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Perturbation of cytosolic calcium by 2‐aminoethoxydiphenyl borate and caffeine affects zebrafish myofibril alignment
Author(s) -
Wu HsinJu,
Fong TsorngHarn,
Chen ShenLiang,
Wei JenCheng,
Wang IJong,
Wen ChiChung,
Chang ChaoYuan,
Chen XingGuang,
Chen WeiYu,
Chen HuiMin,
Horng JuinLin,
Wang YunHsin,
Chen YauHung
Publication year - 2015
Publication title -
journal of applied toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.784
H-Index - 87
eISSN - 1099-1263
pISSN - 0260-437X
DOI - 10.1002/jat.3057
Subject(s) - myofibril , zebrafish , ryanodine receptor , microbiology and biotechnology , caffeine , phalloidin , chemistry , myocyte , myogenesis , actin , biology , medicine , endocrinology , biochemistry , intracellular , cytoskeleton , cell , gene
The objective of the current study was to investigate the effects of Ca 2+ levels on myofibril alignment during zebrafish embryogenesis. To investigate how altered cytoplasmic Ca 2+ levels affect myofibril alignment, we exposed zebrafish embryos to 2‐aminothoxyldiphenyl borate (2‐APB; an inositol 1,4,5‐trisphosphate receptor inhibitor that reduces cytosolic Ca 2+ levels) and caffeine (a ryanodine receptor activator that enhances cytosolic Ca 2+ levels). The results demonstrated that the most evident changes in zebrafish embryos treated with 2‐APB were shorter body length, curved trunk and malformed somite boundary. In contrast, such malformed phenotypes were evident neither in untreated controls nor in caffeine‐treated embryos. Subtle morphological changes, including changes in muscle fibers, F‐actin and ultrastructures were easily observed by staining with specific monoclonal antibodies (F59 and α‐laminin), fluorescent probes (phalloidin) and by transmission electron microscopy. Our data suggested that: (1) the exposure to 2‐APB and/or caffeine led to myofibril misalignment; (2) 2‐APB‐treated embryos displayed split and short myofibril phenotypes, whereas muscle fibers from caffeine‐treated embryos were twisted and wavy; and (3) zebrafish embryos co‐exposed to 2‐APB and caffeine resulted in normal myofibril alignment. In conclusion, we proposed that cytosolic Ca 2+ is important for myogenesis, particularly for myofibril alignment. Copyright © 2014 John Wiley & Sons, Ltd.