Novel mouse embryonic renal marker gene products differentially expressed during kidney development.
Novel mouse embryonic renal marker gene products differentially expressed during kidney development.
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肾脏发育过程中差异表达的新型小鼠胚胎肾脏标记基因产物。
DOI：
10.1152/ajprenal.1996.271.3.f770
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Holzman,LB
中科院分区：
文献类型：
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作者：
Kretzler,M;Fan,G;Rose,D;Arend,LJ;Briggs,JP;Holzman,LB
研究人员接近肾脏器官发生的问题一直受到缺乏适当的分子标记，指定不同的发育表型的阻碍。为了确定这些标记，我们采用差异显示聚合酶链反应（DD-PCR）方法，在受孕后11.5、13.5、15.5和17.5天的小鼠肾脏和成年肾脏中，研究了基因表达的时间模式。鉴定、分离和测序了22个差异表达的扩增产物。17个克隆与先前报道的核苷酸序列没有明显的相似性：两个与两个管家基因产物相似，三个与人类或大鼠表达的序列标签相似。为了证实DD-PCR观察到的差异表达模式，使用序列特异性寡核苷酸引物进行半定量逆转录pcr。22个克隆中有19个在肾脏发育过程中存在差异表达[小鼠胚胎肾标志物（MERM）序列1-19]。在小鼠后肾器官培养中进一步评估了MERM作为发育标志物的价值，其中MERM转录物的表达模式模仿了在体内观察到的。因此，DD-PCR方法允许开发一组标记序列，可用于表征肾脏发育过程，并可能允许鉴定新的、功能相关的基因产物。
Investigators approaching the problem of renal organogenesis have been hampered by a paucity of suitable molecular markers that specify distinct developmental phenotypes. To identify such markers, differential display-polymerase chain reaction (DD-PCR) was used to survey the temporal pattern of gene expression in mouse kidney at 11.5, 13.5, 15.5, and 17.5 days after conception and in the adult kidney. Twenty-two differentially expressed amplification products were identified, isolated, and sequenced. Seventeen clones showed no significant similarity with previously reported nucleotide sequences: two were similar to two housekeeping gene products, and three were similar to human or rat expressed sequence tags. To confirm the differential expression patterns observed by DD-PCR, semiquantitative reverse transcription-PCR was performed using sequence-specific oligonucleotide primers. Nineteen of 22 clones were differentially expressed during kidney development [mouse embryonic renal marker (MERM) sequences 1-19]. The value of MERMs as developmental markers was further assessed in mouse metanephric organ culture, where the pattern of MERM transcript expression mimicked that observed in vivo. Therefore, the DD-PCR method permitted development of a panel of marker sequences that can be used to characterize renal developmental processes and that may allow the identification of novel, functionally relevant gene products.