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Study of some enzyme activities in cultured chick embryo brain nerve cells treated by chick embryo brain extracts

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Abstract

Brain extracts from 8-day-old chick embryos have been shown to influence morphological development of dissociated brain cells from 7-day-old chick embryos in culture. Stimulatory, effects on size of the neuronal somas and on growth of long processes were observed by adding the cytosol of the brain extract or the dialysate of the cytosol. These morphological changes parallel modifications of various enzyme activities according to the age of the cultures. Adenyl cyclase, (Na+, K+)- and Mg2+-ATPase, 5′-nucleotidase, choline acetyltransferase, and acetylcholinesterase activities were studied between 5 and 14 days of culture. Adenyl cyclase activity was strongly stimulated at 8 days by both extracts. (Na+, K+)-and Mg2+-ATPase activities were stimulated in 8-day-old cultures only by the dialysate. 5′-Nucleotidase activity was stimulated in 8-day-old cultures by the dialysate and in 11-day-old cultures by both extracts. Choline acetyltransferase activity was stimulated by the cytosol in 8-day-old cultures and by the dialysate in 11-day-old cultures. The total acetylcholinesterase activity was higher in 8-, 11-, and 14-day-old cultures treated with the cytosol. When the cells were treated with the dialysate, the activity was only higher in 14-day-old cultures. We also found that following the addition of brain extracts, the specific activity of the enzymes we studied was enhanced and became close to the values found in vivo during embryogenesis. Thus in parallel to the morphological modifications observed in nerve cell cultures treated by embryo brain extracts, biochemical variations especially involved in synaptogenesis and membrane development could be measured.

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References

  1. Sensenbrenner, M., Booher, J., andMandel, P. 1971. Cultivation and growth of dissociated neurons from chick embryo cerebral cortex in the presence of different substrates. Z. Zellforschung 98:538–549.

    Google Scholar 

  2. Cam, Y., Sensenbrenner, M., Ledig, M., andMandel, P. 1977. Partial characterization of a brain extract that stimulates nerve cell differentiation in culture. Neuroscience 2:801–805.

    Google Scholar 

  3. Cam, Y., Ledig, M., Sensenbrenner, M., andMandel, P. 1978. Guanyl cyclase activity of nervous cells cultured in presence of brain extracts Brain Res. 144:199–203.

    Google Scholar 

  4. Burkard, W. R., Pieri, L., andHaefely, W. 1976. In vivo changes of guanosine 3′, 5′-cyclic phosphate in rat cerebellum by dopaminergic mechanisms. J. Neurochem. 27:297–298.

    Google Scholar 

  5. Zwiller, J., Ciesielski-Treska, J., andMandel, P. 1975. Effect of lysolecithine on guanylate and adenylate cyclase activities in neuroblastoma cells in culture. FEBS Lett. 69:286–290.

    Google Scholar 

  6. Prasad, K. N., Gilmer, K. N., andSahu, S. K. 1974. Demonstration of acetyl-choline-sensitive adenyl cyclase in malignant neuroblastoma cells in culture. Nature (London) 249:765–767.

    Google Scholar 

  7. Clark, R. B., andPerkins, J. P. 1971. Regulation of, adenosine 3′, 5′-cyclic monophosphate concentration in cultured human astrocytoma cells by catecholamines and histamine. Proc. Natl Acad. Sci. U.S.A. 68:2757–2760.

    Google Scholar 

  8. Schwartz, J. P., Mooris, N. R., andBreckenridge, B. M. 1973. Adenosine 3′, 5′-monophosphate in glial tumor cells. J. Biol. Chem. 248:2699–2704.

    Google Scholar 

  9. Schmidt, M. J., Palmer, E. C., Dettbarn, W., andRobison, G. A. 1970. Cyclic AMP adenyl cyclase in the developing rat brain. Dev. Psychobiol. 3:53–67.

    Google Scholar 

  10. Hosie, R. J. A. 1965. The localization of ATPase in morphologically characterized subcellular fractions of guinea pig brain. Biochem. J. 96:404–419.

    Google Scholar 

  11. Kurokawa, M., Sakamoto, T., andKato, M. 1965. Distribution of Na+-plus-K+ stimulated ATPase activity in isolated nerve ending particles. Biochem. J. 97:833–844.

    Google Scholar 

  12. Cotman, C. W., andMatthews, D. A. 1971. Synaptic plasma membranes from rat brain synaptosomes, isolation and partial characterization. Biochim. Biophys. Acta 249:380–394.

    Google Scholar 

  13. Morgan, I. G., Wolfe, L. S., Mandel, P., andGombos, G. 1971. Isolation of plasma membranes from rat brain. Biochim. Biophys. Acta 241:737–751.

    Google Scholar 

  14. Henn, F. A., Haljamae, H., andHamberger, A. 1972. Glial cell function: Active control of extracellular K+ concentration. Brain Res. 43:437–443.

    Google Scholar 

  15. Nagata, Y., Mikoshiba, K., andTsukada, Y. 1974. Neuronal cell body enriched and glial cell enriched fractions from young and adult rat brains. Preparations and morphological and biochemical properties. J. Neurochem. 22:493–503.

    Google Scholar 

  16. Kimelberg, H. K. 1974. Active potassium transport and (Na++K+) ATPase activity in cultured glioma and neuroblastoma cells. J. Neurochem. 22:971–976.

    Google Scholar 

  17. Ledig M., Ciesielski-Treska, J., Cam, Y., Montagnon, D., andMandel, P. 1975. ATPase activity of neuroblastoma cells in culture. J. Neurochem. 25:636–640.

    Google Scholar 

  18. Logan, J. G., andO'Donovan, D. J. 1976. The effects of ouabain and the activation of neural membrane ATPase by biogenic amines. J. Neurochem. 27:185–189.

    Google Scholar 

  19. Moonen, G., andFranck, G. 1977. Potassium effect on (Na+, K+), ATPase activity of cultured new-born rat astroblasts during differentiation. Neurosci. Lett. 4:263–267.

    Google Scholar 

  20. Kimelberg, H. K., Biddlecome, S., Narumi, S., andBourke, R. S. 1978. ATPase and carbonic anhydrase activities of bulk isolated neuronal, glial and synaptosome fractions from rat brain. Brain Res. 141:305–323.

    Google Scholar 

  21. Trams, E. G., andLauter, C. J. 1974. On the sideness of plasma membrane enzyme. Biochim. Biophys. Acta 245:180–197.

    Google Scholar 

  22. Mandel, P., Stefanovic, V., Ciesielski-Treska, J., andEbel, A. 1974. ATPase activity of the cell surface of astroglia in culture. FEBS Lett. 45:337–339.

    Google Scholar 

  23. Stefanovic, V., Ciesielski-Treska, J., Ebel, A., andMandel, P. 1974. Ca2+ activated ATPase of the external surface of neuroblastoma cells in culture. FEBS Lett. 49:43–46.

    Google Scholar 

  24. Stefanovic, V., Mandel, P., andRosenberg, A. 1975. Concanavalin A inhibition of ecto-5′-nucleotidase of intact cultured C6 cells. J. Biol. Chem. 250:7081–7083.

    Google Scholar 

  25. Cohen, S. R. 1970. Page 87,in Lajtha, A. (ed.), Handbook of neurochemistry, Vol. 3, Plenum Press, New York. USA.

    Google Scholar 

  26. Wechsler, W. 1966. Pages 213–223,in Hassler, R., andStephan, H. (eds.), Evolution of the Forebrain, Georg Thieme Verlag, Stuttgart.

    Google Scholar 

  27. Burdick, C. J., andStrittmatter, C. F. 1965. Appearance of biochemical compounds related to acetylcholine metabolism during the embryonic development of chick brain. Arch. Biochem. Biophys. 109:293–301.

    Google Scholar 

  28. Ebel, A., Massarelli, R., Sensenbrenner, M., andMandel, P. 1974. Choline acetyltransferase and acetylcholinesterase activities in chicken brain hemispheres in vivo and in cell culture. Brain Res. 76:461–472.

    Google Scholar 

  29. Ramirez, G. 1977. Cholinergic development in chick brain reaggregated cell cultures. Neurochem. Res. 2:417–425.

    Google Scholar 

  30. Nachmansohn, D. 1972. Biochemistry a part of my life. Annu. Rev. Biochem. 41:1–28.

    Google Scholar 

  31. Varela, J. M. 1975. Pages 315–342,in Marbert, Clement, L. (ed.), Isozymes II: Physiological Function, Academic Press, New York.

    Google Scholar 

  32. Lowry, O. H., Rosebrough, N. J., Farr, A. L., andRandall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.

    Google Scholar 

  33. Ramachandran, J., andLee, V. 1970. Divergent effects of nitrophenylsulfonyl ACTH on rat and rabbit fat cell adenylcyclase. Biochem. Biophys. Res. Commun. 41:358–366.

    Google Scholar 

  34. Weil-Malherbe, H., andGreen, R. H. 1951. The catalytic effect of molybdate on the hydrolysis of organic phosphate bonds. Biochem. J. 49:286–292.

    Google Scholar 

  35. McCaman, R. E., andHunt, J. M. 1965. Microdetermination of choline acetylase in nervous tissue. J. Neurochem. 12: 253–259.

    Google Scholar 

  36. Goldberg, A. M., Kaita, A. A., andMcCaman, R. E. 1969. Microdetermination of choline acetyltransferase. A comparison of Reinecke vs. periodide precipitation. J. Neurochem. 16:823–824.

    Google Scholar 

  37. McCaman, M. W., Tomey, L. R., andMcCaman, R. E. 1968. Radiometric assay of acetylcholinesterase activity in submicrogram amounts of tissue. Life Sci. 7:233–244.

    Google Scholar 

  38. Koslow, S. H., andGiacobini, E. 1969. An isotopic micromethod for the measurement of cholinesterase activity in individual cells. J. Neurochem. 16:1523–1528.

    Google Scholar 

  39. Cam, Y. 1978. Thèse de Doctorat ès-Sciences, Université Louis Pasteur, Strasbourg.

  40. Zaheer, N., Iqbal, Z., andTalwar, G. P. 1968. Metabolic parameters of ontogenesis of electrical, activity in the brain. J. Neurochem. 15:1217–1224.

    Google Scholar 

  41. Bray, D. 1977. Actin and myosin in neurons: A first review. Biochimie 59:1–6.

    Google Scholar 

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Cam, Y., Ledig, M., Ebel, A. et al. Study of some enzyme activities in cultured chick embryo brain nerve cells treated by chick embryo brain extracts. Neurochem Res 5, 831–845 (1980). https://doi.org/10.1007/BF00965783

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