Skip to main content
Log in

A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system

  • Heart, Circulation, Respiration and Blood; Environmental and Exercise Physiology
  • Instruments and techniques
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

(1) A preparation is described which allows patch clamp recordings to be made on mammalian central nervous system (CNS) neurones in situ. (2) A vibrating tissue slicer was used to cut thin slices in which individual neurones could be identified visually. Localized cleaning of cell somata with physiological saline freed the cell membrane, allowing the formation of a high resistance seal between the membrane and the patch pipette. (3) The various configurations of the patch clamp technique were used to demonstrate recording of membrane potential, whole cell currents and single channel currents from neurones and isolated patches. (4) The patch clamp technique was used to record from neurones filled with fluorescent dyes. Staining was achieved by filling cells during recording or by previous retrograde labelling. (5) Thin slice cleaning and patch clamp techniques were shown to be applicable to the spinal cord and almost any brain region and to various species. These techniques are also applicable to animals of a wide variety of postnatal ages, from newborn to adult.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alger BE, Dhanjal SS, Dingledine R, Garthwaite J, Henderson G, King GL, Lipton P, North A, Schwartzkroin PA, Sears TA, Segal M, Whittingham TS, Williams J (1984) Brain slice methods. In: Dingledine R (ed) Brain slices. Plenum Press, New York, pp 381–437

    Google Scholar 

  • Allen CN, Brady R, Swann J, Hori N, Carpenter DO (1988) N-Methyl-D-aspartate (NMDA) receptors are inactivated by trypsin. Brain Res 458:147–150

    Google Scholar 

  • Åslund N, Liljeborg A, Forsgren, P-O, Wahlsten S (1987) Threedimensional digital microscopy using the PHOIBOS scanner. Scanning 9:227–235

    Google Scholar 

  • Armstrong CM, Bezanilla F (1974) Charge movement associated with the opening and closing of the activation gates of the Na channels. J Gen Physiol 63:533–552

    Google Scholar 

  • Barnes S, Werblin F (1986) Gated currents generate single spike activity in amacrine cells of the tiger salmander retinal Proc Natl Acad Sci USA 83:1509–1512

    Google Scholar 

  • Crunelli V, Forda S, Kelly J (1983) Blockade of amino acid-induced depolarizations and inhibition of excitatory post-synaptic potentials in rat dentate gyrus. J Physiol 341:627–640

    Google Scholar 

  • Fricke RA, Prince DA (1984) Electrophysiology of dentate gyrus granule cells. J Neurophysiol 51:195–209

    Google Scholar 

  • Gray R, Johnston D (1985) Rectification of single GABA-gated chloride channels in adult hippocampal neurons. J Neurophysiol 54:134–142

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch clamp techniques for high-resolution current recording from cells and cell free patches. Pflügers Arch 391:85–100

    Google Scholar 

  • Knowles WD, Schwartzkroin PA (1981) Local circuit, synaptic interactions in hippocampal brain slices. J Neurosci, 1:318–322

    Google Scholar 

  • Konnerth A, Obaid AL, Salzberg BM (1987) Optical recording of electrical activity from parallel fibres and other cell types in skate cerebellar slices in vitro. J Physiol 393:681–702

    Google Scholar 

  • Lorente de Nó (1934) Studies on the structure of the cerebral cortex. II. Continuation of the study of the ammonic system. J Psychol Neurol (Lpz) 46:113–117

    Google Scholar 

  • Llinas R, Sugimori M (1980) Electrophysiological properties of in vitro Purkinje cell somata in mammalian cerebellar slices. J Physiol 305:171–195

    Google Scholar 

  • MacVicar BA, Dudek FE (1980) Local synaptic circuits in rat hippocampus: Interactions between pyramidal cells. Brain Res 184:220–223

    Google Scholar 

  • Marty (1983) Ca2+-dependent K+ channels with large unitary conductance. Trends in Neuroscience 6:262–265

    Google Scholar 

  • McCarren M, Alger BE (1987) Papain effects on rat hippocampal neurons in the slice preparation. Neurosci Lett 78:307–310

    Google Scholar 

  • Miles R, Wong RKS (1984) Umtary inhibitory synaptic potentials in the guinea-pig hippocampus, in vitro. J Physiol 356:97–113

    Google Scholar 

  • Pusch M, Neher E (1988) Rates of diffusional exchange between small cells and a measuring patch pipette. Pflügers Arch 411:204–211

    Google Scholar 

  • Ross WN, Werman R (1987) Mapping calcium transients in the dendrites of Purkinje cells from the guinea-pig cerebellum in vitro. J Physiol 389:319–336

    Google Scholar 

  • Takahashi T (1978) Intracellular recording from visually identified motoneurones in rat spinal cord slices. Proc R Soc Lond B [Biol] 202:417–421

    Google Scholar 

  • Wallén P, Carlsson K, Liljeborg A, Grillner S (1988) Three-dimensional reconstruction of neurons in the lamprey spinal cord in whole-mount, using a confocal laser scanning, microscope. J Neurosci Meth 24:91–100

    Google Scholar 

  • Yamamoto C (1975) Recording of electrical activity from microscopically identified neurons of the mammalian brain. Experientia 31:309–311

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Edwards, F.A., Konnerth, A., Sakmann, B. et al. A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system. Pflugers Arch. 414, 600–612 (1989). https://doi.org/10.1007/BF00580998

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00580998

Key words

Navigation