Characterization of 3-[(123)I]iodo-L-alpha-methyl tyrosine transport in astrocytes of neonatal rats

J Neurochem. 2001 Jan;76(1):97-104. doi: 10.1046/j.1471-4159.2001.00048.x.

Abstract

3-[(123)I]Iodo-L-alpha-methyl tyrosine ((123)I-IMT) is used for diagnosis and monitoring of brain tumours by means of single-photon emission tomography. As recently shown, (123)I-IMT is predominantly mediated into rat C6 glioma cells by sodium-independent system L for large neutral amino acids. Until now, (123)I-IMT transport in non-neoplastic glial cells has not been examined. Therefore, the aim of this study was to examine the cellular pathways and precise transport kinetics of (123)I-IMT uptake into astrocytes of neonatal rats. In particular sodium-independent (123)I-IMT transport into neonatal astrocytes was compared with sodium-independent (123)I-IMT uptake into neoplastic rat C6 glioma cells. Competitive inhibition experiments showed that (123)I-IMT is exclusively transported via sodium-independent system L into the neonatal astrocytes (92%). Kinetic analysis of sodium-independent (123)I-IMT uptake into neonatal astrocytes and into C6 glioma cells revealed apparent Michaelis constants K(M) = 13.9 +/- 0.5 microM and K(M) = 33.9 +/- 4.1 microM, respectively, which are in the same range of K(M) values as those recently determined for amino acid transport into neoplastic and non-neoplastic glial cells. Indeed, the K(M) values in the micromolar range correspond to the expression of the LAT-1 subunit of system L both in the neonatal astrocytes and in C6 glioma cells. However, sodium-independent maximum transport velocities (V(max)) differed significantly between neonatal astrocytes and C6 glioma cells (11.1 +/- 0.3 and 39.9 +/- 3.3 nmol/mg protein/10 min, respectively).

MeSH terms

  • Amino Acid Transport Systems
  • Amino Acids, Cyclic / metabolism
  • Amino Acids, Cyclic / pharmacology
  • Animals
  • Animals, Newborn
  • Astrocytes / cytology
  • Astrocytes / metabolism*
  • Binding, Competitive / drug effects
  • Biological Transport / drug effects
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Glioma / diagnosis
  • Glioma / metabolism*
  • Leucine / metabolism
  • Leucine / pharmacology
  • Methyltyrosines / metabolism*
  • Methyltyrosines / pharmacokinetics*
  • Phenylalanine / metabolism
  • Phenylalanine / pharmacology
  • RNA, Messenger / metabolism
  • Radiopharmaceuticals
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • alpha-Methyltyrosine / metabolism
  • alpha-Methyltyrosine / pharmacology
  • beta-Alanine / analogs & derivatives*
  • beta-Alanine / metabolism
  • beta-Alanine / pharmacology

Substances

  • Amino Acid Transport Systems
  • Amino Acids, Cyclic
  • Carrier Proteins
  • Methyltyrosines
  • RNA, Messenger
  • Radiopharmaceuticals
  • beta-Alanine
  • 2,2-dimethyl-beta-alanine
  • 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid
  • Phenylalanine
  • alpha-Methyltyrosine
  • 3-iodo-alpha-methyltyrosine
  • Leucine