Repeated whole-cell patch-clamp recording from CA1 pyramidal cells in rodent hippocampal slices followed by axon initial segment labeling

Laura S Oliveira; Anna Sumera; Sam A Booker

doi:10.1016/j.xpro.2021.100336

Repeated whole-cell patch-clamp recording from CA1 pyramidal cells in rodent hippocampal slices followed by axon initial segment labeling

STAR Protoc. 2021 Feb 5;2(1):100336. doi: 10.1016/j.xpro.2021.100336. eCollection 2021 Mar 19.

Authors

Laura S Oliveira^{1

2

3}, Anna Sumera^{1

2

3}, Sam A Booker^{1

2

3}

Affiliations

¹ Simons Initiative for the Developing Brain, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK.
² Patrick Wild Centre for Research into Autism, Fragile X Syndome, and Intellectual Disability, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK.
³ Centre for Discovery Brain Sciences, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK.

Abstract

This protocol allows repeated whole-cell patch-clamp recordings from individual rodent CA1 hippocampal neurons, followed by immunohistological labeling of the axon initial segment. This overcomes the need to maintain whole-cell recordings over the timescales required for homeostatic modification to cellular excitability, allowing for correlative analysis of the structure and function of neurons. Moreover, this protocol allows for paired analysis of physiological properties assessed before and after pharmacological treatment, thus providing increased statistical power, despite the relatively low-throughput nature of the recordings. For complete details on the use and execution of this protocol, please refer to Booker et al. (2020a).

Keywords: Microscopy; Neuroscience; Single cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Axon Initial Segment / metabolism*
CA1 Region, Hippocampal / cytology
CA1 Region, Hippocampal / metabolism*
Male
Mice
Patch-Clamp Techniques
Pyramidal Cells / cytology
Pyramidal Cells / metabolism*
Rats
Rats, Long-Evans