Gene expression profiles of E15 ureter (GUDMAP Series ID: 26)

The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing genitourinary tract. The central thesis is straightforward. The combination of microdissected tissues and FACS sorted cells plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing urogenital system. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. The data submitted here delineates the gene expression profiles of the e15 mouse ureter. Comparisons with the same compartments in the neonate will be made to determine changes in gene expression from relatively undifferentiated tissue (e15) to a highly differentiated structure (neonate). FVB/N mice were time mated. At embryonic day 15 mice were euthanized by decapitation and the ureters were microdissected, cut from pelvis to the bladder insertion point and snap frozen in liquid nitrogen. Total RNA was isolated for gene expression analysis using the Affymetrix GeneChip Mouse Genome 430 2.0 Array.本研究的长期目标为构建发育阶段泌尿生殖道各组分的全基因表达水平百科图谱。本研究的核心论点清晰明确：将显微切割组织与荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）得到的纯化细胞结合基因芯片（microarray）分析策略，可为发育中泌尿生殖系统构建全局基因表达图谱提供一种高效、可靠且功能强大的技术方案。覆盖度近乎完整的基因芯片可用于定量检测所有基因的表达水平，该技术所得的快速检测结果，可生成一套相较于单纯原位杂交（in situ hybridization）更为灵敏、经济且完整的表达图谱。 本研究提交的数据集描绘了e15小鼠输尿管的基因表达谱。后续将与新生个体的相同组织组分开展比对，以解析从相对未分化的e15组织到高度分化的新生器官过程中的基因表达变化。 实验通过定时交配获得FVB/N品系小鼠，在胚胎发育第15天时采用断颈法处死小鼠，对其输尿管进行显微切割：从肾盂至膀胱附着点完整分离输尿管，并立即置于液氮中快速冷冻保存。随后提取总RNA，使用Affymetrix GeneChip Mouse Genome 430 2.0 Array芯片完成基因表达分析。