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Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae

Zebrafish are easy to breed in a laboratory setting as they are extremely fertile and produce dozens of eggs per set. Because zebrafish eggs and the skin of the early-stage larvae are transparent, their embryos and the hearts and muscles of their larvae can be easily observed. Multiple rapid analyse...

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Autores principales: Maeda, Hideyuki, Fukushima, Noritoshi, Hasumi, Akihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8389650/
https://www.ncbi.nlm.nih.gov/pubmed/34440088
http://dx.doi.org/10.3390/biomedicines9080884
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author Maeda, Hideyuki
Fukushima, Noritoshi
Hasumi, Akihiro
author_facet Maeda, Hideyuki
Fukushima, Noritoshi
Hasumi, Akihiro
author_sort Maeda, Hideyuki
collection PubMed
description Zebrafish are easy to breed in a laboratory setting as they are extremely fertile and produce dozens of eggs per set. Because zebrafish eggs and the skin of the early-stage larvae are transparent, their embryos and the hearts and muscles of their larvae can be easily observed. Multiple rapid analyses of heart rate and behavior can be performed on these larvae simultaneously, enabling investigation of the influence of neuroactive substances on abnormal behavior, death, and associated pathogenetic mechanisms. Zebrafish larvae are becoming increasingly popular among researchers and are used in laboratories worldwide to study various vertebrate life phenomena; more experimental systems using zebrafish will undoubtedly be developed in the future. However, based on the available literature, we believe that the conceptualization of a protocol based on scientific evidence is necessary to achieve standardization. We exposed zebrafish larvae at 6–7 days post-fertilization to 50 repeated light–dark stimuli at either 15-min or 5-min intervals. We measured the traveled distance and habituation time through a video tracking apparatus. The traveled distance stabilized after the 16th repetition when the zebrafish were exposed to light–dark stimuli at 15-min intervals and after the 5th repetition when exposed at 5-min intervals. Additionally, at 15-min intervals, the peak of the traveled distance was reached within the first minute in a dark environment, whereas at 5-min intervals, it did not reach the peak even after 5 min. The traveled distance was more stable at 5-min intervals of light/dark stimuli than at 15-min intervals. Therefore, if one acclimatizes zebrafish larvae for 1 h and collects data from the 5th repetition of light/dark stimuli at intervals of 5 min in the light/dark test, a stable traveled distance result can be obtained. The establishment of this standardized method would be beneficial for investigating substances of unknown lethal concentration.
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spelling pubmed-83896502021-08-27 Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae Maeda, Hideyuki Fukushima, Noritoshi Hasumi, Akihiro Biomedicines Article Zebrafish are easy to breed in a laboratory setting as they are extremely fertile and produce dozens of eggs per set. Because zebrafish eggs and the skin of the early-stage larvae are transparent, their embryos and the hearts and muscles of their larvae can be easily observed. Multiple rapid analyses of heart rate and behavior can be performed on these larvae simultaneously, enabling investigation of the influence of neuroactive substances on abnormal behavior, death, and associated pathogenetic mechanisms. Zebrafish larvae are becoming increasingly popular among researchers and are used in laboratories worldwide to study various vertebrate life phenomena; more experimental systems using zebrafish will undoubtedly be developed in the future. However, based on the available literature, we believe that the conceptualization of a protocol based on scientific evidence is necessary to achieve standardization. We exposed zebrafish larvae at 6–7 days post-fertilization to 50 repeated light–dark stimuli at either 15-min or 5-min intervals. We measured the traveled distance and habituation time through a video tracking apparatus. The traveled distance stabilized after the 16th repetition when the zebrafish were exposed to light–dark stimuli at 15-min intervals and after the 5th repetition when exposed at 5-min intervals. Additionally, at 15-min intervals, the peak of the traveled distance was reached within the first minute in a dark environment, whereas at 5-min intervals, it did not reach the peak even after 5 min. The traveled distance was more stable at 5-min intervals of light/dark stimuli than at 15-min intervals. Therefore, if one acclimatizes zebrafish larvae for 1 h and collects data from the 5th repetition of light/dark stimuli at intervals of 5 min in the light/dark test, a stable traveled distance result can be obtained. The establishment of this standardized method would be beneficial for investigating substances of unknown lethal concentration. MDPI 2021-07-24 /pmc/articles/PMC8389650/ /pubmed/34440088 http://dx.doi.org/10.3390/biomedicines9080884 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Maeda, Hideyuki
Fukushima, Noritoshi
Hasumi, Akihiro
Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title_full Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title_fullStr Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title_full_unstemmed Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title_short Standardized Method for the Assessment of Behavioral Responses of Zebrafish Larvae
title_sort standardized method for the assessment of behavioral responses of zebrafish larvae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8389650/
https://www.ncbi.nlm.nih.gov/pubmed/34440088
http://dx.doi.org/10.3390/biomedicines9080884
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