Angiogenesis in chicken chorioallantoic membrane under hypoxic incubations

in Animal Biology
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Under hypoxic conditions, angiogenesis in the chorioallantoic membrane exhibits a sensitive response depending on the developmental timing and intensity and duration of the hypoxia. Furthermore, the effects of hypoxia on vascularization in the chorioallantoic membrane are controversial. In this study, we used microscopy to determine the vascular density index of chicken embryonic chorioallantoic membrane and quantitative PCR to examine the expression of the HIF-1α and VEGFA genes. Two levels of hypoxia (15% and 13% O2) and three durations (days 0–10, 11–18, and 0–18) were applied. The results showed that 13% O2 incubation restrained angiogenesis in the chorioallantoic membrane at an early stage of embryonic development as seen on day 6, but incubation under 13% or 15% O2 efficiently stimulated vascularization of the chorioallantoic membrane as seen on days 16 and 18. Notably, 13% O2 incubation caused visible curling of the blood vessels in the chorioallantoic membrane on day 18. The increased vascular density index under hypoxia was accompanied by an increase in the expression of VEGFA and HIF-1α. Curled blood vessels were observed on day 18 under 13% O2 incubation; however, the curling was not a result of VEGFA overexpression. Thus, the present study helps in elucidating vascularization of the chorioallantoic membrane under hypoxic conditions.

Angiogenesis in chicken chorioallantoic membrane under hypoxic incubations

in Animal Biology



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    Vascularity of the CAM during incubation. Panels A to T are representative photomicrographs for days 6, 10, 14, and 18 of incubation, showing CAMs that had been exposed to various hypoxic conditions; panels U and V show quantitative comparisons of VDI under O2 21%, O2 15%, and O2 13% (U) versus O2 21%, Early O2 15%, Late O2 15%, Early O2 13%, and Late O2 13% (V). In panels U and V, all values are represented as the mean ± standard error. Values labeled with the same letter (a, b, c) do not differ significantly; values labeled with different letters show significant differences at P<0.05. The heavy dashed line indicates the control group. CAM, chorioallantoic membrane; VDI, vascular density index.

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    Expression of the HIF-1α (A) and VEGFA (B) genes in the CAM. Values labeled with the same letter (a, b, c) do not differ significantly; those labeled with different letters show significant differences (P<0.05). CAM, chorioallantoic membrane; HIF-1α, hypoxia inducible factor-1α; VEGFA, vascular endothelial growth factor A.


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