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Identification of genes and proteins related to hair growth and analysis of alopecia inducing and preventing factor

毛发生长相关基因和蛋白的鉴定及脱发诱发和预防因素分析

基本信息

批准号：
18590551
负责人：
SAIJOH Kiyofumi
金额：
$ 2.48万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590551/
关键词：
male hair male-type alopecia hair root outer root sheath mRNA DNA tip hair keratin acidic 5 and basic 5 tvrosinase and tyrosinase related protein 1 毛髪 RNA DNAtip ケラチン protease albinism

项目摘要

At first, we succeed to purify mRNA directly from hair root and outer root sheath. Using mRNA from the normal male scalp hair and the scalp hair from male-type alopecia, expression difference among mRNA species were analyzed by DNA tip analysis. We focused on difference of expression of keratin that is supposed to affect the quality of hair. We also focused on genes related to melanin synthesis like tyrosinase (TYR) and tyrosinase related protein 1 (TYRP1) because hair color changes were frequently observed in male-type alopecia. TYR is a rate-limiting enzyme of this cascade and TYRP1 is essential to maintain TYR function. On DNA tip analysis, hair keratin KA36 and keratin 25 showed abundant expression in normal hair. In alopecia pair, hair keratin acidic 5 (KRTA5) and basic 5 (KRTB5) as well as keratin 15 (KRT15) were abundantly expressed but TYR and TYRP I were not. Next, we tried to quantitate their expression using RT-PCR. Expression of KRTA5 and B5 was approx. 3.5 times as much and p-actin expression in normal hair, whereas that in alopecia hair was approx. 5.5 times. KRT15 expression in normal hair and alopecia hair was respectively approx. 0.5 times and 6.5 times as that of β-actin, viz. KRT expression in alopecia was more than 13 times higher than that in normal hair. TYR and TYRP1 expression was approx. 0.1 and 0.3 times as that of (-actin, respectively. It is not always observed in all alopecia hair but some alopecia hair showed almost null expression of them. Even in alopecia patient, hair obtained from the non-bald area displayed the expression pattern similar with normal hair. Thus, we consider that the analysis of these genes can show us the way to distinguish the area affected and non-affected by male-type alopecia and apply to the patent.

首先，我们成功地从毛根和外根鞘中直接纯化mRNA。以正常男性头皮毛发和男性型秃发头皮毛发的mRNA为样本，采用DNA尖端分析方法分析mRNA在不同物种间的表达差异。我们重点研究了角蛋白的表达差异，角蛋白被认为会影响头发的质量。我们还关注了与黑色素合成相关的基因，如酪氨酸酶（TYR）和酪氨酸酶相关蛋白1 (TYRP1)，因为在男性型脱发中经常观察到头发颜色的变化。TYR是该级联的限速酶，TYRP1对维持TYR功能至关重要。DNA尖端分析显示，头发角蛋白KA36和角蛋白25在正常头发中表达丰富。在脱发对中，头发角蛋白酸性5 （KRTA5）和碱性5 （KRTB5）以及角蛋白15 （KRT15）大量表达，而TYR和TYRP I未表达。接下来，我们尝试使用RT-PCR来定量它们的表达。KRTA5和B5的表达量相近。正常头发中p-actin的表达量为正常头发的3.5倍，而脱发头发中p-actin的表达量约为正常头发的3.5倍。5.5倍。KRT15在正常发和脱发发中的表达分别接近。是β-肌动蛋白的0.5倍和6.5倍，即KRT在脱发中的表达比正常头发高13倍以上。TYR和TYRP1表达量相近。分别是（-actin）的0.1倍和0.3倍。并不是所有的秃发都有表达，但有些秃发几乎没有表达。即使在脱发患者中，从非秃顶区域获得的头发也显示出与正常头发相似的表达模式。因此，我们认为对这些基因的分析可以向我们展示区分男性型脱发的影响区域和非影响区域的方法，并申请专利。