Dataset Information

Effects of the chlorophyll derivative 132-hydroxypheophytine a on zebrafish metabolomics

ABSTRACT: Zebrafish larvae were exposed from 3 DPF to 5 DPF to the chlorophyll derivative 13-2-hydroxypheophytine a. Metabolites were extracted with some modification of the protocol published by Lin et al 2019 (10.1016/j.bbrc.2019.04.003.). 3 experimental groups: DMSO 0.1% (solvent control) and hpa at 7.5 ug/ml (hpa1) and at 15 ug/ml (hpa2). All details can be found in the paper that published this data set. Title: Insights into the mechanism of action of the chlorophyll derivative 132-hydroxypheophytine a on reducing neutral lipid reserves in zebrafish larvae and mice adipocytes Authors: Ana Carrasco del Amor, Rene Hernandez Butista, Siegfried Ussar, Susana Cristobal, Ralph Urbatzka European Journal of Pharmacology accepted: 10/24/2023 doi: 10.1016/j.ejphar.2023.176158

INSTRUMENT(S): Orbitrap Exploris 120

ORGANISM(S): Danio Rerio (ncbitaxon:7955)

SUBMITTER: Ralph Urbatzka

PROVIDER: MSV000092187 | MassIVE | Fri Jun 16 04:18:00 BST 2023

REPOSITORIES: MassIVE

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Insights into the mechanism of action of the chlorophyll derivative 13-<sup>2</sup>-hydroxypheophytine a on reducing neutral lipid reserves in zebrafish larvae and mice adipocytes.

Carrasco Del Amor Ana A Bautista Rene Hernandez RH Ussar Siegfried S Cristobal Susana S Urbatzka Ralph R

European journal of pharmacology 20231026

Obesity is a worldwide epidemic and natural products may hold promise in its treatment. The chlorophyll derivative 13-<sup>2</sup>-hydroxypheophytine (hpa) was isolated in a screen with zebrafish larvae to identify lipid reducing molecules from cyanobacteria. However, the mechanisms underlying the lipid-reducing effects of hpa in zebrafish larvae remain poorly understood. Thus, investigating the mechanism of action of hpa and validation in other model organisms such as mice represents important ...[more]

PMID: 37898286

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Project description:Purpose: Bepridil is a commonly used drug for angina pectoris and arrhythmia. However, its toxicity and other potential effects on embryonic development remain unknown. In this study, we revealed the physical and developmental toxicity of bepridil on zebrafish models and human myocardial H9c2 cells. Methods: The AB line zebrafish embryos were treated with different concentrations of bepridil at 24 hpf. The mRNA profiles of zebrafish larvae at 120 hpf in 0, 250, 750 and 1295 ng/mL bepridil groups were generated with deep sequencing, in triplicate, using Illumina sequencing platform (HiSeqTM 2500). RNA integrity was analyzed through the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara). The RNA integrity number ≥ 7 was subjected to the following analysis. Genes with adjusted P-value < 0.05 and foldChange >2 were defined as differentially expressed genes (DEGs). Results: About 45 million sequence reads were mapped to the zebrafish genome in each group. The RNA-seq data confirmed that low-dose of bepridil treatment induced an obvious xenobiotic stimulus and drug catabolic process. The enriched DEGs of GO and KEGG workflow showed negative regulation of proteolysis and peptidase in 250 and 750 ng/mL bepridil groups, which eventually led to the metabolic disorders of lipids and a variety of essential amino acids. As a long-term acting calcium antagonist, bepridil caused partial changes (Generatio, 61/426) of calcium signaling channel in 120 hpf zebrafish larvae after 1295 ng/mL (LC10) treatment. However, the cardiac muscle contraction and adrenergic of cardiomyocytes were affected significantly in the same group. Besides, the overall DEGs were highly correlated to cell proliferation and differentiation of the zebrafish. In brief, the transcriptome results were similar to our in vivo phenotype data. The liver metabolism and cardiovascular function of zebrafish would be affected by bepridil during early embryonic development. Conclusions: Our study reveals the potential embryotoxicity of bepridil, a traditional antiarrhythmic drug, in zebrafish embryos. The transcriptomes of zebrafish larvae were progressed with biological replicates (n = 3) and generated through RNA-seq. Our results reveal the detailed biological and metabolic responses of bepridil to embryonic development, which has guiding significance for the clinical medication of prenatal heart patients.

Project description:This project aimed at identifying developmental stage specific transcript profiles for catecholaminergic neurons in embryos and early larvae of zebrafish (Danio rerio). Catecholaminergic neurons were labeled using transgenic zebrafish strains to drive expression of GFP. At stages 24, 36, 72 and 96 hrs post fertilization, embryos were dissociated and GFP expressing cells sorted by FACS. Isolated RNAs were processed using either polyA selection and libray generation or NanoCAGE. This is the first effort to determine stage specific mRNA profiles of catecholaminergic neurons in zebrafish. Catecholaminergic neurons were labeled by four different strategies: (1) 24 hrs old embryos: we used the ETvmat2:GFP transgenic line (Wen et al. 2007). Visualization of monoaminergic neurons and neurotoxicity of MPTP in live transgenic zebrafish. Dev Biol. 2008 Vol 314 p84-92) which at this early stage labels catecholaminergic neurons in posterior tuberculum and locus coeruleus; (2) 24 hrs old embryos: we used Tg(otpb.A:egfp)zc48 transgenic line (Fujimoto et al. Identification of a dopaminergic enhancer indicates complexity in vertebrate dopamine neuron phenotype specification. Dev Biol 2011, Vol 352, p393–404) which at this stage label ventral diencephalic dopaminergic neurons and some preoptic neurons. (3) For 72 and 96 hrs old zebrafish larvae we used a th:GFP BAC transgenic lines that labels catecholaminergic neurons (Tay et al., Comprehensive catecholaminergic projectome analysis reveals single-neuron integration of zebrafish ascending and descending dopaminergic systems. Nat Comms 2011 Vol 2, 171; also: T. Leng and W. Driever, unpublished). (4) for the 36 and 48 hrs old zebrafish larvae we used a th:Gal4VP16 driver and UAS:EGFP responder transgenic line system to label catecholaminergic cells (Fernandes et al., Deep brain photoreceptors control light-seeking behavior in zebrafish larvae. Curr Biol. 2012 Vol 22 DOI 10.1016/j.cub.2012.08.016). We used the different transgenic lines, because lines (3) and (4) do not efficiently label catecholaminergic neurons at early stages, while lines (1) and (2) also have GFP expression in several other non-catecholaminergic populations at later stages of development. Embryos were dissociated and catecholaminergic neurons were FACS sorted from GFP-tagged zebrafish (Manoli and Driever, 2012, Cold Spring Harbor Protoc. DOI 10.1101/pdb.prot069633). RNA was either processed for NanoCAGE, or mRNA was isolated and amplified. cDNA was sequenced by Illumina technique. This data submission is a series of data files consisting of three independent experiments with diffrent RNA-Seq depth: Samples 1-4 (NanoCage): Samples 5-8 (RNA-Seq high read numbers), and SAmples 9-12 (RNA-Seq low read numbers).

Dataset Information

Effects of the chlorophyll derivative 132-hydroxypheophytine a on zebrafish metabolomics

Publications

Insights into the mechanism of action of the chlorophyll derivative 13-<sup>2</sup>-hydroxypheophytine a on reducing neutral lipid reserves in zebrafish larvae and mice adipocytes.

Similar Datasets

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