Abstract
The activities of monoamine oxidase (MAO), cathechol-O-methyltransferase (COMT) and γ-aminobutyric acid transaminase (GABA-T) were measured in primary cultures from newborn rat cultivated from 6 different brain regions. These primary cultures contained mostly astroglial cells, evaluated by the presence of the glial fibrillary acidic protein (GFAp, α-albumin) and the S-100 protein. The enzyme activities in the corresponding brain areas from adult rat were also quantified. MAO activities were on the same level in 14-day old cultures and in adult rat brain homogenates, with significantly lower values in brain stem as compared to the other brain regions examined. COMT activities were on a higher level in the cultures than in adult rat brain homogenates. Astroglial cells from hippocampus were found to have the highest and those from brain stem the lowest COMT-activities. GABA-T activities were lower in the cultures than in adult rat homogenates. No significant differences were seen in the various astroglial cultures. Accumulation of [3H]dopamine and [3H]γ-aminobutyric acid (GABA) visualized by autoradiography showed only a slight uptake of dopamine in comparison with the uptake of GABA. It is concluded that astroglial cells in culture have enzymatic properties similar to those of astroglial cells in different brain regions of adult rat brain. Studies are in progress to evaluate if the regional heterogeneity observed among cultivated astroglial cells is affected by in vivo differentiation until cultivation and/or time in culture.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bock, E., Møller, M., Nissen, C., andSensenbrenner, M. 1977. Glial fibrillary acidic protein in primary astroglial cell cultures derived from newborn rat brain. FEBS Letters 83:207–211.
Booher, J., andSensenbrenner, M. 1972. Growth and cultivation of dissociated neurons and glial cells from embryonic chick, rat and human brain in flask cultures. Neurobiology 2:97–105.
Burstone, M. S. (ed) 1962. Alkaline phosphatase, naphtol AS phosphate method. Pages 275–276,in Enzyme histochemistry and its application in the study of neoplasms Academic Press, New York and London.
Currie, D. N. 1980. Identification of cell type by immunofluorescence in defined cell cultures of cerebellum, Pages 75–87,in E. Giacobini, A. Vernadakis, andA. Shahar (eds.) Tissue Culture in Neurobiology. Raven Press, New York.
De Boer, Th., andBruinvels, J. 1977. Assay and properties of 4-aminobutyric-2-oxoglutaric acid transaminase and succinic semialdehyde dehydrogenase in rat brain tissue. J. Neurochem. 28:471–478.
Drejer, J., Larsson, O. M., andSchousboe, A. 1982. Characterization ofl-glutamate uptake into and release from astrocytes and neurons cultured from different brain regions. Exp. Brain Res. 47:259–269.
Goldstein, D. J., Weinshilboum, R. M., Dunnette, J. H., andCreveling, C. R. 1980. Developmental patterns of catechol-O-methyltransferase in genetically different rat strains: Enzymatic and immunochemical studies. J. Neurochem. 34:153–162.
Hall, Z. W., andKravitz, E. A. 1967. The metabolism of γ-aminobutyric acid (GABA) in the lobster nervous system. I. GABA-glutamate transaminase. J. Neurochem. 14:45–54.
Hallermayer, K., Harmening, C., andHamprecht, B. 1981. Cellular localization and regulation of glutamine synthetase in primary cultures of brain cells from newborn mice. J. Neurochem. 37:43–52.
Hamberger, A. 1971. Amino acid uptake in neuronal and glial cell fractions from rabbit cerebral cortex. Brain Research 31:169–178.
Hansson, E. 1982. Primary astroglial cultures; aspects of morphology, biochemistry and transmitter metabolism. Thesis, Göteborg.
Hansson, E. 1983. Accumulation of putative amino acid neurotransmitter, monoamines andd-ala2-met-enkephaline-amide in primary astroglial cultures from various brain areas, visualized by autoradiography. Brain Research, in press.
Hansson, E., Sellström, Å., Persson, L. I., andRönnbäck, L. 1980. Brain primary culture—a characterization. Brain Research 188:233–246.
Hansson, E., Rönnbäck, L., Lowenthal, A., Noppe, M., Alling, C., Karlsson, B., andSellström, Å. 1982. Brain primary culture—a characterization (part II). Brain Research 231:173–183.
Hansson, E., Rönnbäck, L., Persson, L. I., Lowenthal, A., Noppe, M., Alling, C., andKarlsson, B. Cellular composition of primary cultures from cerebral cortex, striatum, hippocampus, brain stem and cerebellum. Brain Research, in press.
Hansson, E., andSellström, Å. 1983. MAO, COMT and GABA-T activities in primary astroglial cultures. J. Neurochem. 40:220–225.
Hansson, E., Isacsson, H., andSellström, Å. 1983. Some characteristics of dopamine and GABA tranport in primary cultures of astroglial cells. Submitted.
Hazama, H., Ito, M., Hirano, M., andUchimura, H. 1976. Monoamine oxidase activities in neuronal and glial fractions from regional areas of rat brain. J. Neurochem. 26:417–419.
Henn, F. A. 1975. Glial transport of amino acid neurotransmitter candidates. Pages 91–97in Berl, S., Clarke, D. D., andSchneider, D. (eds.), Metabolic compartmentation and neurotransmission. Plenum Press. New York.
Henn, F. A. 1976. Neurotransmission and glial cells: A functional relationship? J. Neurosci. Res. 2:271–282.
Henn, F. A., andHamberger, A. 1971. Glial cell function: Uptake of transmitter substances. Proc. Nat. Acad. Sci. 68:2686–2690.
Hertz, L., Wu, P. H., andSchousboe, A. 1978. Evidence for net uptake of GABA into mouse astrocytes in primary cultures. Its sodium dependence and potassium independence. Neurochem. Res. 3:313–323.
Hyde, J. C., andRobinson, N. 1974. Appearance of gamma aminobutyrate transaminase activity in developing rat brain. J. Neurochem. 23:365–367.
Hösll, L., Hösli, E., andAndrès, P. F. 1973. Nervous tissue culture—A model to study action and uptake of putative neurotransmitters such as amino acids. Brain Research 62:597–602.
Hösli, E., Bucher, U. M., andHösli, L. 1975. Uptake of [3H]noradrenaline and [3H]5-hydroxytryptamine in cultured rat brain-stem. Experientia 31:354–356.
Hösli, L., Hösli, E., Andrès, P. F., andWolff, J. R. 1975. Amino acid transmitters—action and uptake in neurons and glial cells of human and rat CNS tissue culture. Pages 473–488.in Santini, M. (ed.) Golgi Centennial Symposium Proceedings Raven Press, New York.
Hösli, E., andHösli, L. 1976. Autoradiographic studies on the uptake of [3H]noradrenaline and [3H]GABA in cultured rat cerebellum. Exp. Brain Research 26:319–324.
Hösli, L., andHösli, E. 1978. Action and uptake of neurotransmitters in CNS tissue culture. Rev. Physiol. Biochem. Pharmacol. 81:135–188.
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.
Maitre, M., Ossola, L., andMandel, P. 1979. GABA-transaminase of mammalian brain. Pages 3–20,in Mandel, P., De Feudis F. V. (eds.) GABA—Biochemistry and CNS functions. Adv. Exp. Med. Biol. Vol. 123, Plenum Press, New York & London.
Manthorpe, M., Adler, R., andVaron, S. 1979. Development, reactivity and GFA immunofluorescence of astroglia-containing monolayer cultures from rat cerebrum. J. Neurocytol. 8:605–621.
McCarthy, K. D., andDe Vellis, J. 1980. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J. Cell Biol. 85:890–902.
McGeer, P. L., Eccles, J. C., andMcGeer, E. (eds). 1978. Inhibitory amino acid neurons: GABA and glycine. Pages 199–231,in Molecular neurobiology of the mammalian brain. Plenum Press, New York.
McGeer, P. L., Eccles, J. C., andMcGeer, E. (eds). 1978. Catecholamine neurons. Pages 233–271,in Molecular neurobiology of the mammalian brain. Plenum Press, New York.
Pelton, E. W. II, Kimelberg, K. H., Shipherd, S. V., andBourke, R. S. 1981. Dopamine and norepinephrine uptake and metabolism by astroglial cells in culture. Life Sci. 28:1655–1663.
Romeis, B. (ed). 1948. Mikroskopische Technik, Oldenburg, München, p 472.
Schousboe, A., andDivac, I. 1979. Differences in glutamate uptake in astrocytes cultured from different brain regions. Brain Research 177:407–409.
Schousboe, A., Drejer, J., andDivac, I. 1980. Regional heterogeneity in astroglial cells. Implications of neuron-glia interactions. Trends in Neurosciences 3:XIII-XIV.
Schousboe, A., Fosmark, H., andFormby, B. 1976. Effect of serum withdrawal on Na+−K+ ATPase activity in astrocytes cultured from dissociated brain hemispheres. J. Neurochem. 26:1053–1055.
Schousboe, A., Hertz, L., andSvenneby, G. 1977. Uptake and metabolism of GABA in astrocytes cultured from dissociated mouse brain hemispheres. Neurochem. Res. 2:217–229.
Sellström, Å., andHamberger, A. 1975. Neuronal and glial systems for γ-aminobutyric acid transport. J. Neurochem. 24:847–852.
Silberstein, S. D., Schein, H. M., andBerv, K. R. 1972. Catechol-O-methyltransferase and monoamine oxidase activity in cultured rodent astrocytoma cells. Brain Research 41:245–248.
Van den Berg, C. J., van Kempen, G. M. J., Schadé, J. P., andVeldstra, H. 1965. Levels and intracellular localization of glutamate decarboxylase and γ-aminobutyrate transaminase and other enzymes during the development of the brain. J. Neurochem. 12:863–869.
Waksman, A., Rubinstein, M. K., Kuriyama, K., andRoberts, E. 1968. Localization of γ-aminobutyric-oxoglutaric acid transaminase in mouse brain. J. Neurochem. 15:351–357.
Wurtman, R. J., andAxelrod, J. 1964. A sensitive and specific assay for the estimation of monoamine oxidase. Biochem. Pharmacol. 12:1439–1441.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hansson, E. Enzyme activities of monoamine oxidase, cathechol-o-methyltransferase and γ-aminoubutyric acid transaminase in primary astroglial cultures and adult rat brain from different brain regions. Neurochem Res 9, 45–57 (1984). https://doi.org/10.1007/BF00967658
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00967658