9/8/2010 The role of runt, en, and slp in establishing MP cell fate.

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9/8/2010 The role of runt, en, and slp in establishing MP cell fate

Epidermal regulation of runt, en, slp Wild-typerunt4496 odd en

runt, slp, and en are expressed in distinct patterns in the developing midline Stage 9 Stage 11 slp En Runt

Delta mutant embryos still specify distinct MP fates slp En Runt

Slp1 expression patterns – antibody vs in situ Slp1 En Stage 11Stage 12Stage 14 Stage 11Stage 12Stage 14 Stage 10 slp1 En

MP3 assumes an MP4-like fate in slp1 mutant embryos

Model RuntSlpEn MP3AMG, MP1MP4, PMG tup

Reagent to test this model – Tup – antibody and probe Tup tup En

Examine 4 genotypes to test this model sim-Gal4 uas-slp1 sim-Gal4 uas-enVP16 enA/enE transheterozygote runt 4496 null mutant

Is slp1 sufficient to induce MP3 formation? RuntSlpEn MP3AMG, MP1MP4, PMG Slp sim-gal4 uas-slp1 tup En sim-gal4 uas-slp1 ple En

Is tup a downstream target of engrailed? sim-Gal4 uas-enVP16 enA/enE RuntSlpEn MP3AMG, MP1MP4, PMG tuptup Elav

Is tup a downstream target of engrailed? tup Odd

RuntSlpEn MP3AMG, MP1MP4, PMG What is the role of En in posterior neurons? Does a transcription factor specify MP4 fate? tup Hh

What purpose does en serve in midline neurons? Joe Kearney’s data

Variable number of neurons in en mutant embryos From 43 segments: Average number of neurons: Range: 4-10 wor Elav MNB is missing

Average number of Zfh1 positive cells (stage 13-15, 9 segments) : 2.8

Segment mixing? stage 11 Elav Zfh1 Elav tup Elav Zfh1 Elav Early enA/enE segments are disorganized

MP1s are also present in en mutant embryos HbZfh1 tup MP1smVUMs H-cell wor Elav MNBs

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?

MP1 cell fate Stage 9 slp1 En

MP1s still form in the absence of either runt or slp1 tup Odd slp1 2 runt4496 en+ segmenten- segment Look at additional MP1 mutant animals to determine mechanisms that establish MP1 cell fate hb En

Current Model RuntSlpEn MP3AMG, MP1MP4, PMG tup odd, hb Hh