Your SlideShare is downloading. ×
0
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Talk1 Drosophila Talk 205 L
Upcoming SlideShare
Loading in...5
×

Thanks for flagging this SlideShare!

Oops! An error has occurred.

×
Saving this for later? Get the SlideShare app to save on your phone or tablet. Read anywhere, anytime – even offline.
Text the download link to your phone
Standard text messaging rates apply

Talk1 Drosophila Talk 205 L

599

Published on

Presentation of our data and results from the first antibody staining of the new allele, evePJ4 (created by Dr. Amy Bejsovec\'s lab) and the wgNE2 allele.

Presentation of our data and results from the first antibody staining of the new allele, evePJ4 (created by Dr. Amy Bejsovec\'s lab) and the wgNE2 allele.

Published in: Health & Medicine
0 Comments
1 Like
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total Views
599
On Slideshare
0
From Embeds
0
Number of Embeds
2
Actions
Shares
0
Downloads
4
Comments
0
Likes
1
Embeds 0
No embeds

Report content
Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
No notes for slide
  • Transcript

    • 1. Characterizing eve(PJ4) Biology 205L: Experiments in Developmental Biology Fall 2006 Myung Ko, Jasmine Lau, Joshua Mendoza-Elias
    • 2. Gene Regulatory Network Insert Figure 9.26 (Gilberts) runt odd TF eve wg en hh +
    • 3. Consequences of wg expression Jones & Bejsovec (2003)
    • 4. An anomaly: eve(PJ4)wg(NE2) <ul><li>Gastrulation </li></ul><ul><li>-  wingless protein expression </li></ul><ul><li>Cuticle patterning </li></ul><ul><li>-  wingless expression </li></ul>
    • 5. Questions: <ul><li>Is eve(PJ4) an amorph? </li></ul><ul><li>Role of wg(NE2) in double mutant? </li></ul>Hypothesis: <ul><li>eve(PJ4) is an amorph </li></ul><ul><li>wg(NE2) causes wg expression to persist </li></ul>
    • 6. Materials and Methods <ul><li>Statistical Methods:  2-test, t-test </li></ul>Wild-type eve(3) eve(PJ4) eve(PJ4)wg(NE2) AB-staining: anti-wg antibody Stages: 9, 10, 11 Cuticle Preparations
    • 7. Part I: Is eve(PJ4) an amorph?
    • 8. Are we seeing the right ratio? <ul><li>eve (PJ4) </li></ul><ul><li>eve(3) </li></ul> 2 value = 0.333 p < 0.05  2 value= 0.022 p< 0.01 n Ratio 8 17 1 3 Mutant WT n Ratio 16 45 1 3 Mutant WT
    • 9. Stage 9 eve(PJ4) Mean: 5.89 (n=9) eve(3) Mean:6.21 (n=9) WT Mean: 13.2 (n= 5) P= 0.31 P= 0.00 P= 0.00
    • 10. 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
    • 11. 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
    • 12. 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
    • 13. Cuticles eve(PJ4) eve(3)
    • 14. Summary <ul><li>eve(PJ4) behaves like an amorph </li></ul><ul><li>As embryos mature, </li></ul><ul><li>- decreasing # of stripes </li></ul><ul><li>- stripes are less defined </li></ul><ul><li>- increasing similarity b/w eve(PJ4) and eve(3) </li></ul>
    • 15. Part II: Role of wg(NE2) in eve(PJ4)wg(NE2) wg(NE2) to the rescue??
    • 16. Are we seeing the right ratio? <ul><li>eve(PJ4) </li></ul><ul><li>eve(PJ4)wg(NE2) </li></ul> 2 value = 0.33 p < 0.05  2 value= 0.89 p < 0.01 n Ratio 8 17 1 3 Mutant WT 18 2 +/- n Ratio 10 8 1 1 Mutant WT
    • 17. 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
    • 18. 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
    • 19. Stage 11 eve(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
    • 20. 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)
    • 21. Cuticles eve(PJ4) eve(PJ4)wg (NE2)
    • 22. Summary <ul><li>Effects of wg(NE2) </li></ul><ul><li>- slower decrease in wg expression </li></ul><ul><li>- more defined stripes </li></ul><ul><li>- increasing deviation of eve(PJ4)wg(NE2) from eve(PJ4) </li></ul><ul><li>- double mutant still behaves like an amorph </li></ul>
    • 23. Discussion <ul><li>EVX1, a human homeobox gene homologous to eve </li></ul><ul><li>- Transcriptional repressor: specifies A-P axis in embryos </li></ul><ul><li>- EVX1 -/- : increased oncogenic potential </li></ul>
    • 24. Future Directions <ul><li>0-4 hours Ab staining </li></ul><ul><li>0-4 hours and 4-8 hours In Situ hybridization </li></ul><ul><li>eve(1) mutants </li></ul><ul><li>Stages 12 to 15 (cuticle secretion) </li></ul><ul><li>Sequence eve(PJ4) </li></ul>
    • 25. Acknowledgements <ul><li>Dr. Bejsovec </li></ul><ul><li>Dr. Perz-Edwards </li></ul><ul><li>Wendy Beane </li></ul><ul><li>Monica Zhang </li></ul><ul><li>Our wonderful classmates! </li></ul>
    • 26. Thank you!
    • 27. References <ul><li>Paul M. Mcdonald, Phil Ingham and Gary Struhi. Isolation, Structure, and Expression of even-skipped: A Second Pair-Rule Gene of Drosophila Containing a Homeo Box . Cell: Vol. 47, 721-734 (1986) </li></ul><ul><li>Marcel van den Heuvel, Roel Nusse, Paul Johnston, and Peter A. Lawrence. Distribution of the wingless Gene Product in Drosophila Embryos: A Protein Involved in Cell-Cell Communication . Cell: Vol. 59, 739-749 (1989) </li></ul><ul><li>Herman A. Dierick and Amy Bejsovec. Functional analysis of Wingless reveals a link between intercellular ligand transport and dorsal-cell-specific signaling. Development: 125, 4729-4738 (1998) </li></ul><ul><li>4. Whitney M Jones and Amy Bejsovec. Wingless Signaling: An Axin to Grind. Current Biology: Vol.13, R479-R481 (2003) </li></ul>
    • 28. <ul><li>5. Antonio Faiella, Maurizio D'Esposito, Marco Rambaldi, Dario Acampora, Silvia Balsofiore, Anna Stornaiuolo, Antonio Mallamaci, Enrica Migliaccio, Massimo Gulisano, Antonio Simeone and Edoardo Boncinelli. Isolation and mapping of EVX1, a human homeobox gene homologous to even-skipped, localized at the 5' end ofHOXi locus on chromosome 7. Nucleic Acids Research: Vol. 19, No. 23 6541-6545 (1991). </li></ul><ul><li>6. Chi Li and James Manley. Even-skipped Represses Transcription by Binding TATA Binding Protein and Blocking the TFIID-TATA Box Interaction . Molecular and Cellular Biology: Vol. 18, No.7, p. 3771–3781 (1998). </li></ul><ul><li>7. Paola Briata, Rinke Van De Werken, Irma Airoldi, Cristina Ilengo, Erica Di Blas, Edoardo Boncinelli, and Giorgio Corte. Transcriptional Repression by the Human Homeobox Protein EVX1 in Transfected Mammalian Cells. The Journal of Biological Chemistry: Vol . 270, No. 46, pp. 27695–27701, (1995). </li></ul>

    ×