Conclusion
Obstacles and Limitations
Inconsistent Data Collection & Spring Break
One issue that impacted the ability to collect data is that during the week of March 11-15, no locomotor assays were run since students were not present due to the occurrence of Spring Break. The reason is unclear, but there were no assays run the week after Spring Break either. This leaves a two-week gap in all trials that occurred during that time period.
Small Sample Size
Over the course of life of every vial, there were various reasons that contributed to a decreasing sample size throughout the trial. Some reasons included flies escaping during the transfer of flies from the vials to the graduated cylinder, between vials, or during the transfer from the graduated cylinder to a new vial. Another contributor is that some flies died throughout the course of the trial - some died from getting stuck in the food in the vials, and some died from unknown reasons. This led to some vials only containing 3 or 4 flies in the later weeks of the trial, compared to the 10-15 in the first few weeks.
Lack of Movement
There were some days when the flies simply did not move at first, even though it was earlier in their life. There was one day when, during the actual recorded activity, the flies did not move very much at all, and no flies crossed the 190mL line. However, after the tube was placed to the side, while running another test, there was more fly activity observed where a majority of the flies ended up reaching the top.
Discussion
We didn't see any significant difference between the two groups in CASK or Shrb. We did, however see a longer week length of higher locomotion in GR-80 expressing flies, compared to the GR-control in Vha100.
What we really need to do is compare more types of controls-- like just UAS-RNAi crossed to a vglutGal4, and also we need a lot more data for "typical ALS" expression with just the UAS-GR-80 with no RNAi manipulation. It would be better to compare our data to the UAS-GR80 x vglutGal4 combo rather than comparing each RNAi manipulation between GR-control to the same RNAi of GR-80 expression.
"Weird things are happening"
Another interesting thing to note, around week 6 or 7 there's a spike in activity for all three RNAi lines in GR-control. Whether this was just some crazy coincidence or something to do with how the RNAi is affecting the flies at a later stage and the effect is only able to be measured in GR-control, is yet to be determined.
For the Future
We definitely need to redo some of our data, most notably--next time make sure that we don't let Spring Break mess up our data. Spring Break was in the middle of some of the most important weeks in development for our flies. Additionally, use a larger sample size for each. And finally, control for temperature and humidity during locomotor assays.
It might be worthwhile to retrial the GR-controls for longer to see if the weird spike in activity happens past week 6 and 7, or if that data is re-creatable.