Novel Na+ -independent and adenine-specific transport system for adenine in primary cultured rat cortical neurons.
Endogenous adenine is an important modulator of cell survival and activity in the central nervous system. In the present study, we examined the transport mechanisms for adenine in primary cultured rat cortical neurons and astrocytes. [3H]Adenine was time-dependently taken up into neurons, but not into astrocytes. In kinetic analysis, the [3H]adenine uptake by neurons was observed to be saturable, and an Eadie-Hofstee plot showed that a single component was involved in the uptake, with kinetic parameters of Km=6.09 microM and Vmax=0.340 nmol/mg protein per min. In inhibition assaying by nucleobases and nucleosides, and inhibitors for equilibrative nucleoside transporters, organic ion transporters and peptide transporters, which were reported to transport nucleobases and their analogues, the [3H]adenine uptake by neurons was found to be significantly inhibited by excess concentrations of adenine, hypoxanthine and adenosine, and was greatly reduced only by the addition of adenine. Therefore, it was indicated that adenine in the extracellular fluid in the central nervous system is taken up into neurons, but not into astrocytes, and that neurons may present a novel Na+ -independent and adenine-specific transport system.
Nagai
Katsuhito
K
Department of Environmental Biochemistry, Kyoto Pharmaceutical University, Misasagi, Kyoto, Japan. nagai@mb.kyoto-phu.ac.jp
Nagasawa
Kazuki
K
Matsunaga
Ryoko
R
Yamaji
Masumi
M
Fujimoto
Sadaki
S
D000225:Adenine / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism*; D002414:Cations, Monovalent; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D045604:Extracellular Fluid / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D012964:Sodium / Q000502:physiology*
D002352:Carrier Proteins; D002414:Cations, Monovalent; D012964:Sodium; D000225:Adenine