Biomedical Engineering Reference
In-Depth Information
Celebrex
Genistein
120
120
**
100
100
*
80
80
60
60
40
40
20
20
0
0
0
0.0001
0.001
1.01
-20
10
20
30
40
-20
-40
Concentration (
μ
m)
Concentration (
μ
m)
-40
SU5416
120
**
100
80
60
40
20
0
-20
0.5
1
1.5
2
2.5
-40
Concentration (
μ
m)
Figure 16.4
Drug effects on CNV in CoCl
2
-treated zebrafish. Onedpf zebrafish were co-treated
with 0.1mg/mLCoCl
2
and varying drug concentrations for 4 days. At 5dpf, zebrafishwere processed for
the CNV ELISA, as described in Section 16.2. Each point represents meanSE (
n
¼5-18).
indicates
p
<0.1 and
indicates
p
<0.05 by ANOVA.
concern, using immunohistochemistry, we examined zebrafish eye structure after
Celebrex treatment. Normal 5dpf zebrafish exhibit a distinctive five-layer retinal
structure with no vessels. In contrast, 0.1mg/mL CoCl
2
-treated 5dpf zebrafish
exhibited disrupted inner nuclear layer, indicating degenerated retinal structure, and
abnormal vessel structure. Five dpf zebrafish co-treated with CoCl
2
and DMSO
showed degenerated retinal structure, newly formed vessels invading the retina from
the choroidal layer, and a degenerated photoreceptor layer, indicating that DMSO
alone had no effect on CoCl
2
-induced CNV. Five dpf zebrafish co-treated with CoCl
2
and Celebrex exhibited a normal five-layer retina with no vessels present (data
not shown). These results clearly demonstrate that Celebrex treatment inhibited
CoCl
2
-induced CNV phenotypes.
16.4 DISCUSSION
16.4.1 CNV Zebrafish Model Is Specific for Retinal
and Choroidal Neoangiogenesis
Using a low CoCl
2
concentration (0.1mg/mL), we generated a chemical induced
zebrafish CNV model. As evidenced by overall morphology (data not shown), CoCl
2
treatment did not inhibit zebrafish development. Although a stimulatory effect on
angiogenesis was present throughout zebrafish, the most striking effect of CoCl
2
treatment was the presence of a network of abnormal small vessels in the CVP, often
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