Generation And Behavioral Studies Of Dopamine Receptor D5B Transgenic Rats

We have cloned a new subtype of human dopamine D5 receptor (D5B). The sequence of human D5B receptor is 98% identical to human D5 receptor and 94% identical to two human D5 receptor pseudogenes. Unlike the pseudogenes of D5, the D5B receptor gene contains a long open reading frame that encodes a functional protein, which is one amino acid longer than human D5 receptor. The pharmacological study showed that the D5B receptor had distinct pharmacological properties from D5 receptor. In order to study the function of D5B receptor, we have generated transgenic rats overexpressing human D5B receptors driven by CamKII promoter. The receptors were specifically expressed in the forebrain of transgenic rats by RT-PCR and Western blot. Behavioral studies, including anxiety, depress, learning and memory, were conducted for D5B transgenic rat. Our results showed that D5B might play a complicated role in regulating multiple functions of brain such as mood, learning and memory.1. Validation of the cloning of D5B geneWe have used different DNA templates and primers to clone D5B receptor gene from genomic DNA isolated from human blood and human cell lines, including SH-Sy5Y and HEK293 cell lines. The PCR products were cloned to T vector and sequenced. Twelve D5B clones were identified from 41 PCR clones.2. Generation of transgenic ratsRats have been widely used for pharmacological studies. The first transgenic rat was generated in 1990. Since then, a large number of transgenic rats have been generated and used as human disease models. To generate D5B transgenic rats, we...