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First published online May 30, 2008
Journal of Experimental Biology 211, 1874-1881 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.012690
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Extensive apoptosis and abnormal morphogenesis in pro-caspase-3 transgenic zebrafish during development

Michiaki Yamashita*, Nanami Mizusawa, Misako Hojo and Takeshi Yabu

National Research Institute of Fisheries Science, 2-12-4 Fukuura, Yokohama 236-8648, Japan

* Author for correspondence (e-mail: mic{at}affrc.go.jp)

Accepted 2 April 2008

The pro-apoptotic caspase-3 gene has been shown to have key functions in the execution of apoptosis (programmed cell death) in vertebrate cells. However, the central role of caspase-3 in morphogenesis during development remains unclear. In this study, transgenic zebrafish that overexpress full-length pro-caspase-3 were generated to determine the effects of caspase genes on vertebrate morphogenesis and stress tolerance. The enhanced expression of the full-length pro-caspase-3 cDNA induced extremely high levels of caspase activity and extensive apoptosis in the transgenic embryos, and 33–46% of F2 embyos in the transgenic lines exhibited some form of morphological abnormality. Pro-caspase-3 transgenic zebrafish exhibited abnormal morphogenesis in the eyes, notochord, heart and yolk sac, suggesting that enhanced processing of pro-caspase-3 triggers significant apoptotic responses in the specific target tissues that are undergoing morphogenesis during development. The transgenic fish had reduced eye size and showed degeneration of the retina, including the photoreceptor cell layers, whereas pigmentation and lens formation were not affected. In addition, heart failure due to a weakened heartbeat and reduced circulation was noted in the pro-caspase-3 transgenic embryos. The transgenic embryos were markedly sensitive to stress conditions, such as UV irradiation at 2 or 5 mJ cm–2. On the other hand, caspase-3 deficiency through injection of antisense morpholino oligo into embryos repressed apoptosis and enhanced stress tolerance after UV irradiation. Therefore, the caspase-3-mediated pro-apoptotic signalling pathway and its activation play critical roles in the induction of apoptosis and stress tolerance during zebrafish embryogenesis.

Key words: caspase-3, apoptosis, transgenic zebrafish, eye and heart formation, stress response, antisense morpholino oligo


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