on Monday, June 5th, 2023 12:59 | by Ellie
Things are not going well…
Over the last week I have encountered numerous problems in carrying out the planned experiment, and the biggest of these is definitely the generation of offspring. Since it has been shown that long-term memory decays quickly in the first 24 hours after operant self-learning on the flight simulator (and with that possible epigenetic marker I suppose), I decided to mate the flies immediately after training and separate them again after 16-20 hours. However, I did not get any offspring by doing this. I tried mating virgin males/virgin females and also adult males/virgin females for 16-20 h, but no eggs could be found in the vials. I have not yet found out what prevents the flies from mating and am now trying again with older (3-4 days old) adult male flies.
Another problem is the training protocol for memory consolidation by S Xia, L Liu, C Feng et al. 1997. Although I have practiced gluing and inserting flies into the flight simulator, I don’t think I can train enough flies in one day. To maintain sufficient larval density after mating, I need at least 5 females and 2-3 males, and I would need to train them twice, giving a total of 16-18 training rounds per day (and that doesn’t include failed attempts…). Alternatively, I can use the standard protocol and see if the training is enough to reveal epigenetic inheritance of acquired habits, if any.
For now, I will continue to try to produce offspring and practice gluing and training the flies. I have started to do further research on inheritance mechanisms to see if it might also be possible to work with only the F0 generation and detect epigenetic markers (methylation of genes, e.g. foxP, siRNAs) in the trained flies and compare them with untrained flies. Here (1, 2) you can find a few interesting articles on this topic.
However there is also some good news to report: my ContonS line will be ready soon, so I don’t have to work with the wtb virgin flies anymore, which decease very quickly. I have also made some progress on the statistical background after talking to a friend of mine although I still need to do some research on how to set my effect size so I can finally calculate the sample size I need for the experiment.