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Methods Mol Biol DOI:10.1007/978-1-4939-1096-0_14

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

Publication TypeJournal Article
Year of Publication2014
AuthorsTing, JT, Daigle, TL, Chen, Q, Feng, G
JournalMethods Mol Biol
Volume1183
Pages221-42
Date Published2014
ISSN1940-6029
KeywordsAging, Animals, Brain, Equipment Design, Gene Expression, Histocytological Preparation Techniques, Mice, Mice, Transgenic, Neurons, Optogenetics, Patch-Clamp Techniques, Rhodopsin
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.

DOI10.1007/978-1-4939-1096-0_14
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/25023312?dopt=Abstract

Alternate JournalMethods Mol. Biol.
PubMed ID25023312
PubMed Central IDPMC4219416
Grant ListF32 MH084460 / MH / NIMH NIH HHS / United States
F32-MH084460 / MH / NIMH NIH HHS / United States