ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
Biotechnologia Acta Т. 17, No. 2 , 2024
P. 27-28, Bibliography 8, Engl.
UDC:57.042:547.485+57.052:577.214:595.77+577.213.3
DOI: https://doi.org/10.15407/biotech17.02.027
ALPHA-KETOGLUTARATE INDUCES NUCLEAR RECEPTORS RATHER THAN NRF2 IN THE FRUIT FLY Drosophila melanogaster
O.I. Demianchuk, D.V. Gospodaryov
Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine
Aim. To test whether expression of Nrf2 targets in the fruit fly Drosophila melanogaster is activated by diet supplemented with alpha-ketoglutarate (AKG).
Methods. The Canton-S strain of D. melanogaster was used in the study. Female flies were reared in demographic cages (150 flies per group) on a medium containing 5% sucrose, 5% yeast, 1.2% agar, and 0.18% nipagin. The experimental diet was supplemented with 10 mM disodium salt of AKG. The flies were reared during 21 days and after that were anesthetized with carbon dioxide and snap-frozen in liquid nitrogen for further biochemical studies. Expression of genes Ugt37A2, GstD2, and Cyp6a2, coding for a uridine diphosphate glycosyltransferase family 37 member A2, glutathione S-transferase D2, and cytochrome P450 6a2, respectively, was analyzed using semi-quantitative reverse transcription polymerase chain reaction followed by visualization of the products in agarose gel. Tbp (TATA-box binding protein) was used as a reference gene.
Results. Flies fed an AKG-supplemented diet for 21 days had a 2.8-fold higher level of Cyp6a2 expression than control flies. At the same time, AKG-supplemented food did not affect the expression of Ugt37A2 and GstD2 genes.
Conclusions. Continuous consumption of AKG-supplemented food results in an increase in the levels of messenger ribonucleic acid of the Cyp6a2 gene, a target of transcriptional factors Nrf2 and DHR96, but not Ugt37A2 and GstD2 genes. Since the expression of the latter two genes was unaffected by an AKG-supplemented diet, it indicates that AKG may influence other transcriptional regulators, such as nuclear receptors that have common targets with Nrf2.
Key words: Drosophila melanogaster, alpha-ketoglutarate, mRNA, Cyp6a2, Nrf2.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2024