Analysis of novel chemokine "WHCHE" in development and hematopoies

一种新的趋化因子‘WECHE’(CXCLI5)是从背主动脉来源的内皮基质细胞系中分离得到的。此前有报道称，WECHE抑制卵黄囊来源的内皮细胞的增殖和胎肝来源的造血细胞的分化。为了研究WECHE在体内的作用，利用腺病毒系统和转基因小鼠在体内进行了WECHE的高效表达。(1)腺病毒系统：在鸡β-肌动蛋白启动子的控制下，连接WECHE基因，构建成重组腺病毒。将重组腺病毒感染E13.5胚胎，在E17胎龄时对胎肝进行分析。(2)利用转基因小鼠CAAG启动子构建了WECHE转基因小鼠，通过对50多只小鼠的分析，发现有1只小鼠表达了WECHE基因。然后提取几种器官的mRNA，用RT-PCR分析WECHE基因的表达。在转基因小鼠中没有检测到WECHE基因的表达。我们将建立Flk-1-WECHE转基因小鼠，以检测VVECHE对体内血管内皮细胞发育的影响。

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.