Wang Yue-Si, Li Jian-Yuan, Jin Shao-Hua, Wang Hai-Yan
Department of Biotech, Binzhou Medical College, Yantai 264003, China.
Zhonghua Xue Ye Xue Za Zhi. 2008 Aug;29(8):535-9.
To explore the effect of basic fibroblast growth factor 1 (bFGF1) during embryonic development on hematopoiesis, to study the expression of FGF1, vascular endothelial growth factor receptor (KDR), CD133, CD34 and transcription factors Ihh, SCL, GATA-1, GATA-2 and PU. 1 in the yolk sac, and to learn about the role and relationships of FGF1, hematopoietic cells and transcription factors during embryonic hematopoiesis.
10 microm sections and total RNA were prepared from 107 human embryos aged 3-12 weeks. Immunohischemical SP staining and RT-PCR were performed.
The yolk sac blood islands of human 3 approximately 12 weeks embryos consisted of peripheral vascular endothelial cells and central hematopoietic cells. The expression of FGF1 was firstly found in visceral mesoderm around periphery of yolk sac blood island at day 16, while was little inside it. KDR was not or lowly expressed and CD34 and CD133 were not expressed then. The expression increased, gray value decreased and staining enhanced at day 21. Strong staining of CD34+, CD133+ and KDR+ cells were found in blood island and mesoderm at day 30, their gray values changed from 156 +/- 16, 173 +/- 18 and 160 +/- 14 to 53 +/- 7, 52 +/- 6 and 69 +/- 8 respectively. FGF1 expression was strong positive, the gray value declined dramatically from 161 +/- 13 to 40 +/- 5. Some positive cells formed vessel-like structure along the periphery of blood island. Moderate expression of CD34+, CD133+, KDR+ cells increased at day 45, the cells aggregated into mass in blood island and FGF1+ cells did the same in blood island, while little in mesoderm. Its gray valve was increased. After 7 weeks, CD133+, KDR+, CD34+ cells significantly decreased their gray values increased, the staining became week. FGF1 was weakly expressed in yolk sac and its gray value increased to 179 +/- 22. RT-PCR showed Ihh, SCL, GATA-1 and GATA-2 were expressed at different time in yolk sac. PU. 1 were not expressed at day 16, and then expressed.
The hematopoietic properties of yolk sac may be dependent on signaling through FGF receptors and FGF1 plays an important role in hematopoietic stem cell homeostasis. The FGF pathway regulates primitive hematopoiesis by modulating transcription factors such as Gata1 expression level and activity.
探讨胚胎发育过程中碱性成纤维细胞生长因子1(bFGF1)对造血的影响,研究FGF1、血管内皮生长因子受体(KDR)、CD133、CD34以及转录因子Ihh、SCL、GATA-1、GATA-2和PU.1在卵黄囊中的表达情况,了解FGF1、造血细胞及转录因子在胚胎造血过程中的作用及相互关系。
取107例3至12周龄人胚胎制备10μm切片及总RNA,进行免疫组织化学SP染色及RT-PCR检测。
人3至12周胚胎的卵黄囊血岛由外周血管内皮细胞和中央造血细胞组成。FGF1于第16天首先在卵黄囊血岛周边的脏壁中胚层表达,血岛内表达较少。此时KDR不表达或低表达,CD34和CD133不表达。第21天时表达增加,灰度值降低,染色增强。第30天时血岛及中胚层可见CD34+、CD133+和KDR+细胞强染色,其灰度值分别从156±16、173±18和160±14变为53±7、52±6和69±8。FGF1表达呈强阳性,灰度值从161±13急剧降至40±5。部分阳性细胞沿血岛周边形成血管样结构。第45天时CD34+、CD133+、KDR+细胞中度表达增加,细胞在血岛聚集,FGF1+细胞在血岛也聚集,中胚层较少。其灰度值增加。7周后,CD133+、KDR+、CD34+细胞灰度值显著降低,染色变弱。FGF1在卵黄囊中弱表达,灰度值增至179±22。RT-PCR显示Ihh、SCL、GATA-1和GATA-2在卵黄囊中于不同时间表达。PU.1在第16天时不表达,之后表达。
卵黄囊的造血特性可能依赖于通过FGF受体的信号传导,FGF1在造血干细胞稳态中起重要作用。FGF途径通过调节转录因子如Gata1的表达水平和活性来调控原始造血。