Targeted mutagenesis and functional analysis of adipokinetic hormone-encoding gene in Drosophila

Suresh Sajwan, Roman Sidorov, Tereza Stašková, Anna Žaloudíková, Yoko Takasu, Dalibor Kodrík, Michal Zurovec

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

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 Akh1 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 Akh1 mutants are slightly heavy and had a slow metabolic rate. Furthermore, we showed that the ectopic expression of Dmel\Akh reverses the Akh1 phenotype and restores the wild-type characteristics. Our results confirmed that Akh is an important regulator of metabolic homeostasis in Drosophila.

Original languageEnglish (US)
Pages (from-to)79-86
Number of pages8
JournalInsect Biochemistry and Molecular Biology
Volume61
DOIs
StatePublished - Jun 1 2015
Externally publishedYes

Keywords

  • Carbohydrate metabolism
  • Drome-Akh
  • Neuropeptide
  • NHEJ
  • Stress
  • Trehalose

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

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