Investigation of SLC36 and KAAT Family Genes as Tyrosine Transporters in the Malpighian Tubule

William F. Mueller
Marquette University
Milwaukee, WI
Mentor: Dr. Edward Blumenthal

Tyrosine is an amino acid found Drosophila melanogaster.   It has been found to cause a rapid depolarization of the transepethilial membrane of the malpighian tubule, a vital part of the urinary/excretory system.  However, the means of transport by which that tyrosine is taken into the tubule cells is unknown.   Evidence of a gradient-coupled transporter suggests that two gene families could code for a tyrosine-ion coupled transporter, Pathetic (a.k.a SLC36, proton coupled transporters, 11 genes) and KAAT (potassium coupled transporters, 6 genes) are those two families.   Using real-time PCR the expression levels of the genes with in these families were determined, this lead to the initial steps of in situ hybridization on the highly expressed genes.  The pathetic family showed no expression for genes CG12943 and CG32079, thus those genes were ruled out as possibilities.  Of the remaining nine genes, the most highly expressed genes, verified by microarray data, were CG1139, CG3424 (pathetic), CG6327, and CG8785.  However in comparison to rp49 (control house keeping gene) expression levels and KAAT family levels it was not likely that a pathetic gene coded for the transporter.  The most highly expressed KAAT family gene was CG15088.  Taking a non-conserved section of is transcript; a riboprobe was synthesized for in situ hybridization to localize the expression this gene in the tubule. 
 
 
 

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