title: CCHamide-2 Is an Orexigenic Brain-Gut Peptide in Drosophila.
abstract: The neuroendocrine peptides CCHamide-1 and -2, encoded by the genes ccha1 and -2, are produced by endocrine cells in the midgut and by neurons in the brain of Drosophila melanogaster. Here, we used the CRISPR/Cas9 technique to disrupt the ccha1 and -2 genes and identify mutant phenotypes with a focus on ccha-2 mutants. We found that both larval and adult ccha2 mutants showed a significantly reduced food intake as measured in adult flies by the Capillary Feeding (CAFE) assay (up to 72% reduced food intake compared to wild-type). Locomotion tests in adult flies showed that ccha2 mutants had a significantly reduced locomotor activity especially around 8 a.m. and 8 p.m., where adult Drosophila normally feeds (up to 70% reduced locomotor activity compared to wild-type). Reduced larval feeding is normally coupled to a delayed larval development, a process that is mediated by insulin. Accordingly, we found that the ccha2 mutants had a remarkably delayed development, showing pupariation 70 hours after the pupariation time point of the wild-type. In contrast, the ccha-1 mutants were not developmentally delayed. We also found that the ccha2 mutants had up to 80% reduced mRNA concentrations coding for the Drosophila insulin-like-peptides-2 and -3, while these concentrations were unchanged for the ccha1 mutants. From these experiments we conclude that CCHamide-2 is an orexigenic peptide and an important factor for controlling developmental timing in Drosophila.
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Guilin R
authors.initials: GR
authors.lastname: Ren
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Frank
authors.initials: F
authors.lastname: Hauser
authors.affiliation: Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Kim F
authors.initials: KF
authors.lastname: Rewitz
authors.affiliation: Genetic Strains Research Center, National Institute of Genetics, Mishima, Shizuoka, Japan.
authors.forename: Shu
authors.initials: S
authors.lastname: Kondo
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Engelbrecht
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Anders K
authors.initials: AK
authors.lastname: Didriksen
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Suzanne R
authors.initials: SR
authors.lastname: Schjøtt
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Frederikke E
authors.initials: FE
authors.lastname: Sembach
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Shizhong
authors.initials: S
authors.lastname: Li
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Karen C
authors.initials: KC
authors.lastname: Søgaard
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Leif
authors.initials: L
authors.lastname: Søndergaard
authors.affiliation: Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, DK-Copenhagen, Denmark; Section of Cell and Neurobiology, Department of Biology, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Cornelis J P
authors.initials: CJ
authors.lastname: Grimmelikhuijzen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D004247:DNA; D004330:Drosophila / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D005247:Feeding Behavior; D007413:Intestinal Mucosa / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008124:Locomotion; D008969:Molecular Sequence Data; D009154:Mutation; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism*
chemical_list: C555823:CCH-amide-2 neuropeptide, Drosophila; D029721:Drosophila Proteins; D009479:Neuropeptides; D004247:DNA