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)

Development of the Lateral CNS

Skeath and Thor, 2003

Prospero

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)

Prospero Expression in Midline Stage 11:

Nucleus: 2 MP1s, 2 MP3s, 6 VUMs Cortical: MNB

Prospero Expression in Midline Stage 13:

Nucleus: 3 MNB progeny Cortical: MNB

Prospero Expression in Midline Stage 17:

Nucleus: MNB progeny nucleus (subset 4/8)

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 16 Sagittal vie

w ISH: tbh 3 mVUMs

10X

40X pros17 Mutant Wild type

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

H cell in pros17

Stage 15 Sagittal view ISH: pale One H-cell

10X

40X

pros17 Mutant Wild type (S14)

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

GMC4

Cas

fd59ATkr

CasCas

Cas

fd59ATkr

fd59ATkrfd59ATkr

Wild Type

pros17 Mutant

GMC4

Cas

fd59ATkr

CasCas

Cas

fd59ATkr

fd59ATkrfd59ATkr

Cas

Cas

fd59ATkr

fd59ATkr

Cas Cas

fd59ATkrfd59ATkr

AlternativeMechanism

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