Abstract
Pigment cells in fish species play crucial roles in forming colour patterns of each species and other physiological characteristics, including photoprotection. Research on animal pigment cells’ photoprotection has primarily concentrated on black pigment cells, known as melanophores. However, the roles of other pigment cells on UV protection remain poorly understood. In addition, UV sensitivity/resistance mechanisms have been studied in a small number of selected model organisms, and difference of UV sensitivity has not been fully investigated in different species. In this study, we use the Arabian killifish embryos as a novel model for studying the mechanisms of UV protection by different pigment cells. This species features highly fluorescent pigment cells called fluoroleucophores and black pigment cells melanophores. The fluorescent pteridine pigments and black melanin pigments are generated by genes gch (GTP cyclohydrolase) and tyr (tyrosinase), respectively. We generated gch-/- and gch-/-tyr-/- double mutant lines using CRISPR/Cas9 genome editing and examined the UV sensitivity of WT and these mutant embryos. Firstly, we have observed that Arabian killifish embryos showed higher UV resistance to the one in zebrafish from which majority of UV toxicity studies was conducted in fish species so far. Secondly, both morphology and gene expression data revealed that the gch-/-tyr-/- double mutant line exhibited the highest UV sensitivity, and the gch-/- line also demonstrated a greater stress response compared to wild type (WT). The study revealed a diversity of UV resistance between fish species and also identified the roles of black and fluorescent pigment cells in providing effective UV protection from the early stages of embryonic development.
Data availability
The datasets generated during the current study are available in the Genbank (accession numbers, PQ588425 for *gch* and PQ588426 for *tyr, * respectively).
References
Maan, M. E. & Sefc, K. M. Colour variation in cichlid fish: developmental mechanisms, selective pressures and evolutionary consequences. Semin Cell. Dev. Biol. 24, 516–528. https://doi.org/10.1016/j.semcdb.2013.05.003 (2013).
Miyamoto, K. Effects of body color luminance and behavioral characteristics on predation risk in salmonid fishes. Hydrobiologia 783, 249–256. https://doi.org/10.1007/s10750-015-2573-x (2016).
Hoglund, E., Balm, P. H. & Winberg, S. Behavioural and neuroendocrine effects of environmental background colour and social interaction in Arctic Charr (Salvelinus alpinus). J. Exp. Biol. 205, 2535–2543. https://doi.org/10.1242/jeb.205.16.2535 (2002).
Kelsh, R. N. Genetics and evolution of pigment patterns in fish. Pigment Cell. Res. 17, 326–336. https://doi.org/10.1111/j.1600-0749.2004.00174.x (2004).
Parichy, D. M. & Spiewak, J. E. Origins of adult pigmentation: diversity in pigment stem cell lineages and implications for pattern evolution. Pigment Cell. Melanoma Res. 28, 31–50. https://doi.org/10.1111/pcmr.12332 (2015).
Fujii, R. Cytophysiology of fish chromatophores. Int. Rev. Cytol. 143, 191–255 (1993).
Sugimoto, M. Morphological color changes in fish: regulation of pigment cell density and morphology. Microsc Res. Tech. 58, 496–503. https://doi.org/10.1002/jemt.10168 (2002).
Hirata, M., Nakamura, K., Kanemaru, T., Shibata, Y. & Kondo, S. Pigment cell organization in the hypodermis of zebrafish. Dev. Dyn. 227, 497–503. https://doi.org/10.1002/dvdy.10334 (2003).
Parichy, D. M. Evolution of pigment cells and patterns: recent insights from teleost fishes. Curr. Opin. Genet. Dev. 69, 88–96. https://doi.org/10.1016/j.gde.2021.02.006 (2021).
Fujii, R. The regulation of motile activity in fish chromatophores. Pigment Cell. Res. 13, 300–319. https://doi.org/10.1034/j.1600-0749.2000.130502.x (2000).
Frohnhofer, H. G., Krauss, J., Maischein, H. M. & Nusslein-Volhard, C. Iridophores and their interactions with other chromatophores are required for stripe formation in zebrafish. Development 140, 2997–3007. https://doi.org/10.1242/dev.096719 (2013).
Kimura, T. et al. Leucophores are similar to xanthophores in their specification and differentiation processes in Medaka. P Natl. Acad. Sci. USA. 111, 7343–7348. https://doi.org/10.1073/pnas.1311254111 (2014).
Hamied, A. et al. Identification and characterization of highly fluorescent pigment cells in embryos of the Arabian killifish (Aphanius dispar). iScience 23, 101674. https://doi.org/10.1016/j.isci.2020.101674 (2020).
Frenkel, V. & Goren, M. Factors affecting growth of killifish, Aphanius dispar, a potential biological control of mosquitoes. Aquaculture 184, 255–265. https://doi.org/10.1016/S0044-8486(99)00326-9 (2000).
Esmaeili, H. R., Asrar, T. & Gholamifard, A. Cyprinodontid fishes of the world: an updated list of taxonomy, distribution and conservation status (Teleostei: Cyprinodontoidea). Iran. J. Ichthyol. 5, 1–29 (2018).
Plaut, I. Resting metabolic rate, critical swimming speed, and routine activity of the Euryhaline cyprinodontid, Aphanius dispar, acclimated to a wide range of salinities. Physiol. Biochem. Zool. 73, 590–596. https://doi.org/10.1086/317746 (2000).
Akbarzadeh, A. et al. Responses of the killifish (Aphanius dispar) to long-term exposure to elevated temperatures: growth, survival and microstructure of gill and heart tissues. Mar. Freshw. Behav. Physiol. 47, 429–434 (2014).
Alsakran, A. et al. Stage-by-stage exploration of normal embryonic development in the Arabian killifish, Aphanius dispar. Dev. Dyn. https://doi.org/10.1002/dvdy.738 (2024).
Madronich, S., McKenzie, R. L., Björn, L. O. & Caldwell, M. M. Changes in biologically active ultraviolet radiation reaching the earth’s surface. J. Photoch Photobio B. 46, 5–19. https://doi.org/10.1016/S1011-1344(98)00182-1 (1998).
McKenzie, R. L., Aucamp, P. J., Bais, A. F., Bjorn, L. O. & Ilyas, M. Changes in biologically-active ultraviolet radiation reaching the earth’s surface. Photochem. Photobiol Sci. 6, 218–231. https://doi.org/10.1039/b700017k (2007).
El-Nouby, A. M. Effect of stratospheric Ozone in UVB solar radiation reaching the earth’s surface at Qena, Egypt. Atmos. Pollut. Res. 1, 155–160 (2010).
Diaz, S. et al. Ozone and UV radiation over Southern South america: climatology and anomalies. Photochem. Photobiol. 82, 834–843. https://doi.org/10.1562/2005-09-26-RA-697 (2006).
Dong, Q., Svoboda, K., Tiersch, T. R. & Monroe, W. T. Photobiological effects of UVA and UVB light in zebrafish embryos: evidence for a competent photorepair system. J. Photochem. Photobiol B. 88, 137–146. https://doi.org/10.1016/j.jphotobiol.2007.07.002 (2007).
Mahmoud, U. M. & Mekkawy, I. A. Sayed Ael, D. Ultraviolet radiation-A (366 nm) induced morphological and histological malformations during embryogenesis of Clarias gariepinus (Burchell, 1822). J. Photochem. Photobiol B 95, 117–128. https://doi.org/10.1016/j.jphotobiol.2009.02.003 (2009).
Torres Nunez, E. et al. Molecular response to ultraviolet radiation exposure in fish embryos: implications for survival and morphological development. Photochem. Photobiol. 88, 701–707. https://doi.org/10.1111/j.1751-1097.2012.01088.x (2012).
Zang, L., Shimada, Y., Miyake, H. & Nishimura, N. Transcriptome analysis of molecular response to UVC irradiation in zebrafish embryos. Ecotoxicol. Environ. Saf. 231, 113211. https://doi.org/10.1016/j.ecoenv.2022.113211 (2022).
Lai, Y. S., Chiue, L. F. & Hsu, T. Low-molecular-weight vitellogenin 1-like proteins are components of a UV-damaged-DNA binding activity highly expressed in zebrafish (Danio rerio) embryos. J. Exp. Zool. Comp. Exp. Biol. 305, 215–224. https://doi.org/10.1002/jez.a.222 (2006).
Strahle, U. & Jesuthasan, S. Ultraviolet irradiation impairs epiboly in zebrafish embryos: evidence for a microtubule-dependent mechanism of epiboly. Development 119, 909–919. https://doi.org/10.1242/dev.119.3.909 (1993).
Rice, M. C. et al. Gadusol is a maternally provided sunscreen that protects fish embryos from DNA damage. Curr Biol 33, 3229-3237.e3224. https://doi.org/10.1016/j.cub.2023.06.012 (2023).
Gavriouchkina, D. et al. Thyrotroph embryonic factor regulates light-induced transcription of repair genes in zebrafish embryonic cells. PLoS One. 5, e12542. https://doi.org/10.1371/journal.pone.0012542 (2010).
Costa, S. R., Velasques, R. R., Hoff, M. L. M., Souza, M. M. & Sandrini, J. Z. Characterization of different DNA repair pathways in hepatic cells of zebrafish (Danio rerio). DNA Repair. (Amst). 83, 102695. https://doi.org/10.1016/j.dnarep.2019.102695 (2019).
Winter, M. J. et al. A combined human in Silico and CRISPR/Cas9-Mediated in vivo zebrafish based approach to provide phenotypic data for supporting early target validation. Front. Pharmacol. 13, 827686. https://doi.org/10.3389/fphar.2022.827686 (2022).
Gag, O. et al. UVA/UVB irradiation exerts a distinct phototoxic effect on human keratinocytes compared to human malignant melanoma cells. Life (Basel) https://doi.org/10.3390/life13051144 (2023).
Shih, T. L., Hsiao, C. A., Lin, Z. Y. & Chen, Y. H. An alternative synthesis of 3’,4’-diaminoflavones to evaluate their antioxidant ability and cell apoptosis of zebrafish larvae. Molecules 17, 8206–8216. https://doi.org/10.3390/molecules17078206 (2012).
Chen, R. Y. et al. UVB irradiation induced cell damage and early onset of Junbb expression in zebrafish. Animals (Basel) https://doi.org/10.3390/ani10061096 (2020).
Ojeda, F. et al. Ultraviolet exposure of thymocytes: selective Inhibition of apoptosis. Int. J. Radiat. Biol. 80, 445–450. https://doi.org/10.1080/09553000410001702328 (2004).
Winckler, K. & Fidhiany, L. Significant influence of UVA on the general metabolism in the growing cichlid fish, Cichlasoma nigrofasciatum. J. Photochem. Photobiol., B. 33, 131–135. https://doi.org/10.1016/1011-1344(95)07238-1 (1996).
Svitačová, K., Slavík, O. & Horký, P. Pigmentation potentially influences fish welfare in aquaculture. Appl. Anim. Behav. Sci. 262, 105903. https://doi.org/10.1016/j.applanim.2023.105903 (2023).
Funayama, T., Mitani, H. & Shima, A. Ultraviolet-induced dna damage and its photorepair in tail fin cells of the Medaka, Oryzias lutipes. Photochem. Photobiol. 58, 380–385 (1993).
Fukunishi, Y. et al. Comparison of UV-B tolerance between wild-type and albino Japanese flounder Paralichthys Olivaceus juveniles. Aquaculture Sci. 65, 149–152 (2017).
Aspengren, S., Hedberg, D., Sköld, H. N. & Wallin, M. New insights into melanosome transport in vertebrate pigment cells. Int. Rev. cell. Mol. Biology. 272, 245–302 (2008).
Nilsson Skold, H., Aspengren, S. & Wallin, M. Rapid color change in fish and amphibians: function, regulation, and emerging applications. Pigment Cell. Melanoma Res. 26, 29–38. https://doi.org/10.1111/pcmr.12040 (2013).
Arpigny, J. et al. Factors influencing motile activities of fish chromatophores. Adv. Comp. Environ. Physiology: Volume. 20, 1–54 (1994).
Sayed, A. E. D. H. & Mitani, H. Immunostaining of UVA-induced DNA damage in erythrocytes of Medaka (Oryzias latipes). J. Photochem. Photobiol., B 171, 90–95 (2017).
Wagner, J. T. & Podrabsky, J. E. Extreme tolerance and developmental buffering of UV-C induced DNA damage in embryos of the annual killifish, Austrofundulus limnaeus. J. Exp. Zool. Ecol. Genet. Physiol. 323, 10–30. https://doi.org/10.1002/jez.1890 (2015).
Dutto, I., Tillhon, M., Cazzalini, O., Stivala, L. A. & Prosperi, E. Biology of the cell cycle inhibitor p21(CDKN1A): molecular mechanisms and relevance in chemical toxicology. Arch. Toxicol. 89, 155–178. https://doi.org/10.1007/s00204-014-1430-4 (2015).
Acknowledgements
We also thank to Aquatic Resource Facility staff for fish husbandry, and Aaron Geffery for his support.
Author information
Authors and Affiliations
Contributions
MA generated the resource and data, and wrote the manuscript draft. RM supported both generation of the resource and data collection. TK generated the resource, supervised the project and wrote the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Supplementary Material 1
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Reprints and permissions
About this article
Cite this article
Alenize, M., Minhas, R. & Kudoh, T. Melanophore and fluoroleucophore photo-protect the Arabian killifish, Aphanius dispar, embryo from ultraviolet light.
Sci Rep (2026). https://doi.org/10.1038/s41598-026-37311-6
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-37311-6
Keywords
- Arabian killifish
- Pigments
- UV
- UVC
- Fluoroleucophore
- Melanophore
- Photoprotection
Source: Ecology - nature.com
