期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 111, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/53955
关键词
Developmental Biology; Issue 111; Hemodynamics; chicken; outflow tract; embryonic heart; banding; ex ovo; shell-less chick embryo culture
资金
- SPARC Graduate Research Grant from the Office of the Vice President for Research at the University of South Carolina
- Cook Biotech
- FirstString Research Inc
- NIH [2 P20-RR016434-06]
- NIH IDeA Networks of Biomedical Research Excellence (INBRE) grant [P20GM103499]
The new model presented here can be used to understand the influence of hemodynamics on specific cardiac developmental processes, at the cellular and molecular level. To alter intracardiac hemodynamics, fertilized chicken eggs are incubated in a humidified chamber to obtain embryos of the desired stage (HH17). Once this developmental stage is achieved, the embryo is maintained ex ovo and hemodynamics in the embryonic heart are altered by partially constricting the outflow tract (OFT) with a surgical suture at the junction of the OFT and ventricle (OVJ). Control embryos are also cultured ex ovo but are not subjected to the surgical intervention. Banded and control embryos are then incubated in a humidified incubator for the desired period of time, after which 2D ultrasound is employed to analyze the change in blood flow velocity at the OVJ as a result of OFT banding. Once embryos are maintained ex ovo, it is important to ensure adequate hydration in the incubation chamber so as to prevent drying and eventually embryo death. Using this new banded model, it is now possible to perform analyses of changes in the expression of key players involved in valve development and to understand the role of hemodynamics on cellular responses in vivo, which could not be achieved previously.
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