Tested Flies with the Joystick

Reinforcement

FoxP3955 flies were raised and compared to normal WTB flies. Reportedly, the Foxp mutants have a reduced flight performance as their total flight duration is decreased. This was also something I experienced. The problem seemed to be greater due to the heat in the flight simulator room, initial temperature was 27°C but increased to close to 30°C. I had troubles getting a large sample size enough (same number of Foxp and wtb were loaded into the flight simulator), heat-shock proteins and other stress-related behavior might be an issue. The genotype of the flies were known during the hooking of the flies but was later on concelead and flies were randomly distributed.  

Positive Control: Gr28bd-G4, TrpA1-G4

Parameters: Light: intensity (500 Lux side, 1000 Lux bottom); frequency = 20Hz; Delay = 1 ms; Duration = 9.9 ms; volts = 6.4

Red lines: completed

mb025b: not selected against tubby

Mean trace of the positive control in the Joystick to get to see what are the overall dynamics and maybe to get an idea what might be the best score to pick.Here the standard deviation of the flies along the time axis. This is just to see if all the flies have more similar phenotypes with each other or not at each time.

This is to see if the flies have less degrees of freedom at any segment by measuring the standard deviation at each segment. There does not seem to be any effect. Although this might be mixed with the wiggle scores. I think measuring entropy is a better measure.

All the same plots as above but for TH-D’, the interesting line from the screen.

Standard deviation across flies

Standard deviation across segments

This are the performance indices for the different models performed to estimate the valence of the dopaminergic clusters. AIC: Akaike Information Criteria; BIC: Bayesian information Criteria; LogLikelihood: log Likelihood estimation

lm: linear model

+ int: taking double interaccions into consideration

b lm: bayesian linear model with bayesglm function

b lm MCMC: bayesian linear model with MCMCglm function

nlm: nonlinear model with lm function with splines fitted

b nlm: splines fitted to each cluster and MCMCglm function

GAM: general additive model with gam function

Adding double interactions seems to produce better models, nonlinearities also make models better and frequentist also. To me it seems like this  data might be noise and therefore adding interactions, nonlinearities and frequentist methods is just fitting the noise better (overfitting) and that is why I get better scores with them. In addition, care needs to be taken since I use different functions that calculate the model performance scores differently (although the formulas are theoretically the same for all!)

This is the correlation from the T-maze experiments from Gaia and Naman. Neither ranked nor regular correlation show any significant effect. This means that these effects seem to be random, at least for most of them, is this an overfitting result?

I would say blue 1 is a line that was negative for all the tests I have so far seen. So this might be an interesting line. What to do next?

I would unblind the blue1, which is TH-D’. It was shown to be required for classical conditionning in shock and temperature learning (Galili et al. 2014). Another interesting observation is that th-g4+th-g80 seems to have like zero PI scores in all of the experiments (Naman and Gaia in the Tmaze, Joystick and Y-mazes). So could it be that all of these neurons have indeed a meaning, but is depending every time in the context?? Maybe Vanessa Ruta´s work might be interesting for that.

Results of the T-maze screen analysis, both individual and combined.

Yellow 1 (Positive Control): Gr28bd-G4, TrpA1-G4

Parameters: Light: intensity (500 Lux side, 1000 Lux bottom); frequency = 20Hz; Delay = 1 ms; Duration = 9.9 ms; volts = 6.4

The mean ratio of the flies that stay in the middle during the experiments.

Correlation plot between the mean ratio of the flies that stay in the middle versus the Weighted PIs

Slope = 0.0053
Intercept = 0.240
R square value = -0.03834

contrary to the expectations, there seems to be no correlation .

Yellow 1 (Positive Control): Gr28bd-G4, TrpA1-G4

Parameters:
Light: intensity (500 Lux side, 1000 Lux bottom)

frequency = 20Hz

Delay = 1 ms

Duration = 9.9 ms

volts = 6.4

Dopaminergic clusters are differently targeted by the different Gal4s. Some of the express faintly, others stronger. Here I try to see if the dose-response curve (or expression-PI curve) seems to be linear or not. Here I put two examples from the 17 clusters, where the first two seem to have nonlinear curves, with and optimal expression level, and the last two seem to have a linear response curve.

This will be important for the modelling in order to decide to make a linear/nonlinear model. Down below I show the results from a linear model and it´s statistics. From Aso et al. 2012, one could see that activating the lines with TrpA1 shows a linear response curve. But in this case it does not necessarily seem to be the case. Therefore, light intensities might have an effect, as well as the expression level, and conclusion needs to be taken carefully.

In addition it is difficult to calculate this for all the clusters with just one single light intensity test, because not all clusters are expressed in several Gal4s to different level, so that we can estimate from there. So for the interesting lines we might need to make several experiments at different intensities, and see the dose response curve.

The G4s I have used for the modelling are the ones shown here.