Transcriptional and Behavioral Responses of Zebrafish Larvae to Microcystin-LR Exposure. 2017

Eleni Tzima, and Iliana Serifi, and Ioanna Tsikari, and Ainhoa Alzualde, and Ioannis Leonardos, and Thomais Papamarcaki
Laboratory of Biological Chemistry, Medical School, University of Ioannina, 45110 Ioannina, Greece. elenatzima@gmail.com.

Microcystins are cyclic heptapeptides that constitute a diverse group of toxins produced by cyanobacteria. One of the most toxic variants of this family is microcystin-LR (MCLR) which is a potent inhibitor of protein phosphatase 2A (PP2A) and induces cytoskeleton alterations. In this study, zebrafish larvae exposed to 500 μg/L of MCLR for four days exhibited a 40% reduction of PP2A activity compared to the controls, indicating early effects of the toxin. Gene expression profiling of the MCLR-exposed larvae using microarray analysis revealed that keratin 96 (krt96) was the most downregulated gene, consistent with the well-documented effects of MCLR on cytoskeleton structure. In addition, our analysis revealed upregulation in all genes encoding for the enzymes of the retinal visual cycle, including rpe65a (retinal pigment epithelium-specific protein 65a), which is critical for the larval vision. Quantitative real-time PCR (qPCR) analysis confirmed the microarray data, showing that rpe65a was significantly upregulated at 50 μg/L and 500 μg/L MCLR in a dose-dependent manner. Consistent with the microarray data, MCLR-treated larvae displayed behavioral alterations such as weakening response to the sudden darkness and hypoactivity in the dark. Our work reveals new molecular targets for MCLR and provides further insights into the molecular mechanisms of MCLR toxicity during early development.

UI MeSH Term Description Entries
D007814 Larva Wormlike or grublike stage, following the egg in the life cycle of insects, worms, and other metamorphosing animals. Maggots,Tadpoles,Larvae,Maggot,Tadpole
D008387 Marine Toxins Toxic or poisonous substances elaborated by marine flora or fauna. They include also specific, characterized poisons or toxins for which there is no more specific heading, like those from poisonous FISHES. Marine Biotoxins,Phycotoxins
D010766 Phosphorylation The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety. Phosphorylations
D005786 Gene Expression Regulation Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation. Gene Action Regulation,Regulation of Gene Expression,Expression Regulation, Gene,Regulation, Gene Action,Regulation, Gene Expression
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001522 Behavior, Animal The observable response an animal makes to any situation. Autotomy Animal,Animal Behavior,Animal Behaviors
D014158 Transcription, Genetic The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION. Genetic Transcription
D015027 Zebrafish An exotic species of the family CYPRINIDAE, originally from Asia, that has been introduced in North America. Zebrafish is a model organism for drug assay and cancer research. Brachydanio rerio,Danio rerio,B. rerio,D. rerio,Zebra Fish,Zebra Fishes,Zebra danio,Zebrafishes,D. rerios,Fishes, Zebra,Zebra danios,danio, Zebra
D052998 Microcystins Cyclic heptapeptides found in MICROCYSTIS and other CYANOBACTERIA. Hepatotoxic and carcinogenic effects have been noted. They are sometimes called cyanotoxins, which should not be confused with chemicals containing a cyano group (CN) which are toxic. Cyanoginosins
D054648 Protein Phosphatase 2 A phosphoprotein phosphatase subtype that is comprised of a catalytic subunit and two different regulatory subunits. At least two genes encode isoforms of the protein phosphatase catalytic subunit, while several isoforms of regulatory subunits exist due to the presence of multiple genes and the alternative splicing of their mRNAs. Protein phosphatase 2 acts on a broad variety of cellular proteins and may play a role as a regulator of intracellular signaling processes. PPP2CA Phosphatase,PPP2CB Phosphatase,Protein Phosphatase 2, Catalytic Subunit,Protein Phosphatase 2, Catalytic Subunit, alpha Isoform,Protein Phosphatase 2, Catalytic Subunit, beta Isoform,Protein Phosphatase 2, Regulatory Subunit,Protein Phosphatase 2, Regulatory Subunit A, alpha Isoform,Protein Phosphatase 2, Regulatory Subunit A, beta Isoform,Protein Phosphatase 2, Regulatory Subunit B, alpha Isoform,Protein Phosphatase 2, Regulatory Subunit B, beta Isoform,Protein Phosphatase 2, Regulatory Subunit B, delta Isoform,Protein Phosphatase 2, Regulatory Subunit B, gamma Isoform,Protein Phosphatase 2A, Catalytic Subunit, alpha Isoform,Protein Phosphatase 2A, Catalytic Subunit, beta Isoform,Protein Phosphatase 2A, Regulatory Subunit A , alpha Isoform,Protein Phosphatase 2A, Regulatory Subunit A, beta Isoform,Protein Phosphatase 2A, Regulatory Subunit B, alpha Isoform,Protein Phosphatase 2A, Regulatory Subunit B, beta Isoform,Protein Phosphatase 2A, Regulatory Subunit B, delta Isoform,Protein Phosphatase 2A, Regulatory Subunit B, gamma Isoform,Protein Phosphatase-2A,Serine Threonine Protein Phosphatase 2A Catalytic Subunit beta Isoform,Serine-Threonine-Protein Phosphatase 2A Catalytic Subunit alpha Isoform,Phosphatase, PPP2CA,Phosphatase, PPP2CB,Protein Phosphatase 2A,Serine Threonine Protein Phosphatase 2A Catalytic Subunit alpha Isoform

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