2. Academic Validation
  3. Malformations and mortality in zebrafish early stages associated with elevated caspase activity after 24 h exposure to MS-222

Malformations and mortality in zebrafish early stages associated with elevated caspase activity after 24 h exposure to MS-222

  • Toxicol Appl Pharmacol. 2021 Feb 1;412:115385. doi: 10.1016/j.taap.2020.115385.
Luís M Félix 1,  Ana Luzio 2,  Luís Antunes 3,  Ana M Coimbra 2,  Ana M Valentim 4
Affiliations

Affiliations

  • 1 i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; Laboratory Animal Science, IBMC - Instituto de Biologia Molecular Celular, Universidade do Porto, Porto, Portugal; Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal. Electronic address: luis.felix@ibmc.up.pt.
  • 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal; School of Life and Environmental Sciences (ECVA), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
  • 3 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal; School of Agrarian and Veterinary Sciences (ECAV), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
  • 4 i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; Laboratory Animal Science, IBMC - Instituto de Biologia Molecular Celular, Universidade do Porto, Porto, Portugal; Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
PMID: 33370555 DOI: 10.1016/j.taap.2020.115385
Abstract

Tricaine methanesulfonate (MS-222) is a commonly used anaesthetic agent for immobilization of aquatic species. However, delayed development and malformations have been observed in 24 hpf (hours post-fertilization) zebrafish embryos after long-term immobilization. Still, no comprehensive study has been described regarding zebrafish exposure to MS-222 during the first hours of development, which are one of the most sensitive life stages to toxicants. Therefore, this research aimed to assess the toxicity of a 24 h exposure to MS-222 on zebrafish embryonic development. Based on the MS-222 LC50, early blastula stage embryos (~2 hpf) were exposed to 0, 12.5, 25 and 50 mg L-1 for 24 h and then allowed to develop up to 144 hpf. The chromatographic analysis showed that this anaesthetic agent bioaccumulates in 26 hpf zebrafish larvae in a concentration-dependent manner. In addition, increased mortalities and skeletal abnormalities were observed at 144 hpf, namely in the highest tested concentration. Yet, no craniofacial anomalies were observed either by alcian blue or calcein staining methods. Independently of the tested concentration, decreased speed and distance travelled were perceived in 144 hpf larvae. At the biochemical level, decreased in vivo Reactive Oxygen Species (ROS) generation and Apoptosis was observed. Additionally, catalase activity was increased at 26 hpf while results of mRNA expression showed a decreased gclc transcript content at the same time-point. Overall, data obtained highlight the toxicological risk of MS-222 and support ROS-mediated cell death signalling changes through the elevation of catalase activity as an adaptative or protective response.

Keywords

Behaviour; Development; MS-222; Molecular Responses; Morphology; Zebrafish.

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