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.

从秃头病变中获得的切除的毛囊和非秃头枕骨病变培养真皮乳头状细胞，该毛囊患有雄性脱发性脱发。然后从这些细胞（DPC）中提取总RNA，并使用Oligo-DT Tailed Puc 19作为底漆构建了3'定向的cDNA libraly。随机拾取大约2000个菌落，并确定每个cDNA的核苷酸序列。在序列的硅分析中表明，每个文库都包含数百种基因，因为它们的冗余，表明其中一些被克隆了两次或更多。 I型胶原蛋白，纤连蛋白和骨蛋白的基因被克隆了十次或更多，这意味着DPC大量产生那些细胞外基质。我们将高度表达的基因分为（1）在DPC中高度表达的新基因，（2）已知的基因，其表达在两种类型的皮肤乳头细胞之间高度不同。我们选择了8种候选新基因，其DNA序列是固定的，并使用单克隆非略微对这些基因产物的特定肽进行了基因产物的定位。不幸的是，所有这些基因不仅在DPC中表达，而且在体内（不是DPC）中的其他类型的皮肤细胞表达。尚未确定那些基因的动物。未知基因可能与雄激素脱发的头发生长和发育有关。目前正在进行对这些基因的进一步分析。我们证实，真皮乳头状细胞产生的趋化因子，通过趋化性吸引毛囊角质形成细胞。我们还发现了一些现象，这表明米诺克尼直接与DPC的硫酸盐受体结合并最终增强了VEGF VEVGF VEVEGF VEGF VEVENS VEVES VEVES VEVES VEVINCES VEVINCES VEVINCES VEVINCES VEVINCES VEVINCES VEVINCES VEVES VENINCES VEVINES VEVINCES VEVES VEVINESS。通过DPC中的腺苷 - 腺苷受体途径产生。