Figure 1.

Fly and human spastin transgenes have similar subcellular distributions. (A) Constructs containing GAL4 binding sites (UAS) followed by genes encoding a fluorescent tag (CFP or Venus YFP) and either the spastin full-length wild-type fly genomic (D^WT) or human cDNA (H^WT) sequence were generated using the Drosophila Gateway system. Transgenic flies bearing either construct could then be used to express fluorescently tagged fly or human Spastin under the spatiotemporal control of a promoter-GAL4 line. Numbers denote amino acid position. (B) en-GAL4 was used to drive expression of the constructs in larval epidermal cells to reveal their subcellular distribution. Anti-GFP immunostaining shows, in both cases, diffuse cytoplasmic signal with scattered aggregates. No expression is detected in the nucleus. (C) A single copy of wild-type Drosophila or human spastin driven by GS-elav-GAL4 in the spastin null background (genotypes D^WT,Ø and H^WT,Ø, respectively) induced pan-neuronal spastin transgene expression. Anti-GFP reveals cytoplasmic expression of both proteins in neuronal cell bodies as well as their processes, including the long photoreceptor axons (arrows).