Biomedical Engineering Reference
In-Depth Information
Chapter
16
Development of a
Hypoxia-Induced Zebrafish
Choroidal Neovascularization
Model
Wen Lin Seng, Yingxin Lin, Susie Tang, and Lisa Zhong
Phylonix, Cambridge, MA, USA
16.1 INTRODUCTION
In this research, using CoCl
2
, a hypoxia mimetic, we describe development of a
zebrafish choroidal neovascularization (CNV) model for drug screening. We dem-
onstrate that abnormal CNV can be observed 4 days after adding CoCl
2
directly to the
fish water, facilitating rapid screening of compound libraries. To eliminate nonspe-
cific staining from angiogenesis present in other sites, we describe methods for
removing intact eyes fromwhole zebrafish using collagenase enzyme treatment. This
method facilitates both visual assessment of CNV, which occurs in the back of the eye,
and quantitative assessment of compound effects on isolated eyes using a compar-
atively high-throughput microplate format. This convenient model will facilitate drug
discovery and assist in elucidating the pathogenesis of choroidal neovascularization.
16.1.1 Hypoxia and Ocular Neovascularization
Oxygen homeostasis plays a crucial role in supporting normal development and
physiology in all animals. Pathological conditions can develop if the balance between
oxidative phosphorylation and oxidative stress is disrupted. In eye diseases, hypoxia
has been shown to be a major mechanism of abnormal neovascularization (NV)
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