I retried the gene-switch method with a new solution of RU486. Since it did not worked before I just tried one line. The flies were left for two days on instant food mixed with a sugar solution of RU486. This time I saw normal GFP expression. So I will test all 4 line in parallel after the break.

I finished my cross for the colocalisation of FoxP with the 6 Gal4-lines i ordered.

Two lines show a nice colocalisation and will be tested.

One would show some overlap, but a completly diffrent expression pattern than it should have. So i will try this line again from the stock to exclude a mixup of the line.

The next one dont seem to have any overlap, but also the pattern looks not like it should.

This line seems to have coexpression but the pattern also looks a bit odd, this may be due to a general weak signal and a bad dissection.

The final line seems to have no overlap with the FoxP expression.

Comparison between Control (elavGal4 x CD8GFP) and hsGla4 x CD8GFP. Hs for 3h, fixation and dissection after 24h.

Recently I measured the optomotor response in T4/T5 flies. As expected, they did not respond to the optomotor stimulus as seen in the left chart below. However, statistical evaluation struggles to quantify this difference. It might be that this is because of the low sample size, or that we are using the wrong statistical analysis?

1. ato-Gal4 x conditional KO —> Removing FoxP in Dorsal Cluster Neurons for Buridan
2. cmpy-Gal4 x conditional KO —> Removing FoxP in Protocerebral Bridge for Buridan
3. temperature switch at embryo stage but shorter time —> for replicating the KO phenotype
4. LexA + Gal4 together to analyze larval brain (I have always considered larvae except for this experiment, so I am doing it to be more complete)

Furthermore, for not loosing time this week I did:

1. Leipzig presentation
2. Wrote a bit of results for paper/thesis

1. Protocerebral bridge lines X CD8-GFP
2. C380-Gal4 X UAS-gFOXP;UAS-Cas9 (+ 2 controls)
3. stinger;Foxp-iB X FoxP-KO

Last week (1.-7.Apr 2019) we noted how many flies want to fly more than 8 min and here is the result (only the ones we noted):
– Wild type: 8/9 (88%)
– ElavGal4 x UAS-Ryr 28919: 3/12 (25%)
– ElavGal4 x UAS-Ryr 65885: 6/35 (17%)

Trace (Downsampled 5) Graphs of Recordings

Wild type:

ElavGal4 x UAS-Ryr 28919:

ElavGal4 x UAS-Ryr 65885:

New Findings:

Elav-Gal4 x UAS-SERCA 44581 flies => Lethal before adulthood! (in all 5 vials so far)

A flight performance test to see how the flies are behaving while flying. They have a slightly lower flight duration compared to the genetic controls but nothing that should impair them in the experiment.

The learning experiments are ongoing. I am doing n=15 for each line and each experiment, both Composite Control and also a Isolated Operant experiment. The Composite control experiment is done but I only got 13 flies for Isolated Operant. I do not want to preanalyze the data and will therefor post it when all the flies are done.