In this project, one of the two experimental groups inhibits the ellipsoid body ring neurons using TNT-E. The second uses RNAi to knock down the DopR2 receptor in these neurons and the third is the driver control. As I am blind to the groups and it’s only been one week, the data are not terribly revealing, yet.

The 1st cross produced eggs, but they did not hatch or become pupae. In all of the 10 vials, eggs were observed, but no pupae. On the other hand, the 2nd cross did not produce any eggs. (The first trial was kept up for 10 days. After 10 days, a new trial began from the same stock. ) Control and the other two crosses produced eggs and hatched new flies.

The same crosses were repeated to ensure the lethality. In the second trial, ‘T4/T5xWTB’ cross produced eggs and pupae. But there is still the possibility that the T4/T5xWTB is also a lethal crossing. ‘T4/T5xTNTE’ cross again produced only eggs but no pupae. The other two crosses produced pupae again and became dark, but haven’t hatched yet. The second attempt is 10 days old.

Control groups fed with ATR (Gr28bdTrpA1_Suc_ATR and Gr28bdTrpA1_Suc_ATR) look fine and for now the avoidance by activating heat sensing neurons does not seem to depend on dopamine. The negative control (Gr28bdTrpA1_Suc_EtOH) shows stronger avoidance than we would expect. Also PIs of PPM2 lies do not follow the same pattern as we previosly observed… My current hypothesis is that some sort of mutation led to the Chrimson channel being more sensitive to light, even without ATR. Since we rear the PPM2 flies in light until they receive the ATR this might affect their behavior.

Luisa’s results for yellow light:

After the 10th day, cross 2 and the control group hatched new flies. On the 13th day, all of the groups have hatched flies. None of the crosses seems to be lethal.

Confirmation of 3IY+ATR

After figuring our how to apply 3IY to the flies and confirming that we can simply mix in the ATR with the sucrose to apply it, I stumbled upon another problem: when 3IY and ATR are used together the tissue paper will go from yellow to orange:

Since we cannot know if this affects the action of 3IY I conducted a final trial in the open field, measuring locomotion of WTB flies after treating them with 3IY and ATR for 48h.

N_{WTB_3IY_ATR} = 16 ; N_{WTB_3IY_EtOH} = 13; N_{WTB_SUC_EtOH} = 8

It seems, that ATR does not affect the action of 3IY and we can proceed with our experiments.

T-Maze results

These are the first results from the set of T-Maze experiments with 3IY treatment. I used red light (1600 Lux) with a decision time of one minute. P-values above plots indicate results of Wilcoxon’s test.