Motion blindness in T4/T5 using Optomotor Response

on Tuesday, March 31st, 2020 1:48 | by

T4/T5 driver line was crossed TNT line to test for motion blindness using the optomoter response assay. While the controls respond perfectly fine to the rotating striped arena the T4/T5 flies are seemingly unaware of it. Sample size for T4/T5 flies is still a bit low. However, as this is only a trial experiment for the drosophila course I consider it successful.
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Positive controls in joystick optogenetics

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I’ve started rescreening Christian’s most consistent dopaminergic lines with the positive controls at the joystick machine, to see if everything works. These flies express the optogenetic channel Chrimson in Gr28bd and TrpA1 heat sensitive neurons, so they should use the joystick to switch the light off.

I tested three rounds of three flies each and I got the impression that it worked while I was looking at the flies. The data didn’t look that great, however:

Experimental time course of positive control flies. PIs (Y-axis) should be mostly negative.

Setting up the experiments, I noticed that the middle setup didn’t really focus the light on the head of the fly very well. This shows in the raw performance index data:

The two middle flies (row #2 and #8) have positive preferences, but never avoid the light, while nearly all other flies do avoid the light to various extents, even if they start out with a positive preference. So I deleted these two flies to see how the data would look then:

As above, only this time with the training periods averaged at the end (“Means”).

I decided that this should be good enough. I improved the middle setup to focus the light better on the fly’s head. Hopefully it will improve the PIs of the flies placed there now.

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FoxP-KO at embryo stage

on Monday, March 30th, 2020 12:57 | by

The results again replicate the ones I had before.. FoxP flies where the temperature switch has been done for 12hrs during embyo stage can barely walk and surely can not fly.

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Making sense of dwelling data

on Tuesday, March 24th, 2020 12:54 | by

Mean or median?

UnpunishedPunished
1001
13
1,21
0,52
0,35
5
Mean182,4
Median1,12

A highly hypothetical scenario of how the distribution of dwelling times could be. Even if unrealistic, it still illustrates the problem of using median instead of mean.

Plotting the dwelling times as means

An example fly of how the dwelling times are distributed over the different periods.

Grouped analysis of dwelling times

Cumming estimation plot

Used the R package Dabestr to plot the estimation statistics. The effect size and 95% CIs are plotted a separate axes
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Potential Gal4-lines

on Monday, March 23rd, 2020 5:15 | by

Potential Gal4-lines matching parts of the FoxP expression.

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Search for neurons matching the FoxP expression pattern

I started looking at the FoxP expression pattern from Ottavia to see if i can identify the neurons.

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Ongoing experiments

on Sunday, March 22nd, 2020 10:50 | by

  1. PCB and ato crossed to GFP-RFP
  2. I crossed the tub-Gal80 again and I will temperature-induce in adulthood, then I will test at 5 days old, to see if I have any difference if compared to the ones tested at 3 days old
  3. Did the tub-Gal80 again with the induction during embryo, for andy to test together with his one
  4. Wrote almost all of the captions for the figures (exception made for the ones I need to finish experiments)
  5. Writing the results, I think I can manage to finish them for next monday.
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Optomotor response works consistently with old platform

on Friday, March 20th, 2020 3:23 | by

The old platform works a lot more consistently compared to the new one. Unless major improvements are seen I do not see any reason to update it.

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Updates on optomotor response software

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  • Added a timer
  • Previous version generated faulty xml files. This bug has been squashed (see image below)
  • Display window of the software now also prints which direction the striped arena turns
The highlighted red areas are the ones that caused issues. Contingency has also been added to the xml file from the optomotor response (not shown). This did not cause any errors when running the DTS analysis Rscript but I thought it looked nicer having it there.
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New optomotor platform not yet optimized

on Monday, March 16th, 2020 1:42 | by

A new platform have been designed for the optomotor response. It is placed on a pivot with the axis attached on the back. In contrast to the previous design witch only monitored movement in the left/right direction, the new design in theory allows the fly to rotate the platform.

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