FIGURE SUMMARY
Title

Computational and functional analysis of biopharmaceutical drugs in zebrafish: erythropoietin as a test model

Authors
Guarienti, M., Giacopuzzi, E., Gianoncelli, A., Sigala, S., Spano, P., Pecorelli, S., Pani, L., Memo, M.
Source
Full text @ Pharmacol. Res.
Fig. 4

Negative control (Neg ctrl), Eprex®, Binocrit® or Retacrit® injected in 48 hpf tg (kdrl:EGFP; gata1:ds-red) casper embryos and photographed after 4 h. (A) Double channel fluorescence: in green blood vessels, expressing kdrl:EGFP and in red circulating erythrocytes, expressing gata1:ds-red. Lateral views, anterior to the left, 11.5X magnification; (B) single red-channel fluorescence shows circulating erythrocytes, expressing gata1:ds-red. Lateral views, anterior to the left, 4X magnification. Quantification of (C) red fluorescence signal and (D) green fluorescence signal, measured with ImageJ 1.45 s image analysis software on a mean of 25 embryos for each experimental point. Asterisk indicates statistically significant increase of the positive area (p < 0.05), data are the mean ± S.D. of 3 independent experiments. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Fig. 5

O-dianisidine staining and Drabkin assay quantification of 48 hpf wild type AB embryos injected with negative control (Neg ctrl), Eprex®, Binocrit® or Retacrit®. (A) Representative images of embryos stained with o-dianisidine 4 h after treatment. Lateral views, anterior to the left, 4X magnification; (B) quantification of o-dianisidine positive area in the region of trunk and tail, measured with ImageJ 1.45 s image analysis software on a mean of 25 embryos for each experimental point. Asterisk indicates statistically significant increase of the positive area (p < 0.05), data are the mean ± S.D. of 3 measurements; (C) cyanomethemoglobin absorbance measured at 540 nm, proportional to the amount of red blood cells in groups of 20 embryos. Asterisk indicates statistically significant increase of absorbance (p < 0.05), data are the mean ± S.D. of 3 measurements.
Fig. 6

In situ hybridization with pu1, lplastin and mpx probes to detect leukocytes precursors, monocytes/macrophages and granulocytes neutrophils, respectively. (A) Representative images of 72 hpf embryos injected into the otic capsule with Eprex®, Binocrit® or Retacrit® and positive or negative control (Pos ctrl and Neg ctrl, respectively). Lateral views, anterior to the left, 11.5X magnification; (B) quantification of pu1, lplastin and mpx positive area, measured with ImageJ 1.45 s image analysis software on a mean of 40 embryos for each experimental point. Asterisk indicates statistically significant increase of the positive area (p < 0.05), data are the mean ± S.D. of 3 measurements.
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Pharmacol. Res.
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