The 1st cross produced eggs, but they did not hatch or become pupae. In all of the 10 vials, eggs were observed, but no pupae. On the other hand, the 2nd cross did not produce any eggs. (The first trial was kept up for 10 days. After 10 days, a new trial began from the same stock. ) Control and the other two crosses produced eggs and hatched new flies.

The same crosses were repeated to ensure the lethality. In the second trial, ‘T4/T5xWTB’ cross produced eggs and pupae. But there is still the possibility that the T4/T5xWTB is also a lethal crossing. ‘T4/T5xTNTE’ cross again produced only eggs but no pupae. The other two crosses produced pupae again and became dark, but haven’t hatched yet. The second attempt is 10 days old.

After the 10th day, cross 2 and the control group hatched new flies. On the 13th day, all of the groups have hatched flies. None of the crosses seems to be lethal.

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: