I post these figures here ; we discussed in last meeting about these figures

GRIP on Inter event intervals of Buridan output data ,,, time threshold method,

Inter activity interval

Inter pause interval

I am receiving GRIP results with NaN (no number available for at least 28 flies and Infinitive for the few more) and I checked twice. I

Basically I am extracting inter event intervals and had it run by GRIP to see the level of randomness.GRIP is based on decimal extension of pi.

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:

–          1^st cross

–          2^nd cross

Flies:

–          1^st cross (with heterozygous controls):

                  males                         females                                          females

w1118,tßh/y ;; HStßh  x  w+,tßh//FM7   ->    w1118,tßh//w+,tßh ;; HStßh//+
w1118,tßh/y                    x  w+,tßh//FM7   ->    w1118,tßh//w+,tßh
w1118,tßh/y ;; HStßh  x  w+                        ->   w1118,tßh//w+ ;; HStßh//+
w1118,tßh/y                    x  w+                        ->    w1118,tßh//w+

– 2^st cross (only mutants):

w1118,tßh/y ;; HStßh  x  w+,tßh//FM7    ->    w1118,tßh//w+,tßh ;; HStßh//+
w1118,tßh/y                    x  w+,tßh//FM7    ->    w1118,tßh//w+,tßh

Heat shock timing:

I started to work with the data I got from Li Liu. I have no metadata file, so I am not sure what will finally come out of the analysis.
What is already clear is that the number of female and male they tested seem different for each group and there is a big difference in speed between male and female… The value reported are also huge (>20 mm/s, no distinction male/female) while I get the normal 15mm/s (if the data I got is in mm and not in pixel, of course).
I do not know what GAK and ‘PAN stands for…

speedplot from liu data. X axis is not explained yet. homogenous environment, platform is 86 mm.

Remember that from our data, there is no significant difference in male versus female speed.

Chrisi’s day…

1 Collected virgins for our ill post-doc, made some crosses:

d42-GAL4 males x tubGAL80-UAS-PKCi and x CS

d42-chaGAL80 males x tubGAL80-UAS-PKCi and x CS

2 Buridan: Prepared colored food (10mg/ml, 2.5ml total in the very little vials, ~ 5 flies in each vial). Denise tested in the afternoon females in the Buridan. Data (4n now) will be shown in the lab meeting.

3 Imaging Made a new cross for the Imaging-project:

TDC2-GAL4 males x UAS-GCaMP3(2)

replaced Neloy’s flies by flies from stock

Florian’s larval preparation (good!) showed that GCaMP-expression is much less than GFP-expression. They hope for the homozygous line.

4 Flight: Performed flight experiments with the tbh,TDC,UAStbh flies. Raw data, see below. Thinking about analysis (probably tomorrow)

Protocol: glued the day before, sugar in wheel chambers, little EVIAN-water.

5s filter on the legs before starting, aspirated, stopped when no response 3 times in a row, stopped time when 30min, stopped after 15 aspiration trials

5 Trehalose: Found time to try out Gérard’s idea (no pigment in the probes to not disturb the NanoDrop measurement). 4 groups: 10 flies with or without heads, 50 flies with or without heads. Hilde showed me how to remove heads with liquid nitrogen and vortex. The other flies (with heads) were frozen at -20° as usual. Trehalase incubation over night. Tomorrow glucose oxidase and measurement

Hi all, I forgot about the lab book email, because it reached me in New Orleans… My day:

1 Collected virgins for our ill post-doc

2 Prepared colored food (10mg/ml, 2.5ml total in the very little vials, ~ 7 flies in each vial). Vicky tested in the afternoon females in the Buridan. The the duration of drug application was increased to 3h  because there was no effect after 1 hour. Data will be shown in the lab meeting.

3 Answered to Dennis Pauls to make an appointment for the Trehalose measurement in Würzburg

4 Glued hooks for a flight experiment I did not do because I did not know, how… (idea was to test whether the UAS-tbh cross is ok)

5 Checked my TDCxGCaMP flies for expression in the fluorescence dissection scope (very cool!) in Marco’s room (borrowed from Konstantin). Then realized that there are pupa in the virgin vials :(

6 Temperature-dependent experiments in Buridan: wanted to test the infrared lamp with the dimmer (measure the temperature change). Was impossible because the thermometer itself heated up. So, I finally looked for an infrared thermometer (you can point on the area you want to measure).

7 boring stuff… Dienstreiseabrechnung was wrong… Email to the Chinese guys again about the cylinder for Buridan… Explained Florian (Marco’s master student) some fly stuff…

1. Working on the buridan speed threshold activity data.

2. I mailed to Dr.Thomas Binz who supplied TNT antobody to Heisenberg lab and awaiting for response.

3. C105-Gal4 will be out of quarantine within this week.

4. Working on the thesis writing, gathering information about tetanus toxin expression studies done by sweeney.

Here I post the figures belong to,

I. Boxplot of one fly input data to GRIP.

Legend: 1.Time interval between each activity (Pause length)

2.Difference between subsequent activity (subtracting the subsequent activity)

3. Times between start of activity

4.  Time duration of each Activity (activity data itself)

II. GRIP on Matlab generated pseudo-Poisson process (on the modified code )

III.Times between animal starts

Bar: 1.c105;c232-wtb

2.c105232-TNT

3. Wtb-TNT

IV. Length of activity

V. Total distance travelled during activity period

Grip analysis on:

1. Interval between activities (temporal)

2.Difference between the subsequent activity (temporal) like  [X2-X1,X3-X2]

3.Difference between the subsequent distance travelled during the activity period (spatial)

4.Pysolo midline crossing data. Data sampling at 1 min interval. number of time fly crosses the virtual midline over the period of 60 s.

Bar: 1.c105;c232-wtb

2.c105232-TNT

3. Wtb-TNT

Inter-activity interval of buridan activity

2.Difference between the subsequent activity

Difference between the subsequent distance travelled during the activity period

pysolo midlines crossing

I worked on CeTrAn, corrected some bugs and added boxplot visualization of each variable. As an examples see how the stripe deviation variable vary in computer generated data, quite impressive: