Now that I’ve reached a sample size of near 30 animals in each group, it was time to break the code to see which group was wich. To my surprise, the worst performing group in term,s of learning was the control group:

It is unusual for between-groups comparisons to become statistically significant at the 0.005 level with a sample size of less than 50, let alone 30, but these DopR2 knock-downs in ellipsoid body ring neurons are just a class of their own:

The DopR2 receptor knock-down not only performed exceedingly well in these leaning experiments, it was also the cross with the strongest optomotor response:

And also that difference is statistically significant:

The TNT-E flies had a slight positive preference, worse avoidance (esp. in the first training period) and flew really bad. One could say they did have some learning deficits, but it doesn’t look as if learning was completely abolished (which is roughly what Andreas had found before):

Andi’s results:

Unfortunately I am still dealing with the problem that flies from my negative control group avoided optogenetic activation, even when their optogenetic channel should not work without ATR supplementation. To tackle this problem I used a “new” effector line (I prepared a new stock from our stock collection) for crossings and tested the offspring, without any improvement concerning the avoidance. Unfortunately the “new” effector line I tested turned out to have lost its “NorpA” mutation which would ensure that the male offspring is blind, thus should rule out any phototaxis-bias. Since flies still avoided the light, a) the effector line does not seem to be the problem and b) the ability to see does not seem to affect the flies behavior in the JoyStick setting.
As a next step I targeted the driver line, as maybe a mutation might have lead to an increase in Gal4. Since we always observed a slightly negative values for our negative control group, hinting that at least some residual activation of the CsChrimson channel is possible even without ATR, an increase in Gal4 might lead to a higher expression of the optogenetic channel.

Residual activation + More channels = More residual activation = Our observed avoidance???

So positive control looks good, negative control is still slightly negative but to an extend that I would consider neglectable.
The additional group here called “OA” are OA;VuMA2 x NorpA, 20xUAS-Chrimson flies. It’s way too early to make any assumptions from the current data but I am looking forward to the results for this group.

Getting close to a sample size of 30! One, maybe two weeks left, depending on how much time I find to measure. Time to break the code is drawing nearer. Here the current results:

Some interesting statistics:

2.1 OMRs at start

2.2 OMRs at end

2.3 PI( preference subtractted)

2.4 statistics

1.1 OMRs at start

1.2 OMRs at end

1.3 Asymmetry Index at start

1.4 Asymmetry Index at end

1.5 PI (preference subtracted)

1.6 Statistics

2.