Abstract
Two synaptosomal fractions could be obtained from bovine caudate nucleus on sucrose density gradients one of which had a much greater capacity for ‘high affinity’ choline uptake than the other but comparable amounts of CAT and choline kinase activity. Specific binding of QNB was widely distributed among all the subcellular fractions except the mitochondrial fraction and in quantitative terms by far the greatest amount was in the microsomal fraction. Only the microsomal fraction contained measurable amounts of glycerophosphocholine phosphodiesterase.
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Cheney, D. L., LeFevre, H. F., andRagagni, G. 1975. Choline acetyltransferase activity and mass fragmentation measurement of acetylcholine in specific nuclei and tracts of rat brain. Neuropharmacology 14:801–809.
Wastek, G. J., Stern, L. Z., Johnson, P. C., andYamamura, H. I. 1976. Huntington's disease: regional alteration in muscarinic cholinergic receptor binding in human brain. Life Sci. 19:1033–1040.
Hiley, C. R., andBurgen, A. S. V. 1974. The distribution of muscarinic receptor sites in the nervous system of the dog. J. Neurochem. 22:159–162.
Laverty, R., Michaelson, I. A., Sharman, D. F., andWhittaker, V. P. 1963. The subcellular localisation of dopamine and acetylcholine in the dog caudate nucleus. Brit. J. Pharmacol. 21:482–490.
Hickey, S. M., Ansell, G. B., Mitchell, K., andPearce, G. W. 1976. Subcellular fractions of normal human substantia nigra and caudate nucleus: a study of their morphology and some enzymes including glutamic decarboxylase and choline acetyltransferase. J. Neurochem. 27:957–962.
Bajgar, J., Patocka, J., Ornst, F., Žižkovský, V., andLacina, P. 1971. Subcellular distribution of acetylcholinesterase activity in human brain caudate nucleus. J. Neurochem. 18:529–530.
Gray, E. G., andWhittaker, V. P. 1962. Isolation of nerve endings from brain: an electron microscope study of cell fragments. J. Anat. 96:79–88.
DeRobertis, E., Pellegrino, de Iraldi, A., Rodriguez, de Lores Arnaiz, G. andSalganicoff, L. 1962. Cholinergic and non-cholinergic nerve endings rat brain. 1. Isolation and subcellular distribution of acetylcholine and acetylcholinesterase. J. Neurochem. 9:23–35.
Gregg, M. R., Spanner, S., andAnsell, G. B. 1977. The binding of a specific muscarinic antagonist to subcellular fractions of the bovine caudate nucleus. Biochem. Soc. Trans. 5:182–184.
Yamamura, H. I. andSnyder, S. H. 1974. Muscarinic cholinergic binding in rat brain. Proc. Natl. Acad. Sci. 71:1725–1729.
Bull, G., andOderfeld-Novak, B. 1971. Standardisation of a radiochemical assay of choline acetyltransferase and a study of the activation of the enzyme in the rabbit brain. J. Neurochem. 18:936–941.
Fonnum, F. 1969. Isolation of choline esters from aqueous solutions by extraction with sodium tetraphenylboron in organic solvents. Biochem. J. 113:291–298.
Fonnum, F. 1969. Radiochemical micro assays for the determination of choline acetyltransferase and acetylcholinesterase activities. Biochem. J. 115:465–472.
Simon, J. R., Atweh, S., andKuhar, M. J. 1976. Sodium-dependent high affinity choline uptake. A regulatory step in the synthesis of acetylcholine. J. Neurochem. 26:909–922.
Spanner, S., andAnsell, G. B. 1979. Choline kinase and ethanolamine kinase activity in the cytosol of nerve endings from rat forebrain. Biochem. J. 178:753–760.
Ansell, G. B., andSpanner, S. 1981. The activity of glycerophosphocholine phosphodiesterase in brain tissue. Pages 393–403,in Pepeu, G. andLadinsky, H. (ed.), Advances in Behavioural biology 25: Plenum Press, N. Y.
Weichselbaum, T. E. 1946. An accurate and rapid method for the determination of proteins in small amounts of blood serum and plasma. Am. J. Clin. Path. 16: suppl. P40.
Ellman, G. L., Courtney, D. K., Andres, V., andFeatherstone, R. M. 1961. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7:88–95.
Porteus, J. W., andClark, B. 1965. The isolation and characterisation of subcellular components of the epithelial cells of rabbit small intestine. Biochem. J. 96:159–171.
Michell, R. H., andHawthorne, J. N. 1965. The site of diphosphoinositide synthesis in rat liver. Biochem. Biophys. Res. Commun. 21:333–338.
Hökfelt, T., Jonsson, G., andLidbrink, P. 1970. Electron microscopic identification of monoamine nerve ending particles in rat brain homogenates. Brain Res. 22:147–151.
Cotman, C. W., andMatthews, D. A. 1971. Synaptic plasma membranes from rat brain synaptsomes: isolation and partial characterisation. Biochim. Biophys. Acta. 249:380–394.
Lapetina, E. G., Soto, E. F., andde Robertis, E. 1967. Gangliosides and acetylcholinesterase in isolated membranes of the rat brain cortex. Biochim. Biophys. Acta. 135:33–43.
Lehmann, J., andFibiger, H. C. 1978. Acetylcholinesterase in the substantia nigra and caudate-putamen of the rat: properties and localisation in dopaminergic neurones. J. Neurochem. 30:615–624.
Silver, A. 1974. Subcellular localisation, synthesis and transport of acetylcholinesterase. Pages 99–131,in Neuberger, A. andTatum, E. L. (ed.), The Biology of Cholinesterases. Frontiers of Biology Vol 36, North Holland Publ. Co. Amsterdam.
Steck, T. L., andWallach, O. F. H. 1970. The isolation of plasma membranes. Pages 93–153,in Busch, H. (ed.), Methods in Cancer Research Vol. 5, Academic Press, N. Y.
Bernstein, H. G., Weiss, J., andLuppa, H. 1973. Cytochemical investigations on the localisation of 5′nucleotidase in the rat hippocampus with specific reference to synaptic regions. Histochemistry 55:261–267.
Farrow, J. T. andO'Brien, R. D. 1973. Binding of atropine and muscarine to rat brain fractions and its relation to the acetylcholine receptor. Mol. Pharmacol. 9:33–40.
Ben-Barak, J., andDudai, Y. 1980. Scopolamine induces an increase in muscarinic receptor level in rat hippocampus. Brain Res. 193:309–313.
Fonnum, F. 1968. Choline acetyltransferase binding to and release from membranes. Biochem. J. 109:389–398.
Yamamura, H. I., andSnyder, S. H. 1973. High affinity transport of choline into synaptosomes of rat brain. J. Neurochem. 21:1355–1374.
Sorimachi, M., andKataoka, K. 1974. Choline uptake by nerve terminals: A sensitive and specific marker of cholinergic innervation. Brain Res. 72:350–353.
Kuhar, M. J., Haven, R. M. de, Yamamura, H. I., Rommelspracher, H., andSimon, J. R. 1975. Further evidence for cholinergic habenulo interpeduncular neurones. Pharmacological and functional characteristics. Brain Res. 97:265–275.
Wheeler, D. D. 1979. A model of high affinity choline transport in rat cortical synaptosomes. J. Neurochem. 32:1197–1213.
Atweh, S., Simon, J. R. andKuhar, M. J. 1975. Utilization of sodium-dependent high affinity choline uptake in vitro as a measure of the activity of cholinergic neurones in vivo. Life Sci. 17:1535–1544.
Gregg, M. R., Spanner, S., andAnsell, G. B. 1981. Muscarinic receptor binding sites in the synaptosomal and microsomal fractions of bovine caudate nucleus. Biochem. Soc. Trans. 9:416–417.
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This paper is dedicated to Dr. Derek Richter on his seventy-fifth birthday.
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Gregg, M.R., Spanner, S. & Ansell, G.B. The subcellular fractionation of the bovine caudate nucleus. Neurochem Res 7, 1045–1058 (1982). https://doi.org/10.1007/BF00964885
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DOI: https://doi.org/10.1007/BF00964885