candidate genes interact with sim::tgo yu-chi chen december 19 th, 2006 part 1 (august-october)
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Candidate Genes Interact with Sim::Tgo
Yu-Chi ChenDecember 19th, 2006
PART 1 PART 1 (August-October)(August-October)
Sim Ectopic Expression Sim::Tgo bind to CNS midline element (ACGTG) Only neuroectodermal cells exhibited midline mor
phology
Nambu et al., 1991
Screen for Strains with deletions
Ect3 : 7642, 7641, 7966, 7967, 7968 cact and midline genes: 3588, 1491, 2583, 4
20, 7833, 8945, 7522, 7834, 7835, 7838, 7836, 7837, 7839, 7840
sim phenotype? 6647, 3467, 7896, 6609, 3689
sim: 9088 slit: 3519
BP102 staining Stock# 3519 (slit)
10X
40X
McGovern, et al., 2003
Wild type (regular ladder-like structure)
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.
Drosophila CNS Midline Development in pros17
mutant
Yu-Chi ChenDecember 19th, 2006
PART 2 PART 2 (October-December)(October-December)
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 Mira
nda (adaptor protein)
Questions Previously, much more En+ cells were found
in the pros17 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)
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
Normal iVUMs in pros17
Stage 16 Sagittal View ISH: CG15236 (red S
tain En (blue) More CG15236- En+
Cells Wild type:
1 MNB 7 MNB progeny
CG15236+ cells? (3 iVUMs)
6
More NB progeny in pros17
Stage 16 Sagittal View Antibody staining E
lav (red) and Castor (blue)
~15 Cas+ and Elav+ Cells
Wild type: 2 of 3 MNB progeny 2 iVUMs, anterior 2 mVUMs, anterior
2
Normal mVUMs in pros17
Stage 15 Ventral View ISH: fd59A (re
d) Stain: En (blue)
~3 fd59A+ and En+ Cells
Wild type: 1 of 3 MNB progeny
Normal fd59A+ and En- Cells (3 mVUMs)3
Number of Neurons Increased in pros17
Stage 15 Ventral View ISH: tkr (red) St
ain: En (blue) ~5 tkr+ / En+ Cel
ls Wild type:
1 MNB progeny 1 post iVUM
4
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
Discussion Prospero has a different role in midlin
e than lateral CNS (Additional En+ cells are also Elav+)
Neural differentiation? # of GMCs: specific marker? Abnormal position for some cells in pr
os17
More dividing cells
pros17 Mutant
GMC4
Cas
fd59ATkr
CasCas
Cas
fd59ATkr
fd59ATkrfd59ATkr
Cas
Cas
fd59ATkr
fd59ATkr
Cas Cas
fd59ATkrfd59ATkr
AlternativeMechanism