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4_PKC53_RNAi _ Heatshock_32d_3days__25dfor30-90min.pdf (25.22 kB)

PKC53e RNAi preliminary experiments

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posted on 2015-09-30, 09:51 authored by Julien ColombJulien Colomb, Björn BrembsBjörn Brembs

Description of the experiment and the figures

 

 

 

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.

 

 

 

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).

 

 

 

 

Detailed information.

Genetics: parental lines UAS-PKC53e.RNAi lines (V: from VDRC, T: Trip line) males were crossed to elavGal4; tubgal80ts females.

Only females were tested, Flies were grown at 25 degrees and put for two days at 32 degrees after eclosion before being tested..

NB: data where the pretest score was higher than 0.6 were excluded from the analysis.

Full genotype of the flies tested is indicated below:

elavG4;tG80 x CS : UAS_PKC53eRNAi_vienna/+ .

CS x 53eRi_V : elavG4/+;tubGal80ts/+;UAS_PKC53eRNAi_vienna/+ .

CS x 53eRi_T : elavG4/+;tubGal80ts/+ .

elavG4;tG80 x 53eRi_T : elavG4/+;tubGal80ts/UAS_PKC53eRNAi_Trip .

elavG4;tG80 x 53eRi_V : UAS_PKC53eRNAi_Trip/+ .

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flybase numbers:

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

elavGal4= FBti0002575

UAS-PKC53e.RNAi_V= FBti0159711

UAS-PKC53e.RNAi_T= FBti0128750

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