Acute brain slice methods for adult and aging animals: application of targeted patch clamp analysis and optogenetics.

Methods Mol Biol
Authors
Keywords
Abstract

The development of the living acute brain slice preparation for analyzing synaptic function roughly a half century ago was a pivotal achievement that greatly influenced the landscape of modern neuroscience. Indeed, many neuroscientists regard brain slices as the gold-standard model system for detailed cellular, molecular, and circuitry level analysis and perturbation of neuronal function. A critical limitation of this model system is the difficulty in preparing slices from adult and aging animals, and over the past several decades few substantial methodological improvements have emerged to facilitate patch clamp analysis in the mature adult stage. In this chapter we describe a robust and practical protocol for preparing brain slices from mature adult mice that are suitable for patch clamp analysis. This method reduces swelling and damage in superficial layers of the slices and improves the success rate for targeted patch clamp recordings, including recordings from fluorescently labeled populations in slices derived from transgenic mice. This adult brain slice method is suitable for diverse experimental applications, including both monitoring and manipulating neuronal activity with genetically encoded calcium indicators and optogenetic actuators, respectively. We describe the application of this adult brain slice platform and associated methods for screening kinetic properties of Channelrhodopsin (ChR) variants expressed in genetically defined neuronal subtypes.

Year of Publication
2014
Journal
Methods Mol Biol
Volume
1183
Pages
221-42
Date Published
2014
ISSN
1940-6029
DOI
10.1007/978-1-4939-1096-0_14
PubMed ID
25023312
PubMed Central ID
PMC4219416
Links
Grant list
F32 MH084460 / MH / NIMH NIH HHS / United States
F32-MH084460 / MH / NIMH NIH HHS / United States