Vol. 295, Issue 1, 392-403, October 2000

Transport of N-Acetylaspartate by the Na⁺-Dependent High-Affinity Dicarboxylate Transporter NaDC3 and Its Relevance to the Expression of the Transporter in the Brain¹

Wei Huang, Haiping Wang, Ramesh Kekuda, You-Jun Fei, Anne Friedrich, Jian Wang, Simon J. Conway, Richard S. Cameron, Frederick H. Leibach and Vadivel Ganapathy

Departments of Biochemistry and Molecular Biology (W.H., H.W., R.K., Y.-J.F., A.F., F.H.L., V.G.), and the Institute of Molecular Medicine and Genetics (J.W., S.J.C., R.S.C.), Medical College of Georgia, Augusta, Georgia

N-Acetylaspartate is a highly specific marker for neurons and is present at high concentrations in the central nervous system. It is not present at detectable levels anywhere else in the body other than brain. Glial cells express a high-affinity transporter for N-acetylaspartate, but the molecular identity of the transporter has not been established. The transport of N-acetylaspartate into glial cells is obligatory for its intracellular hydrolysis, a process intimately involved in myelination. N-Acetylaspartate is a dicarboxylate structurally related to succinate. We investigated in the present study the ability of NaDC3, a Na⁺-coupled high-affinity dicarboxylate transporter, to transport N-acetylaspartate. The cloned rat and human NaDC3s were found to transport N-acetylaspartate in a Na⁺-coupled manner in two different heterologous expression systems. The Michaelis-Menten constant for N-acetylaspartate was ~60 µM for rat NaDC3 and ~250 µM for human NaDC3. The transport process was electrogenic and the Na⁺:N-acetylaspartate stoichiometry was 3:1. The functional expression of NaDC3 in the brain was demonstrated by in situ hybridization and reverse transcription-polymerase chain reaction as well as by isolation of a full-length functional NaDC3 from a rat brain cDNA library. In addition, the expression of a Na⁺-coupled high-affinity dicarboxylate transporter and the interaction of the transporter with N-acetylaspartate were demonstrable in rat primary astrocyte cultures. These studies establish NaDC3 as the transporter responsible for the Na⁺-coupled transport of N-acetylaspartate in the brain. This transporter is likely to be an essential component in the metabolic role of N-acetylaspartate in the process of myelination.