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PART 1
(August-October)
Candidate Genes Interact
with Sim::Tgo
Yu-Chi Chen
December 19th, 2006
Sim Ectopic Expression
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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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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
BP102 staining
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Stock# 3519 (slit)
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Wild type
(regular ladder-like structure)
10X
McGovern, et al., 2003
40X
BP102 staining
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Stock# 3467
(56F9-17; 57D11-12)
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Stock# 7641 (Ect3)
(87A1; B5)
10X
40X
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The gene interacting with Sim::Tgo might be
essential for earlier embryonic development.
PART 2
(October-December)
Drosophila CNS Midline
Development in pros17 mutant
Yu-Chi Chen
December 19th, 2006
Development of the Lateral CNS
Prospero
Skeath and Thor, 2003
Prospero (Pros)
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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)
Prospero Expression in Midline
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Stage 11:
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Nucleus: 2 MP1s, 2 MP3s, 6 VUMs
Cortical: MNB
Prospero Expression in Midline
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Stage 13:
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Nucleus: 3 MNB progeny
Cortical: MNB
Prospero Expression in Midline
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Stage 17:
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Nucleus: MNB progeny nucleus (subset 4/8)
Questions
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Previously, much more En+ cells were found
in the pros17 mutant
En+ cells
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S11: VUMs, MNB, PMGs
S13: MNB, MNB progeny, iVUMs, PMGs
S17: MNB, MNB progeny, iVUMs
Other Pros+ cells:
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MP1, MP3 (stage 11)
Number of Neurons Increased in pros17
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Stage 13
Sagittal View
Antibody staining for
Engrailed (red) and
Elav (blue)
~25 Engrailed+ and
Elav+ Cells
Wild type
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1
3 MNB progeny
3 iVUMs
Normal En+ Elav- Cells
(MNB and 4PMGs)
Normal iVUMs in pros17
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Stage 16
Sagittal View
ISH: CG15236 (red
Stain En (blue)
More CG15236- En+
Cells
Wild type:
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6
1 MNB
7 MNB progeny
CG15236+ cells? (3
iVUMs)
More NB progeny in pros17
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Stage 16
Sagittal View
Antibody staining
Elav (red) and
Castor (blue)
~15 Cas+ and
Elav+ Cells
Wild type:
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2
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2 of 3 MNB
progeny
2 iVUMs, anterior
2 mVUMs, anterior
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Normal mVUMs in pros17
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Stage 16
Sagittal view
ISH: tbh
3 mVUMs
10X
40X
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pros17 Mutant
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Wild type
Normal mVUMs in pros17
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3
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)
Number of Neurons Increased in pros17
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Stage 15
Ventral View
ISH: tkr (red)
Stain: En (blue)
~5 tkr+ / En+
Cells
Wild type:
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4
1 MNB progeny
1 post iVUM
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H cell in pros17
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Stage 15
Sagittal view
ISH: pale
One H-cell
10X
40X
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pros17 Mutant
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Wild type
(S14)
Summary
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More differentiated MNB progeny
at Stage 13 to 17
Normal # of iVUMs, mVUMs
Normal # of MP1s and MP3s?
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Normal # of MNB?
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Only one H-cell
H-cell sib: ISH for fkh
MP1: ISH for odd
Normal En+ Elav- cells
ISH for wor and ase
Normal # of MGs
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ISH for wrapper or Cad74A
Discussion
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Prospero has a different role in midline
than lateral CNS (Additional En+ cells are also
Elav+)
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Neural differentiation?
# of GMCs: specific marker?
Abnormal position for some cells in
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More dividing cells
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pros17
Wild Type
Cas
fd59A
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Cas
Cas
Cas
fd59A
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GMC
4
fd59A
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fd59A
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pros17 Mutant
Cas
Alternative
Mechanism
Cas
Cas
Cas
Cas
Cas
Cas
Cas
fd59A
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fd59A
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fd59A
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fd59A
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GMC
4
fd59A
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fd59A
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fd59A
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fd59A
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