Characterization of uptake and release processes ford- andl-aspartate in primary cultures of astrocytes and cerebellar granule cells

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 sameKm- andVmax-values in bouth astrocytes (l-aspartate:Km 77 μM;Vmax 11.8 nmol×min−1×mg−1;d-aspartate:Km 83 μM;Vmax 14.0 nmol×min−1×mg−1) and granule cells (l-aspartate:Km 32 μM;Vmax 2.8 nmol ×min−1×mg−1;d-aspartate:Km 26 μM;Vmax 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.

[1]  P. Beart,et al.  COMPETITIVE INHIBITION OF GABA UPTAKE IN RAT BRAIN SLICES BY SOME GABA ANALOGUES OF RESTRICTED CONFORMATION , 1972, Journal of neurochemistry.

[2]  A. Schousboe,et al.  Differences between GABA receptor binding to membranes from cerebellum during postnatal development and from cultured cerebellar granule cells. , 1982, Developmental neuroscience.

[3]  A. Yu,et al.  Uptake of glutamate, GABA, and glutamine into a predominatly GABA‐ergic and a predominantly glutamatergic nerve cell population in culture , 1982, Journal of neuroscience research.

[4]  V. Balcar,et al.  THE STRUCTURAL SPECIFICITY OF THE HIGH AFFINITY UPTAKE OF l‐GLUTAMATE AND l‐ASPARTATE BY RAT BRAIN SLICES , 1972, Journal of neurochemistry.

[5]  L. P. Davies,et al.  UPTAKE AND RELEASE OF D‐ AND L‐ASPARTATE BY RAT BRAIN SLICES , 1976, Journal of neurochemistry.

[6]  G. N. Wilkinson.  Statistical estimations in enzyme kinetics. , 1961, The Biochemical journal.

[7]  A. M. Benjamin,et al.  CEREBRAL UPTAKES AND EXCHANGE DIFFUSION IN VITRO OF l‐ AND d‐GLUTAMATES , 1976, Journal of neurochemistry.

[8]  V. Balcar,et al.  Stereospecificity of the inhibition of l‐glutamate and l‐aspartate high affinity uptake in rat brain slices by threo‐3‐hydroxyaspartate , 1977, Journal of neurochemistry.

[9]  Roberts Pj,et al.  Radioreceptor binding studies with glutamate and aspartate. , 1981 .

[10]  F. V. Defeudis,et al.  CNS receptors : from molecular pharmacology to behavior , 1983 .

[11]  V. Gallo,et al.  Glutamate as a Putative Transmitter in the Cerebellum: Stimulation by GABA of Glutamic Acid Release from Specific Pools , 1981, Journal of neurochemistry.

[12]  A. Schousboe,et al.  UPTAKE AND METABOLISM OF GLUTAMATE IN ASTROCYTES CULTURED FROM DISSOCIATED MOUSE BRAIN HEMISPHERES , 1977, Journal of neurochemistry.

[13]  A. Schousboe,et al.  Trophic actions of GABA on the development of physiologically active GABA receptors. , 1983, Advances in biochemical psychopharmacology.

[14]  A. Schousboe,et al.  Characterization of L-glutamate uptake into and release from astrocytes and neurons cultured from different brain regions , 2004, Experimental Brain Research.

[15]  G. G. Járos,et al.  Neuronal cells from chick embryo cerebral hemispheres cultivated on polylysine-coated surfaces. , 1978, Developmental neuroscience.

[16]  C. Cotman,et al.  An overview of glutamate as a neurotransmitter. , 1981, Advances in biochemical psychopharmacology.

[17]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[18]  J. Watkins,et al.  Pharmacology of receptors for excitatory amino acids. , 1981, Advances in biochemical psychopharmacology.

[19]  A. Lajtha,et al.  Heterogeneity of the mediated transport systems of amino acid uptake in brain. , 1966, Brain research.

[20]  G. Johnston,et al.  HIGH AFFINITY UPTAKE OF TRANSMITTERS: STUDIES ON THE UPTAKE OF l‐ASPARTATE, GABA, l‐GLUTAMATE AND GLYCINE IN CAT SPINAL CORD , 1973, Journal of neurochemistry.

[21]  A. Messer.  The maintenance and identification of mouse cerebellar granule cells in monolayer culture , 1977, Brain Research.

[22]  J. L. Johnson.  The excitant amino acids glutamic and aspartic acid as transmitter candidates in the vertebrate central nervous system , 1978, Progress in Neurobiology.

[23]  H. Glaumann,et al.  Structural Analogues of Ibotenic Acid. Syntheses of 4-Methylhomoibotenic Acid and AMPA, Including the Crystal Structure of AMPA, Monohydrate. , 1980 .

[24]  K. Krnjević,et al.  Chemical Nature of Synaptic Transmission in Vertebrates , 1974 .

[25]  I. H. Segel.  Enzyme Kinetics: Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems , 1975 .

[26]  P. Mandel,et al.  HIGH AFFINITY UPTAKE OF L‐GLUTAMATE AND L‐ASPARTATE BY GLIAL CELLS , 1977, Journal of neurochemistry.

[27]  S. Snyder,et al.  Gamma-aminobutyric acid (GABA) receptor binding selectively depleted by viral induced granule cell loss in hamster cerebellum , 1976, Brain Research.

[28]  R. Shank,et al.  Glutamate and GABA uptake by cerebellar granule and glial cell enriched populations , 1978, Brain Research.