Summary
Crude synaptosomal preparations of rat brain were separated by concomitant flotation and sedimentation in a sucrose gradient into fractions containing myelin, nerve endings, and mitochondria, respectively. Two myelin-free nerve ending fractions were obtained which differ in the serotonin content and the activities of the enzymes 5-hydroxytryptophan decarboxylase, monoamine oxidase, choline acetylase, and acetylcholin-esterase. In glucose medium sodium ions stimulate serotonin uptake into the nerve endings while potassium ions in sucrose solution induce the release of this amine.
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References
Baldessarini, R. J., andM. Karobath: Biochemical physiology of central synapses. Ann. Rev. Physiol.35, 273–304 (1973).
Berg, S. W., andR. P. Maickel: A simplified method for determination of brain acetylcholinesterase activity. Life Sci.7, 1197–1202 (1968).
Berneis, K. H., M. Da Prada, andA. Pletscher: Micelle formation between 5-hydroxytryptamine and adenosine triphosphate in platelet storage organelles. Science165, 913–914 (1969).
Bogdanski, D. F., A. Pletscher, B. B. Brodie, andS. Udenfriend: Identification and assay of serotonin in brain. J. Pharmacol. Exptl. Therap.117, 82–88 (1956).
Bogdanski, D. F., A. Tissari, andB. B. Brodie: Role of sodium, potassium, ouabain, and reserpine in uptake, storage, and metabolism of biogenic amines in synaptsomes. Life Sci.7, 419–428 (1968).
Bogdanski, D. F., T. P. Blaszkowski, andA. H. Tissari: Mechanism of biogenic amine transport and storage. IV. Relationship between K÷ and the Na+ requirement for transport and storage of 5-hydroxytryptamine and norepinephrine in synaptosomes. Biochim. Biophys. Acta211, 521 to 532 (1970).
Bradford, H. F.: Respiration in vitro of synaptosomes from mammalian cerebral cortex. J. Neurochem.16, 675–684 (1969).
Carnegie, P. R.: Properties, structure and possible neuroreceptor role of the encephalitogenic protein of human brain. Nature (London)229, 25–28 (1971).
Chase, T. N., R. I. Katz, andI. J. Kopin: Release of [3H] serotonin from brain slices. J. Neurochem.16, 607–615 (1969).
Cotman, C. W., andH. R. Mahler: Resolution of insoluble proteins in rat brain subcellular fractions. Arch. Biochem. Biophys.120, 384–396 (1967).
De Belleroche, J. S., andH. F. Bradford: Metabolism of beds of mammalian cortical synaptosomes: response to depolarizing influences. J. Neurochem.19, 585–602 (1972).
De Robertis, E., A. Pellegrino de Iraldi, G. Rodriguez de Lores Arnaiz, andL. Salganicoff: Cholinergic and non-cholinergic nerve endings in rat brain. I. Isolation and subcellular distribution of acetylcholine and acetylcholinesterase. J. Neurochem.9, 23–35 (1962).
Diamond, I., andR. A. Fishman: Development of glucose oxidation in isolated nerve endings. Nature (London)243, 519–520 (1973 a).
Diamond, I., andR. A. Fishman: Development of Na+-stimulated glucose oxidation in synaptosomes. J. Neurochem.21, 1043–1050 (1973 b).
Glover, V., andD. P. L. Green: A simple quick microassay for choline acetyltransferase. J. Neurochem.19, 2465–2466 (1972).
Gray, E. G., andV. P. Whittaker: The isolation of nerve endings from brain: An electron microscopic study of cell fragments derived by homogenization and centrifugation. J. Anat. (London)96, 79–88 (1962).
Gurd, J. W., L. R. Jones, H. R. Mahler, andW. J. Moore: Isolation and partial characterization of rat brain synaptic plasma membranes. J. Neurochem.22, 281–290 (1974).
Hill, R. M., andV. E. Bruzek: Apparatus for collecting fractions from density differential interfaces and its use in gravimetric measurement of total myelin. Clin. Chem.16, 171–176 (1970).
Israël, M., andV. P. Whittaker: The isolation of mossy fibre endings from the granular layer of the cerebellar cortex. Experientia21, 325–326 (1965).
Jones, D. H., andA. I. Matus: Isolation of synaptic plasma membrane from brain by combined flotation-sedimentation density centrifugation. Biochim. Biophys. Acta356, 276–287 (1974).
Kuhar, M. J.: Neurotransmitter uptake: a tool in identifying neurotransmitter specific pathways. Life Sci.13, 1623–1634 (1973).
Kuhar, M. J., R. H. Roth, andG. K. Aghajanian: Synaptosomes from forebrains of rats with midbrain raphe lesions: selective reduction of serotonin uptake. J. Pharmacol. Exptl. Therap.181, 36–45 (1972).
Lowry, O. H., N. J. Rosebrough, A. L. Farr, andR. J. Randall: Protein measurement with the folin phenol reagent. J. Biol. Chem.193, 265 to 275 (1951).
McCaman, R. E., M. W. McCaman, J. H. Hunt, andM. S. Smith: Micro-determination of monoamine oxidase and 5-hydroxytryptophan decarboxylase activities in nervous tissues. J. Neurochem.12, 15–23 (1965).
McCaman, M. W., R. E. McCaman, andG. J. Lees: Liquid cation exchange -a basis for sensitive radiometric assay for aromatic amino acid decarboxylases. Anal. Biochem.45, 242–252 (1972).
McGovern, S., M. E. Maguire, R. S. Gurd, H. R. Mahler, andW. J. Moore: Separation of adrenergic and cholinergic synaptosomes from immature rat brain. FEBS Letters31, 193–198 (1973).
Rajan, K. S., J. M. Davis, R. W. Calburn, andF. H. Jarke: Metal chelates in the storage and transport of neurotransmitters: formation of mixed ligand chelates of Mg2+ -ATP with biogenic amines. J. Neurochem.19, 1099–1116 (1972).
Shields, P. J., andD. Eccleston: Effects of electrical stimulation of rat midbrain on 5-hydroxytryptamine synthesis as determined by a sensitive radioisotope method. J. Neurochem.19, 265–272 (1972).
Shields, P. J., andD. Eccleston: Evidence for the synthesis and storage of 5-hydroxytryptamine in two separate pools in the brain. J. Neurochem.20, 881–888 (1973).
Smythies, J. R., F. Bennington, andR. D. Morin: Encephalitogenic protein: aβ-pleated sheet conformation (102–120) yields a possible molecular form of a serotonin receptor. Experientia (Basel)28, 23–24 (1972).
Tamir, H., andY. L. Huang: Binding of serotonin to soluble protein from synaptosomes. Life Sci.14, 83–93 (1974).
Tamir, H., M. M. Rapport, andL. Roizin: Preparation of synaptosomes and vesicles with sodium diatrizoate. J. Neurochem.23, 943–949 (1974).
Verity, M. A.: Cation modulation of synaptosomal respiration. J. Neurochem.79, 1305–1317 (1972).
Wesemann, W.: Isolation of 5-hydroxytryptamine containing vesicles and synaptic membranes from rat brain. FEBS Letters3, 80–84 (1969).
Wesemann, W., J. D. Schollmeyer, andD. Lathia: Studies on the retrieval of 5-hydroxytryptamine by isolated nerve endings. Proc. of the 9th Internat. Congress of Biochemistry, Stockholm 1973, p. 444.
Whittaker, V. P., J. A. Michaelson, andR. J. A. Kirkland: The separation of synaptic vesicles from nerve ending particles (“synaptosomes”). Biochem. J.90, 293–303 (1964).
Wirz-Justice, A.: The influence of various amines and amino acids on K+-stimulated release of dopamine and serotonin from rat striatum. Eur. J. Pharmacol.19, 281–284 (1972).
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Lathia, D., Wesemann, W. Serotonin uptake and release by biochemically characterized nerve endings isolated from rat brain by concomitant flotation and sedimentation centrifugation. J. Neural Transmission 37, 111–126 (1975). https://doi.org/10.1007/BF01663628
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DOI: https://doi.org/10.1007/BF01663628