Format

Send to

Choose Destination
Insect Biochem Mol Biol. 2015 Jun;61:79-86. doi: 10.1016/j.ibmb.2015.01.011. Epub 2015 Jan 30.

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

Author information

1
Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05 Ceské Budejovice, Czech Republic.
2
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.
3
National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan.
4
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.
5
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.

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.

KEYWORDS:

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

PMID:
25641265
DOI:
10.1016/j.ibmb.2015.01.011
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center