Abstract
Although synaptosomes are a widely accepted preparation for studying regulation of transmitter release at the isolated presynaptic level (see also Chapter 4), they are not suited for the investigation of synaptic events mediated by postsynaptic mechanisms. In the past, isolated membrane fractions enriched in postsynaptic densities have been used for this purpose (1). However, this fraction of postsynaptic densities is not appropriate to study receptor-mediated signal transduction because of the absence of intact sealed structures in this preparation. For this reason, a subcellular preparation enriched in resealed presynaptic structures (synaptosomes) with attached sealed postsynaptic entities (neurosomes) has been developed. These composite structures are called synaptoneurosomes. Biochemical characterization of synaptoneurosome preparations has revealed the presence of a number of receptor-mediated properties and a maintained electrochemical gradient (2,3). Since its first description, this preparation has been used for the investigation of numerous phenomena, including neurotransmitter release (4), inositol phospholipid turnover (5,6), CAMP accumulation (2), as well as in functional studies of various neurotransmitter receptors (7).
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References
Siekevitz, P. (1981) Isolation of postsynaptic densities from cerebral cortex. Res. Meth. Neurochem. 5, 75–89.
Hollingsworth, E. B., McNeal, E. T., Burton, J. L., Williams, R. J., Daly, J. W., and Creveling, C. R. (1985) Biochemical characterization of a filtered synaptoneurosome preparation from guinea pig cerebral cortex: cyclic adenosine 3′,5′-monophosphate-generating systems, receptors, and enzymes. J. Neurosci. 5, 2240–2253.
Schwartz, R. D., Jackson, J. A., Weigert, D., Skolnick, P., and Paul, S. M. (1985) GABA-and barbiturate-stimulated chloride efflux from rat brain synaptoneurosomes. J. Neurosci. 5, 2963–2970.
Ebstein, R. P., Seamon, K., Creveling, C. R., and Daly, J. W. (1982) Release of norepinephrine from brain vesicular preparations: effects of an adenylate cyclase activator, forskolin, and a phosphodiesterase inhibitor. Cell Mol. Neurobiol. 2, 179–192.
Chandler, J. L. and Crews, F. T. (1990) Calcium-versus G protein-mediated phosphoinositide hydrolysis in rat cerebral cortical synaptoneurosomes. J. Neurochem. 55, 1022–1030.
Gusovsky, F. and Daly, J. W. (1988) Formation of inositol phosphates in synaptoneurosomes of guinea pig brain: stimulatory effects of receptor agonists, sodium channel agents and sodium ionophores. Neuropharmacology 27, 95–105.
Harris, R. A. and Allan, A. M. (1985) Functional coupling of γ-amino-butyric acid receptor in chloride channels in brain membranes. Science 228, 1108,1109.
Schofield, P. R. (1989) The GABAA receptor: molecular biology reveals a complex picture. TIPS 10, 476–478.
Persohn, E., Malherbe, P., and Richards, J. G. (1992) Comparative molecular neuroanatomy of cloned GABAA receptor subunits in the rat CNS. J. Comp. Neurol. 326, 193–216.
Schónrock, B. and Bormann, J. (1993) Functional heterogeneity of hippocampal GABAA receptors. Eur. J. Neurosci. 5, 1042–1049.
Lopes da Silva, F. H., Kamphuis, W., and Wadman, W. J. (1992) Epileptogenesis as a plastic phenomenon of the brain, a short review. Acta Neurol. Scan. 86, 34–40.
Kamphuis, W. and Lopes da Silva, F. H. (1990) The kindling model of epilepsy: the role of GABAergic inhibition. Neurosci. Res. Comm. 6, 1–10.
Titulaer, M. N. G., Kamphuis, W., Pool, C. W., van Heerikhuize, J. J., and Lopes da Silva, F. H. (1994) Kindling induces time-dependent and regional specific changes in the [3H]muscimol binding in the rat hippocampus: a quantitative autoradiographic study. Neuroscience 59, 817–826.
Titulaer, M. N. G., Ghijsen, W. E. J. M., Kamphuis, W., De Rijk, T. C., and Lopes da Silva, F. H. (1995) Opposite changes in GABAA receptor function in the CA1–3 area and fascia dentata of kindled rat hippocampus. J. Neurochem. 64, 2615–2621.
Bradford, M. M. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72, 248–252.
Schwartz, R. D., Suzdak, P. D., and Paul, S. M. (1986) γ-Aminobutyrtc (GABA)-and barbiturate-mediated 36Cl− uptake in rat brain synaptoneurosomes evidence for rapid desensitization of the GABA receptor-coupled chloride ion channel. Mol. Pharm. 30, 419–426.
Verheul, H. B., de Leeuw, F.-E., Scholten, G., Tulleken, C. A. F., Lopes da Silva, F. H., and Ghijsen, W. E. J. M. (1993) GABAA receptor function in the early period after transient forebrain ischaemia in the rat. Eur. J. Neurosci. 5, 955–960.
Cupello, A. and Rapallino, M. V. (1992) Components of basal and GABA activated 36Cl− influx in rat cerebral cortex microsacs. Int. J. Neurosci. 62, 35–43.
Thallmann, R. H. and Hershkowitz, N. (1985) Some factors that influence the decrement in the response to GABA during its continuous iontophoretic application to hippocampal neurons. Brain Res. 342, 219–233.
Engblom, A. C. and Åkerman, K. E. O. (1991) Effect of ethanol on γ-amino-butyric acid and glycine receptor-coupled Cl− fluxes in rat brain synaptoneurosomes. J. Neurochem., 57, 384–390.
Starke, K, Gothert, M., and Kilbinger, H. (1989) Modulation of neurotransmitter release by presynaptic autoreceptors. Physiol. Rev. 69, 864–989.
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© 1997 Humana Press Inc., Totowa, NJ
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Titulaer, M.N.G., Ghijsen, W.E.J.M. (1997). Synaptoneurosomes. In: Rayne, R.C. (eds) Neurotransmitter Methods. Methods in Molecular Biology, vol 72. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-394-5:49
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DOI: https://doi.org/10.1385/0-89603-394-5:49
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