Confocal images and boxplots from my results in strokelitude

on Tuesday, May 15th, 2018 12:26 | by

Confocal image MAX stack of one of the brains at 20x

 

 

and at 40x

 

In this link we have a video of a 3D stainning pattern          zoomed_CC

 

In addition I add here teh boxplots from the final results of the Ping Pong ball setup with these experiments

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Stainning c105;;c232

on Monday, May 14th, 2018 11:22 | by

 

The first figure shows each of the central complex ring neurons types (Martín Pena et al., 2014). The c105-G4 targets the R1 neurons and the c232-G4 targets the R2 and the R4d neurons

This is the c105-G4 stainning from Martín Pena et al., 2014

232-G4 stainning from Kahsai et al., 2012

 

Axel stainning from c232-G4 alone

Axel stainning from c105-G4 alone

Axel stainning from both drivers together. I would say it really contains both driver lines.

This are both driver lines together as well from Axel. To me it seems that only c105 is present

This are my stainnings at the fluorescence microscope (no confocal). This is to show that in all of the 10-12 brains I have looked at, they all had the c232 pattern present

In addition, they had many more neurons outside from the central complex which I believe belong to the c105-G4 line. This is my only proof to show that c105 is also present, since the R1 neurons seem to be hidden when R2 neurons are stained.

I was also looking to the youtube video you have online, Björn. To me it seems I can only see the R1 ring neuron from the c105

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Testing CaLexA

on Monday, October 30th, 2017 1:32 | by

Here I used the CaLexA tool with the elav driver, and reared the flies on constant darkness or under L-D cycles.

L-D Cycle

D-D

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TbH MIMIC

on Monday, October 9th, 2017 2:28 | by

 

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TbH-LexAs and TDC2-Gal4 comparison

on Monday, March 20th, 2017 2:17 | by

I recently combined the two TbH-lexA  lines (54954 & 54075) with CD8GFP, and the TDC2-GAL4 line with CD8RFP, in order to compare their expression patterns. Here I present some of the dissections. The confocal is not working properly, but it is relatively good to draw some conclusions.

 

TDC2>GFP and anti-TβH (Scholz’s Lab)

TDC2>GFP (Gerber’s Lab)

anti-TDC2 (Goodwin’s Lab)

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DA neuronal populations and photopreference (counting neurons)

on Monday, March 13th, 2017 3:01 | by

After refining my DA screening, I end up having three interesting GAL4s which lead to changes in photopreference after expressing Shibire and rising the temperature. What I am trying to do now is to understand if they label the same neuronal population or not.

 

Genotype PAM PAL PPM1 PPM2 PPM3 PPM4 PPL1 PPL2 VUM
thF1>GFP 0 0 0,25 3,75 4,25 0 3 0,75 0
thF1;C’>GFP 0 0 1 7 5 0 2,5 5,5 1,5
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TbH QF system

on Monday, September 12th, 2016 1:37 | by

In order to study the role of OA and DA in photopreference I am constantly looking for new drivers that label subpopulations of these groups. I recently found a TbH driver from the QF system, and wanted to know how representative of the TDC2-G4 neurons was. I have established a QUAS-mCherry;TbH-QFs line (Magenta) and crossed it with TDC2-G4;UAS-GFP (Green). In both cases, what is shown is the endogenous expression.

tdc2-gfptbhqfs-cherrytbhqftdc2-merge-fixation-protocoltdc2-gfp-vnctbhqfs-cherry-vnctbhqftdc2-vnc-merge-fixation-protocol

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Octopaminergic neurons and phototactic flexibility

on Monday, February 1st, 2016 12:46 | by

In previous experiments, I found one TβH(lexA)>shiTS  combination that recapitulated TDC2>shiTS T-Maze results (http://lab.brembs.net/2015/09/looking-for-the-da-oa-neurons-involved-in-phototactic-flexibility/). Here I present the expression pattern of those two TβH-lexA drivers used.

TβH54954-LexA (Brain)

TbHlexA54954 2-1 (RGB)

MAX_TbHLexA.lif - TbHlexA54954 2-1

TβH54954-LexA (VNC)

TbHlexA54954 VNCs (RGB)

 

MAX_TbHLexA.lif - TbHlexA54954 VNCs-1

TβH54075-LexA (Brain & VNC)

MAX_TbHLexA.lif - TbHlexA54075 1 (RGB)

 

MAX_TbHLexA.lif - TbHlexA54075 1-1

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TH-F1 anatomy

on Monday, December 7th, 2015 2:22 | by

I am starting to study in more detail the genotypes that were positive in the DA screen. One was the TH-F1. Here I show the anatomy. I had several problems with the old confocal. I will try to use only the new one.

MAX_TH-F1nGFP4 MAX_TH-F1nGFP3.lsm (RGB) MAX_TH-F1nGFP2.lsm (RGB)MAX_TH-F1nGFPVNC2.lsm (RGB)MAX_TH-F1nGFPVNC1.lsm (RGB)MAX_TH-F1nGFPbrain1

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Phototactic flexibility – Neural substrates

on Monday, November 23rd, 2015 2:29 | by

In order to find which dopaminergic and octopaminergic neurons are related to light preference and the switch on it seen after clipping the wings, I decided to use a tool called CaLexA.

Here we can see my first attempt to use it. We can see the CNS from TH>CaLexA flies with and without wings.

I did not see any special signal in the brains, but I still have to play around a little bit more.

With Wings

TH CaLexA With Wings

Without Wings

TH CaLexA No wings C

TH CaLexA No wings B TH CaLexA No wings

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