Analysis of novel chemokine "WHCHE" in development and hematopoies

从背主动脉来源的内皮基质细胞系中分离出一种新的趋化因子WECHE （CXCLI 5）。先前有报道称，WECHE抑制卵黄囊来源的内皮细胞的增殖和胎儿肝来源的造血细胞的分化。为了研究WECHE在体内的作用，我们利用腺病毒系统和转基因小鼠在体内过表达WECHE。(1)腺病毒系统在鸡β -肌动蛋白启动子控制下，通过连接WECHE cDNA构建腺病毒。将腺- weche感染至E13.5胚胎，并在E17对胎儿肝脏进行分析。与对照组（Adeno-LacZ）相比，WECHE感染的胎儿肝脏含有2倍以上的CFU-E。(2)利用转基因小鼠secaag启动子制备WECHE转基因小鼠。对50多只小鼠进行了分析，经PCR分析发现1只小鼠携带该基因。然后提取不同脏器的mRNA， RT-PCR分析WECHE的表达情况。在转基因小鼠中未观察到WECHE mRNA。为了验证VVECHE对体内内皮细胞发育的影响，我们将制备Flk-1-WECHE转基因小鼠。

A novel chemokine 'WECHE' (CXCLI 5) was isolated from a dorsal aorta-derived endothelial stromal cell line. It was reported previously that WECHE inhibits the proliferation of yolk sac-derived endothelial cells and the differentiation of fetal liver-derived hematopoletic cells. In order to investigate a role of WECHE in vivo, WECHE was overexpressed in vivo by using Adenovirus system and transgenic mouse.(1) Adenovirus systemAdenovirus was constructed by the ligation of WECHE cDNA under the controlled of chicken beta-actin promoter. The Adeno-WECHE was infected to E13.5 embryos and fetal liver was analyzed at E17. WECHE infected fetal liver contains two-fold higher number of CFU-E compared to the control (Adeno-LacZ).(2) Transgenit mouseCAAG 'promoter was used to generate WECHE transgenic mouse. Over fifty numbers of mice were analyzed, and it was found that one mouse has the transgene by PCR analysis. Then, several kinds of organs were extracted for mRNA, and WECHE expression was analyzed by RT-PCR. None of WECHE mRNA was observed in the transgenic mouse. We are going to generate Flk-1-WECHE transgenic mouse in order to examine the effect of VVECHE for endothelial cell development in vivo.