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Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos

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Castro, PL, Ferraz, ALJ, Patil, JG and Ribeiro, RP 2021 , 'Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos' , Brazilian Journal of Biology, vol. 82 , pp. 1-10 , doi: 10.1590/1519-6984.241081.

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Abstract

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio)embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified(Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 μM (T2) and 1 mM of melatonin(T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos storedin -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2)and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentationanalyses were performed. No live embryos were obtained from vitrified treatments, observing a rapiddegeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followedby breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 hadthe highest relative gene expression, for the three genes (P 0.05). The inclusion of 1 μM of melatonin in thevitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utilityin cryopreserving fish embryos.

Item Type: Article
Authors/Creators:Castro, PL and Ferraz, ALJ and Patil, JG and Ribeiro, RP
Keywords: cryopreservation, zebrafish, melatonin
Journal or Publication Title: Brazilian Journal of Biology
Publisher: Instituto Internacional de Ecologia,International Institute of Ecology
ISSN: 1519-6984
DOI / ID Number: 10.1590/1519-6984.241081
Copyright Information:

This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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