Changes in gene expression patterns in the ureteric bud and metanephric mesenchyme in models of kidney development
Changes in gene expression patterns in the ureteric bud and metanephric mesenchyme in models of kidney development
复制标题
DOI：
10.1046/j.1523-1755.2003.00383.x
复制
发表时间：
2003-12-01
影响因子：
19.6
通讯作者：
Nigam, SK
中科院分区：
文献类型：
--
作者：
Stuart, RO;Bush, KT;Nigam, SK
背景资料。在最近的一项研究中，使用高密度DNA阵列技术(Stuart RO，Bush KT，NigamSK，Proc Natl Acad Sci USA 98：5649-5654,2001)分析了大鼠肾脏发育过程中的基因表达模式。这种方法虽然揭示了发育中肾脏基因表达的整体模式，但并没有提供可能是输尿管芽(UB)和后肾间充质(MM)形态发生程序一部分的基因的贡献，这两个组织在肾单位形成过程中密切相互作用。我们现在已经使用高密度DNA阵列和双重体外转录(DIVT)方法来检测大鼠胚胎肾脏形态发生的体外模型中的基因表达模式，并将这些数据与整个胚胎肾脏在不同发育阶段的基因表达模式进行比较。结果表明，随着形态发生的发生，不同的基因在UB和MM中表达。来自体外UB和MM培养模型的dIVT数据与从整个胚胎肾脏在不同发育阶段获得的单个IVT数据进行了层次聚类，基因表达的整体模式显著兼容，支持该方法的有效性。研究了其表达与单个组织相关的基因的潜在作用，并确定了几条可能在肾脏发育中发挥作用的途径。例如，肝细胞核因子-6(HNF-6)，一种可能位于KSP-钙粘蛋白表达途径上游的转录因子，在UB中高表达。Embigin是一种在细胞/细胞外基质(ECM)相互作用中起重要作用的细胞黏附分子，在UB中也被发现，并可能在肾脏中作为Dolichos biflorus结合蛋白。ADAM10是一种参与Delta-Notch信号转导和Sit-Robo信号转导的去整合素-金属蛋白水解酶，在UB晚期也有高表达。研究发现，CELSR-3蛋白在MM的分化过程中高表达，该蛋白可能与WNT卷曲转导级联的成员一起参与形成肾单位的极化。DDR2是盘状结构域受体家族的成员，被认为在MM的分化过程中起作用。这项研究提供的数据表明，将肾脏发育的体外模型与高密度DNA阵列相结合，可以识别参与形态发生过程的基因。在形态发生的不同阶段，UB和MM表达的基因模式明显不同，其中许多与神经元生长和/或分化有关。总之，高密度微阵列数据不仅开始表明UB和MM中独立的遗传程序如何协调整个肾脏的形成，而且还表明迄今为止大部分未被探索的发育途径(涉及HNF-6、ADAM-10、Celsr-3、DDR2和其他基因)参与了肾脏的形成。
Background. In a recent study, the pattern of gene expression during development of the rat kidney was analyzed using high-density DNA array technology (Stuart RO, Bush KT, NigamSK, Proc Natl Acad Sci USA 98: 5649-5654, 2001). This approach, while shedding light on global patterns of gene expression in the developing kidney, does not provide insight into the contributions of genes that might be part of the morphogenetic program of the ureteric bud (UB) and metanephric mesenchyme (MM), the two tissues that interact closely during nephron formation.Methods. We have now used high-density DNA arrays together with a double in vitro transcription (dIVT) approach to examine gene expression patterns in in vitro models for morphogenesis of the rat UB( isolated UB culture) and MM(coculture with embryonic spinal cord) and compared this data with patterns of gene expression in the whole embryonic kidney at different stages of development.Results. The results indicate that different sets of genes are expressed in the UB and MM as morphogenesis occurs. The dIVT data from the in vitro UB and MM culture models was clustered hierarchically with single IVT data from the whole embryonic kidney obtained at different stages of development, and the global patterns of gene expression were remarkably compatible, supporting the validity of the approach. The potential roles of genes whose expression was associated with the individual tissues were examined, and several pathways were identified that could have roles in kidney development. For example, hepatocyte nuclear factor-6 (HNF-6), a transcription factor potentially upstream in a pathway leading to the expression of KSP-cadherin was highly expressed in the UB. Embigin, a cell adhesion molecule important in cell/extracellular matrix (ECM) interactions, was also found in the UB and may serve as a Dolichos biflorus binding protein in the kidney. ADAM10, a disintegrin-metalloprotease involved in Delta-Notch signaling and perhaps Slit-Robo signaling, was also highly expressed in late UB. Celsr-3, a protein, which along with members of the Wnt-frizzled transduction cascade, might be involved in the polarization of the forming nephron, was found to be highly expressed in differentiating MM. DDR2, a member of the discoidin domain receptor family, which is thought to function in the activation of matrix metalloproteinase- 2 (MMP-2), was also found to be highly expressed in differentiating MM. It is also interesting to note that almost 10% of the highly expressed genes in both tissues were associated with neuronal growth and/or differentiation.Conclusion. The data presented in this study point to the power of combining in vitro models of kidney development with high-density DNA arrays to identify the genes involved in the morphogenetic process. Clear differences were found between patterns of genes expressed by the UB and MM at different stages of morphogenesis, and many of these were associated with neuronal growth and/or differentiation. Together, the high-density microarray data not only begin to suggest how separate genetic programs in the UB and MM orchestrate the formation of the whole kidney, but also suggest the involvement of heretofore largely unexplored developmental pathways (involving HNF-6, ADAM-10, Celsr-3, DDR2, and other genes) in nephrogenesis.