Update: Phototactic Flexibility (OA-Neurons)
Figure 1 shows the results of the tbh-subgroups screen, tested at RT and 24 h later at 32°C.
All genotypes with n=8, expect tbh(39942) x Shi^TS (n=4).
Fig 1: tbh subgroups screen
Update: Phototactic Flexibility (OA Neurons) & “Stainings”
Figure 1 shows the newest results of the screen of the tbh-subgroups.
Fig. 1: tbh-subgroups tested at RT and 32°C
In addition I dissected brains of these flies.
Fig 2: 39940 x GFP (nucl)
Fig 3: 41306 x GFP (nucl)
Fig 4: 48333 x GFP (nucl)
Fig 5: 45904 x GFP (nucl)
Fig 6: 46972 x GFP (nucl)
Fig 7: 47755 x GFP (nucl)
As a next step I’ll compare my “stainings” to the ones from flybase.
Update: Phototactic Flexibility (OA Neurons) & “Stainings”
I started testing some more tbh-GAL4-subgroups. In Fig. 1 you see all tbh-subgroups, the last 5 in the graph are the newest trials (n=2).
Fig. 1: tbh-subgroups screen
Moreover I dissected some more brains.
Fig 2: tbh(46970) x GFP (nucl.)
Fig 3: tbh(48332) x GFP nucl
Update: Phototactic Flexibility (OA Neurons) & “Stainings”
In Fig 1 you see the results of the tbh – subgroups screen.
Furthermore I plotted them in a dot plot. (Fig. 2 and Fig. 3)
Fig.1 tbh subgroups screen
Fig 2. tbh subgroups tested in September and October
Fig. 3 tbh subgroups tested in November and December
Moreover I dissected some brains which I crossed to GFP (nucl.)
Fig4. tbh-48331 x GFP (nucl)
Fig 5. tbh-39941 x GFP (nucl)
Fig 6. tbh-39939 x GFP (nucl)
Fig. 7 tbh-39942 x GFP (nucl)
Fig 8. 47756 x GFP (nucl)
Phototactic Flexibility (OA neurons) – Update & ‘Staining’ of tbh
Phototactic Flexibility
These are the newest results of the tbh-subgroups screen.
Fig 1: tbh-subgroups tested at RT and 24 h later at 32°C
‘Staining’ of tbh subgroup 46970
I crossed tbh-46970 to GFP (nucl), dissected the brains and mounted them (No antibodies were used).
Fig 1: 46970 x GFP
Fig 2: 46970 x GFP
Phototactic flexibility (OA neurons)
I tested some more tbh-GAL4 subgroups. Figure 1 shows the newest results of the screen.
Because of the weird results, we decided to make crossbreedings the other way around. For the last crossbreedings I took virgins of the UAS-Shi^TS and males from the tbh-GAL4 subgroups, now I take virgins of the tbh-GAL4s. So in two weeks the first crossbreedings will hatch and I will test them.
Fig. 1: tbh-subgroups tested at RT and 24 h later at 32°C. All tbh-GAL4 were crossed to Shi^TS virgins
Phototactic flexibility (OA neurons)
I tested some more tbh-GAL4 crossed to shibire^TS. These are the newest results of the screen.
Figure 1: tbh-GAL4 subgroups tested at RT and at 32°C (n(39939, 46970) = 4; n(47756,48331) = 3; n(39941,39942) = 2)
Phototactic flexibility (OA neurons) & Staining of ChaG80;ThG4
Phototactic flexibility
I continue testing the tbh-Gal4 subgroups and tbh-lexA.
Figure 1 shows the newest results of tbh-lexA (n=6)
Figure 2 shows the newest results of tbh-Gal4 (tbh48331, tbh46970 n=3, tbh39942 n=1, rest with a n of 2)
Figure1: tbh-lex A tested at RT -> 32°C -> RT
Figure 2: tbh subgroups tested at RT and 32°C
Stainings
I practiced the dissections of brains for stainings. So I fixed the flies for one hour in PFA 4% , then I dissected the brains, washed them a few times with PBST and after it I mount them with vectashield.
Figure 3 shows a picture of a brain of a Cha-Gal80;TH-Gal4 x GFP (nucl.) flie.
Figure 3: Staining: Cha-Gal80;TH-Gal4 x GFP (nucl.)
Phototatic flexibility
I want to know which octopaminergic neurons are involved in the phototactic switch after wing clipping. So I started with a screen for some subgroubs of tbh-GAL 4. After testing the flies in RT with a T-Maze, I tested the same flies again in 32 degree. I expect, that wingless flies go to the light, so the Choice Index should increase. The Figure 1 shows the results of only 2 experiments (39942-tbh-GAL4 only one experiment).
Figure 1: tbh-subgroups tested at RT and 32°C