And the journey begins…

ElavGal4 x UASRyr65 “the wrong vials”
– > Graphs of the Trace.png (RightTrace – LeftTrace):

Data 1:

Monica:

Anokhi:

This week we worked on perfecting the fly hooking. Some of the data we got is as follows (Data from just one fly).

> Worked on the ping pong ball machine in the last week.
IMG_2907 (1)

The text file looks something like this: Not very sure how to interpret it because there is no column header.

> Did not find any RU486 fly lines in the Brembs fly stock.
What is RU486 and why are we using it? It is a conditional transactivation method that gets activated when introduced with Mifepristone/RU486 and works on the UAS promoter (Roman et al, 2001).

The genes we are knocking down:
1) SERCA gene
2) Ryr gene

 

 

This is a picture of the supplemental figure from Maye et al. 2007

 

Below the results from the sathish scripts running on the data from Maye et al. 2007. It matches, so Sathish scripts in my hands work fine

Confocal image MAX stack of one of the brains at 20x

 

 

and at 40x

 

In this link we have a video of a 3D stainning pattern          zoomed_CC

 

In addition I add here teh boxplots from the final results of the Ping Pong ball setup with these experiments

 

The first figure shows each of the central complex ring neurons types (Martín Pena et al., 2014). The c105-G4 targets the R1 neurons and the c232-G4 targets the R2 and the R4d neurons

This is the c105-G4 stainning from Martín Pena et al., 2014

232-G4 stainning from Kahsai et al., 2012

 

Axel stainning from c232-G4 alone

Axel stainning from c105-G4 alone

Axel stainning from both drivers together. I would say it really contains both driver lines.

This are both driver lines together as well from Axel. To me it seems that only c105 is present

This are my stainnings at the fluorescence microscope (no confocal). This is to show that in all of the 10-12 brains I have looked at, they all had the c232 pattern present

In addition, they had many more neurons outside from the central complex which I believe belong to the c105-G4 line. This is my only proof to show that c105 is also present, since the R1 neurons seem to be hidden when R2 neurons are stained.

I was also looking to the youtube video you have online, Björn. To me it seems I can only see the R1 ring neuron from the c105

After performing EMD to 6 fly traces of 20000 data points (that is 1000 sec flight) for each group (tntXwtb; c105;c232>tnt; c105;c232Xwtb). This data size was chosen to reduce computing time of the SMAP procedure. The EMD decomposes the trace into different time scales in nonstationary data. It seems that the nonlinear behavior occurs at the first IMF (the fastest time scale) and a bit in the second IMF. The potential conclusion to this is that the behavior of the the fly is only unpredictable at the fast movements whereas slow movements are very predictable. Nevertheless, to be cautious it could be that this fastest timescale is just noise, and that this noise is nonlinear. I would say that there is no difference at any time scale between groups (pay attention to the different ranges in the Y-axes), so the ring neurons R1, R3, R4d do not have any effect.

As a groundtruth I have used the same analysis pipeline for the traces in the uniform arena from Maye et al. 2007. Here the effect is even more pronounced at the fastest time scales. So I will conclude that this is real fly behavior and not noise that is shared among both setups: the Ping pong ball machine and the torquemeter.In order to gain more insights into the underlying flight structure I took one random flight trace to explain a few observations. The x-axis is the theta (that actually goes from 0-4 in steps of 0.2 and therefore we see the 21 points), in the y-axis is the correlation of the prediction to groundtruth. We see that IMF has a bigger slope, but not only that, also that its prediction correlation is around 0.88, whereas lower timescales prediction is basically perfect. That is, fast time scales are not only more nonlinear but also less unpredictable. This pattern is repeated in every fly measured

 

To have an impression of how these IMF resultant traces look like: IMF1, IMF2 and IMF8

This is for the sake of playing and curiosity. I made out of these two traces a modelling of their flying trace in a 2D world. Direction is right wing amplitude – left wing amplitude and distance flown is dependent on the sum of both (more amplitude of both, more forward thrust). Funny enough, the second one looks kind of fractal, which is characteristic of chaotic behaviors. If there is any comment to add to this new visualization, all ears!

Trace segment from Maye et al. 2007 in the uniform arena

 

Trace after filtering by selecting the first 8 IMFs (intrinsic mode functions) from EMD (empirical mode decomposition). Since the signal should be quite clean I do not take out the first IMFs. The last IMFs, however, are too slow and change the baseline to much

Some examples of the IMFs obtained from EMD:

Since this is separating behavior adapting to the data intrinsic time scales I am now thinking of analysing with the SMAP algorithm to see if the behavior is more or less nonlinear at certain time scales.

In addition, I have thought of using ICA (Independent component analysis), that is an algorithm famous for the blind source separation problem by extracting the most independent signals from the input signals (in this case the IMFs which are different time scales of the behavior). So the ICs should consist of mixes of different time scales that are correlated together and thus belong (but not necessarily) to the same action/movement module/… Here a few ICs (from 10). My idea is that muscles might coordinate independently between ICs but coordinated within ICs. However to prove that is not that easy I guess