Fig 4.

The prt¹ mutation. A) The relative position of the prt gene is shown in cytological region 95A8 on the 3^rd chromosome. The shaded box at the top of the figure represents an approximately 850 bp deletion created with imprecise excision of a P-element (“P” in the inverted triangle), including the initiating methionine ( ). The numbers 4 and 2 indicate primers used to detect the deletion. The lines below show deficiencies that uncover prt (Df(3R)mbc-30, Df(3R)Exel6195). The shaded boxes represent the deletion. The cross-hatched boxes indicate breakpoints approximated by polytene chromosome squash. Scale bars: 50 kb for deficiencies and 0.2 kb for inset. B) PCR was used to detect the prt¹ mutation. Wild-type CS flies show a major band at 1.2 kb, prt¹ flies show a major band at 400 bp, and prt¹/+ heterozygotes show both. (−) indicates a no-template control. A 1 kb standard is shown on the left. C) Single confocal slices through the mushroom body lobes of whole-mount brains labeled with anti-PRT. A w¹¹¹⁸ brain is on top with labeling of the MB lobes (α, α′, β, β′, and γ) and heel (h) and a prt¹ brain is on bottom, showing no detectable anti-PRT labeling. Scale bar: 20 μm. D) H&E stained paraffin sections show that prt¹ mutants have grossly intact MB morphology (α, α′, β, and γ lobes and heel (h), peduncle (ped), and calyx (ca)) and central complex (fan-shaped body (fsb), noduli (nod) and protocerebral bridge (pb)). The β′ lobe is also grossly intact in prt¹ (not shown). Scale bar: 20 μm. E, F) Volumetric analyses of the MB calyx (E) and CCX (F) in CS and prt¹ did not significantly differ (Student’s t-test, p = 0.1934 (E) and p = 0.2496 (F)). Bars represent means ± SEMs, n = 10 for all groups.