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

黑素细胞在创面愈合过程中可以增殖和迁移，但其机制尚不清楚。黑素合成和黑素细胞的树突状结构为角质形成细胞提供黑色素以抵御紫外线损伤，受抑制角质形成细胞产生的内异素-1(ET-1)的影响。然后，我们研究了ET-1对黑素细胞迁移、树突性和p125 FAK表达的影响，p125 FAK是参与局灶性接触形成的局部接触蛋白之一。用时间推移图像分析方法测量了在含有ET-1的培养液中培养的人新生黑素细胞在纤维连接蛋白覆盖片上的平均迁移率和树突形态。用抗p125 FAK荧光素和罗丹明双染法检测p125 FAK的表达，实时RT-PCR法检测p125 FAK m-RNA在ET-1作用下的表达，细胞在无ET-1作用下7h细胞迁移速率最高，细胞迁移减少依赖于ET-1的浓度。黑素细胞胞壁内肌动蛋白应激纤维周边可见p125FAK斑块，但ET-1浓度可使p125FAK斑块数目减少。ET-1还可使p125FAK m-RNA减少。另一方面，随着ET-1浓度的增加，黑素细胞的树突长度、树突度百分比和树突数增加。我们共同证明了角质形成细胞旁分泌的ET-1在黑素细胞运动和树突状中的作用。我们的结果进一步支持了皮肤中黑素细胞的增殖或分化由角质形成细胞控制的观点，并可用于治疗寻常型白癜风和炎症后色素脱失。

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.