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Patch clamp analysis of excitatory synapses in mammalian spinal cord slices

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Abstract

Excitatory synaptic transmission to visually identified α-moto neurones was studied in thin slice preparations of the neonatal rat spinal cord. Excitatory postsynaptic currents (EPSCs) elicited by stimulation of intraspinal presynaptic fibres were recorded using the whole-cell patch clamp technique, following blockade of inhibitory transmission by bath application of strychnine and bicuculline. The EPSCs could be separated pharmacologically into N-methyl-d-aspartate- (NMDA) and non-NMDA-receptor-mediated components, where the contribution of the NMDA-mediated component was significant only at holding potentials more positive than −50 mV. Graded stimulation of intraspinal fibres showed that the NMDA- and the non-NMDA-mediated EPSCs were evoked by activation of presynaptic fibres with similar sensitivities to the stimulation intensity, suggesting that the same presynaptic fibres released the excitatory amino-acid (EAA) activating the two sub-sets of receptors. Studies of the amplitude fluctuations of EPSCs elicited by stimulation of a presumed single fibre revealed similar proportions of transmission failures and similar distributions of both the NMDA- and the non-NMDA-mediated components. These similarities suggest that the EAA transmitter activating the two sub-types of receptors is released from the same set of synaptic boutons and that the receptors are therefore post-synaptically co-localized. In addition the gamma aminobutyric acidB (GABAB) receptor agonist l-baclofen, which is known to decrease transmitter release, changed the amplitude distributions of non-NMDA- and NMDA-receptor-mediated EPSCs into unimodal distributions without affecting the amplitude of the presumed unitary event. The similarity between the transmitter release profiles of the two EAA components further supports the notion of postsynaptic receptor co-localization.

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References

  • Andreasen M, Lambert JDC, Jensen MS (1989) Effects of new non-N-methyl-d-aspartate antagonists on synaptic transmission in the in vitro rat hippocampus. J Physiol (Lond) 414:317–336

    Google Scholar 

  • Ascher P, Nowak L (1987) Electrophysiological studies on NMDA receptors. Trends Neurosci 10:284–288

    Google Scholar 

  • Baldissera F, Hultborn H, Illert M (1981) Integration in spinal neuroneal systems. In: Brooks VB (ed) Handbook of physiology, vol II, part 1. American Physiological Society, Bethesda, Md, pp 509–595

    Google Scholar 

  • Bekkers JM, Stevens CF (1989) NMDA and non-NMDA receptors are co-localized at individual excitatory synapses in cultured rat hippocampus. Nature 341:230–233

    Google Scholar 

  • Bormann J (1988) Electrophysiology of GABAA and GABAB receptor subtypes. Trends Neurosci 11:112–116

    Google Scholar 

  • Burke RE, Rudomin P (1977) Spinal neurones and synapses. In: Kandel ER (ed) Handbook of physiology, vol 1, part 2. American Physiological Society, Bethesda, Md, pp 877–944

    Google Scholar 

  • Cotman CW, Monaghan DT, Ottersen OP, Storm-Mathiesen J (1987) Anatomical organization of excitatory amino acid receptors and their pathways. Trends Neurosci 7:273–279

    Google Scholar 

  • Dale N, Roberts AJ (1985) Dual-component amino-acid mediated synaptic potentials: excitatory drive for swimming in Xenopus embryos. J Physiol 363:35–59

    Google Scholar 

  • Edwards FA, Konnerth A, Sakmann B, Takahashi T (1989) A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system. Pflügers Arch 414:600–612

    Google Scholar 

  • Edwards FA, Konnerth A, Sakmann B (1990) Quantal synaptic transmission in the central nervous system. A patch clamp study of IPSCs in rat hippocampal slices. J Physiol (Lond) (in press)

  • Edwards FR, Redman SJ, Walmsley B (1976) Statistical fluctuations in charge transfer at Ia synapses on spinal motoneurones. J Physiol (Lond) 259:665–688

    Google Scholar 

  • Edwards FR, Harrison PJ, Jack JJB, Kullmann DM (1989) Reduction by baclofen of monosynaptic EPSCs in lumbosacral motoneurones of the anaesthetized cat. J Physiol (Lond) 416:539–556

    Google Scholar 

  • Finkel AS, Redman SJ (1983) The synaptic current evoked in cat spinal motoneurones by impulses in single group 1a axons. J Physiol (Lond) 342:615–632

    Google Scholar 

  • Forsythe ID, Westbrook GL (1988) Slow excitatory postsynaptic curents mediated by N-methyl-d-aspartate receptors on cultured mouse central neurones. J Physiol (Lond) 396:515–533

    Google Scholar 

  • Fulton BP, Walton K (1986) Electrophysiological properties of neonatal rat motoneurons studied in vitro. J Physiol 370:651–678

    Google Scholar 

  • Glenn II, Burke RF, Fleshman JW, Lev-Tov A (1983) Estimates of electrotonic distance of group Ia contacts on rat alpha motoneurons. An HRP morphological study. Soc Neurosci Abstr 8:995

    Google Scholar 

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

    Google Scholar 

  • Hestrin S, Nicoll RA, Perkel DJ, Sah P (1990) Analysis of excitatory synaptic action in pyramidal cells using whole cell recording from rat hippocampal slices. J Physiol (Lond) 422:203–225

    Google Scholar 

  • Honoré T, Davies SN, Drejer J, Fletcher EJ, Jacobsen P, Lodge D, Nielsen FE (1988) Quinoxalinediones: potent competitive non-NMDA glutamate receptor antagonist. Science 241:701–703

    Google Scholar 

  • Katz B (1969) The release of neuronal transmitter substances. The Sherrington Lecture no. 10. Liverpool University Press, Liverpool

    Google Scholar 

  • Konnerth A, Llano I, Armstrong CM (1990a) Synaptic currents in cerebellar Purkinje cells. Proc Natl Acad Sci USA 87:2662–2665

    Google Scholar 

  • Konnerth A, Keller BU, Ballani K, Yaari Y (1990b) Voltage sensitivity of NMDA-receptor mediated postsynaptic currents. Exp Brain Res 81:209–212

    Google Scholar 

  • Korn H, Faber DS (1987) Regulation and significance of probabilistic release mechanisms at central synapses. In: Edelman GM, Gall WA, Cowan WM (eds) Synaptic function. Wiley, New York, pp 57–108

    Google Scholar 

  • Lev-Tov A, Meyers DER, Burke RE (1988) Activation of type B gamma-aminobutyric acid receptors in the intact mammalian spinal cord mimics the effects of reduced presynaptic Ca++ influx. Proc Natl Acad Sci USA 85:5530–5534

    Google Scholar 

  • Martin AR (1977) Junctional transmission. II. Presynaptic mechanisms. In: Kandel ER (ed) Handbook of physiology, vol I, part 1. American Physiological Society, Bethesda, Md, pp 329–355

    Google Scholar 

  • McDermott AB, Dale N (1987) Receptors, ion channels and synaptic potentials underlying the integrative actions of excitatory amino acids. Trends Neurosci 10:280–284

    Google Scholar 

  • O'Brien RJ, Fischbach GD (1986) Excitatory synaptic transmission between interneurons and motoneurons in chick spinal cord cell cultures. J Neurosci 6:3284–3289

    Google Scholar 

  • Peng Y-Y, Frank E (1989) Activation of GABAB receptors causes presynaptic inhibition at synapses between muscle spindle afferents and motoneurons in the spinal cord of bullfrogs. J Neurosci 9:1502–1515

    Google Scholar 

  • Pun RYK, Westbrook GL (1983) Baclofen: Physiological evidence for a presynaptic site of action in spinal cord cultures. Soc Neurosci Abstr 9:1143

    Google Scholar 

  • Shapovalov AI, Shiriaev BI (1982) Selective modulation of chemical transmission at a dual-action synapse (with special reference to baclofen). Gen Physiol Biophys 1:423–433

    Google Scholar 

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

    Google Scholar 

  • Takahashi T (1990) Membrane currents in visually identified motoneurones of neonatal rat spinal cord. J Physiol (Lond) 423:27–46

    Google Scholar 

  • Walmsley B, Edwards FR, Tracey DJ (1988) Nonuniform release probabilities underlie quantal synaptic transmission at a mammalian excitatory central synapse. J Neurophysiol 60:889–908

    Google Scholar 

  • Wong K, Redman SJ (1980) The recovery of a random variable from a noisy record with application to the study of fluctuations in synaptic potentials. J Neurosci Methods 2:389–409

    Google Scholar 

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Konnerth, A., Keller, B.U. & Lev-Tov, A. Patch clamp analysis of excitatory synapses in mammalian spinal cord slices. Pflugers Arch. 417, 285–290 (1990). https://doi.org/10.1007/BF00370994

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  • DOI: https://doi.org/10.1007/BF00370994

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