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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) pdrm's GAL4 expression pattern. UAS-GFP reveals GAL4 expression pattern of the pdrm enhancer-trap insertion in MB lobes and calyces, ALs, the large cell bodies of the peptidergic LN_vs, and neurons of the pars intercerebralis (PI).

(B) Lmo expression restores wild-type cocaine responses. In the absence of a UAS transgene (“no UAS” columns), hemizygous male pdrm flies (hatched bar) are more sensitive than controls (Ctl GAL4 is line 8.142, solid bar), as shown before in Figure 1A. Male flies hemizygous for pdrm and heterozygous for either of two UAS-Lmo transgenes (UAS-Lmo¹ and UAS-Lmo²; hatched black bars), show normal cocaine sensitivity when compared to either UAS-Lmo transgene alone (white bars) or UAS-Lmo transgenes in the presence of a control GAL4 line (Ctl GAL4, line 8.142; black bars). To control for non-specific effects of transgene overexpression, UAS-GFP and UAS-lacZ transgenes were also driven by pdrm GAL4. Male flies were hemizygous for pdrm (or heterozygous for the control GAL4 insertion) and heterozygous for the specific UAS transgene. One-way ANOVA revealed a significant effect of genotype in the UAS-lacZ (F = 17.4, p < 0.001), UAS-GFP (F = 19.47, p < 0.001), or no UAS transgene (F = 4.1, p < 0.001) groups, but not in either of the UAS-Lmo transgene groups (F = 1.58, p = 0.22 and F = 1.21, p = 0.31 for UAS-Lmo¹ and UAS-LMO², respectively); thus, UAS-Lmo expression specifically restores normal cocaine sensitivity to pdrm flies. Post hoc pairwise planned comparisons, with the critical p-value adjusted to 0.025, revealed significant differences between the “non-rescued” pdrm/UAS-GFP flies and the appropriate controls (UAS-GFP/+ or 8.142/UAS-GFP, p < 0.002); similarly, pdrm/UAS-lacZ flies are significantly different from their controls (UAS-lacZ/+ and 8.142/UAS-lacZ, p < 0.002). Pairwise comparisons revealed no significant differences between “rescued” pdrm/UAS-Lmo¹ flies and their “normal” controls (8.142/UAS-Lmo¹ and UAS-Lmo¹/+, p = 0.99 and p = 0.14, respectively) or pdrm/UAS-Lmo² flies and their controls (8.142/UAS-Lmo²/+ and UAS-Lmo²/+, p = 0.09 and p = 0.99, respectively), indicating full rescue of pdrm cocaine sensitivity. For all genotypes, n = 16–20 experiments.