1 / 34

10,000,000,000 cells 7,000,000,000 people

10,000,000,000 cells 7,000,000,000 people. 1984 Conservation of developmental mechanisms. Transcription factors. Downstream effectors. Pax6. 5’UTR Paired linker Homeo PST 3’UTR

kaylee
Télécharger la présentation

10,000,000,000 cells 7,000,000,000 people

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. 10,000,000,000 cells 7,000,000,000 people

  2. 1984 Conservation of developmental mechanisms

  3. Transcription factors Downstream effectors

  4. Pax6 5’UTR Paired linker Homeo PST 3’UTR domain domain transactivation domain Alternatively spliced exon 5a

  5. Pax6 Paired domain Homeo domain

  6. Mutant Control Mutant Mutant Sisodya et al Nature Genetics 2001

  7. Pax6 gene expression in the developing cortex Caudal Medial Rostral Lateral

  8. Wild-type Mutant

  9. Pax6 expression reporter FACS sorted Pax6-expressing cells

  10. Controls Pax6-/- control -/- +/+ No -/- +/+ Collaboration with Pathway Medicine Mutant -/- +/+

  11. Mi et al 2013 Neuron

  12. Martine Manuel Mike Molinek Da Mi John Mason New Study Delete Pax6 specifically in the cortex at a chosen time Look for changes in gene expression that occur rapidly after loss of Pax6 Primary defects Direct targets

  13. Pax6flox allele Ian Simpson Conditional EGFP reporter loxP loxP Stop EGFP Cre EGFP

  14. Emx1-CreERT2 CreERT2 OFF CreER Tamoxifen CreER ON

  15. Give tamoxifen to pregnant females at E9.5 Collect embryos at E11.5, E12.5 and E13.5 Pax6+/flox Emx1creER Pax6flox/flox Emx1creER control Pax6 cKO Dissect cortex Dissect cortex Extract RNA Extract RNA Deep Sequencing Deep Sequencing

  16. Tamoxifen given at E9.5 E11.5 E12.5 E13.5

  17. E12.5 GFP / Pax6 immuno control Pax6 cKO control Pax6 cKO

  18. E12.5 control Pax6 cKO

  19. E12.5 control1 Pax6cKO1 control2 Pax6cKO2 C R C R C R C R Pax6 bactin

  20. Tamoxifen given at E9.5 E9.5 E11.5 E12.5 E13.5

  21. Tamoxifen given at E9.5 * * E11.5 E12.5 E13.5 E11.5 E12.5 E13.5 E9.5 * E11.5 E12.5 E13.5 E9.5 E11.5 E12.5 E13.5

  22. Tamoxifen given at E9.5 * E11.5 E12.5 E13.5 E9.5 E11.5 E12.5 E13.5 E9.5 * E11.5 E12.5 E13.5 E9.5

  23. Tamoxifen given at E9.5 E9.5 E11.5 E12.5 E13.5 E9.5 E11.5 E12.5 E13.5 Also no significant change in Smad4, Cdk4, cyclin D2, MCM3 and MCM6 expression.

  24. Tamoxifen given at E9.5 R CeM CeL C Caudal Rostral E12.5

  25. Collection of samples for RNAseq Rostral cortex Caudal cortex Pax6 immuno RNA extraction

  26. controls Pax6 cKO Collaboration with Genepool Control Pax6cKO Control Pax6cKO Control Pax6cKO

  27. E13.5 control Pax6 cKO

  28. Pax6 immuno. E13.5. Sagittal. Rostral Rostral Control Pax6 cKO

  29. E13.5 control Pax6cKO C R C R Pax6 bactin

More Related