[Frontiers in Bioscience 14, 3680-3687, January 1, 2009]

Teratogenic effects of sodium thiosulfate on developing zebrafish embryos

Wei Hu1,2, Luyang Cheng1,2, Hongfei Xia2, Daguang Sun1,2, Dan Li1,2, Peng Li2, Yuntao Song3, Xu Ma1,2

1Graduate School, Peking Union Medical College, Beijing 100005, China, 2Department of Genetics, National Research Institute for Family Planning, Beijing 100081, China, 3Peking University Health Science Center, Beijing 100083, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Zebrafish maintenance and collection of embryos
3.2. Stock solutions and exposure protocols
3.3. Morphological observation and statistics
3.4. Whole-Mount immunohistochemistry
3.5. Whole-Mount in situ hybridization
4. Results
4.1. Effects of STS exposure on early embryo development
4.2. Effects of STS on the development of the nervous system
4.3. Effects of STS on cell proliferation
4.4. Effects of STS on the expression pattern of central nervous system- and muscle-specific microRNAs
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

Sulfuric derivatives are potentially hazardous to human health, especially during embryogenesis. Zebrafish were used to study the toxic effect of sodium thiosulfate (STS) (1~1×10-6 mol/L) on embryo development with real-time in vivo imaging. Motor neuron differentiation and proliferation were analyzed by detecting the dynamics of acetylated tubulin (alpha-tubulin) and of proliferating cell nuclear antigen (PCNA). The expression pattern of brain- and muscle-specific microRNAs was detected by whole-mount in situ hybridization. The development of embryos exposed to 0.1~1 mol/L STS was severely retarded and was accompanied by malformation of multiple organs; embryos exposed to 10 μmol/L~10 mmol/L STS had circulatory, nervous and maxillofacial malformations. Embryos were more sensitive to STS at 48 hours post fertilization (hpf) compared with 24 and 96 hpf. STS can destroy the normal development of motor neurons and can affect cell proliferation. We also found differential expression of miR-124a and miR-133a in STS-treated embryos. STS interferes with the normal cytoskeleton structure, inhibits cell proliferation and leads to nervous, cardiac and maxillofacial malformations. MiR-124a and miR-133a were involved in STS malformation induction.