New methods for chicken embryo manipulations
Abstract
The capacity to image a growing embryo while simultaneously studying the developmental function of specific molecules provides invaluable information on embryogenesis. However, until recently, this approach was accomplished with difficulty both because of the advanced technology needed and because an easy method of minimizing damage to the embryo was unavailable. Here, we present a novel way of adapting the well-known EC culture of whole chick embryos to time-lapse imaging and to functional molecular studies using blocking agents. The novelty of our method stems from the ability to apply blocking agents ex ovo as well as in ovo. We were able to study the function of a set of molecules by culturing developing embryos ex ovo in tissue culture media containing these molecules or by injecting them underneath the live embryo in ovo. The in ovo preparation is particularly valuable, because it extends the period of time during which the developmental function of the molecule can be studied and it provides an easy, reproducible method for screening a batch of molecules. These new techniques will prove very helpful in visualizing and understanding the role of specific molecules during embryonic morphogenesis, including blood vessel formation.
Additional Information
© 2009 Wiley-Liss, Inc. Received 8 April 2009; accepted in revised form 27 May 2009. Published online 6 July 2009. Contract grant sponsor: NIH; Contract grant numbers: 3R37-036585 and 3R37-036585-06S1; Contract grant sponsor: NIH/NINDS AREA ; Contract grant number: 1R15-NS060099-01; Contract grant sponsor: NIH-MBRS; Contract grant number: SCORE-5S06GM048680-13. The authors thank Helen McBride for encouraging comments during the conception of this work and Cindy Malone for reviewing it. The authors thank the members of the Bronner-Fraser and Fraser lab who tried our technique successfully, confirming its reproducibility. The authors thank Scott E. Fraser and Kathryn McCabe for suggestions to improve this technique.Attached Files
Published - ElGhali2010p6982Microsc_Res_Techniq.pdf
Supplemental Material - sm001.avi
Supplemental Material - sm002.avi
Supplemental Material - sm003.tiff
Files
Name | Size | Download all |
---|---|---|
md5:53afb4e3dcba73b04acac366cdc4bba7
|
284.5 kB | Preview Download |
md5:2d5652b273b6c65743da1460d2e2f0db
|
15.3 kB | Preview Download |
md5:d73ed3f8c61f0f6479648931a9013566
|
101.0 kB | Preview Download |
md5:c046ab82e7c939f5a09905245b578180
|
1.1 MB | Download |
md5:114c9f1e8e9e4451dd8b145744f43101
|
4.4 MB | Preview Download |
md5:df6d4528f4d856f92f567257e9bb56c1
|
10.7 MB | Download |
Additional details
- Eprint ID
- 17438
- Resolver ID
- CaltechAUTHORS:20100210-090324146
- 31137-036585
- NIH
- 3R37-036585-06S1
- NIH
- 1R15-NS060099-01
- NIH National Institute of Neurological Disorders and Stroke (NINDS)
- SCORE-5SOE;GM048680-13
- NIH Minority Biomedical Research Support (MBRS)
- Created
-
2010-02-16Created from EPrint's datestamp field
- Updated
-
2021-11-08Created from EPrint's last_modified field