Abstract
The uptake ofl-andd-aspartate was studied in astrocytes cultured from prefrontal cortex and in granule cells cultured from cerebellum. A high affinity uptake system forl- andd-aspartate was found in both cell types, and the two stereoisomers exhibited essentially the sameK m - andV max -values in bouth astrocytes (l-aspartate:K m 77 μM;V max 11.8 nmol×min−1×mg−1;d-aspartate:K m 83 μM;V max 14.0 nmol×min−1×mg−1) and granule cells (l-aspartate:K m 32 μM;V max 2.8 nmol ×min−1×mg−1;d-aspartate:K m 26 μM;V max 3.0 nmol×min−1×mg−1). To investigate whetherl-glutamate,l-aspartate andd-aspartate use the same uptake system a detailed kenetic analysis was performed. The uptake kinetics of each one of the three amino acids was studied in the presence of the two other amino acids, and no essential differences between the uptake characteristics of the amino acids were found. In addition to the uptake studies the release ofD-aspartate from cerebellar granule cells was investigated and compared withl-glutamate release. A Ca2+-dependent, K+-induced release was found for both amino acids.
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References
Krnjevic, K. 1974. Chemical nature of synaptic transmission in vertebrates. Physiol. Rev. 54:418–540.
Cotman, C. W., Foster, A., andLanthorn, T. 1981. An overview of glutamate as a neurotransmitter. Pages 1–27,in DiChiara, G., andGessa, G. L. (eds.), Glutamate as a Neurotransmitter, Adv. Biochem. Psychopharmacol. Vol. 27, Raven Press, New York.
Schousboe, A. 1982. Chap. 6. Pages 107–141,in Pfeiffer, S. E. (ed.), Neuroscience Approached Through Cell Culture. Vol. I. CRC Press, Boca Raton, Florida.
Johnson, J. L. 1978. The excitant amino acids glutamic and aspartic acid as transmitter candidates in the vertebrate central nervous system. Prog. Neurobiol. 10:155–202.
Watkins, J. C., Davies, J., Evans, R. H., Francis, A., andJones, A. W. 1981. Pharmacology of receptors of excitatory amino acids. Pages 263–273,in DiChiara, G., andGessa, G. L. (eds.), Glutamate as a Neurotransmitter, Adv. Biochem. Psychopharmacol. Vol. 27. Raven Press, New York.
Balcar, V. J., andJohnston, G. A. R. 1972. The structural specificity of the high affinity uptake ofl-glutamate andl-aspartate by rat brain slices. J. Neurochem. 19:2657–2666.
Balcar, V. J., andJohnston, G. A. R. 1973. High affinity uptake of transmitters: Studies on the uptake ofl-aspartate, GABA,l-glutamate and glycine in cat spinal cord. J. Neurochem. 20:529–539.
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.
Davies, L. P., andJohnston, G. A. R. 1976. Uptake and release ofd-andl-aspartate by rat brain slices. J. Neurochem. 26:1007–1014.
Levi, G., andGallo, V. 1981. Glutamate as a putative transmitter in cerebellum: Stimulation by GABA of glutamic acid release from specific pools. J. Neurochem. 37:22–31.
Honoré, T., andLauridsen, J. 1980. Structural analogues of ibotenic acid. Syntheses of 4-methylhomoibotenic acid and AMPA, including the crystal structure of AMPA, monohydrate. Acta Chem. Scand. B34:235–240.
Hertz, L., Juurlink, B. H. J., Fosmark, H., andSchousboe, A. 1982. Pages 175–186. Chap 9in Pfeiffer, S. E. (ed.), Neuroscience Approached Through Cell Culture. Vol. I. CRC Press, Boca Raton, Florida.
Messer, A. 1977. The maintenance and identification of mouse cerebellar granule cells in monolayer cultures. Brain Res. 130:1–12.
Yu., A. C. H., andHertz, L. 1982. Uptake of glutamate, GABA, and glutamine into a predominantly GABA-ergic and a predominantly glutamatergic nerve cell population in culture. J. Neurosci. Res. 7:23–35.
Sensenbrenner, M., Maderspach, K., Latzkovits, L., andJaros, G. G. 1978. Neuronal cells from chick embryo cerebral hemispheres cultivated on polylysine-coated surfaces. Dev. Neurosci. 1:90–101.
Currie, D. N. 1980. Identification of cell types by immunofluorescence in defined cell cultures of cerebellum. Pages 75–87,in Giacobini, E., Vernadakis, A., andShahar, A. (eds.), Tissue culture in neurobiology. Raven Press, New York.
Balázs, R., Regan, C., Meier, E., Woodhams, P. L., Wilkin, G. P., Patel, N. J., Gordon, R. D. 1980. Biochemical properties of neural cell types from rat cerebellum. Pages 155–168,in Giacobini, E., Vernadakis, A., andShahar, A. (eds.), Tissue Culture in Neurooiology. Raven Press, New York.
Schousboe, A., Larsson, O. M., Meier, E., Hertz, L., Krogssgaard-Larsen, P. 1982. Tissue culture studies of GABA-Neurotransmission. Pages 249–260,in Okada, Y., andRoberts, E. (eds.), Problems in GABA Research. Excerpta Medica, Amsterdam.
Meier, E., andSchousboe, A. 1982. Differences between GABA receptor binding to membranes from cerebellum during postnatal development and from cultured cerebellar granule cells. Dev. Neurosci. 5:546–553.
Simantov, R., Oster-Granite, M. L., Herndon, R. M., Snyder, S. H. 1976. Gammaaminobutyric aicd (GABA) receptor binding selectively depleted by viral induced granule cell loss in hamster cerebellum. Brain Res. 105:365–371.
Campbell, G. L. M., andShank, R. P. 1978. Glutamate and GABA uptake by cerebellar granule and glial cell enriched populations. Brain Res. 153:618–622.
Seil, F. J. 1979. Cerebellum in tissue culture. Rev. Neurosci. 4:105–177.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., andRandall, R. L. 1951. Protein measurement with the phenol reagent. J. Biol. Chem. 193:265–275.
Beart, P. M., Johnston, G. A. R., Uhr, M. L. 1972. Competitive inhibition of GABA uptake in rat brain slices by some GABA analogues of restricted conformation. J. Neurochem. 19:1855–1861.
Balcar, V. J., Borg, J., andMandel, P. 1977. High affinity uptake ofl-glutamate andl-aspartate by glial cells. J. Neurochem. 28:87–93.
Schousboe, A., Svenneby, G., andHertz, L. 1977. Uptake and metabolism of glutamate in astrocytes cultured from dissociated mouse brain hemispheres. J. Neurochem. 29:999–1005.
Blasberg, R., andLajtha, A. 1966. Heterogeneity of the mediated transport systems of amino acid uptake in brain. Brain Res 1:86–104.
Benjamin, A. M., andQuastel, J. H. 1976. Cerebral uptakes and exchange diffusion in vitro ofl-andd-glutamates. J. Neurochem. 26:431–441.
Balcar, V. J., Johnston, G. A. R., andTwitchin, B. 1977. Stereospecific inhibition ofl-glutamate andl-aspartate high affinity uptake in rat brain slices by threo-3-hydroxyaspartate. J. Neurochem. 28:1145–1146.
Meier, E., Drejer, J., andSchousboe, A. 1983. Trophic actions of GABA on the development of physiologically active GABA receptors,in DeFeudis, F. V. andMandel, P. (eds.), CNS Receptors—From Molecular Pharmacology to Behaviour. Raven Press, New York in press.
Roberts, P. J., andSharif, N. A. 1981. Radioreceptor binding studies with glutamate and aspartate. Pages 295–305,in DiChiara, G., andGessa, G. L. (eds.), Glutamate as a Neurotransmitter. Adv. Biochem. Psychopharmacology, Vol. 27, Raven Press, New York.
Wilkinson, G. N. 1961. Statistical estimations in enzyme kinetics. Biochem. J. 80:324–332.
Segel, I. H. 1975. Enzyme kinetics. Behaviour and analysis of rapid equilibrium and steady-state enzyme systems. Pages 170–176. Wiley & Sons, New York.
Drejer, J. 1982. Glutamate-mediated neurotransmission. A review. Studies in neurons and astrocytes in primary cultures. Ph.D. Thesis, Pages 1–81, Royal Danish School of Pharmacy, Copenhagen.
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Drejer, J., Larsson, O.M. & Schousboe, A. Characterization of uptake and release processes ford- andl-aspartate in primary cultures of astrocytes and cerebellar granule cells. Neurochem Res 8, 231–243 (1983). https://doi.org/10.1007/BF00963923
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DOI: https://doi.org/10.1007/BF00963923