Gene expression profiles of E13 bladder compartments. (GUDMAP Series ID: 24)

The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing bladder. 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 mesenchymal and epithelial compartments of the e13 mouse bladder. Keywords: Mouse embryonic day 13 bladder mesenchyme and epithelium. FVB/N mice were time mated. At embryonic day 13 mice were euthanized by decapitation and the bladders were microdissected, cut just above the ureters, and treated with 20 micromolar EDTA for 20 minutes. Mesenchymal and epithelial compartments were then separated by rimming the bladder with a needle and harvested in RLT. Total RNA was isolated for gene expression analysis using the Affymetrix MOE430 microarray chip本研究的长期目标为构建发育中膀胱多组织组分的全基因表达水平百科全书。本研究的核心论点清晰直白：将显微切割组织、荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）所得细胞与基因微阵列分析相结合，可为发育中的泌尿生殖系统构建全域基因表达图谱提供一种高效、强力且可靠的技术方案。覆盖全基因组的基因微阵列可用于定量检测所有基因的表达水平，其快速检出结果可生成一套相较于单纯原位杂交技术更为灵敏、经济且全面的基因表达图谱。本次提交的数据详细描绘了胚胎期13天（e13）小鼠膀胱的间充质与上皮细胞组分的基因表达谱。关键词：胚胎期13天小鼠膀胱、间充质、上皮细胞。实验采用定时交配的FVB/N品系小鼠：于胚胎发育第13天时，通过断头法处死孕鼠，显微切割获取膀胱组织，在输尿管上方截断膀胱后，以20微摩尔浓度的乙二胺四乙酸（EDTA）处理20分钟；随后通过针头刮擦膀胱壁的方式分离间充质与上皮细胞组分，并将其收集于RLT裂解液中；最终提取总RNA，采用Affymetrix MOE430基因微阵列芯片开展基因表达分析。