Local FoxP knock out
Set-Up test
New review of the shiming-setup
Recent wtb test
There often seems to be a strong preference for a side. The laser was off.
Flies often seem to have a strong preferenc for a side. In the imiage below the top fly was punished for left torque and survived, the fly below was punished for the right torque side and died.
Trails with drift
Testing of the repaired torque-meter
I’ve started again to test the set-up after repairs. It does not seem to be perfectly stable ,with some drift in the positive direction. Most often, it seems that the amount of drifting would be fine for measuring.
Therefor I tried measuring some flies. A problem now is a very negative baseline, when the fly stops flying. Also the response for left and right seems not to be equal.
So now I’m trying to get the line close to 0, so that I have conclusive results if the set-up if usable at the current state.
2 examples of drift:
Gene switch finally worked
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.
Testing of possible Gal4-lines
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.
FoxP knock out in adult.
The repetition of the knockout experiment in adult flies indicates that FoxP is not required for learning in adult flies. (Exp = elav tubGal80>Cas9gFoxP)
Test for hsGal4
Comparison between Control (elavGal4 x CD8GFP) and hsGla4 x CD8GFP. Hs for 3h, fixation and dissection after 24h.
Ato x Cas9FoxP
Knockout of FoxP in Ato.
