Development What causes cell differentiation? Molecular Markers.
-
date post
19-Dec-2015 -
Category
Documents
-
view
218 -
download
1
Transcript of Development What causes cell differentiation? Molecular Markers.
![Page 1: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/1.jpg)
![Page 2: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/2.jpg)
Development
What causes cell differentiation?
![Page 3: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/3.jpg)
Molecular Markers
![Page 4: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/4.jpg)
Homeobox Genes
1. Transcription factors encoding the DNA binding homeodomain.
2. Expressed in temporal and spatial specific patterns during development.
3. Cell differentiation is disrupted when they are mutated.
![Page 5: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/5.jpg)
Adult Spinal Cord
![Page 6: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/6.jpg)
Neural Tube Development
How are the expression patterns for homeodomain proteins created?
![Page 7: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/7.jpg)
Lbx1 Expression Pattern
Pax3•Lbx1
![Page 8: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/8.jpg)
Project Goal
Identify the tissue specific enhancer that creates the Lbx1 expression pattern in the dorsal horn of the spinal cord.
![Page 9: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/9.jpg)
Mouse Lbx1 Locus
![Page 10: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/10.jpg)
Delivery of Reporter Constructs to Embryonic Cells
• Different experimental models can be used.
– Mouse common but difficult, expensive.
– Chick easy, inexpensive.
• Our method of choice: electroporation of chick embryos.
• Method is relatively new.
![Page 11: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/11.jpg)
In Ovo Electroporation: GFPß-actin
EnhancerGFP
![Page 12: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/12.jpg)
Instruments
![Page 13: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/13.jpg)
Positive Control: ß-Actin GFP Expression Vector
![Page 14: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/14.jpg)
Asymmetric Expression of GFP
![Page 15: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/15.jpg)
In Ovo Electroporation: LacZCytomeglavirus
EnhancerLacZ
![Page 16: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/16.jpg)
Positive Control: ß-Galactosidase Reporter Gene
![Page 17: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/17.jpg)
Results
![Page 18: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/18.jpg)
Conclusion
• Tissue specific enhancer does not exist within our search area.
• Can now start searching upstream and downstream.
![Page 19: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/19.jpg)
Acknowledgements
Dr. Michael Gross
Dr. Kevin Ahern
Howard Hughes Medical Institute
![Page 20: Development What causes cell differentiation? Molecular Markers.](https://reader036.fdocuments.net/reader036/viewer/2022062516/56649d3e5503460f94a17d01/html5/thumbnails/20.jpg)
Questions?