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Sodium-dependent release of exogenous glycine from preloaded rat hippocampal synaptosomes

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Summary

The effect of high potassium, veratridine, and ouabain stimulation upon the release of exogenously-loaded [3H]glycine was evaluated in crude synaptosomal preparations from rat hippocampi by means of a superfusion technique in the presence of media with different ionic compositions and of tetrodotoxin (TTX). Four minute superfusion of synaptosomes with 30 mM KC1, 10 μM veratridine or 0.4 mM ouabain caused a significant increase in the [3H]glycine efflux which averaged 6.6±0.2, 25.5±1.0, and 8.9±1.0% of the total radioactivity present in the synaptosomes, respectively. The omission of Ca2+ ions in the superfusion medium markedly decreased K+-evoked [3H]glycine efflux (2.5±0.5%), did not appreciably modify that evoked by veratridine (24.2±2.0%) and significantly increased that evoked by ouabain (18.5±0.5%). The superfusion of synaptosomes with Na+-free media always resulted in a drastic decrease of the depolarization-stimulated [3H]glycine efflux, whereas the omission of Cl− generally resulted in a moderate increase of [3H]glycine efflux. TTX (0.8 μM) markedly affected the stimulatory effect of veratridine (2.5±0.9%) and ouabain (2.2±0.5%), but failed to modify significantly that evoked by high potassium (6.5±0.7%). Finally, [3H]glycine was seen readily to exchange in a partially sodium-dependent way with unlabelled glycine present in the medium. On the whole these findings appear to be consistent with the neurotransmitter character of the glycine release from hippocampal synaptosomes.

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References

  • Aragòn MC, Giménez C (1986) Efflux and exchange of glycine by synaptic plasma membrane vesicles derived from rat brain. Biochim Biophys Acta 855: 257–264

    Google Scholar 

  • Archibald JT, White TD (1974) Rapid reversal of internal Na+ and K+ contents of synaptosomes by ouabain. Nature 252: 595–596

    Google Scholar 

  • Bashir ZI, Tam B, Collingridge GL (1990) Activation of the glycine site in the NMDA receptor is necessary for the induction of LTP. Neurosci Lett 108: 261–266

    Google Scholar 

  • Benjamin AM, Quastel JH (1972) Locations of amino acids in brain slices from the rat. Biochem J 128: 631–646

    Google Scholar 

  • Blaustein MP (1975) Effects of potassium, veratridine, and scorpion venom on calcium accumulation and transmitter release by nerve terminals in vitro. J Physiol (Lond) 247: 617–655

    Google Scholar 

  • Blaustein MP, Goldring JM (1975) Membrane potentials in pinched-off presynaptic nerve terminals monitored with a fluorescent probe: evidence that synaptosomes have potassium diffusion potentials. J Physiol (Lond) 247: 589–615

    Google Scholar 

  • Bonhaus DW, Yeh GC, Skaryak L, McNamara JO (1989) Glycine regulation of the N-methyl-D-aspartate receptor-gated ion channel in hippocampal membranes. Mol Pharmacol 36: 273–279

    Google Scholar 

  • Bradford HF, Bennet GW, Thomas AJ (1973) Depolarizing stimuli and the release of physiologically active amino acids from suspensions of mammalian synaptosomes. J Neurochem 21: 495–505

    Google Scholar 

  • Bristow DR, Bowery NG, Woodruff G (1986) Light microscopic autoradiographic localisation of [3H]glycine and [3H]strychnine binding sites in rat brain. Eur J Pharmacol 126: 303–307

    Google Scholar 

  • Cunningham J, Neal MJ (1981) On the mechanism by which veratridine causes a calcium-independent release of gamma-aminobutyric acid from brain slices. Br J Pharmacol 73: 655–667

    Google Scholar 

  • Curtis DR, Hosli L, Johnston GAR (1968) A pharmacological study of the depression of spinal neurones by glycine and related amino acids. Exp Brain Res 6: 1–18

    Google Scholar 

  • Douglas WW (1968) Stimulus-secretion coupling: the concept and clues from chromaffm and other cells. Br J Pharmacol Chemother 34: 451–474

    Google Scholar 

  • Erecinska M (1987) The neurotransmitter amino acid transport systems. A fresh outlook on an old problem. Biochem Pharmacol 36: 3547–3555

    Google Scholar 

  • Fedele E, Foster AC (1992) [3H]Glycine uptake in rat hippocampus: kinetic analysis and autoradiographic localization. Brain Res 572: 154–163

    Google Scholar 

  • Fletcher EJ, Beart PM, Lodge D (1990) Involvement of glycine in excitatory neurotransmission. In: Ottersen OP, Storm-Mathisen J (eds) Glycine neurotransmission. Wiley, New York, pp 193–218

    Google Scholar 

  • Frankenhaeuser B, Hodgkin AL (1957) Action of calcium on the electrical properties of squid taxons. J Physiol (Lond) 137: 218–244

    Google Scholar 

  • Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electronmicroscopic study of cell fragments derived by homogenization and centrifugation. J Anat 96: 79–87

    Google Scholar 

  • Hammerstad JP, Murray J, Cutler RWP (1971) Efflux of amino acid neurotransmitters from rat spinal cord slices. II. Factors influencing the electrically induced efflux of [14C]glycine and [3H]GABA. Brain Res 35: 357–367

    Google Scholar 

  • Holopainen I, Kontro P (1989) Uptake and release of glycine in cerebellar granule cells and astrocytes in primary culture: potassium-stimulated release from granule cells is calcium-dependent. J Neurosci Res 24: 374–383

    Google Scholar 

  • Hopkin J, Neal MJ (1971) Effect of electrical stimulation and high potassium concentrations on the efflux of [14C]glycine from slices of spinal cord. Br J Pharmacol 42: 215–223

    Google Scholar 

  • Johnson JW, Ascher P (1987) Glycine potentiates the NMDA response in cultured mouse brain neurons. Nature 325: 529–531

    Google Scholar 

  • Kleckner NW, Dingledine R (1988) Requirement for glycine in activation of NMDA receptors expressed in Xenopus oocytes. Science 241: 835–837

    Google Scholar 

  • Levi G, Raiteri M (1974) Exchange of neurotransmitter amino acid at nerve endings can stimulate high affinity uptake. Nature 250: 735–737

    Google Scholar 

  • Levi G, Bernardi G, Cherubini E, Gallo V, Marciani MG, Stanzone P (1982) Evidence in favour of a neurotransmitter role of glycine in the rat cerebral cortex. Brain Res 236: 121–131

    Google Scholar 

  • Levi G, Patrizio M, Gallo V (1991) Release of endogenous and newly synthesized glutamate and other amino acids induced by non-N-methyl-D-aspartate receptor activation in cerebellar granule cell cultures. J Neurochem 56: 199–206

    Google Scholar 

  • Lianas R, Steinberg IZ, Walton K (1976) Presynaptic calcium currents and their relation to synaptic transmission: voltage clamp study in squid giant synapse and theoretical model for the calcium gate. Proc Natl Acad Sci USA 73: 2918–2922

    Google Scholar 

  • Lowe DA, Richardson BP, Taylor P, Donatsch P (1976) Increasing intracellular sodium triggers calcium release from bound pools. Nature 260: 337–338

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275

    Google Scholar 

  • Monaghan DT, Bridges RJ, Cotman CW (1989) The excitatory amino acid receptors: their classes, pharmacology, and distinct properties in the function of the central nervous system. Ann Rev Pharmacol Toxicol 29: 365–402

    Google Scholar 

  • Mulder AH, Snyder S (1974) Potassium-induced release of amino acids from cerebral cortex and spinal cord slices of the rat. Brain Res 76: 297–308

    Google Scholar 

  • Nadler JV, White WF, Vaca KW, Redburn DA, Cotman CW (1977) Characterization of putative amino acid transmitter release from slices of rat dentate gyrus. J Neurochem 29: 279–290

    Google Scholar 

  • Narahashi T (1974) Chemicals as tools in the study of excitable membranes. Physiol Rev 54: 813–889

    Google Scholar 

  • Neal MJ, Bowery NG (1979) Differential effects of veratridine and potassium depolarization on neuronal and glial GABA release. Brain Res 167: 337–343

    Google Scholar 

  • Nicholls DG (1989) Release of glutamate, aspartate, and gamma-aminobutyric acid from isolated nerve terminals. J Neurochem 52: 331–341

    Google Scholar 

  • Rahamimoff R, Erulkar SD, Lev-Tov A, Meiri H (1978) Intracellular and extracellular ions in transmitter release at the neuromuscular synapse. Ann NY Acad Sci 307: 583–598

    Google Scholar 

  • Raiteri M, Angelini F, Levi G (1974) A simple apparatus for studying the release of neurotransmitters from synaptosomes. Eur J Pharmacol 25: 411–414

    Google Scholar 

  • Raiteri M, Fontana G, Fedele E (1990) Glycine stimulates [3H]noradrenaline release by activating a strychnine-sensitive receptor present in rat hippocampus. Eur J Pharmacol 184: 239–250

    Google Scholar 

  • Raiteri M, Bonanno G, Pende M (1992) Gamma-aminobutyric acid and glycine modulate each other's release through heterocarriers sited on the releasing axon terminals of rat CNS. J Neurochem 59: 1481–1489

    Google Scholar 

  • Richelson E (1977) Lithium ion entry through the sodium channel of cultured mouse neuroblastoma cells: a biochemical study. Science 196: 1001–1002

    Google Scholar 

  • Sandoval ME (1980) Sodium-dependent efflux of [3H]GABA from synaptosomes probably related to mitochondrial calcium mobilization. J Neurochem 35: 915–921

    Google Scholar 

  • Sandoval ME, Horch P, Cotman CW (1978) Evaluation of glutamate as a hippocampal neurotransmitter: glutamate uptake and release from synaptosomes. Brain Res 142: 285–299

    Google Scholar 

  • Simon JR, Martin DL, Kroll M (1974) Sodium-dependent efflux and exchange of GABA in synaptosomes. J Neurochem 23: 981–991

    Google Scholar 

  • Smith KE, Borden LA, Hartig PR, Branchek T, Weinshank RL (1992) Cloning and expression of a glycine transporter reveal colocalization with NMDA receptors. Neuron 8: 927–935

    Google Scholar 

  • Weiss S, Kemp DE, Bauce L (1989) Kainate evokes the release of endogenous glycine from striatal neurons in primary culture. Neurosci Lett 107: 205–210

    Google Scholar 

  • Werman R, Davidoff RA, Aprison MH (1967) Inhibition of motoneurones by iontophoresis of glycine. Nature 214: 681–683

    Google Scholar 

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  1. A. Galli

    Present address: Department of Preclinical and Clinical Pharmacology, University of Florence, Italy

Authors and Affiliations

  1. Department of Preclinical and Clinical Pharmacology, University of Florence, Italy

    F. Mori, M. Bargellini & L. Coppini

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  1. A. Galli
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  2. F. Mori
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  3. M. Bargellini
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  4. L. Coppini
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Galli, A., Mori, F., Bargellini, M. et al. Sodium-dependent release of exogenous glycine from preloaded rat hippocampal synaptosomes. J. Neural Transmission 93, 167–179 (1993). https://doi.org/10.1007/BF01244994

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  • DOI: https://doi.org/10.1007/BF01244994

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