A highly conservative insulin signaling pathway, stable work of which is indicated by carbohydrates metabolism, is also known to play an important role in the control of stress resistance. Here we demonstrate that exposure to heat stress leads to a rise in the levels of trehalose and glucose in females of Drosophila melanogaster, but does not affect the expression level of the trehalase (Treh) gene. We have shown that the rise in juvenile hormone (JH) and dopamine decreases levels of both carbohydrates under the normal conditions but brings them to values close to normal following the stress exposure. The data obtained suggest that (a) dopamine and JH involved in the neuroendocrine stress reaction in D. melanogaster also take part in the regulation of carbohydrates metabolism, tending to normalize it after stress; (b) the regulation of trehalose content under stress does not occur at the level of transcription of the degrading enzyme.
The insulin/insulin-like growth factor signaling (IIS) pathway is one of the key elements in an organism’s response to unfavourable conditions. The deep homology of this pathway and its evolutionary conservative role in controlling the carbohydrate and lipid metabolism make it possible to use Drosophila melanogaster for studying its functioning. To identify the properties of interaction of two key IIS pathway components under heat stress in D. melanogaster (the forkhead box O transcription factor (dFOXO) and insulin-like peptide 6 (DILP6), which intermediates the dFOXO signal sent from the fat body to the insulin-producing cells of the brain where DILPs1–5 are synthesized), we analysed the expression of the genes dilp6, dfoxo and insulin-like receptor gene (dInR) in females of strains carrying the hypomorphic mutation dilp641 and hypofunctional mutation foxoBG01018. We found that neither mutation influenced dfoxo expression and its uprise under short-term heat stress, but both of them disrupted the stress response of the dilp6 and dInR genes. To reveal the role of identified disruptions in metabolism control and feeding behaviour, we analysed the effect of the dilp641 and foxoBG01018 mutations on total lipids content and capillary feeding intensity in imago under normal conditions and under short-term heat stress. Both mutations caused an increase in these parameters under normal conditions and prevented decrease in total lipids content following heat stress observed in the control strain. In mutants, feeding intensity was increased under normal conditions; and decreased following short-term heat stress in all studied strains for the first 24 h of observation, and in dilp641 strain, for 48 h. Thus, we may conclude that dFOXO takes part in regulating the IIS pathway response to heat stress as well as the changes in lipids content caused by heat stress, and this regulation is mediated by DILP6. At the same time, the feeding behaviour of imago might be controlled by dFOXO and DILP6 under normal conditions, but not under heat stress.
A method for automation of imago quantifying and fecundity assessment in Drosophila with the use of mobile devices running Android operating system is proposed. The traditional manual method of counting the progeny takes a long time and limits the opportunity of making large-scale experiments. Thus, the development of computerized methods that would allow us to automatically make a quantitative estimate of D. melanogaster fecundity is an urgent requirement. We offer a modification of the mobile application SeedCounter that analyzes images of objects placed on a standard sheet of paper for an automatic calculation of D. melanogaster offspring or quantification of adult flies in any other kind of experiment. The relative average error in estimates of the number of flies by mobile app is about 2% in comparison with the manual counting and the processing time is 6 times shorter. Study of the effects of imaging conditions on accuracy of flies counting showed that lighting conditions do not significantly affect this parameter, and higher accuracy can be achieved using high-resolution smartphone cameras (8 Mpx and more). These results indicate the high accuracy and efficiency of the method suggested.
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