Dopamine Lab Report

Locating the Dopamine Transporter via Immunostaining and Colocalization

Taryn Nurse+, Kadija King+, Dasia McClain+, Jahmel Fowler+, Phyllis Freeman+, and Shawn Goodwin*.

Life and Physical Science Department, Fisk University+ and Department of Biology. Meharry Medical College School of Graduate Studies and Research*, Nashville, TN, 37208.

Introduction (Dasia)

The focus of this experiment was to identify the Dopamine Active Transporter (DAT), known as SLC6A3, which is located on the 5th chromosome. By using a plasmid, the gene that encodes for DAT was attached to the gene for a Yellow Fluorescent Protein (YFP) in Human Embryonic Kidney (HEK) cells. This resulted in the production of the DAT and YFP proteins in the cells. Two antibodies were used to cause fluorescence in the cells.

Background (Taryn)

Dopamine Active Transporter (DAT)

The DAT is a 12-membrane spanning protein that clears away excess dopamine from the synaptic cleft between neurons.

Dopamine

Dopamine acts to inhibit acetylcholine, which is responsible for muscular contractions. The gradual loss of dopamine is attributed to the abnormal nerve-firing patterns associated with Parkinson’s disease. Parkinson’s disease is a progressive disease caused by the deterioration of the central nervous system. In treatment, dopamine is presented to patients in the form of a precursor, L-DOPA. Dopamine is biosynthesized from one of 20 the essential amino acids. This occurs when tyrosine is converted to L-DOPA with hydroxylase. L-DOPA is then converted to dopamine via a decarboxylation