Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel.

Autor:	Pérez-Atehortúa M; Doctoral student in Agricultural Sciences, Faculty of Natural Resources, Catholic University of Temuco, Temuco 4781312 Chile.; Nucleus of Research in Food Production, Faculty of Natural Resources, Catholic University of Temuco, Temuco 4781312 Chile., Short SE; Department of Chemical Engineering, Faculty of Engineering and Sciences, La Frontera University, Temuco, Chile., Aranzaez-Rios C; Department of Chemical Engineering, Faculty of Engineering and Sciences, La Frontera University, Temuco, Chile., Farías J; Department of Chemical Engineering, Faculty of Engineering and Sciences, La Frontera University, Temuco, Chile., Oliveira RPS; Laboratory of Microbial Biomolecules, School of Pharmaceutical Sciences, University of São Paulo, Rua Do Lago, 250, Cidade Universitária, São Paulo 05508 000, Brazil., Pereira WA; Laboratory of Microbial Biomolecules, School of Pharmaceutical Sciences, University of São Paulo, Rua Do Lago, 250, Cidade Universitária, São Paulo 05508 000, Brazil., Risopatrón J; Center of Excellence in Biotechnology on Reproduction (BIOREN-CEBIOR), Faculty of Medicine, La Frontera University, Temuco 4811230, Chile., Valdebenito I; Nucleus of Research in Food Production, Faculty of Natural Resources, Catholic University of Temuco, Temuco 4781312 Chile., Villalobos EF; Nucleus of Research in Food Production, Faculty of Natural Resources, Catholic University of Temuco, Temuco 4781312 Chile.
Jazyk:	angličtina
Zdroj:	MethodsX [MethodsX] 2023 Dec 23; Vol. 12, pp. 102533. Date of Electronic Publication: 2023 Dec 23 (Print Publication: 2024).
DOI:	10.1016/j.mex.2023.102533
Abstrakt:	The chorion fulfills important functions in fish embryos, including protecting the embryo during development. The characterization of the protein profile of this envelope could be used as a bioindicator in the evaluation of the quality of embryonic development. The object of this work was to validate a standardized protocol for protein extraction from chorion of Salmo salar embryos at 280 accumulated thermal units (ATU) by comparing and combining existing methods. The protocol consists of consecutive washing of the chorion samples followed by protein extraction with the solution that was named SDS solution (Tris-HCl 100 mM (pH 8), Urea 8 M, 1% SDS, β-mercaptoethanol 300 mM and EGTA 10 Mm, and 1% protease inhibitor cocktail) and mechanical methods. Protein extraction is enhanced by a working temperature of 75 °C and use of a disperser. The protein concentration was quantified by Bradford Assay. After extraction, the samples were diluted (dilution factor 10) before reading against the calibration curve. After gel electrophoresis with a load of 3 µg of protein, staining showed more than 4 bands, with molecular weights between 25 kDa and 180 kDa.•The protein profile of fish chorion was between 25 kDa and 180 kDa.•Solution containing 1% SDS allows a higher extraction of proteins from the chorion of Atlantic salmon embryos with 280 ATU.•Chorion protein identification is a valuable tool in determining gamete and embryo quality in fish. Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (© 2023 The Authors. Published by Elsevier B.V.)
Databáze:	MEDLINE
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