Paramor for comments on the protocol, Kimberley Evans for comments on the protocol and assistance with the graphics, Dr Omar de Faria Jr for assistance with the figures, and our following funders: the European Research Council (ERC: the European Union's Horizon 2020 research and innovation programme grant agreement No 771411; R.T.K.); the Wellcome (Studentship award 102160/Z/13/Z; Y.K.); the Fonds de recherche du Qu?bec-Sant? (a scholarship, Y.K.); The Cambridge Commonwealth European & International Trust (a scholarship, Y.K.); and the Lister Institute of Preventive Medicine (a research prize, R.T.K.). The funders had no role in decision to publish or preparation of the manuscript. Y.K. and R.T.K. wrote the manuscript. The authors declare no competing interests. Publisher Copyright: © 2021 The Author(s)

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10.1016/j.xpro.2021.100439

https://opinvisindi.is/bitstreams/054aeb7e-1cc2-4220-b02e-2f07534ae791/downloadLeyfi: CC BY

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title = "Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice",

abstract = "Single-cell electrophysiological recordings combined with dye loading and immunohistochemistry provide unparalleled single-cell resolution of cell physiology, morphology, location, and protein expression. When correlated with bulk RNA sequencing, these data can define cell identity and function. Here, we describe a protocol to prepare acute brain slices from embryonic and postnatal mice for whole-cell patch clamp, dye loading and post-hoc immunohistochemistry, and cell isolation for bulk RNA sequencing. While we focus on oligodendrocyte precursor cells, this protocol is applicable to other brain cells. For complete details on the use and execution of this protocol, please refer to Spitzer et al. (2019).",

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doi = "10.1016/j.xpro.2021.100439",

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Y1 - 2021/6/18

N2 - Single-cell electrophysiological recordings combined with dye loading and immunohistochemistry provide unparalleled single-cell resolution of cell physiology, morphology, location, and protein expression. When correlated with bulk RNA sequencing, these data can define cell identity and function. Here, we describe a protocol to prepare acute brain slices from embryonic and postnatal mice for whole-cell patch clamp, dye loading and post-hoc immunohistochemistry, and cell isolation for bulk RNA sequencing. While we focus on oligodendrocyte precursor cells, this protocol is applicable to other brain cells. For complete details on the use and execution of this protocol, please refer to Spitzer et al. (2019).

AB - Single-cell electrophysiological recordings combined with dye loading and immunohistochemistry provide unparalleled single-cell resolution of cell physiology, morphology, location, and protein expression. When correlated with bulk RNA sequencing, these data can define cell identity and function. Here, we describe a protocol to prepare acute brain slices from embryonic and postnatal mice for whole-cell patch clamp, dye loading and post-hoc immunohistochemistry, and cell isolation for bulk RNA sequencing. While we focus on oligodendrocyte precursor cells, this protocol is applicable to other brain cells. For complete details on the use and execution of this protocol, please refer to Spitzer et al. (2019).

KW - Cell isolation

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KW - Molecular Biology

KW - Neuroscience

KW - RNA-seq

KW - Stem Cells

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Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice

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