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. 2016 Mar 17;11(3):e0151279. doi: 10.1371/journal.pone.0151279

Fig 3. Lqf/epsin in PNs is required for the odor induced increase in glomeruli volume, while Ach release from ORNs and nAchR activation in PNs are required to restrict the extent of the increase.

Fig 3

(A) Schematics of experimental protocols. (B) Cartoons of the olfactory circuit depicting the relevant genotypes. (C and D) Scatterplots of VA6 volumes. The data were normalized as described in Fig 2. Magenta circles, controls; blue circles, Dα7-DN; brown circles, tetanus toxin; green circles, lqf RNAi; pale blue circles, both lqf and Dl RNAi. Empty circles, air exposed; filled circles, GA exposed. Black p values compare air and GA exposed samples of the same genotype. Red p values compare GA exposed samples between genotypes. In C, lanes 1–4, one day old female flies were exposed to 1% GA in paraffin oil or paraffin oil alone and the volumes of VA6 determined. Flies, as illustrated in B(i), were MZ612-GAL4; Or82a-LexAGAD LexOP.dsRED carrying either UAS.Val20 (control) or UAS.Dα7-DN (Dα7-DN). In C, lanes 5–8, flies which had been grown and then maintained at 17°C for eight days were exposed to air or GA at 27.5°C. Flies, as illustrated in B(ii), were Or82a-GAL4/tubP-gal80ts20; UAS.mCD8-GFP with UAS.IMP (inactive tetanus toxin, control) or UAS.TNT (active tetanus toxin, TNT). The temporal protocol was chosen, because we found empirically that younger flies were more susceptible to axonal damage induced by the expression of tetanus toxin. The lower temperature used to inactivate the GAL80ts reduced the amount of tetanus toxin expressed and and reduced the amount of axonal damage. In D, flies that had been grown and then maintained at 17°C for 2 days, were shifted to 30°C for 4 days prior to being exposed to 1% GA or air for a further 4 days. Flies, as illustrated in B(iii), were MZ612-GAL4/tubP-gal80ts10; Or82a-LexAGAD LexOP.dsRED/UAS.dicer-2 with both UAS.epsin RNAi and UAS.Dl shRNA (lqf & Dl RNAi, lanes 7 and 8), just UAS.epsin RNAi (lqf RNAi, lanes 3–6), or neither (control, lanes 1 and 2). (E) Diagram of the non-canonical and canonical N pathways together with the results described in this figure and their relationship to odor induced glomeruli volume change.