Quantal analysis of inhibitory synaptic transmission in the dentate gyrus of rat hippocampal slices: a patch‐clamp study.

1. Synaptically connected neurones were identified in the granule cell layer of slices of 17‐ to 21‐day‐old rat hippocampus. Whole‐cell current recording using the patch‐clamp technique revealed synaptic currents ranging from less than 10 to 200 pA in symmetrical Cl‐ conditions, at a holding potential of ‐50 mV. These currents were blocked by 2 microM‐bicuculline, indicating that they result from the activation of postsynaptic gamma‐aminobutyric acid receptor (GABAA‐receptor) channels. 2. Addition of tetrodotoxin (TTX, 1 microM) resulted in the loss of most currents of more than 40 pA in amplitude. Currents which disappeared after TTX treatment were assumed to be the result of spontaneous presynaptic action potentials. The currents seen in the absence of TTX are referred to as spontaneously occurring inhibitory postsynaptic currents (IPSCs); those remaining in the presence of TTX were defined as miniature IPSCs. 3. Similar currents were observed when recording in the whole‐cell configuration while extracellular stimulation was applied to a nearby neurone. These currents were also completely blocked by 2 microM‐bicuculline and by 0.5 microM‐TTX. They were thus defined as stimulus‐evoked IPSCs. 4. The half rise time of both miniature and stimulus‐evoked IPSCs was fast (less than 1 ms). The time course of decay of both miniature IPSCs and stimulus‐evoked IPSCs could be well fitted with the sum of two exponentials. At a membrane potential of ‐50 mV, the mean decay time constants of the two components were 2.0 +/‐ 0.38 and 54.4 +/‐ 18 ms (mean +/‐ S.D.) for miniature IPSCs (six cells) and 2.2 +/‐ 1.3 and 66 +/‐ 20 ms (three cells) for stimulus‐evoked IPSCs. 5. Stimulus‐evoked IPSCs varied in amplitude from less than ten to hundreds of picoamperes. In eight of eleven cells histograms of IPSC amplitudes showed several clear peaks which, when fitted with the sum of Gaussian curves, were found to be equidistant. This is consistent with the view that stimulus‐evoked IPSC amplitudes vary in a quantal fashion. The quantal size varied between 7 and 20 pA, at a membrane potential of ‐50 mV. 6. Decreasing the Ca2+ and increasing the Mg2+ concentration in the extracellular solution decreased the number of peaks in the IPSC amplitude histogram but did not affect the size of the quantal event.(ABSTRACT TRUNCATED AT 400 WORDS)

[1]  J. Eccles,et al.  The ionic mechanisms concerned in generating the i. p. s. ps of hippocampal pyramidal cells , 1977, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[2]  P. Seeburg,et al.  Sequence and functional expression of the GABAA receptor shows a ligand-gated receptor super-family , 1987, Nature.

[3]  M. Salpeter,et al.  Acetylcholine receptor site density affects the rising phase of miniature endplate currents. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Twyman,et al.  Kinetic properties of the GABAA receptor main conductance state of mouse spinal cord neurones in culture. , 1989, The Journal of physiology.

[5]  H. Atwood,et al.  Short-term and long-term plasticity and physiological differentiation of crustacean motor synapses. , 1986, International review of neurobiology.

[6]  D. R. Curtis,et al.  GABA, Bicuculline and Central Inhibition , 1970, Nature.

[7]  S. Schuetze,et al.  A post‐natal decrease in acetylcholine channel open time at rat end‐plates. , 1980, The Journal of physiology.

[8]  J. Bornstein Multiquantal release of acetylcholine in mammalian ganglia , 1974, Nature.

[9]  D. Faber,et al.  Spontaneous quantal currents in a central neuron match predictions from binomial analysis of evoked responses. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. W. Kuffler,et al.  Post‐synaptic potentiation: interaction between quanta of acetylcholine at the skeletal neuromuscular synapse. , 1975, The Journal of physiology.

[11]  J. Barker,et al.  Phenobarbitone modulation of postsynaptic GABA receptor function on cultured mammalian neurons , 1979, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[12]  A. Takeuchi,et al.  Further analysis of relationship between end-plate potential and end-plate current. , 1960, Journal of neurophysiology.

[13]  B. Walmsley,et al.  Statistical fluctuations in charge transfer at Ia synapses on spinal motoneurones. , 1976, The Journal of physiology.

[14]  C. Erxleben,et al.  Sub-MEPPs, Skew-MEPPs and the Subunit Hypothesis of Quantal Transmitter Release at the Neuromuscular Junction , 1986 .

[15]  B Sakmann,et al.  Synaptic transmission in hippocampal neurons: numerical reconstruction of quantal IPSCs. , 1990, Cold Spring Harbor symposia on quantitative biology.

[16]  D. Faber,et al.  Transmission at a central inhibitory synapse. I. Magnitude of unitary postsynaptic conductance change and kinetics of channel activation. , 1982, Journal of neurophysiology.

[17]  A. W. Liley Spontaneous release of transmitter substance in multiquantal units , 1957, The Journal of physiology.

[18]  H Korn,et al.  Transmission at a central inhibitory synapse. II. Quantal description of release, with a physical correlate for binomial n. , 1982, Journal of neurophysiology.

[19]  E. McLachlan An analysis of the release of acetylcholine from preganglionic nerve terminals. , 1975, The Journal of physiology.

[20]  I. D. Hill,et al.  An Efficient and Portable Pseudo‐Random Number Generator , 1982 .

[21]  S J Redman,et al.  The synaptic current evoked in cat spinal motoneurones by impulses in single group 1a axons. , 1983, The Journal of physiology.

[22]  B. L. Ginsborg,et al.  Spontaneous synaptic activity in sympathetic ganglion cells of the frog , 1963, The Journal of physiology.

[23]  J. Simpson THE RELEASE OF NEURAL TRANSMITTER SUBSTANCES , 1969 .

[24]  A. R. Martin,et al.  Quantal Nature of Synaptic Transmission , 1966 .

[25]  B. Sakmann,et al.  Change in synaptic channel gating during neuromuscular development , 1978, Nature.

[26]  S. Redman Quantal analysis of synaptic potentials in neurons of the central nervous system. , 1990, Physiological reviews.

[27]  L. Henderson,et al.  The single-channel basis for the slow kinetics of synaptic currents in vertebrate slow muscle fibers , 1989, Neuron.

[28]  B. Sakmann,et al.  Functional properties of recombinant rat GABAA receptors depend upon subunit composition , 1990, Neuron.

[29]  J. Weakly,et al.  Quantal components of the inhibitory synaptic potential in spinal motoneurones of the cat , 1972, The Journal of physiology.

[30]  S. J. Smith,et al.  Calcium action in synaptic transmitter release. , 1987, Annual review of neuroscience.

[31]  P W Gage,et al.  Inhibitory post‐synaptic currents in rat hippocampal CA1 neurones. , 1984, The Journal of physiology.

[32]  A. R. Martin,et al.  The end‐plate potential in mammalian muscle , 1956, The Journal of physiology.

[33]  H. Rang The characteristics of synaptic currents and responses to acetylcholine of rat submandibular ganglion cells , 1981, The Journal of physiology.

[34]  K. Magleby,et al.  Is the quantum of transmitter release composed of subunits? A critical analysis in the mouse and frog , 1981, Nature.

[35]  A. Konnerth,et al.  Patch clamp techniques used for studying synaptic transmission in slices of mammalian brain. , 1989, Quarterly journal of experimental physiology.

[36]  S. W. Kuffler,et al.  Synaptic transmission and its duplication by focally applied acetylcholine in parasympathetic neurons in the heart of the frog , 1971, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[37]  H. Kojima,et al.  Characterization of miniature inhibitory post‐synaptic potentials in rat spinal motoneurones. , 1985, The Journal of physiology.

[38]  P. Seeburg,et al.  Two novel GABAA receptor subunits exist in distinct neuronal subpopulations , 1989, Neuron.

[39]  J. Jack,et al.  The components of synaptic potentials evoked in cat spinal motoneurones by impulses in single group Ia afferents. , 1981, The Journal of physiology.

[40]  G. Pilar,et al.  Quantal components of the synaptic potential in the ciliary ganglion of the chick , 1964, The Journal of physiology.

[41]  M. Gold,et al.  Characteristics of inhibitory post‐synaptic currents in brain‐stem neurones of the lamprey. , 1983, The Journal of physiology.

[42]  B. Sakmann,et al.  Mechanism of anion permeation through channels gated by glycine and gamma‐aminobutyric acid in mouse cultured spinal neurones. , 1987, The Journal of physiology.

[43]  R K Wong,et al.  Unitary inhibitory synaptic potentials in the guinea‐pig hippocampus in vitro. , 1984, The Journal of physiology.