Challen Grant A, Martinez Gemma, Davis Melissa J, Taylor Darrin F, Crowe Mark, Teasdale Rohan D, Grimmond Sean M, Little Melissa H
Institute For Molecular Bioscience, Queensland Bioscience Precinct, 306 Carmody Road, The University of Queensland, St. Lucia, Brisbane, QLD, 4072, Australia.
J Am Soc Nephrol. 2004 Sep;15(9):2344-57. doi: 10.1097/01.ASN.0000136779.17837.8F.
Although many of the molecular interactions in kidney development are now well understood, the molecules involved in the specification of the metanephric mesenchyme from surrounding intermediate mesoderm and, hence, the formation of the renal progenitor population are poorly characterized. In this study, cDNA microarrays were used to identify genes enriched in the murine embryonic day 10.5 (E10.5) uninduced metanephric mesenchyme, the renal progenitor population, in comparison with more rostral derivatives of the intermediate mesoderm. Microarray data were analyzed using R statistical software to determine accurately genes differentially expressed between these populations. Microarray outliers were biologically verified, and the spatial expression pattern of these genes at E10.5 and subsequent stages of early kidney development was determined by RNA in situ hybridization. This approach identified 21 genes preferentially expressed by the E10.5 metanephric mesenchyme, including Ewing sarcoma homolog, 14-3-3 theta, retinoic acid receptor-alpha, stearoyl-CoA desaturase 2, CD24, and cadherin-11, that may be important in formation of renal progenitor cells. Cell surface proteins such as CD24 and cadherin-11 that were strongly and specifically expressed in the uninduced metanephric mesenchyme and mark the renal progenitor population may prove useful in the purification of renal progenitor cells by FACS. These findings may assist in the isolation and characterization of potential renal stem cells for use in cellular therapies for kidney disease.
尽管肾脏发育过程中的许多分子相互作用现已得到充分了解，但参与从周围中间中胚层分化出后肾间充质，进而形成肾祖细胞群体的分子却鲜为人知。在本研究中，利用cDNA微阵列来鉴定在小鼠胚胎第10.5天（E10.5）未诱导的后肾间充质（即肾祖细胞群体）中富集的基因，并与中间中胚层更靠前的衍生物进行比较。使用R统计软件分析微阵列数据，以准确确定这些群体之间差异表达的基因。对微阵列异常值进行生物学验证，并通过RNA原位杂交确定这些基因在E10.5及早期肾脏发育后续阶段的空间表达模式。该方法鉴定出21个在E10.5后肾间充质中优先表达的基因，包括尤因肉瘤同源物、14-3-3θ、视黄酸受体α、硬脂酰辅酶A去饱和酶2、CD24和钙黏蛋白-11，这些基因可能在肾祖细胞的形成中起重要作用。像CD24和钙黏蛋白-11这样在未诱导的后肾间充质中强烈且特异性表达并标记肾祖细胞群体的细胞表面蛋白，可能在通过荧光激活细胞分选术（FACS）纯化肾祖细胞方面很有用。这些发现可能有助于分离和鉴定潜在的肾干细胞，用于肾脏疾病的细胞治疗。
