Protocol

Patch-Clamp Methods and Protocols

Volume 1183 of the series Methods in Molecular Biology pp 221-242

Date:

Acute Brain Slice Methods for Adult and Aging Animals: Application of Targeted Patch Clamp Analysis and Optogenetics

  • Jonathan T. TingAffiliated withHuman Cell Types Department, Allen Institute for Brain Science Email author 
  • , Tanya L. DaigleAffiliated withDepartment of Cell Biology, Duke University Medical CenterMcGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, MIT
  • , Qian ChenAffiliated withMcGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, MIT
  • , Guoping FengAffiliated withMcGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, MIT

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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.

Key words

Acute brain slice Adult animals Patch clamp recording Protective recovery method NMDG aCSF Optogenetics GCaMP Channelrhodopsin