文昌鱼胚胎整装荧光原位杂交技术的建立

Whole - mounted fluorescent in situ hybridization of amphioxus embryos

文昌鱼(Branchiostoma belcheri tsingtauense)是进化发育生物学研究的重要模式生物,文昌鱼胚胎整装荧光原位杂交(WFISH,whole-mounted fluorescent in situ hybridization)技术将有助于鉴定文昌鱼胚胎发育过程中具有重要调控作用的功能基因。报告了文昌鱼胚胎整装荧光原位杂交技术,用以快速灵敏分析特定基因在文昌鱼胚胎发育过程中的时空表达图式。用体外转录合成的地高辛标记文昌鱼FGFR基因的反义RNA探针,检测到FGFR在文昌鱼原肠胚中表达于发生内陷的中内胚层细胞中,而预定发育为外胚层的细胞不表达FGFR。

Cephalochordate amphioxus(Branchiostoma belcheri tsingtauense) has emerged as an important animal model in evolutionary developmental biology research in recent years. The whole - mounted fluorescent in situ hybridization (WFISH) of amphioxus embryos contributes to the identification of critical genes functioning in amphioxus embryogenesis. In present study, we report the development of WFISH to analyze rapidly and sensitively the spatial and temporal expression pattern of a specific gene during the development of amphioxus embryos. Ogoxigenin - labelled antisense RNA probes a-gainst FGFR was synthesized through in vitro transcription in the present of Digoxigenin (Dig) -UIP and it was detected that during amphioxus gastrulation, FGFR expression was restrict to the invaginating mesendoderm cells but no positive signals was detected in ectoderm cells.