Bullock S L, Fletcher J M, Beddington R S, Wilson V A
Laboratory of Mammalian Development, Medical Research Council (MRC) National Institute for Medical Research, The Ridgeway, London NW7 1AA, UK.
Genes Dev. 1998 Jun 15;12(12):1894-906. doi: 10.1101/gad.12.12.1894.
Heparan sulfate proteoglycans have been implicated in the presentation of a number of secreted signaling molecules to their signal-transducing receptors. We have characterized a gene trap mutation in the gene encoding a heparan sulfate biosynthetic enzyme, heparan sulfate 2-sulfotransferase (HS2ST). Transgenic mice were generated from embryonic stem cells harboring this insertion. lacZ reporter gene activity in heterozygous embryos demonstrates that the gene is expressed differentially during embryogenesis, presumably directing dynamic changes in heparan sulfate structure. Moreover, mice homozygous for the Hs2st gene trap allele die in the neonatal period, exhibiting bilateral renal agenesis and defects of the eye and the skeleton. Analysis of kidney development in Hs2st mutants reveals that the gene is not required for two early events-ureteric bud outgrowth from the Wolffian duct and initial induction of Pax-2 expression in the metanephric mesenchyme. It is required, however, for mesenchymal condensation around the ureteric bud and initiation of branching morphogenesis. Because 2-O-sulfation has been shown to influence the functional interactions of ligands with heparan sulfate in vitro, we discuss the possibility that the Hs2st mutant phenotype is a consequence of compromised interactions between growth factors and their signal-transducing receptors. These data provide the first genetic evidence that the regulated synthesis of differentially glycosylated proteoglycans can affect morphogenesis during vertebrate development.
硫酸乙酰肝素蛋白聚糖与多种分泌型信号分子向其信号转导受体的呈递有关。我们对编码硫酸乙酰肝素生物合成酶硫酸乙酰肝素2 - O -磺基转移酶(HS2ST)的基因中的一个基因陷阱突变进行了表征。从携带该插入片段的胚胎干细胞中培育出了转基因小鼠。杂合子胚胎中的lacZ报告基因活性表明该基因在胚胎发育过程中差异表达,推测其指导硫酸乙酰肝素结构的动态变化。此外,Hs2st基因陷阱等位基因的纯合子小鼠在新生儿期死亡,表现出双侧肾缺如以及眼睛和骨骼缺陷。对Hs2st突变体肾脏发育的分析表明,该基因对于两个早期事件并非必需——输尿管芽从沃尔夫管长出以及在中肾间充质中Pax - 2表达的初始诱导。然而,它对于输尿管芽周围的间充质凝聚和分支形态发生的起始是必需的。因为在体外已表明2 - O -硫酸化会影响配体与硫酸乙酰肝素的功能相互作用,所以我们讨论了Hs2st突变体表型是生长因子与其信号转导受体之间相互作用受损的结果的可能性。这些数据提供了首个遗传学证据,即差异糖基化蛋白聚糖的调控合成可影响脊椎动物发育过程中的形态发生。