دورية أكاديمية
Optimization of assay conditions to quantify ECOD activity in vivo in individual Daphnia magna. Assay performance evaluation with model CYP 450 inducers/inhibitors.
| العنوان: | Optimization of assay conditions to quantify ECOD activity in vivo in individual Daphnia magna. Assay performance evaluation with model CYP 450 inducers/inhibitors. |
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| المؤلفون: | Ács A; Balaton Limnological Research Institute, Hungarian Research Network, Klebelsberg Kuno u. 3., Tihany H-8237, Hungary. Electronic address: acs.andras@blki.hu., Kovács AW; Balaton Limnological Research Institute, Hungarian Research Network, Klebelsberg Kuno u. 3., Tihany H-8237, Hungary., Győri J; Balaton Limnological Research Institute, Hungarian Research Network, Klebelsberg Kuno u. 3., Tihany H-8237, Hungary., Farkas A; Balaton Limnological Research Institute, Hungarian Research Network, Klebelsberg Kuno u. 3., Tihany H-8237, Hungary. |
| المصدر: | Ecotoxicology and environmental safety [Ecotoxicol Environ Saf] 2024 Mar 15; Vol. 273, pp. 116159. Date of Electronic Publication: 2024 Feb 27. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 7805381 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2414 (Electronic) Linking ISSN: 01476513 NLM ISO Abbreviation: Ecotoxicol Environ Saf Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam, Netherlands : Elsevier |
| مواضيع طبية MeSH: | Daphnia magna* , Cytochrome P-450 Enzyme System*/metabolism, Humans ; Animals ; 7-Alkoxycoumarin O-Dealkylase ; beta-Naphthoflavone/toxicity ; Daphnia |
| مستخلص: | Screening the activity of the cytochrome P450 (CYP450) mixed function oxidase system in aquatic invertebrates received seldom applications in ecotoxicology due to low baseline enzymatic activities characteristic for these organisms. In this study, an existing in vivo spectrofluorometric assay method based on quantifying the cytochrome P450 mediated conversion of 7-ethocycoumarin (EtC) used as substrate to the product 7-hydroxycoumarin (HCm) called: ethoxycoumarin-O-deethylase (ECOD) activity, initially applicable on pooled samples of Daphnia magna, was optimized for use on individual organisms. Optimal assay conditions have been established for as small as 3- and 6 days old individuals, and the limits of spectrofluorometric detection of HCm excreted by daphnids in the incubation media were defined. The modified assay was tested by screening the modulation of ECOD activity in daphnids following 24 h exposure to β-naphthoflavone (β-NF, reference CYP450 inducer) and to prochloraz (PCZ), a potent CYP450 inhibitor. Maximal ECOD activity levels in daphnids were recorded following 2 hours of incubation to 200 nM EtC. The limit of spectrofluorometric detection of HCm in the incubation media was 6.25 nM, achieved by more than 80% of three days old daphnids and all six days old individuals. Exposure of daphnids to β-NF demonstrated a bell-shaped ECOD activity induction potential, while PCZ elicited partial (60%) inhibition of ECOD activity. This optimized in vivo ECOD activity assay may serve as a cost-effective tool to study the responsiveness of Phase-I metabolism in D. magna to toxic pressure and its applicability to other aquatic invertebrates is also worth for consideration. Competing Interests: Declaration of Competing Interest 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. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Beta-naphthoflavone; ECOD activity; Optimized in vivo assay; Prochloraz; Single Daphnia |
| المشرفين على المادة: | EC 1.14.13.- (7-Alkoxycoumarin O-Dealkylase) 9035-51-2 (Cytochrome P-450 Enzyme System) 6051-87-2 (beta-Naphthoflavone) |
| تواريخ الأحداث: | Date Created: 20240228 Date Completed: 20240318 Latest Revision: 20240318 |
| رمز التحديث: | 20240318 |
| DOI: | 10.1016/j.ecoenv.2024.116159 |
| PMID: | 38417318 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1090-2414 |
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| DOI: | 10.1016/j.ecoenv.2024.116159 |