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Candidate Genes Interact with Sim::Tgo

PART 1 (August-October). Candidate Genes Interact with Sim::Tgo. Yu-Chi Chen December 19 th , 2006. Sim Ectopic Expression. Sim::Tgo bind to CNS midline element (ACGTG) Only neuroectodermal cells exhibited midline morphology. Nambu et al., 1991. Screen for Strains with deletions.

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Candidate Genes Interact with Sim::Tgo

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

  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) • Wild type (regular ladder-like structure) 10X McGovern, et al., 2003 40X

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

  6. PART 2 (October-December) Drosophila CNS Midline Development in pros17mutant Yu-Chi Chen December 19th, 2006

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

  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 pros17mutant • 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 pros17 • 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 pros17 • 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 pros17 • 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 pros17 • Stage 16 • Sagittal view • ISH: tbh • 3 mVUMs 10X 40X • pros17 Mutant • Wild type

  17. Normal mVUMs in pros17 • 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 pros17 • 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 pros17 • Stage 15 • Sagittal view • ISH: pale • One H-cell 10X 40X • pros17 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 pros17 • More dividing cells

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

  23. Cas Cas Cas Cas Alternative Mechanism Cas Cas Cas Cas fd59A Tkr fd59A Tkr fd59A Tkr fd59A Tkr GMC 4 fd59A Tkr fd59A Tkr fd59A Tkr fd59A Tkr pros17 Mutant

  24. Thank for Your Attention

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