Drosophila melanogaster imaginal disc assays to study the polyamine transport system

Polyamine metabolism in higher eukaryotes is well studied; however, the mechanism of how the polyamines putrescine, spermidine and spermine enter the cell remains unclear. An effective approach to investigate potential players that function in the uptake of polyamines involves using the Drosophila melanogaster imaginal disc assay. Leg imaginal discs dissected from Drosophila melanogaster wandering third star larvae can be assessed for leg development after 18 h of treatment with hormones to induce this process. The protocol described here details how to use genetically manipulated Drosophila melanogaster to test candidate genes involved in the polyamine transport system, how to dissect leg imaginal discs and how to assess the entry of polyamines into the cells of the imaginal disc.