Generating Transparent Zebrafish: A Refined Method to Improve Detection of Gene Expression During Embryonic Development
Abstract
In zebrafish (Danio rerio) pigmentation is initiated during embryogenesis and begins in the retinal epithelium and in the melanophores. The pigment cells develop rapidly, and within hours they constitute a prominent feature of the embryo. In order to improve signal detection by whole mount in situ hybridization, confocal microscopy, or expression of GFP, embryos may be treated with 1-phenyl 2-thiourea (PTU) during embryogenesis. PTU inhibits melanogenesis by blocking all tyrosinase-dependent steps in the melanin pathway but can be toxic at high concentrations. The embryos remain transparent as long as the PTU treatment is continued. However, PTU treatment must be initiated before the initial pigmentation because it does not remove already formed pigment. Here we provide a protocol for generating transparent zebrafish while avoiding the toxic and teratogenic effects of PTU treatment.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
To date, none of the typical strategies to improve zebrafish larval transparency produce normally behaving larvae that are transparent in both the body and the eyes.
“To date none of these strategies produce normally behaving larvae that are transparent in both the body and the eyes.”
From A crystal-clear zebrafish for in vivo imaging (2016) · cites this paper as [8]
The PTU strategy is associated with toxic side effects that impair morphogenesis, behaviour and survival.
“The first strategy, despite being straightforward (i.e., embryos can be simply raised in medium containing 200 μM PTU), is associated with toxic side effects that impair morphogenesis, behaviour and survival.”
From A crystal-clear zebrafish for in vivo imaging (2016) · cites this paper as [8]
Common strategies to reduce zebrafish skin pigmentation can be grouped into two categories: chemical compounds interfering with melanin synthesis, most popularly PTU, and pigmentation mutants.
“Common strategies to reduce zebrafish skin pigmentation can be grouped in two categories: 1) use of chemical compounds interfering with the synthesis of melanin, the most popular one being 1-phenyl-2-thiourea (PTU)8; 2) use of pigmentation mutants in which genes controlling either pigment formation, chromatophore formation or interactions between chromatophores have been inactivated469.”
From A crystal-clear zebrafish for in vivo imaging (2016) · cites this paper as [8]
The optical clarity of crystal larvae is superior to that of larvae treated with 200 μM PTU, because PTU inhibits melanin production but does not interfere with iridophore function.
“Importantly, the optical clarity of crystal larvae is even superior to that of larvae treated with 200 μM PTU (Fig. 1c,d), since PTU inhibits melanin production but does not interfere with iridophore function8.”
From A crystal-clear zebrafish for in vivo imaging (2016) · cites this paper as [8]
Cited by
Sorted by citations
Zebrafish Differentially Process Color across Visual Space to Match Natural Scenes
Zimmermann, M. J. Y., Nevala, N. E., Yoshimatsu, T., Osorio, D., Nilsson, D.-E., Berens, P., & Baden, T. (2018) · Current Biology · doi:10.1016/j.cub.2018.04.075 · Cited by 227 · see citing works
A crystal-clear zebrafish for in vivo imaging
Antinucci, P., & Hindges, R. (2016) · Scientific Reports · doi:10.1038/srep29490 · Cited by 165 · see citing works
Vopalensky, P., Pergner, J., Liegertova, M., Benito-Gutierrez, E., Arendt, D., & Kozmik, Z. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1207580109 · Cited by 129 · see citing works
- Authors
- Karlsson, J., von Hofsten, J., & Olsson, P.-E.
- Affiliations
- Umeå University (SE)
- ORCID
- Olsson, Per-Erik
- Venue
- Marine Biotechnology
- Published
- 2001-11-01
- Volume
- 3
- Issue
- 6
- Pages
- 0522-0527
- ISSN
- 1436-2228 (ISSN-L)
- OpenAlex
- W2050087790
- Cited by
- 328
- References
- 0
- Citations by year
- 2026: 16 · 2025: 19 · 2024: 24 · 2023: 17 · 2022: 19 · 2021: 26 · 2020: 22 · 2019: 19 · 2018: 22 · 2017: 25 · 2016: 26 · 2015: 15 · 2014: 10 · 2013: 13 · 2012: 11
- FWCI
- 0.32
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Marine Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Zebrafish Biomedical Research Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › melanin and skin pigmentation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- zebrafish, propylthiouracil, melanogenesis inhibition, pigment cell development, melanophores, gene expression detection, teratogenicity, tyrosinase inhibition
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
