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9/8/2010

9/8/2010. The role of runt, en , and slp in establishing MP cell fate. Epidermal regulation of runt, en, slp. Wild-type. runt4496. odd en. runt, slp , and en are expressed in distinct patterns in the developing midline. slp. En. Stage 9. Runt. Stage 11.

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9/8/2010

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  1. 9/8/2010 The role of runt, en, and slp in establishing MP cell fate

  2. Epidermal regulation of runt, en, slp Wild-type runt4496 odden

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

  4. Delta mutant embryos still specify distinct MP fates slp En Runt

  5. Slp1 expression patterns – antibody vs in situ Stage 11 Stage 12 Stage 14 Slp1En Stage 10 Stage 11 Stage 12 Stage 14 slp1En

  6. MP3 assumes an MP4-like fate in slp1 mutant embryos

  7. Model AMG, MP1 MP3 MP4, PMG Runt Slp En tup

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

  9. Examine 4 genotypes to test this model sim-Gal4 uas-slp1 enA/enE transheterozygote sim-Gal4 uas-enVP16 runt4496null mutant

  10. Is slp1 sufficient to induce MP3 formation? AMG, MP1 MP3 MP4, PMG Slp Slp Runt Slp En sim-gal4 uas-slp1 sim-gal4 uas-slp1 tup En ple En

  11. Is tup a downstream target of engrailed? AMG, MP1 MP3 MP4, PMG Runt Slp En enA/enE sim-Gal4 uas-enVP16 tup tup Elav

  12. Is tup a downstream target of engrailed? tup Odd tup Odd

  13. Hh AMG, MP1 MP3 MP4, PMG Runt Slp En tup What is the role of En in posterior neurons? Does a transcription factor specify MP4 fate?

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

  15. Variable number of neurons in en mutant embryos From 43 segments: Average number of neurons: 7.2 +- 1.9 Range: 4-10 worElav MNB is missing

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

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

  18. MP1s are also present in en mutant embryos Zfh1 Hb mVUMs MP1s tup worElav H-cell MNBs

  19. Future en experiments Examine iVUM and mVUM markers – Lim1, vglut, gad1, tkr, and cas Ectopically express En and EnVP16 in all midline neurons – consequences? Rescue En in midline and in paired stripes in an enA/enE mutant background Drive Runt in midline neurons?

  20. MP1 cell fate Stage 9 slp1 En

  21. MP1s still form in the absence of either runt or slp1 slp12 runt4496 hbEn tupOdd tupOdd en+ segment en- segment Look at additional MP1 mutant animals to determine mechanisms that establish MP1 cell fate

  22. Current Model Hh AMG, MP1 MP3 MP4, PMG Runt Slp En odd, hb tup

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