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. 2004 Nov 23;2(12):e408. doi: 10.1371/journal.pbio.0020408

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Copyright: © 2004 Tsai et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) pdf-GAL4-driven expression of Lmo using the EP1306 element rescues the EP1306 insertional phenotype. Lmo mutants EP1306, hdp, and pdrm, as well as a control EP line (Ctl-1 = EP1413), were tested in the absence (−pdf-GAL4; white bars) and presence (+ pdf-GAL4; black bars) of pdf-GAL4. All male flies are hemizygous for the Lmo mutation (or Ctl-1) and heterozygous for pdf-GAL4 (when carrying the transgene). One-way ANOVAs with post hoc planned comparisons (critical p-value adjusted to 0.0125) confirmed that EP1306, pdrm, and hdp flies (in the absence of pdf-GAL4) had significantly increased sensitivity to cocaine compared to control flies (Ctl-1 = EP1413) (p ≤ 0.003, n = 15–26 experiments). One-way ANOVA with post hoc planned comparisons (critical p-value adjusted to 0.01) revealed a significant difference between pdrm/pdf-GAL4 and hdp/pdf-GAL4 flies and their controls (EP1413/pdf-GAL4, pdf-GAL4/+, or EP1413/+, p < 0.003, n = 24–27 experiments), showing that the presence of pdf-GAL4 does not rescue the cocaine sensitivity of pdrm or hdp flies. In contrast, similar comparisons for “rescued” EP1306/pdf-GAL4 flies revealed no significant differences from their “normal” controls (p ≥ 0.026, n = 27–36). Furthermore, within-group comparisons (+/− pdf-GAL4) using t-tests indicate a significant difference only in the EP1306 group (p = 0.002). Asterisk denotes significant difference between −pdf-GAL4 and + pdf-GAL4 phenotype.

(B) Flies overexpressing Lmo in PDF cells show decreased sensitivity to cocaine. Flies heterozygous for both pdf-GAL4 and either one of two UAS-Lmo transgenes (black bars) were compared to flies carrying UAS-Lmo (white bars) or pdf-GAL4 (gray bar) alone. One-way ANOVA revealed a significant effect of genotype for both UAS-Lmo transgene groups. Post-test planned comparisons, with the critical p-value adjusted to 0.025, showed significant differences between the pdf-GAL4/UAS-Lmo flies and either pdf-GAL4/+ (p < 0.02) or UAS-Lmo/+ controls (p < 0.005). Asterisks denote significant differences, n = 16 experiments.

(C) Confocal images demonstrate overlap between pdrm and PDF expression in the LN_vs. In the left panel, UAS-mCD8GFP reveals the pdrm-driven GAL4 expression pattern (green) in the adult brain, and α-PAP staining (magenta) reveals PDF-expressing LN_vs. Right panels are close-ups of the cell bodies of the LN_vs; white areas correspond to regions of overlap between GFP (green) and PAP (magenta) expression.