Synaptic Plasticity in the Hippocampal Slice Preparation

Zuner A. Bortolotto1, Mascia Amici1, William W. Anderson1, John T.R. Isaac2, Graham L. Collingridge3

1 MRC Centre for Synaptic Plasticity, School of Physiology and Pharmacology, University of Bristol, Bristol, United Kingdom, 2 Eli Lilly & Co., Lilly Research Centre, Windlesham, Surrey, United Kingdom, 3 Department of Brain and Cognitive Sciences, Seoul National University, Seoul, Korea
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 6.13
DOI:  10.1002/0471142301.ns0613s54
Online Posting Date:  January, 2011
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Abstract

Synaptic plasticity is the process by which the brain alters the strength of its synaptic connections, a fundamental function of the brain that enables individuals to learn from experience. The study of synaptic plasticity often involves the application of standard in vitro electrophysiological techniques to hippocampal slice preparations. This unit discusses many of the special considerations that are applicable for the optimal study of synaptic plasticity in this system. Most of these principles also apply to the study of synaptic plasticity in other brain slice preparations. Curr. Protoc. Neurosci. 54:6.13.1‐6.13.26. © 2011 by John Wiley & Sons, Inc.

Keywords: LTP; LTD; hippocampus; synaptic plasticity; glutamate receptors; organotypic slice; WinLTP

     
 
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Table of Contents

  • Introduction
  • Types of Synaptic Plasticity
  • The Use of Field Potential Recording to Study LTP
  • The Use of Field Potential Recording to Study LTD
  • Metaplasticity
  • Basic Protocol 1: Extracellular LTP in the Hippocampal Slice Preparation
  • Reagents and Solutions
  • Studying LTP and LTD Using Intracellular Recording
  • Special Considerations when Working with Transgenic Mice
  • Studying Synaptic Plasticity in Organotypic Slices
  • Data Acquisition and Analysis
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Extracellular LTP in the Hippocampal Slice Preparation

  Materials
  • Laboratory animal (e.g., rat, mouse, guinea pig, hamster)
  • Anesthetic (e.g., a mixture of oxygen and halothane)
  • Artificial cerebrospinal fluid (aCSF; see recipe), 4° and 30°C
  • Super glue
  • Extracellular recording pipets (3 to 6 MΩ; unit 6.3) filled with 4 M NaCl or aCSF
  • 0.05‐ to 0.08‐mm diameter insulated NiCr wire (e.g., Advent Research Materials)
  • Borosilicate glass capillary: e.g., 1.2‐mm o.d./0.69‐mm i.d. (e.g., Harvard Apparatus)
  • Glass microelectrode puller (e.g., model PP‐830, Narishige Scientific Instruments)
  • Micromanipulators (e.g., Narishige Scientific Instruments)
  • Constant current stimulus isolation unit for extracellular stimulation (e.g., DS3, Digitimer)
  • Amplifier (either a voltage amplifier or patch clamp amplifier)
  • Surgical instruments for decapitation or cervical dislocation and dissection
  • Filter paper
  • Tissue slicer (e.g., Campden Instruments or Leica)
  • Recording chamber (e.g., Fine Science Tools or Warner Instrument)
  • Perfusion system consisting of a container of aCSF placed in a temperature‐controlled water bath (30°C) and polyethylene tubing that circulates the prewarmed aCSF through a peristaltic pump (e.g., Harvard Apparatus) and the recording chamber
  • Computer with software for on‐line acquisition and on‐ and off‐line analysis of synaptic potentials (e.g., the free basic version of WinLTP; see ); other appropriate software is also available from Axograph Scientific or Molecular Devices; also see Data Acquisition and Analysis
  • Dissecting microscope (e.g., Leica or Nikon)
NOTE: All protocols using live animals must first be reviewed and approved by an Institutional Animal Care and Use Committee (IACUC) and must follow officially approved procedures for the care and use of laboratory animals.
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Figures

Videos

Literature Cited

   Anderson, W.W. and Collingridge, G.L. 2007. Capabilities of the WinLTP data acquisition program extending beyond basic LTP experimental functions. J. Neurosci. Methods 162:346‐356.
   Bashir, Z.I., Alford, S., Davies, S.N., Randall, A.D., and Collingridge, G.L. 1991. Long‐term potentiation of NMDA receptor‐mediated synaptic transmission in the hippocampus. Nature 349:156‐158.
   Blake, J.F., Brown, M.W., and Collingridge, G.L. 1988. A quantitative study of the actions of excitatory amino acids and antagonists in rat hippocampal slices. Br. J. Pharmacol. 95:291‐299.
   Bortolotto, Z.A. and Collingridge, G.L. 1993. Characterisation of LTP induced by the activation of glutamate metabotropic receptors in area CA1 of the hippocampus. Neuropharmacology 32:1‐9.
   Cavus, I. and Teyler, T. 1996. Two forms of long‐term potentiation in area CA1 activate different signal transduction cascades. J. Neurophysiol. 76:3038‐3047.
   Davies, C.H., Starkey, S.J., Pozza, M.F., and Collingridge, G.L. 1991. GABAB autoreceptors regulate the induction of LTP. Nature 349:609‐611.
   Isaac, J.T.R., Lüthi, A., Palmer, M.J., Anderson, W.W., Benke, T.A., and Collingridge, G.L. 1998. An investigation of the expression mechanism of LTP of AMPA receptor‐mediated synaptic transmission at hippocampal CA1 synapses using failures analysis and dendritic recordings. Neuropharmacology 37:1399‐2410.
   Paxinos, G. and Watson, C. 1997. The Rat Brain in Stereotaxic Coordinates, 4th ed. Academic Press, San Diego.
Key References
   Bear, M.F., Connors, B.W., and Paradiso, M.A. 1996. Synaptic mechanisms of memory. In Neuroscience: Exploring the Brain. 1st ed. (T.S. Satterfield, ed.) pp. 546‐575. Williams & Wilkins, Baltimore.
  This chapter brings together combined experimental approaches about the relationship between synaptic plasticity and learning and related mechanisms.
   Bliss, T.V.P. and Collingridge, G.L. 1993. A synaptic model of memory: Long‐term potentiation in the hippocampus. Nature 361:724‐729.
  This is a comprehensive review about LTP as a model for memory formation.
   Fazeli, M.S. and Collingridge, G.L. 1996. Cortical Plasticity LTP and LTD. Molecular and Cellular Neurobiology Series. 1st ed. BIOS Scientific Publishers, Oxford.
  This book provides insight into synaptic plasticity by covering areas of research as diverse as molecular biology and neuronal networks.
Internet Resources
  http://www.bris.ac.uk/synaptic
  The MRC Centre for Synaptic Plasticity's Web site.
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