Cardinali Giorgia, Bolasco Giulia, Aspite Nicaela, Lucania Giuseppe, Lotti Lavinia V, Torrisi Maria R, Picardo Mauro
Istituto Dermatologico San Gallicano, IRCCS, Rome, Italy.
J Invest Dermatol. 2008 Mar;128(3):558-67. doi: 10.1038/sj.jid.5701063. Epub 2007 Sep 20.
The transfer of melanin from melanocytes to keratinocytes is upregulated by UV radiation and modulated by autocrine and paracrine factors. Among them, the keratinocyte growth factor (KGF/FGF7) promotes melanosome transfer acting on the recipient keratinocytes through stimulation of the phagocytic process. To search for possible differences in the melanosome uptake of keratinocytes from different skin color, we analyzed the uptake kinetics and distribution pattern of fluorescent latex beads in primary cultures of light and dark skin-derived keratinocytes stimulated with KGF and we compared the direct effect of KGF on the melanosome transfer in co-cultures of human primary melanocytes with light and dark keratinocytes. KGF-promoted melanosome transfer was more significant in light keratinocytes compared to dark, due to an increased expression of KGF receptor in light skin keratinocytes. Colocalization studies performed by confocal microscopy using FITC-dextran as a phagocytic marker and fluorescent beads as well as inhibition of particle uptake by cytochalasin D, revealed that beads internalization induced by KGF occurs via actin-dependent phagocytosis. 3D image reconstruction by fluorescence microscopy and ultrastructural analysis through transmission electron microscopy showed differences in the distribution pattern of the beads in light and dark keratinocytes, consistent with the different melanosome distribution in human skin.
黑色素从黑素细胞向角质形成细胞的转移受紫外线辐射上调,并受自分泌和旁分泌因子调节。其中,角质形成细胞生长因子(KGF/FGF7)通过刺激吞噬过程作用于受体角质形成细胞,促进黑素小体转移。为了探究不同肤色角质形成细胞在黑素小体摄取方面可能存在的差异,我们分析了用KGF刺激的浅色和深色皮肤来源的角质形成细胞原代培养物中荧光乳胶珠的摄取动力学和分布模式,并比较了KGF对人原代黑素细胞与浅色和深色角质形成细胞共培养物中黑素小体转移的直接影响。与深色角质形成细胞相比,KGF促进的黑素小体转移在浅色角质形成细胞中更显著,这是由于浅色皮肤角质形成细胞中KGF受体的表达增加。使用FITC-葡聚糖作为吞噬标记物和荧光珠通过共聚焦显微镜进行的共定位研究,以及细胞松弛素D对颗粒摄取的抑制作用,表明KGF诱导的珠子内化通过肌动蛋白依赖性吞噬作用发生。通过荧光显微镜进行的三维图像重建和通过透射电子显微镜进行的超微结构分析显示,浅色和深色角质形成细胞中珠子的分布模式存在差异,这与人类皮肤中不同的黑素小体分布一致。