Expression and distribution of <i>Drosophila mnb</i> in adults in relation to <i>sNPF</i>, <i>sNPFR1</i>, and feeding.

<p>(A) <i>mnb</i> mRNA prepared from fly heads was measured by RT-qPCR. <i>mnb</i> mRNA was increased relative to <i>sNPF-Gal4</i> and <i>sNPFR1-Gal4</i> controls when <i>sNPF</i> and <i>sNPFR1</i> was overexpressed in sNPFnergic neurons (<i>sNPF>sNPF, sNPF>2XsNPF</i>) and in sNPFR1 neurons (<i>sNPFR1>sNPFR1</i>). <i>mnb</i> mRNA was decreased when <i>sNPF</i> and <i>sNPFR1</i> were inhibited (<i>sNPF>sNPF-Ri, sNPF^c00448, sNPFR1>sNPFR1-Ri, sNPFR1>sNPFR1-DN</i>). (B) Food consumption measured by the colormetric assay. Relative to <i>sNPFR1-Gal4</i> control, <i>mnb</i> overexpression in sNPFR1 neurons (<i>sNPFR1>mnb, sNPFR1>2Xmnb</i>) increased feeding whereas <i>mnb</i> suppression (<i>sNPFR1>mnb-Ri, mnb^G1767</i>) decreased feeding. Overexpression or inhibition of <i>mnb</i> in the insulin producing cells with the <i>Dilp2-Gal4</i> driver (<i>Dilp2>mnb, Dilp2>2Xmnb, Dilp2>mnb-Ri</i>) did not change the feeding. (C) Food consumption measured by CAFÉ assay. Relative to the <i>sNPFR1-Gal4</i> (open triangle) control, <i>sNPFR1>mnb</i> (closed triangle) increased while <i>sNPFR1>mnb-Ri</i> (closed circle) decreased cumulative food consumption. Data are presented as means ± s.e.m. from three independent experiments. <i>*P</i><0.05, <i>**P</i><0.001 (One-way ANOVA analysis). (D-G) Neurons of the <i>Drosophila</i> adult brain expressing Mnb protein (green) do not overlap with insulin producing cells marked with <i>Dilp2>DsRed</i> (red). (H-M) Mnb protein expression neurons (H, K, red) and sNPFR1 protein expression neurons (I, L, green) were overlapped in the median neurons (J, dot box; M, arrows). Scale bars are 100 µm (D, H) and 50 µm (G, K).