Verification TH-C-AD;TH-D-DBD

on Monday, September 11th, 2023 9:52 | by

Confocal images with anti-GFP and anti-Brp staining of TH-C-AD;TH-D-DBD > mCD::GFP fly brains: A: 4 PPM 2 DANs per hemisphere marked with blue circles, three clusters of Kenyon cell bodies per hemisphere marked with white arrows. B: Five visible cell bodies of unidentified neurons marked with a blue circle in the left hemisphere. The neurons project into the lobula plate and the medulla. Strong fluorescence of cell bodies of the Kenyon cells projecting into the Mushroom bodies is visible. C: Unidentified neurons projecting into structures outside of the optic lobes especially the ventral lateral protocerebrum.

Role of dopaminergic neurons in operant behaviour

on Friday, July 27th, 2018 3:54 | by

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

T-Maze experiments : screen results as on 16-07-2018

on Monday, July 16th, 2018 1:32 | by

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

T-Maze experiments : screen results as on 25-06-2018

on Monday, June 25th, 2018 1:13 | by

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

Usage of TrpA with infrared lamp

on Wednesday, February 13th, 2013 11:40 | by

As a pilot for future experiments I tried to reproduce results from Keene et al., 2012. There, they substituted sugar by a laser to provoke PER after 24h of starvation (females, one week old). In TH-GAL4 they found 100% response when they pointed the laser to head or thorax. In tdc2-GAL4 indeed they found 50% of flies responding to heat but only when the laser was pointed to the thorax but not to the head.
Since we do not have this kind of laser, they used, I played around with an infrared light. Flies were fixed to hooks and a clamp as always. First, there was given a filter paper soaked with EVIAN-water (negative control), then there were 10s of heat (36-38°C), afterwards two positive controls: 30% sucrose on the legs and finally on the labellum.
In the figure, you can see that I am able to reproduce the methods with an infrared lamp instead of a laser. The TH-GAL4 flies crossed to TrpA respond much more to heat compared to the controls (see figure in blue and red). The proboscis extension appears already after a few seconds.
Unfortunately, I could not reproduce the results for tdc2-GAL4 (see figure in green). There was no response to heat found at all. (tdc2-GAL4 has to be ok, because I see fluorescence when crossed to GCaMP). Interestingly, the flies did not respond as strong to the 30% sucrose as expected. The starvation may have to be longer to increase motivation?
Next step will be to see a phenotype that can be seen by activating tdc2-GAL4. That may be PER after longer starvation or flight behavior.

TH-TDC2

Heat shock rescue of tßh-gene in Buridan’s paradigm

on Thursday, November 29th, 2012 12:50 | by

The loss of tßh in adult flies leads to decreased walking speed and an increase in stripe fixation. Rescuing the gene by a heat shock construct (flies from Henrike Scholz, Cologne) increases walking speed back to wild type level but cannot change stripe fixation.

It is possible that the phenotype in stripe fixation is not exclusively tßh-dependent but more due to other problems the tßh-mutants have, e.g. developmental defects. That would ask for another heat shock timing.

Results:

–          1st cross

 

 

 

 

 

 

 

 

 

 

 

 

 

–          2nd cross