title: Effect of PMA-induced protein kinase C activation on development and apoptosis in early zebrafish embryos.
abstract: Protein kinase C (PKC) isoforms have been implicated in several key steps during early development, but the consequences of xenobiotic-induced PKC activation during early embryogenesis are still unknown. In this study, zebrafish embryos were exposed to a range of phorbol 12-myristate 13-acetate (PMA) concentrations (0-200μg/L) at different time points after fertilization. Results showed that 200μgPMA/L caused development of yolk bags, cardiac edema, slow blood flow, pulsating blood flow, slow pulse, elongated heart, lack of tail fins, curved tail, and coagulation. PMA exposure decreased survival rate of the embryos starting within the first 24h and becoming more pronounced after prolonged exposure (96h). PMA increased the number of apoptotic cells in the brain region as demonstrated by acridine orange staining and caused up-regulation of caspase 9 (casp9) and p53 up-regulated modulator of apoptosis (puma) mRNA in whole embryos. PMA caused oxidative stress in the embryos as demonstrated by decreased mRNA expression of catalase and superoxide dismutase 2. Inhibition of Pkc with GF109203X improved overall survival rate, reduced apoptosis in the brain and decreased expression of casp9 and puma in the PMA-exposed embryos. However, Pkc inhibition neither prevented development of deformities nor reversed oxidative stress in the PMA-exposed embryos. These data suggest that direct over-activation of Pkc during early embryogenesis of zebrafish is associated with apoptosis and decreased survival rate of the embryos.
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia.
authors: Jelena
authors: J
authors: Hrubik
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia.
authors: Branka
authors: B
authors: Glisic
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia.
authors: Dragana
authors: D
authors: Samardzija
authors: University of Novi Sad, Faculty of Technical Sciences, Department of Environmental Engineering and Occupational Safety and Health, Novi Sad, Serbia.
authors: Bojana
authors: B
authors: Stanic
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia.
authors: Kristina
authors: K
authors: Pogrmic-Majkic
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia.
authors: Svetlana
authors: S
authors: Fa
authors: University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Ecotoxicology, Novi Sad, Serbia. Electronic address: nebojsa.andric@dbe.uns.ac.rs.
authors: Nebojsa
authors: N
authors: Andric
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D047108:Embryonic Development / Q000187:drug effects; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000633:toxicity*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D020536:Enzyme Activators; D047428:Protein Kinase Inhibitors; D029961:Zebrafish Proteins; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate
keywords: Apoptosis; PMA; Pkc; Zebrafish embryos