Li Dan, Sun Hua-qin, Zhao Jun, Lu Yi-lu, Deng Wen-qian, Ma Yong-xin
Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2010 Nov;41(6):923-6.
To study the role for piwil2 gene (zili) in the development of the ectoderm, mesoderm and endoderm during early embryogenesis of zebrafish.
zili morpholino antisense oligonucleotide and 5 mis-pair control morpholino were used in this study. zili was cloned into expression vector. zili mRNA was synthesized in vitro. The antisense RNA probes of gsc, evel and sox17 were synthesized. zili-MO, zili-cMO and zili mRNA was microinjected into one-cell embryos, respectively. Whole-mount in situ hybridization was used to monitor the expressions of marker genes.
Microinjection of zili-MO, which knocked down the expression of zili, downregulated the expression of the ectodermal and mesodermal marker gene gsc, promoting the expression of the ectodermal marker gene evel and resulting in the decrease of endodermal cell expressed sox17. The overexpression of zili, promoting the expression of gsc, inhibiting the expression of eve1 and resulting in the decrease of endodermal cell expressed sox17 were observed after microinjection of zili-mRNA.
zili might have some effect on the formation of the ectoderm, mesoderm and endoderm during early embryogenesis and might be important for normal embryonic development.
研究piwil2基因（zili）在斑马鱼早期胚胎发育过程中外胚层、中胚层和内胚层形成中的作用。
本研究使用zili吗啉代反义寡核苷酸和5个错配对照吗啉代。将zili克隆到表达载体中。体外合成zili mRNA。合成gsc、evel和sox17的反义RNA探针。分别将zili-MO、zili-cMO和zili mRNA显微注射到单细胞胚胎中。采用整胚原位杂交技术监测标记基因的表达。
显微注射敲低zili表达的zili-MO后，外胚层和中胚层标记基因gsc的表达下调，外胚层标记基因evel的表达上调，内胚层细胞表达的sox17减少。显微注射zili-mRNA后，观察到zili过表达，促进gsc表达，抑制eve1表达，内胚层细胞表达的sox17减少。
zili可能在早期胚胎发育过程中对外胚层、中胚层和内胚层的形成有一定作用，可能对正常胚胎发育很重要。
