Blakely Lab

My primary responsibility is the oversight of the Blakely lab cell culture facility. Duties here involve upkeep of cell incubators and laminar flow hoods, oversight and ordering of stock cell culture media and supplies, the preparation of stable cell lines, transient transfections of transporter genes, and general oversight of banked cell lines. I also participate in the development and characterization of transporter antibodies and the analysis of transporter protein expression via Western blots and immunoprecipitations.

My duties involve support for multiple aspects of transporter genetic analyses using both rodent model systems and human genetic analyses. My current activities with transgenic mice focus on the serotonin transporter and include genotyping and colony management of novel “knock-in” transgenic mice. Additionally, I support the screening of genomic DNA from human subjects utilizing REVEAL TGCE technology, presently involving a search for functional coding variation in the norepinephrine transporter gene in ADHD subjects. Finally, I manage the lab research paper database and image archive.

My lab responsibilities include the oversight and maintenance of human genomic DNA stocks used by various lab members, equipment repair and services, and provide back up for purchasing consumable reagents. I am also involved in general molecular biology support centered on projects related to human transporter genetic variation. Additionally, I support the lab by developing and implementing assays for screening human genomic DNA from various clinical syndromes for single nucleotide polymorphisms (SNPs) relating to norepinephrine transporter dysfunction. Methods include PCR, DNA sequencing, pyrosequencing and REVEAL analysis..

As Laboratory Manager for the Blakely Lab, I am responsible for the general oversight and smooth functioning of the lab. I assist in experimental problem solving, work to develop and improve laboratory protocols and oversee the cohesive activities of the laboratory staff. I also serve as a liaison with other labs within and without Vanderbilt. My research examines the impact of genetic mutations and regulatory signaling on the association of SERT interacting proteins. I exploit both cell culture and transgenic models to ascertain the functional contributions of SERT regulatory molecules in vitro and in vivo.

My research is primarily oriented around the use of the powerful genetic model system Caenorhabditis elegans and involves animal husbandry, transgenic animal production, crosses and archiving, and behavioral/pharmacological challenges that relate to the function of the dopamine and choline transporters. I utilize microscopy and computer-assisted motion detection systems for phenotyping mutant nematodes and also assist with research on the choline transporter in mammalian model systems.

My primary effort in the Blakely lab is to maintain the neuroscience core microscopes and histology equipment and help CMN investigators with cellular/anatomic studies as needed. Using immunohistochemical and in situ hybridization techniques, I am involved in antibody purification and in identifying sites of expression of biogenic amine and a newly clones choline transporter in native tissue and cell lines. I am also responsible for maintaining the colony of various genetically modified mice established in, and used by, the Blakely lab. Finally, I facilitate the development of transgenic or viral constructs to modify transporter expression in vitro and in vivo.