Prevention of PKCi effect by the presence of a UAS-GFP construct

2015-09-30T09:52:12Z (GMT) by julien colomb


Description of the experiment and the figures




Groups represent a cross between the tubGal80ts;UAS-PKCi line (which is effective in blocking self-learning when expressed pan neuronal) and 2 lines with specific combination of 3 construct: d42Gal4 or c380Gal4, as well as chaGal80 (inhibit Gal4 expression in cholinergic neutrons) and UAS-GFP (used to localize expression). In both cases, we have no effect on self learning. While we know that if the GFP construct is taken out, the prevention works (Colomb et al., in prep), we conclude that the presence of a second UAS element prevents the expression of the PKC inhibitor.

Representation legend .

The score during the final test is depicted in boxplots (bar is the median, boxes are quartiles, bars are 100% of the data not considering the outliers presented by points).





Drosophila self-learning.

The flies are tethered to a torque meter which measures the angular momentum around the fly's vertical body axis (yaw torque), caused by attempted turning maneuvers. Attempted left (or right) turning maneuvers are punished during training phases and no predictive stimuli are present. Thus, the only predictor of punishment is the behavior of the fly. If the fly continue to avoid the punished behavior when laser is absent during test phases, we talk about self-learning.




Scoring and learning protocol.

 A score corresponding to the proportion of time the fly is doing the safe turning attempt is calculated for each 2 minutes period. Before training the score is around 0 (no preference). During training phases, the score rises since the fly avoid doing the laser punishment. After two rounds of 4 minutes training separated by a 2min test, a final 2 minutes test round is performed. The performance during these two minutes is used for statistical analysis. After the experiment, the torque force of the fly is tested again using its optomotor response and the accuracy of the laser position (the fly is sacrificed by letting the laser on for about 15s). If any of these two tests is not satisfactory, the data is not included in the analysis.




Detailed information.

Only females were tested, flies were grown at 25° and did receive an heat shock (35° for 4 hours) 0.5 to 4 hours before being tested at 25°.

NB: data where the pretest score was higher than 0.65 were excluded from the analysis.

Full genotype of the flies tested:

CS x T_PKCi : tubGal80ts/+;UASPKCi/+ .

GFP, c380;chaG80 x T_PKCi : UAS-GFP,C380-GAL4/+;;cha-Gal80/+ .

GFP; d42G4,chaG80 x T_PKCi : UAS-GFP;tubGal80ts/+;UASPKCi/d42Gal4,chaGal80 .

GFP, c380G4;chaG80 x CS : UAS-GFP,C380-GAL4;tubGal80ts/+;UASPKCi/cha-Gal80 .

GFP; d42G4,chaG80 x CS : UAS-GFP;;d42Gal4,chaGal80/+ .





flybase numbers:

tubGal80ts= P{tubP-GAL80ts} (FBtp0017264)[on the 2nd chromosome],

d42GAl4= FBti0002759

c380Gal4 = FBti0016294

UAS-GFP = unknown

UASPKCi = FBti0010565

cha-Gal80 = unknown







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