Abstract
In order to investigate the influence of light/dark cycle on the biosynthesis of metabolites during oogenesis, here we demonstrate a simple experimental protocol which combines in-vivo isotopic labeling of primary metabolites with mass spectrometric analysis of single eggs of fruit fly (Drosophila melanogaster). First, fruit flies were adapted to light/dark cycle using artificial white light. Second, female flies were incubated with an isotopically labeled sugar (13C6-glucose) for 12 h – either during the circadian day or the circadian night, at light or at dark. Third, eggs were obtained from the incubated female flies, and analyzed individually by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS): this yielded information about the extent of labeling with carbon-13. Since the incorporation of carbon-13 to uridine diphosphate glucose (UDP-glucose) in fruit fly eggs is very fast, the labeling of this metabolite was used as an indicator of the biosynthesis of metabolites flies/eggs during 12-h periods, which correspond to circadian day or circadian night. The results reveal that once the flies adapted to the 12-h-light/12-h-dark cycle, the incorporation of carbon-13 to UDP-glucose present in fruit fly eggs was not markedly altered by an acute perturbation to this cycle. This effect may be due to a relationship between biosynthesis of primary metabolites in developing eggs and an alteration to the intake of the labeled substrate – possibly related to the change of the feeding habit. Overall, the study shows the possibility of using MALDI-MS in conjunction with isotopic labeling of small metazoans to unravel the influence of environmental cues on primary metabolism.
Highlights
Circadian clock helps biological organisms to control their physiological and developmental processes [1]
Fruit fly has served as a model organism in several studies of circadian rhythms [8,9,10], limited data is currently available on the influence of adaptation of fruit fly to light/dark cycle on primary metabolism in this species
The study has shown the feasibility of isotopic labeling of fruit flies with the purpose of pursuing metabolic effects of environmental cues by mass spectrometry
Summary
Circadian clock helps biological organisms to control their physiological and developmental processes [1]. Studying relationships between circadian rhythms and metabolism may contribute to a better understanding of the mechanism and robustness of the biological clock. Small metazoa are convenient models for studying circadian rhythms; one of them is fruit fly (Drosophila melanogaster) [5]. Fruit fly has served as a model organism in several studies of circadian rhythms [8,9,10], limited data is currently available on the influence of adaptation of fruit fly to light/dark cycle on primary metabolism in this species. Feeding and metabolic activities of fruit fly are affected by the existence of circadian rhythms [11]. The small volumes of samples obtained from individual flies disable the possibility of analyzing metabolites using conventional analytical tools, and following the effects of light/dark cycle on biosynthesis of metabolites in single fly organs
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
