1. Candidate Genes Interact with Sim::Tgo Yu-Chi Chen December 19 th, 2006 PART 1 (August-October)

2. Sim Ectopic Expression Sim::Tgo bind to CNS midline element (ACGTG) Only neuroectodermal cells exhibited midline morphology Nambu et al., 1991

3. Screen for Strains with deletions Ect3 : 7642, 7641, 7966, 7967, 7968 cact and midline genes: 3588, 1491, 2583, 420, 7833, 8945, 7522, 7834, 7835, 7838, 7836, 7837, 7839, 7840 sim phenotype? 6647, 3467, 7896, 6609, 3689 sim: 9088 slit: 3519

4. BP102 staining Stock# 3519 (slit) 10X 40X McGovern, et al., 2003 Wild type (regular ladder-like structure)

5. BP102 staining Stock# 3467 (56F9-17; 57D11-12) 10X 40X Stock# 7641 (Ect3) (87A1; B5) The gene interacting with Sim::Tgo might be essential for earlier embryonic development.

6. Drosophila CNS Midline Development in pros 17 mutant Yu-Chi Chen December 19 th, 2006 PART 2 (October-December)

7. Development of the Lateral CNS Skeath and Thor, 2003 Prospero

8. Prospero (Pros) A transcription factor acts in GMC  a single division Represses cell-cycle genes: cyclin A, cyclin E, E2F, and string (Li and Vaessin, 2000) Only transcribed in NB Asymmetric localization requires Miranda (adaptor protein)

9. Prospero Expression in Midline Stage 11: Nucleus: 2 MP1s, 2 MP3s, 6 VUMs Cortical: MNB

10. Prospero Expression in Midline Stage 13: Nucleus: 3 MNB progeny Cortical: MNB

11. Prospero Expression in Midline Stage 17: Nucleus: MNB progeny nucleus (subset 4/8)

12. Questions Previously, much more En + cells were found in the pros 17 mutant En + cells S11: VUMs, MNB, PMGs S13: MNB, MNB progeny, iVUMs, PMGs S17: MNB, MNB progeny, iVUMs Other Pros + cells: MP1, MP3 (stage 11)

13. Number of Neurons Increased in pros 17 Stage 13 Sagittal View Antibody staining for Engrailed (red) and Elav (blue) ~25 Engrailed + and Elav + Cells Wild type 3 MNB progeny 3 iVUMs Normal En + Elav - Cells (MNB and 4PMGs) 1

14. Normal iVUMs in pros 17 Stage 16 Sagittal View ISH: CG15236 (red Stain En (blue) More CG15236 - En + Cells Wild type: 1 MNB 7 MNB progeny CG15236 + cells? (3 iVUMs) 6

15. More NB progeny in pros 17 Stage 16 Sagittal View Antibody staining Elav (red) and Castor (blue) ~15 Cas + and Elav + Cells Wild type: 2 of 3 MNB progeny 2 iVUMs, anterior 2 mVUMs, anterior 2

16. Normal mVUMs in pros 17 Stage 16 Sagittal view ISH: tbh 3 mVUMs 10X 40X pros 17 Mutant Wild type

17. Normal mVUMs in pros 17 Stage 15 Ventral View ISH: fd59A (red) Stain: En (blue) ~3 fd59A + and En + Cells Wild type: 1 of 3 MNB progeny Normal fd59A + and En - Cells (3 mVUMs) 3

18. Number of Neurons Increased in pros 17 Stage 15 Ventral View ISH: tkr (red) Stain: En (blue) ~5 tkr + / En + Cells Wild type: 1 MNB progeny 1 post iVUM 4

19. H cell in pros 17 Stage 15 Sagittal view ISH: pale One H-cell 10X 40X pros 17 Mutant Wild type (S14)

20. Summary More differentiated MNB progeny at Stage 13 to 17 Normal # of iVUMs, mVUMs Normal # of MP1s and MP3s? Only one H-cell H-cell sib: ISH for fkh MP1: ISH for odd Normal # of MNB? Normal En + Elav - cells ISH for wor and ase Normal # of MGs ISH for wrapper or Cad74A

21. Discussion Prospero has a different role in midline than lateral CNS (Additional En + cells are also Elav +) Neural differentiation? # of GMCs: specific marker? Abnormal position for some cells in pros 17 More dividing cells

22. GMC 4 Cas fd59A Tkr Cas fd59A Tkr fd59A Tkr fd59A Tkr Wild Type

23. pros 17 Mutant GMC 4 Cas fd59A Tkr Cas fd59A Tkr fd59A Tkr fd59A Tkr Cas fd59A Tkr fd59A Tkr Cas fd59A Tkr fd59A Tkr Alternative Mechanism

24. Thank for Your Attention