Deguelin down-regulates the expression of cyclin D1 gene in zebrafish embroys through the whole mount in situ hybridization

doi:10.3969/j.issn.2095-4344.2015.15.017

Abstract

Abstract:

BACKGROUND: In the early development of zebrafish embryos, cells divide and proliferate rapidly, but low concentration of deguelin can delay the development of zebrafish embryos.
OBJECTIVE: To observe the effect of different concentrations of deguelin on cyclin D1 gene expression in zebrafish embryos.
METHODS: Though normal fertilization, zebrafish embryos that incubated to the 2-cell stage (about 0.75 hour after fertilization) and shield stage (6 hours after fertilization) were collected and put into 12-well plates treated with 100, 200, 400 nmol/L deguelin at 28.5 ℃ in an incubator till the shield period and 24 hours after fertilization, respectively. Simultaneously embroys treated with 1% dimethyl sulfoxide solution were as a control group, cultured in the same conditions. Cyclin D1 RNA probes were prepared for the whole mount in situ hybridization, observing staining by an upright fluorescent microscope camera to detect the effect of deguelin on cyclin D1 expression in zebrafish embryos.
RESULTS AND CONCLUSION: Deguelin showed significant negative regulation on the expression of cyclin D1 gene in zebrafish embryos. Cyclin D1 expressed in outsourcing cells in embryos of shield stage, and a significant reduction in the expression of cyclin D1 came up with the increasing concentrations of deguelin. In the 400 nmol/L deguelin treatment group, there was nearly no expression of cyclin D1. Cyclin D1 expressed in the brain, central nervous system, immature eye, somites, trunk, and tail of embryos at 24 hours after fertilization, and reduced significantly in the 100 nmol/L deguelin treatment group, especially in the proliferative area. In the 200 and 400 nmol/L treatment groups, the embryonic development slowed down signficantly, and cyclin D1 gene mainly expressed in the dorsal ectoderm cells.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: Zebrafish, Derris, Cell Cycle

CLC Number:

R318

Hai Yang, Wu Xin-rong. Deguelin down-regulates the expression of cyclin D1 gene in zebrafish embroys through the whole mount in situ hybridization[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(15): 2382-2386.

Figures/Tables 1

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