Update: DA and OA in photopreference
This is an update of the T-maze experiments for flies with OA and DA neurons manipulated.
- TH-G4s x shiTS. Except for 2 groups the N is at least 5.
2. TbH-lexAs x shiTS. Last year I tried to complete this experiment and several problems appear. Now I am starting to do it again and it seams that it works. N=4.
3. I am starting to test the same TbH lines crossed to TrpA1. So far, not much to say.
Photopreference TH subgroups (protocol changes & new data)
Since I finished the first round of the screen to find out the DA substrate of the photopreference, I was testing the candidate lines in a WTB background. I had some unexpected results with the controls, so now I’m continuing the experiments, while adjusting some things to understand the problem.
In this case, I reared and tested the flies as before (reared at 18°C, and tested at 25 & 32 °C in consecutive days). But, I also made a copy of those crosses and placed them at 25 during development. After that I moved them to 18°C until the experiment was done.
Update (25°C):
Phototactic variability and individual choices
I am starting to look the data generated by Isabelle to generate the final graphs and the definitive analysis of it. This is a sample of a possible plot.
Photopreference TH subgroups
After finishing the first round of the screen to find out the DA substrate of the photopreference, I decided to change the genetic background of the GAL4s and UAS. Now all of them are in WTB background.
Here are the first results. It is only an n of 1.
Update:
Flying ability with DA or OA neurons silenced
As a control for my T-Maze experiments I decided to test if treated flies were able to fly at the same conditions they were tested in the T-Maze (32°C).
As in the T-Maze experiments, I put the flies 15 min at 32°C before testing them at the same temperature. The, I measure the duration of the first spontaeous flight (Blue). If within 5min the fly doesn’t start to flight, I gently remove the steroform ball from the legs (Red), simulating a takeoff.
Details from 0 s to 200 s.
Flying capability of flies with DA neurons silenced
As a control for my T-Maze experiments I decided to test if treated flies were able to fly at the same conditions they were tested in the T-Maze (32°C).
As in the T-Maze experiments, I put the flies 15 min at 32°C before testing them at the same temperature.
The dashed line at 11 min indicates when a T-Maze experiment would have ended.
Almost all flies tested flew more than the time required to perform a T-Maze experiment.
Octopaminergic neurons and phototactic flexibility
In previous experiments, I found one TβH(lexA)>shiTS combination that recapitulated TDC2>shiTS T-Maze results (https://lab.brembs.net/2015/09/looking-for-the-da-oa-neurons-involved-in-phototactic-flexibility/). Here I present the expression pattern of those two TβH-lexA drivers used.
TβH54954-LexA (Brain)
TβH54954-LexA (VNC)
TβH54075-LexA (Brain & VNC)
Flying capability of flies with DA or OA neurons silenced
As a control for my T-Maze experiments I decided to test if treated flies were able to fly at the same conditions they were tested in the T-Maze (32°C).
As in the T-Maze experiments, I put the flies 15 min at 32°C before testing them at the same temperature.
The dashed line at 15 min indicates when a T-Maze experiment would have ended.
All flies tested flew almost twice the time required to perform a T-Maze experiment.
T-Maze problems
I am trying to spot source of the current behavioral problem we are having. I tested some crosses that I know how they behave. I did the crosses in both directions and tested to different lights sources (cold and warm) at 25°C.
TH-F1 anatomy
I am starting to study in more detail the genotypes that were positive in the DA screen. One was the TH-F1. Here I show the anatomy. I had several problems with the old confocal. I will try to use only the new one.
