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Influence of endothelin to cell morphology of dendrites and cell motility in cultured normal human melanocytes

内皮素对培养正常人黑素细胞树突细胞形态和细胞运动的影响

基本信息

批准号：
10670802
负责人：
AKASAKA Toshihide
金额：
$ 1.34万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

Melanocyte can proliferate and migrate in wound healing, however, the mechanism of it remains unknown Synthesis of melanin and dendricity of melanocytes, which supply melanin for keratinocytes to defense against UV damageis, influenced by endotherin-1 (ET-1) that produced by surrunded keratinocytes. Then we study the influence of ET-1 to melanocytes migration, dendricity and expression of p125 FAK, which is known as one of the focal contct localized protein involved in formation of focal contact.Mean migration rates and dendrites morphology of human newborn melanocytes cultured in ET-1 contained medium on a cover-slips coated fibronectin were mesured using the time-lapse image analysis. Expression of p125 FAK by double stained with anti-p125 FAK fluolescein and rhodamine, and of p125 FAK m-RNA by real time RT-PCR.under the ET-1 contained medium were also examined.Highest migration rate of melanocyte under no ET-1 was noted in 7 hrs after cell seeding.Reduced cell migration dependent on concentration of endothelin-1 was exibited. Melanocytes exhibited p125 FAK containing plaques localized in the periphery of actin stress fiber within the cell wall, however, plaque number of p125 FAK was reduced by ET-1 concentration. p125 FAK m-RNA was also reduced by ET-1. The otherhand length of dendrites, %dendricity, and number of dendrites of the melanocytes were increased dependent on ET-1 concentration.Taken together, we demonstrated roles of ET-1, which is paracrined by keratinocytes, in melanocyte motility and dendricity. Our data provide further support for the concept that proliferation or differentiation of melanocyte in the skin is controlled by keratinocyte, and can be usefull for the treatment for the vitiligo vulugalis, postinflammatory depigmentation.

黑素细胞在伤口愈合过程中可以增殖和迁移，但其机制尚不清楚。黑色素的合成和黑素细胞的树突性受周围角质形成细胞产生的内皮素-1 （ET-1）的影响，为角质形成细胞提供黑色素以抵御紫外线损伤。然后研究ET-1对黑素细胞迁移、树突和p125 FAK表达的影响，p125 FAK是参与局灶接触形成的定位蛋白之一。用延时图像分析方法测量新生黑素细胞在含有ET-1的培养基中培养的平均迁移率和树突形态。通过抗p125 FAK荧光素和罗丹明双染色检测p125 FAK的表达，实时RT-PCR检测p125 FAK m-RNA的表达。在ET-1所含介质下也进行了检查。未使用ET-1的黑素细胞在细胞播种后7小时的迁移率最高。内皮素-1的浓度降低了细胞迁移。黑素细胞表现出位于细胞壁内肌动蛋白应激纤维周围的含有p125 FAK的斑块，然而，ET-1浓度降低了p125 FAK的斑块数量。ET-1也减少了p125 FAK m-RNA。另一方面，黑素细胞的树突长度、树突%和树突数量随ET-1浓度的增加而增加。综上所述，我们证明了ET-1在黑素细胞运动和树突中的作用，ET-1是角质形成细胞旁分泌的。我们的数据进一步支持了皮肤中黑素细胞的增殖或分化受角质细胞控制的概念，并可用于治疗白癜风，炎症后色素沉着。