小鼠早期胚胎的全胚原位杂交技术

Whole-mount in situ hybridization for analysis of gene expression in mouse embryos of early developmental-stage embryos

目的:探索一种适合于早期胚胎基因表达型检测的全胚原位杂交技术。方法:收集小鼠囊胚、固定;制备VASP、IL1R2基因特异性地高辛标记的cRNA探针;对囊胚进行杂交前处理、全胚原位杂交、抗体处理、显色以及显色后处理与镜检。结果:VASP、IL1R2特异性表达在囊胚的内细胞团细胞和滋养层细胞中。结论:小鼠早期胚胎全胚原位杂交技术适用于早期胚胎基因时空表达型的检测。

Aim： To establish a powerful technique as a whole-mount in situ hybridization for determi-nation of the temporal and spatial gene expression patterns in early developmental-stage embryos. Meth-otis： Murine blastocysts were collected and fixed. Whole-mount in situ hybridization was performed as：preparation of VASP and IL1R2 gene specific digoxigenin-labeled cRNA probe, pre-treatment and whole-mount in situ hybridization of the blastocysts, visualization of the labeled cRNA and treatment after visual-ization, microscope analysis. Results： Expression of VASP and ILIR2 was detected in both inner cell massand trophoblast of blastocysts. Conclusion：The present protocol described here is suitable for detection ofthe gene expression patterns in early developmental-stage embryos.