Astroglia contain a specific transport mechanism for N-acetyl-L-aspartate.
N-Acetylaspartate (NAA) is the second most abundant amino acid in the adult brain. It is located and synthesized in neurons and probably degraded in the glia compartment, but the transport mechanisms are unknown. Rat primary neuron and astrocyte cell cultures were exposed to the L isomer of [3H]NAA and demonstrated concentration-dependent uptake of [3H]NAA with a Km approximately 80 microM. However, Vmax was 23+/-6.4 pmol/mg of protein/min in astrocytes but only 1.13+/-0.4 pmol/mg of protein/min in neurons. The fact that neuron cultures contain 3-5% astrocytes suggests that the uptake mechanism is expressed only in glial cells. The astrocyte uptake was temperature and sodium chloride dependent and specific for L-NAA. The affinity for structural analogues was (IC50 in mM) as follows: L-NAA (0.12) > N-acetylaspartylglutamate (0.4) > N-acetylglutamate (0.42) > L-aspartate (>1) > L-glutamate (>1) > or = DL-threo-beta-hydroxyaspartate > N-acetyl-L-histidine. The naturally occurring amino acids showed no inhibitory effect at 1 mM. The glutamate transport blocker trans-pyrrolidine-2,4-dicarboxylate exhibited an IC50 of 0.57 mM, whereas another specific glutamate transport inhibitor, DL-threo-beta-hydroxyaspartate, had an IC50 of >1 mM. The experiments suggest that NAA transport in brain parenchyma occurs by a novel type of sodium-dependent carrier that is present only in glial cells.
Sager
T N
TN
Department of Neuroendocrine Pharmacology, Novo Nordisk A/S, Maaloev, Denmark.
Thomsen
C
C
Valsborg
J S
JS
Laursen
H
H
Hansen
A J
AJ
D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D001253:Astrocytes / Q000737:chemistry / Q000201:enzymology*; D001692:Biological Transport / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D002712:Chlorides / Q000493:pharmacokinetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005333:Fetus / Q000166:cytology; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D009474:Neurons / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology; D051381:Rats; D012964:Sodium / Q000493:pharmacokinetics; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013379:Substrate Specificity; D014316:Tritium; D014746:Vimentin / Q000032:analysis
D002712:Chlorides; D004791:Enzyme Inhibitors; D005904:Glial Fibrillary Acidic Protein; D014746:Vimentin; D014316:Tritium; D001224:Aspartic Acid; D010042:Ouabain; C000179:N-acetylaspartate; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase