Summary
Explants of 10–12 day chick embryo spinal cord were cultured by coverslip-roller tube method for 3–80 days. The cellular and subcellular localization of acetylcholinesterase activity in cultured neurons was studied by the thiocholine techniques of Karnovsky and Roots and Lewis and Shute.
At the light microscopic level, acetylcholinesterase was demonstrated in the neurons of both ventral and dorsal horn regions. Occasionally neurons migrated in the outgrowth zone exhibited strong intracellular activity.
At the electron microscopic level, acetylcholinesterase activity was found in the nuclear envelope, granular endoplasmic reticulum and the Golgi apparatus of the neurons. No enzyme reaction was detected in the glial cell cytoplasm.
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References
Bornstein, M.B., Murray, M.R.: Serial observations on patterns of growth, myelin formation, maintenance and degeneration in cultures of newborn rat and kitten cerebellum. J. biophys. biochem. Cytol. 4, 499–504 (1958)
Burdick, C.J., Strittmatter, C.F.: Appearance of biochemical components related to acetylcholine metabolism during the embryonic development of chick brain. Arch. Biochem. Biophys. 190, 293–301 (1965)
Burt, A.: Acetylcholinesterase and choline acetyltransferase activity in the developing chick spinal cord. J. exp. Zool. 169, 107–112 (1968)
Gwyn, D.G., Flumerfelt, B.A.: Acetylcholinesterase in non-cholinergic neurons: A histochemical study of dorsal root ganglion in the rat. Brain Res. 34, 193–198 (1971)
Hansson, H.: The distribution of acetylcholinesterase and other hydrolytic enzymes in retinal cultures. Acta physiol. scand. Suppl. 288, 1–30 (1966)
Hervonen, H., Rechardt, L.: Light and electron microscopic demonstration of cholinesterases of the cultured sympathetic ganglia of the chick embryo. Histochemistry 39, 129–142 (1974)
Hösli, E., Hösli, L.: Acetylcholinesterase in cultured rat spinal cord. Brain Res. 30, 193–196 (1971)
Karnovsky, M.J., Roots, L.: A “direct-coloring” thiocholine method for cholinesterase. J. Histochem. Cytochem. 12, 219–221 (1964)
Kim, S.U.: Localization of acetylcholinesterase in chick spinal cord culture in vitro. J. Histochem. Cytochem. 18, 688 (1970)
Kim, S.U.: Quantitative and cytochemical studies of acetylcholinesterase in organotypic cultures of chick spinal cord. In: Proceedings of 4th International Congress of Histochemistry and Cytochemistry, pp. 479–480. Kyoto: Japan Society of Histochemistry 1972
Kim, S.U., Munkacsi, I.: Cytochemical demonstration of catecholamines and acetylcholinesterase in cultures of chick sympathetic ganglia. Experientia (Basel) 28, 824–825 (1972)
Kim, S.U., Munkacsi, I.: Morphological and cytochemical characterization of neurons in cultures of mouse sympathetic ganglia. Exp. Neurol. 45, 94–103 (1974)
Kim, S.U., Murray, M.R.: Histochemical demonstration of acetylcholinesterase in organized cultures of mouse cerebellum and sensory ganglia. Anat. Rec. 163, 310 (1969)
Kim, S.U., Oh, T.H., Johnson, D.D.: Developmental changes of acetylcholinesterase and pseudocho-linesterase in organotypic cultures of spinal cord. Exp. Neurol. 35, 274–281 (1972)
Kim, S.U., Oh, T.H., Johnson, D.D.: Increased activity of choline acetyltransferase and acetylcho-linesterase in developing cultures of chick spinal cord: A correlation with morphological development. Neurobiology 5, 119–227 (1975)
Kim, S.U., Tunnicliff, G.: Morphological and biochemical development of chick cerebrum cultured in vitro. Exp. Neurol. 43, 515–526 (1974)
Klingman, G.I., Klingman, J.D., Policzczuk, A.: Acetyl- and pseudocholinesterase activities in sympathetic ganglia of rats. J. Neurochem. 15, 1121–1130 (1968)
Leuzinger, W., Baker, A.L.: Acetylcholinesterase. I. Large-scale purification, homogeneity, and amino acid analysis. Proc. nat. Aced. Sci. (Wash.) 57, 446–451 (1967)
Lewis, P.R., Shute, C.D.: The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscope. J. Cell Sci. 1, 381–390 (1966)
Lumsden, C.E.: Nervous tissue in culture. In: The structure and function of nervous tissue (G.H. Bourne, ed.), Vol. 1, pp. 68–140. New York: Academic Press 1968
Maletta, G.J., Vernadakis, A., Timiras, P.S.: Pre- and postnatal development of the spinal cord: Increased acetylcholinesterase activity. Proc. Soc. exp. Biol. (N.Y.) 121, 1210–1211 (1966)
Mazza, J.P., Hanker, J.S., Dixon, A.S.: Ultrastructural localization of cholinesterase activity in the trigeminal ganglion of the rat. J. Anat. (Lond.) 115, 65–78 (1973)
McCaman, R.E., Aprison, M.H.: The synthetic and catabolic systems for acetylcholine and serotonin in several discrete areas of the developing rabbit brain. Progr. Brain Res. 9, 220–223 (1971)
Minelli, G., Ciani, F., Contestabile, A.: The occurrence of some enzymatic activities in the differentiation of nerve tissue of gallus embryos in cultures in vitro. Histochemie 28, 160–169 (1971)
Murray, M.R.: Nervous tissues isolated in culture. In: Handbook of neurochemistry (A. Lajtha, ed.), Vol. 5, pp. 373–438. New York: Plenum 1971
Nelson, P.G.: Nerve and muscle cells in culture. Physiol. Rev. 55, 1–61 (1975)
Pannese, E., Luciano, L., Iurato, S., Reale, E.: Cholinesterase activity in spinal ganglia neuroblasts: A histochemical study at the electron microscope. J. Ultrastruct. Res. 36, 46–67 (1971)
Rechardt, L., Hervonen, H.: Light and electron microscopic demonstration of acetylcholinesterase activity in cultured spinal ganglia of the chick embryo. Histochemistry 40, 371–376 (1974)
Sensenbrenner, M., Booher, J., Mandel, P.: Histochemical study of dissociated nerve cells from embryonic chick cerebral hemispheres in flask cultures. Experientia (Basel) 29, 699–701 (1973)
Silver, A.: Cholinesterases of the central nervous system with special reference to the cerebellum. Int. Rev. Neurobiol 10, 57–109 (1967)
Silver, A.: The significance of cholinesterase in the developing nervous system. Progr. Brain Res. 34, 345–355 (1971)
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Kim, S.U. Acetylcholinesterase distribution in chick spinal cord cultures. Histochemistry 48, 205–217 (1976). https://doi.org/10.1007/BF00497456
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DOI: https://doi.org/10.1007/BF00497456