9/8/2010 The role of runt, en, and slp in establishing MP cell fate.
-
date post
19-Dec-2015 -
Category
Documents
-
view
216 -
download
1
Transcript of 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.
![Page 1: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/1.jpg)
9/8/2010
The role of runt, en, and slp in establishing MP cell fate
![Page 2: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/2.jpg)
Epidermal regulation of runt, en, slp
Wild-type runt4496
odd
en
![Page 3: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/3.jpg)
runt, slp, and en are expressed in distinct patterns in the developing midline
Stage 9
Stage 11
slp
En
Runt
![Page 4: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/4.jpg)
Delta mutant embryos still specify distinct MP fates
slp
En
Runt
![Page 5: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/5.jpg)
Slp1 expression patterns – antibody vs in situ
Slp1 En
Stage 11 Stage 12 Stage 14
Stage 11 Stage 12 Stage 14
Stage 10
slp1 En
![Page 6: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/6.jpg)
MP3 assumes an MP4-like fate in slp1 mutant embryos
![Page 7: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/7.jpg)
Model
Runt Slp En
MP3AMG, MP1 MP4, PMG
tup
![Page 8: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/8.jpg)
Reagent to test this model – Tup – antibody and probe
Tup
tup En
![Page 9: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/9.jpg)
Examine 4 genotypes to test this model
sim-Gal4 uas-slp1
sim-Gal4 uas-enVP16
enA/enE transheterozygote
runt4496 null mutant
![Page 10: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/10.jpg)
Is slp1 sufficient to induce MP3 formation?
Runt Slp En
MP3AMG, MP1 MP4, PMG
SlpSlp
sim-gal4 uas-slp1
tup En
sim-gal4 uas-slp1
ple En
![Page 11: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/11.jpg)
Is tup a downstream target of engrailed?
sim-Gal4 uas-enVP16 enA/enE
Runt Slp En
MP3AMG, MP1 MP4, PMG
tup tup Elav
![Page 12: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/12.jpg)
Is tup a downstream target of engrailed?
tup Odd tup Odd
![Page 13: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/13.jpg)
Runt Slp En
MP3AMG, MP1 MP4, PMG
What is the role of En in posterior neurons?
Does a transcription factor specify MP4 fate?
tup
Hh
![Page 14: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/14.jpg)
What purpose does en serve in midline neurons?
Joe Kearney’s data
![Page 15: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/15.jpg)
Variable number of neurons in en mutant embryos
From 43 segments: Average number of neurons: 7.2 +- 1.9
Range: 4-10
wor Elav
MNB is missing
![Page 16: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/16.jpg)
Average number of Zfh1 positive cells (stage 13-15, 9 segments) : 2.8
![Page 17: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/17.jpg)
Segment mixing?
stage 11
Elav
Zfh1 Elav
tup Elav
Zfh1 Elav
Early enA/enE segments are disorganized
![Page 18: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/18.jpg)
MP1s are also present in en mutant embryos
HbZfh1
tup
MP1smVUMs
H-cell
wor Elav
MNBs
![Page 19: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/19.jpg)
Examine iVUM and mVUM markers – Lim1, vglut, gad1, tkr, and cas
Future en experiments
Rescue En in midline and in paired stripes in an enA/enE mutant background
Ectopically express En and EnVP16 in all midline neurons – consequences?
Drive Runt in midline neurons?
![Page 20: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/20.jpg)
MP1 cell fate
Stage 9
slp1 En
![Page 21: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/21.jpg)
MP1s still form in the absence of either runt or slp1
tup Oddtup Odd
slp12
runt4496
en+ segment en- segment
Look at additional MP1 mutant animals to determine mechanisms that establish MP1 cell fate
hb En
![Page 22: 9/8/2010 The role of runt, en, and slp in establishing MP cell fate.](https://reader035.fdocuments.net/reader035/viewer/2022081519/56649d2d5503460f94a03867/html5/thumbnails/22.jpg)
Current Model
Runt Slp En
MP3AMG, MP1 MP4, PMG
tup
odd, hb
Hh