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Identification of dermal papilla cell specific genes

真皮乳头细胞特异性基因的鉴定

基本信息

批准号：
11670833
负责人：
ARASE Seiji
金额：
$ 2.11万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Dermal papilla cells were cultured from the excised hair follicles obtained from the bald lesion and non bald occipital lesion of a male subject afflicted with an androgenic alopecia. Then total RNAs were extracted from those cells (DPCs), and 3'-directed cDNA libraly were constructed using oligo-dT tailed pUC 19 as a primer. Approximately 2000 colonies were picked up randomly and nucleotide sequence of each cDNA was determined. In silico analisis of the sequences revealed that each library contained hundreds kinds of genes because of their redunduncy, indicating that some of them were cloned twice or more. Genes of type I collagen, fibronectin, and osteonectin were cloned ten times or more, which means that DPCs abundantly produce those extracellular matrix. We categorize highly expressed genes into (1) new genes highly expressed in DPC, (2) known genes whose expression is highly different between the two types of dermal papilla cells. We chose 8 candidate new genes and whose DNA sequences were fixed, and localization of gene products were studied using monoclonal untibodies against specific peptides of those gene products. Unfortunately, all of those genes were expressed not only in DPCs but also other types of cutaneous cells in vivo (not specific to DPCs). Knock out animals of those genes were not yet established. The unknown genes may be involved in the maintenance of hair growth and development of androgenic alopecia. Further analysis of these genes are currently in progress.We confirmed that dermal papilla cells produce a chemotactic factors which attracts hair follicle keratinocytes by chemotaxis.We also found some lines of evidense which suggest that Minoxidil directly binds to sulfonylurea receptor of DPCs and finally enhances VEGF production through the adenosin-adenosine receptor pathway in DPCs.

毛乳头细胞是从一位患有雄激素性脱发的男性患者的秃发和非秃头枕部病变中取出的毛囊中培养出来的。然后从这些细胞(DPC)中提取总RNA，以寡聚脱氧核糖核酸末端的PUC19为引子，构建3‘-DNA文库。随机抽取约2000个菌落，测定了每个基因的核苷酸序列。序列的电子分析表明，每个文库包含数百种基因，因为它们是冗余的，表明有些基因被克隆了两次或更多。I型胶原、纤维连接蛋白和骨连接蛋白的基因被克隆了10次或更多，这意味着DPC大量地产生这些细胞外基质。我们将高表达的基因分为(1)在DPC中高表达的新基因，(2)在两种类型的毛乳头细胞中表达高度不同的已知基因。我们选择了8个DNA序列固定的候选新基因，并利用针对这些基因产物的特异性多肽的单抗对基因产物进行了定位研究。不幸的是，所有这些基因不仅在DPC中表达，而且在体内其他类型的皮肤细胞中也表达(不是DPC所特有的)。这些基因敲除的动物还没有确定。这些未知基因可能参与了雄激素性脱发毛发生长发育的维持。对这些基因的进一步分析目前正在进行中。我们证实了毛乳头细胞产生一种趋化因子，通过趋化作用吸引毛囊角质形成细胞。我们还发现了一些证据，表明米诺地尔直接与DPC的磺脲受体结合，最终通过腺苷-腺苷受体途径促进DPC中VEGF的产生。