Perturbation of cytosolic calcium by 2-aminoethoxydiphenyl borate and caffeine affects zebrafish myofibril alignment

Hsin Ju Wu, Tsorng Harn Fong, Shen Liang Chen, Jen Cheng Wei, I. Jong Wang, Chi Chung Wen, Chao Yuan Chang, Xing Guang Chen, Wei Yu Chen, Hui Min Chen, Juin Lin Horng, Yun Hsin Wang, Yau Hung Chen

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The objective of the current study was to investigate the effects of Ca2+ levels on myofibril alignment during zebrafish embryogenesis. To investigate how altered cytoplasmic Ca2+ 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 Ca2+ levels) and caffeine (a ryanodine receptor activator that enhances cytosolic Ca2+ 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 Ca2+ is important for myogenesis, particularly for myofibril alignment.

Original languageEnglish
Pages (from-to)287-294
Number of pages8
JournalJournal of Applied Toxicology
Volume35
Issue number3
DOIs
Publication statusPublished - Mar 1 2015

Fingerprint

Myofibrils
Zebrafish
Caffeine
Embryonic Structures
Calcium
Muscle
Inositol 1,4,5-Trisphosphate Receptors
Phalloidine
Borates
Ryanodine Receptor Calcium Release Channel
Fibers
Phenotype
Laminin
Muscles
Somites
Fluorescent Dyes
Muscle Development
Actins
Transmission Electron Microscopy
Embryonic Development

Keywords

  • Caffeine
  • Calcium
  • Embryogenesis
  • Myogenesis
  • Zebrafish

ASJC Scopus subject areas

  • Toxicology
  • Medicine(all)

Cite this

Perturbation of cytosolic calcium by 2-aminoethoxydiphenyl borate and caffeine affects zebrafish myofibril alignment. / Wu, Hsin Ju; Fong, Tsorng Harn; Chen, Shen Liang; Wei, Jen Cheng; Wang, I. Jong; Wen, Chi Chung; Chang, Chao Yuan; Chen, Xing Guang; Chen, Wei Yu; Chen, Hui Min; Horng, Juin Lin; Wang, Yun Hsin; Chen, Yau Hung.

In: Journal of Applied Toxicology, Vol. 35, No. 3, 01.03.2015, p. 287-294.

Research output: Contribution to journalArticle

Wu, Hsin Ju ; Fong, Tsorng Harn ; Chen, Shen Liang ; Wei, Jen Cheng ; Wang, I. Jong ; Wen, Chi Chung ; Chang, Chao Yuan ; Chen, Xing Guang ; Chen, Wei Yu ; Chen, Hui Min ; Horng, Juin Lin ; Wang, Yun Hsin ; Chen, Yau Hung. / Perturbation of cytosolic calcium by 2-aminoethoxydiphenyl borate and caffeine affects zebrafish myofibril alignment. In: Journal of Applied Toxicology. 2015 ; Vol. 35, No. 3. pp. 287-294.
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AU - Wu, Hsin Ju

AU - Fong, Tsorng Harn

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AU - Wei, Jen Cheng

AU - Wang, I. Jong

AU - Wen, Chi Chung

AU - Chang, Chao Yuan

AU - Chen, Xing Guang

AU - Chen, Wei Yu

AU - Chen, Hui Min

AU - Horng, Juin Lin

AU - Wang, Yun Hsin

AU - Chen, Yau Hung

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N2 - The objective of the current study was to investigate the effects of Ca2+ levels on myofibril alignment during zebrafish embryogenesis. To investigate how altered cytoplasmic Ca2+ 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 Ca2+ levels) and caffeine (a ryanodine receptor activator that enhances cytosolic Ca2+ 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 Ca2+ is important for myogenesis, particularly for myofibril alignment.

AB - The objective of the current study was to investigate the effects of Ca2+ levels on myofibril alignment during zebrafish embryogenesis. To investigate how altered cytoplasmic Ca2+ 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 Ca2+ levels) and caffeine (a ryanodine receptor activator that enhances cytosolic Ca2+ 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 Ca2+ is important for myogenesis, particularly for myofibril alignment.

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