Identification of kidney mesenchymal genes by a combination of microarray analysis and Sall1-GFP knockin mice
Identification of kidney mesenchymal genes by a combination of microarray analysis and Sall1-GFP knockin mice
复制标题
DOI：
10.1016/j.mod.2004.04.007
复制
发表时间：
2004-06-01
影响因子：
2.6
通讯作者：
Nishinakamura, R
中科院分区：
文献类型：
--
作者：
Takasato, M;Osafune, K;Nishinakamura, R
SALL1 是 Townes-Brocks 综合征的致病基因，编码锌指蛋白，其小鼠同源物 (Sall1) 对于后肾发育至关重要，正如基因靶向过程中所指出的那样。在胚胎肾中，Sall1 在间充质衍生结构中大量表达，从浓缩间充质、S-、逗号形小体到肾小管和足细胞。我们生成了将绿色荧光蛋白 (GFP) 基因插入 Sall1 基因座的小鼠，并通过荧光素激活细胞分选从这些小鼠的胚胎肾脏中分离出 GFP 阳性群体。 GFP阳性群体确实表达间充质基因，而阴性群体则表达输尿管芽中的基因。为了系统地寻找间充质来源的细胞中表达的基因，我们使用微阵列分析比较了 GFP 阳性和阴性群体的基因表达谱，然后进行原位杂交。我们检测到许多已知对后肾发育重要的基因，包括 Sall1、GDNF、Raldh2、Pax8 和 FoxD1，以及在后肾间充质中大量表达的基因，如 Unc4.1、Six2、Osr-2 和 PDGFc。我们还发现了包括 SSB-4、Smarcd3、mu-Crystallin、TRB-2 在内的几组基因，这些基因在后肾间充质中尚未表达。因此，微阵列技术和 Sall1-GFP 小鼠的结合可用于系统鉴定发育中肾脏中表达的基因。 (C) 2004 Elsevier Ireland Ltd. 保留所有权利。
SALL1, a causative gene for Townes-Brocks syndrome, encodes a zinc finger protein, and its mouse homolog (Sall1) is essential for metanephros development, as noted during gene targeting. In the embryonic kidney, Sall1 is expressed abundantly in mesenchyme-derived structures from condensed mesenchyme, S-, comma-shaped bodies, to renal tubules and podocytes. We generated mice in which a green fluorescent protein (GFP) gene was inserted into the Sall1 locus and we isolated the GFP-positive population from embryonic kidneys of these mice by fluorescein-activated cell sorting. The GFP-positive population indeed expressed mesenchymal genes, while the negative population expressed genes in the ureteric bud. To systematically search for genes expressed in the mesenchyme-derived cells, we compared gene expression profiles in the GFP-positive and -negative populations using microarray analysis, followed by in situ hybridization. We detected many genes known to be important for metanephros development including Sall1, GDNF, Raldh2, Pax8 and FoxD1, and genes expressed abundantly in the metanephric mesenchyme such as Unc4.1, Six2, Osr-2 and PDGFc. We also found groups of genes including SSB-4, Smarcd3, mu-Crystallin, TRB-2, which are not known to be expressed in the metanephric mesenchyme. Therefore a combination of microarray technology and Sall1-GFP mice is useful for systematic identification of genes expressed in the developing kidney. (C) 2004 Elsevier Ireland Ltd. All rights reserved.