Preparation of Hippocampal Brain Slices

Daniel V. Madison1, Eleanore B. Edson1

1 Stanford University School of Medicine, Stanford, California
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 6.4
DOI:  10.1002/0471142301.ns0604s00
Online Posting Date:  May, 2001
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Abstract

This unit presents a procedure for the preparation of acute mammalian hippocampal slices for electrophysiological recording. Although this protocol should not be taken as the only means of making brain slices, it is a widely‐used typical procedure. It is simple and straightforward. It can be used on a variety of mammalian species, though it is discussed here for use in rats.

     
 
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Table of Contents

  • Basic Protocol 1: Preparation of Acute Mammalian Hippocampal Slices
  • Reagents and Solutions
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Preparation of Acute Mammalian Hippocampal Slices

  Materials
  • Dissection buffer (see recipe), chilled to 4°C and carbogenated with 95% O 2/5% CO 2 gas mixture
  • 95% O 2/5% CO 2 gas mixture (carbogen)
  • Animal
  • 19 × 16 × 10–cm. Nalgene utility box
  • Tygon tubing (of an appropriate diameter to fit the connector on the air stone)
  • Soldering iron
  • Silicon bathtub sealant (Home Depot)
  • Aquarium air stone
  • Large rubber stoppers
  • Rectangle of ⅛‐in.‐thick Lucite (see step 1b)
  • 35‐mm plastic petri dishes
  • Whatman no. 1 or no. 2 filter paper cut to 37 × 37–mm square
  • Time tape
  • Rongeurs
  • Tissue sectioner (Stoelting) and double‐edged razor blade
  • Small‐animal decapitator (Harvard Apparatus)
  • 2 small Teflon‐coated pointed weighing spatulas
  • Small sharp‐nosed dissecting scissors
  • No. 2 soft artist's paintbrush (e.g., white sable)
  • Wide‐bore pipet (plastic transfer pipet cut off at ∼⅔ of length from the tip, or Pasteur pipet with taper cut off and with cut end fire polished)
  • Additional reagents and equipment for whole cell recording (unit 6.6)
NOTE: Most of the materials for preparing hippocampal slices are commonly found around any laboratory. The materials listed are those preferred by the author; substitutions can be made as desired.
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Figures

Videos

Literature Cited

Literature Cited
   Blanton, M.G., Lo Turco, J.J., and Kriegstein, A.R. 1989. Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex. J. Neurosci. Methods 30:203‐210.
   Buchs, P.A., Stoppini, L., and Muller, D. 1993. Structural modifications associated with synaptic development in area CA1 of rat hippocampal organotypic cultures. Dev. Brain Res. 71:81‐91.
   Gahwiler, B.H. 1988 Organotypic cultures of neural tissue. Trends Neurosci. 11:484‐489.
   Muller, D., Buchs, P.A., and Stoppini, L. 1993. Time course of synaptic development in hippocampal organotypic cultures. Dev. Brain Res. 71:93‐100.
   Yamamoto, C. and McIlwain, H. 1966a. Electrical activities in thin sections from the mammalian brain maintained in chemically‐defined media in vitro. J. Neurochem. 13:1333‐1343.
   Yamamoto, C. and McIlwain, H. 1966b. Potentials evoked in vitro in preparations from the mammalian brain. Nature 210:1055‐1056.
Key References
   Alger, B.E., Dhanjal, S.S., Dingledine, R., Garthwaite, J., Henderson, G., King, G.L., Lipton, P., North, A., Schwartzkroin, P.A., Sears, T.A., Segal, M., Whittingham, T.S., and Williams, J. 1984. Appendix: Brain slice methods. In Brain Slices (R. Dingledine, ed.) pp. 381‐437. Plenum, New York.
  An earlier, and still excellent, description of brain slice cutting technique.
   Madison, D.V. 1992. Whole‐cell voltage‐clamp techniques applied to the study of synaptic function in hippocampal slices. In Cellular Neurobiology: A Practical Approach (J. Chad and H. Wheal, eds.) pp. 137‐149. IRL Press, Oxford.
  A description of brain slice technique, including discussion of whole‐cell recording in slices.
   Yamamoto, C. and McIlwain, H. 1966a and 1996b. See above.
  The earliest reports, to the authors' knowledge, of electrophysiological recording in brain slices.
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