Atlantic Salmon Gill Epithelial Cell Line (ASG-10) As a Suitable Model for Xenobiotic Biotransformation

Overview

Journal Metabolites

Publisher MDPI

Date 2023 Jun 27

PMID 37367928

Authors

Lada Ivanova

Christiane Kruse Faeste

Anita Solhaug

Affiliations

Soon will be listed here.

Abstract

Fish are exposed to xenobiotics in the water. Uptake occurs mainly through the gills, which function as an exchange point with the environment. The gills' ability to detoxify harmful compounds by biotransformation is an essential protection mechanism. The enormous numbers of waterborne xenobiotics requiring ecotoxicological assessment makes it necessary to replace in vivo fish studies with predictive in vitro models. Here, we have characterized the metabolic capacity of the ASG-10 gill epithelial cell line from Atlantic salmon. Inducible CYP1A expression was confirmed by enzymatic assays and immunoblotting. The activities of important cytochrome P450 (CYP) and uridine 5'-diphospho-glucuronosyltransferase (UGT) enzymes were established using specific substrates and metabolite analysis by liquid chromatography (LC) triple quadrupole mass spectrometry (TQMS). Metabolism of the fish anesthetic benzocaine (BZ) in ASG-10 confirmed esterase and acetyl transferase activities through the production of N-acetylbenzocaine (AcBZ), -aminobenzoic acid (PABA) and p-acetaminobenzoic acid (AcPABA). Moreover, we were able to determine hydroxylamine benzocaine (BZOH), benzocaine glucuronide (BZGlcA) and hydroxylamine benzocaine glucuronide (BZ(O)GlcA) by LC high-resolution tandem mass spectrometry (HRMS/MS) fragment pattern analysis for the first time. Comparison to metabolite profiles in hepatic fractions, and in plasma of BZ-euthanized salmon, confirmed the suitability of the ASG-10 cell line for investigating biotransformation in gills.

Citing Articles

Isolation and characterisation of two epithelial-like cell lines from the gills of Chrysophrys auratus (Australasian snapper) and Oncorhynchus tshawytscha (Chinook salmon) and their use in aquatic toxicology.

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PMID: 38987436 DOI: 10.1007/s11626-024-00941-z.

Functional and molecular characterization of the Atlantic salmon gill epithelium cell line ASG-10; a tool for gill research.

Slattery O, Dahle M, Sundaram A, Nowak B, Gjessing M, Solhaug A Front Mol Biosci. 2023; 10:1242879.

PMID: 37916189 PMC: 10616884. DOI: 10.3389/fmolb.2023.1242879.

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