title: Targeted mutagenesis and functional analysis of adipokinetic hormone-encoding gene in Drosophila.
abstract: Adipokinetic hormones (Akhs) are small peptides (8-10 amino acid [aa] residues long) found in insects that regulate metabolic responses to stress by stimulating catabolic reactions and mobilizing energy stores. We employed Transcription activator-like effector nuclease (TALEN) mutagenesis and isolated an Akh(1) mutant carrying a small deletion in the gene that resulted in a truncated peptide; the second aa (Leu) was missing from the functional octapeptide. This null Dmel/Akh mutant is suitable to study Akh function without any effect on the C-terminal associated peptide encoded by the same gene. The mutant flies were fully viable and compared to the control flies, had significantly low levels of hemolymph saccharides including trehalose and were resistant to starvation. These characteristics are similar to those obtained from the flies carrying targeted ablation of Akh-expressing neurons (reported earlier). We also found that the Akh(1) mutants are slightly heavy and had a slow metabolic rate. Furthermore, we showed that the ectopic expression of Dmel∖Akh reverses the Akh(1) phenotype and restores the wild-type characteristics. Our results confirmed that Akh is an important regulator of metabolic homeostasis in Drosophila.
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic.
authors: Suresh
authors: S
authors: Sajwan
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic.
authors: Roman
authors: R
authors: Sidorov
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic; Faculty of Science, University of South Bohemia, Branisovska 31, 370 05 Ceske Budejovice, Czech Republic.
authors: Tereza
authors: T
authors: Stašková
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic.
authors: Anna
authors: A
authors: Žaloudíková
authors: National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan.
authors: Yoko
authors: Y
authors: Takasu
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic; Faculty of Science, University of South Bohemia, Branisovska 31, 370 05 Ceske Budejovice, Czech Republic. Electronic address: kodrik@entu.cas.cz.
authors: Dalibor
authors: D
authors: Kodrík
authors: Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic; Faculty of Science, University of South Bohemia, Branisovska 31, 370 05 Ceske Budejovice, Czech Republic. Electronic address: zurovec@entu.cas.cz.
authors: Michal
authors: M
authors: Zurovec
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001835:Body Weight; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D004734:Energy Metabolism; D005508:Food Deprivation; D006458:Hemolymph / Q000737:chemistry; D006706:Homeostasis; D007301:Insect Hormones / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009842:Oligopeptides / Q000235:genetics* / Q000378:metabolism; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D014199:Trehalose / Q000378:metabolism
chemical_list: D007301:Insect Hormones; D009842:Oligopeptides; C044833:adipokinetic hormone; D014199:Trehalose; D011761:Pyrrolidonecarboxylic Acid
keywords: Carbohydrate metabolism; Drome-Akh; NHEJ; Neuropeptide; Stress; Trehalose