Minibrain/Dyrk1a Regulates Food Intake through the Sir2-FOXO-sNPF/NPY Pathway in Drosophila and Mammals
Figure 2
sNPF/NPY-sNPFR1/NPYR1-PKA-CREB-mnb/Dyrk1a signaling in Drosophila neuronal BG2-c6 cells and mouse hypothalamic GT1-7 cells.
(A) mnb mRNA in Drosophila neuronal BG2-c6 cells increased in response to treatment with sNPF peptide, but not when co-treated with H89 PKA inhibitor. The ERK inhibitor PD98059 and PKC inhibitor CC did not suppress sNPF-induced mnb expression. (B) Dyrk1a mRNA in mouse hypothalamic GT1-7 cells increased in response to treatment with NPY peptide, but not when co-treated with the PKA inhibitor H89. (C) In Drosophila BG2-c6 cells, sNPF peptide induced cAMP, while transfection of cells with Gαs siRNA but not Gαi siRNA repressed this effect. (D) In mouse GT1-7 cells, NPY peptide induced cAMP, while co-treatment with NPYR1 inhibitor BIBO3304 but not NPYR2 and NPYR5 inhibitors strongly decreased this effect. (E) Western blot to detect activated CREB (pCREB) in Drosophila BG2-c6 cells. sNPF peptide increasd pCREB but not in cells transfected with Gαs siRNA. (F) Western blot to detect activated CREB (pCREB) in mouse GT1-7 cells. NPY peptide increased pCREB but not when cells are co-treated with PKA inhibitor H89 or NPYR1 inhibitor BIBO3304. (G) In Drosophila BG2-c6 cells, sNPF peptide induced mnb mRNA, while transfection of cells with Gαs siRNA but not Gαi siRNA repressed this effect. (H) In mouse GT1-7 cells, NPY peptide induced Dyrk1a mRNA, while co-treatment with NPYR1 inhibitor BIBO3304 but not NPYR2 and NPYR5 inhibitors strongly decreased this effect. Data are presented as means ± s.e.m. from three independent experiments. *P<0.05 (One-way ANOVA analysis).