Cloning Of Sert And The Elucidation Of Sert Gene Structure

Following the scheme we outline above for polymorphism identification and having chosen our candidate gene (SERT) for investigation, we now must understand the structure of the human gene and its reported patterns of genetic variation. Rodent cDNA cloning studies (Blakely et al., 1991 Hoffman et al., 1991) first revealed that SERTs exhibit a 12-transmembrane domain structure with intracellular amino and carboxy termini, as predicted for aminobutyric acid (GABA) (Guastella et al., 1990) and NETs...

Annotated References

Amara SG, Kuhar MJ (1993) Neurotransmitter transporters Recent progress. Annu Rev Neurosci 16 73-93. Bieber AJ (1994) Analysis of cellular adhesion in cultured cells. Methods Cell Biol 44 683-695. Bellen HJ, O'Kane CJ, Wilson C, Grossniklaus U, Pearson RK, Gehring WJ (1989) P-element-me-diated enhancer detection A versatile method to study development in Drosophila. Genes Dev 3 1288-1300. Besson M-T, Soustelle L, Birman S (1999) Identification and structural characterization of two genes...

Cocaine Effects In Dat Ko Mice

Because the DAT is thought to be the primary target of the rewarding actions of cocaine, it has been of interest as to why mice with no DAT still self-administer cocaine (Rocha et al., 1998). The other remaining targets of cocaine, the norepinephrine transporter (NET) and the serotonin transporter (SERT), seemed likely candidates as mediators of cocaine reward in mice without DAT. Two studies have attempted to answer the question, with differing results. One group focused on the SERT. First,...

Searching For Rare Genetic Variants In Candidate Genes

For complex disorders in general, as for the mental illnesses where disrupted serotonergic signaling is suspected in particular, two different strategies have dominated the attempt to understand genetic underpinnings. The first involves a genome-wide scan in affected subjects or pedigrees for the presence or segregation of polymorphic DNA markers. This top-down approach has the advantage of making no assumptions as to which gene contributes variation in a trait or the risk for occurrence of a...

Mechanism of DAT Function

Dopamine transporters operate by coupling the transmembrane translocation of substrate to the movement of driving ions down pre-established electrochemical gradients. Substrates of the DAT include DA, AMPH, and tyramine. The most widely held concept of how co-transporters function is founded on the alternating access model (Jardetzky, 1966), in which the binding sites for substrates and co-substrates are alternately exposed to extracellular or cytoplasmic environments via conformational changes...

References

Abi-Dargham A, Gandelman MS, DeErausquin GA, et al. (1996) SPECT imaging of dopamine transporters in human brain with iodine-123-fluoroalkyl analogs of beta-CIT. J Nucl Med 37 1129-1133. Acton PD, Kung MP, Mu M, et al. (1999a) Single-photon emission tomography imaging of serotonin transporters in the non-human primate brain with the selective radioligand 123I IDAM. Eur J Nucl Med 26 854-861. Acton PD, Kushner SA, Kung MP, et al. (1999b) Simplified reference region model for the kinetic analysis...

Fluorescence labeling protocol

Cysteines are introduced by site-directed mutagenesis into putative extracellular loops or pore regions of glutamate transporters, one at a time, and are analyzed after reaction with cysteine-specific fluorescent probes. The cysteines can be introduced into a wild-type rather than Cys-less EAAT3 background, because EAAT3 does not react with any of the fluorescence probes tested to date. This is verified by comparing fluorescence in oocytes expressing transporters before and after labeling and...

Dopamine Transporter Dat Imaging Agents Development of DAT Imaging Agents

Ligands for dopamine transporters DAT are useful in evaluating changes in presynaptic DAT sites in vivo and in vitro, especially for patients with movement disorders such as Parkinson's disease PD , which is characterized by a selective loss of DA neurons in the basal ganglia and substantia nigra. The first successful DAT imaging agent for PET, 11C CFT WIN35,428 , was reported in 1993 for evaluation of patients with PD Frost et al., 1993 . Numerous other 11C phenyltropanes have been reported...