Characterizing eve(PJ4)

Biology 205L: Experiments in Developmental Biology Fall 2006

Myung Ko, Jasmine Lau, Joshua Mendoza-Elias

Gene Regulatory Network

Insert Figure 9.26 (Gilberts)

eve

wg

runt odd

en

hh

TF

+

Consequences of wg expression

Jones & Bejsovec (2003)

An anomaly: eve(PJ4)wg(NE2)

• Gastrulation- wingless protein expression

• Cuticle patterning- wingless expression

Questions:

• Is eve(PJ4) an amorph? • Role of wg(NE2) in double mutant?

Hypothesis:

• eve(PJ4) is an amorph• wg(NE2) causes wg expression to persist

Materials and Methods

Statistical Methods: 2-test, t-test

Wild-type

eve(3)

eve(PJ4) eve(PJ4)wg(NE2)

AB-staining: anti-wg antibody Stages: 9, 10,

11

Cuticle Preparations

Part I: Is eve(PJ4) an amorph?

Are we seeing the right ratio?

• eve (PJ4) • eve(3)

WT Mutant

Ratio

3 1

n 17 8

WT Mutant

Ratio

3 1

n 45 162 value = 0.333 p < 0.05

2 value= 0.022 p< 0.01

Stage 9eve(PJ4)

Mean: 5.89

(n=9)

eve(3)

Mean:6.21

(n=9)

WTWT

WT

Mean: 13.2

(n= 5)

P= 0.31

P= 0.00

P= 0.00

Stage 10

eve (PJ4)

Mean: 5.86

(n=7)

eve(3)

Mean: 5.5

(n=12)

WT

Mean:13.5

(n= 17)

P= 0.47

P= 0.00

P= 0.00

Stage 11

eve(PJ4)

Mean: 5.3

(n=3)

eve(3)

Mean: 5

(n=3)

WT

Mean: 14.3

(n=3)

P= 0.65

P= 0.00

P= 0.00

Mean: 5.89

Mean:6.21

Mean: 5.86

Mean: 5.5

Mean: 5.3

Mean: 5

eve(PJ4)

Stage 9 Stage 10 Stage 11

eve (3)

P= 0.31 P= 0.47 P= 0.65

Cuticles

eve(PJ4) eve(3)

Summary

• eve(PJ4) behaves like an amorph

• As embryos mature,

- decreasing # of stripes

- stripes are less defined

- increasing similarity b/w eve(PJ4) and eve(3)

Part II: Role of wg(NE2) in eve(PJ4)wg(NE2)

wg(NE2) to the rescue??

Are we seeing the right ratio?

• eve(PJ4) • eve(PJ4)wg(NE2)

WT Mutant

Ratio

3 1

n 17 8

WT +/- Mutant

Ratio

1 2 1

n 8 18 10

2 value = 0.33 p < 0.05

2 value= 0.89 p < 0.01

Stage 9

eve(PJ4)

Mean: 5.89

(n=9)

eve(PJ4)wg(NE2)

Mean:5.86

(n=7)

WT

Mean: 13.2

(n= 5)

P= 0.95

P= 0.00

P= 0.00

Stage 10

eve(PJ4)

Mean: 5.86

(n= 7)

eve(PJ4)wg(NE2)

Mean:5.7

(n= 10)

WT

Mean: 13.5

(n= 17)

P= 0.76

P= 0.00

P= 0.00

Stage 11eve(PJ4)

Mean: 5.3

(n=3)

eve(PJ4)wg(NE2)

Mean: 6.14

(n= 7)

WT

Mean: 14.3

(n= 3)

P= 0.12

P= 0.00

P= 0.00

Mean: 5.89

Mean:5.86

Mean: 5.86

Mean:5.7

Mean: 5.3

Mean: 6.14

P= 0.95 P= 0.76 P= 0.12

eve(PJ4)

Stage 9 Stage 10 Stage 11

eve(PJ4) wg(NE2)

Cuticles

eve(PJ4) eve(PJ4)wg (NE2)

Summary

• Effects of wg(NE2)

- slower decrease in wg expression

- more defined stripes

- increasing deviation of eve(PJ4)wg(NE2) from eve(PJ4)

- double mutant still behaves like an amorph

Discussion

• EVX1, a human homeobox gene homologous to eve

- Transcriptional repressor: specifies A-P axis in embryos

- EVX1 -/- : increased oncogenic potential

Future Directions

• 0-4 hours Ab staining• 0-4 hours and 4-8 hours In Situ

hybridization• eve(1) mutants• Stages 12 to 15 (cuticle secretion)• Sequence eve(PJ4)

Acknowledgements

• Dr. Bejsovec

• Dr. Perz-Edwards

• Wendy Beane

• Monica Zhang

• Our wonderful classmates!

Thank you!

References

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