Localization of GABA transporter and its associated gene at the blood-brain barrier / 中国药理学通报

ABSTRACT

Aim To identify the GABA transporter subtypes and to find its new associated member at the blood brain barrier.Methods Labeled by in vitro infusion of magnetic beads through carotid arteries,the brain microvessels without intact neural cells were sorted in magnetic fields,and identified by RT PCR.The homologous primer of GAT superfamily and the tRNA from isolated brain microvessels were used in the RT PCR to amplify target DNA.The PCR products were isolated by polyacrylamide gel electrophoresis(PAGE) and cloned, sequenced rospectively.The sequences were screened in dbEST of Genbank by Blast.Results Seven DNA bands were isolated from RT PCR products of isolated brain microvessels by PAGE. B3,B5 complete sequences were highly homologous with rat partial GAT 2 and BGT 1 respectively,B7 complete sequence was highly homologous with rat partial TAUT. The other 4 EST of B1(Accession NoCF358965), B2(CD568346), B4(CF358966) and B6(CD568347) were submitted to dbEST,they were homologous with some sequences in Genbank,but were not homologous with GAT members.Conclusion GAT 2 and BGT 1 of GAT and TAUT were localized at the blood brain barrier which might be responsible for the GABA transport across the blood brain barrier.The genes and their functions of 4 EST associated with GAT need to be clarified.