Spatial expressions of fibronectin and integrins by human and rodent dermal fibroblasts
Spatial expressions of fibronectin and integrins by human and rodent dermal fibroblasts
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DOI：
10.1111/j.1365-2133.2006.07399.x
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发表时间：
2006-09-01
影响因子：
10.3
通讯作者：
Takahashi, K.
中科院分区：
文献类型：
--
作者：
Yasuda, M.;Miyachi, Y.;Takahashi, K.
背景：人类皮肤表现出不同的形态学特征，这取决于身体部位。由于这些不同的表型已被解释的基础上的差异表皮角质形成细胞和皮肤附件的存在下，空间的区别，真皮成分还没有得到充分阐明。目的：确定和表征的档案，丰富或特异性表达的成纤维细胞来源于躯干皮肤，但不是从掌跖皮肤或口腔粘膜。方法：为了鉴定躯干皮肤成纤维细胞的不同mRNA表达，首先进行消减cDNA筛选，然后对培养的人和大鼠真皮成纤维细胞和这些皮肤组织进行北方印迹、Western印迹和免疫组织化学。最后，整体安装在原位杂交（WISH）进行检查相应的基因在小鼠胚胎发育阶段的表达差异。结果：我们确定了三个cDNA克隆编码纤连蛋白（FN），妊娠特异性β 1-糖蛋白5和β-肌动蛋白，分别是丰富或特异性表达的主干皮肤成纤维细胞的mRNA。进一步证实FN和一些整合素在人和大鼠躯干皮肤成纤维细胞中更有选择性地表达，无论是在RNA还是蛋白质水平上，与来自足底皮肤和口腔粘膜的成纤维细胞相比。结论：FN是最强的细胞外基质分子之一，其转录的空间依赖于体细胞部位。FN的高表达提示皮肤发育和毛胚附属器的形态发生过程中的基本定型。FN在掌跖皮肤和口腔粘膜中的缺乏可能解释了这些皮肤表型的特征。
Background: Human skin shows various morphological characteristics, depending on the body site. As these distinct phenotypes have been explained on the basis of the variance in epidermal keratinocytes and the presence of skin appendages, the spatial distinction of the dermal components has not been fully elucidated.Objectives: To identify and characterize the profiles of mRNAs that are abundantly or specifically expressed by fibroblasts derived from trunk skin, but not from palmoplantar skin or oral mucosa.Methods: In order to identify the distinct mRNA expression by trunk skin fibroblasts, a subtraction cDNA screening was performed first, followed by Northern blotting, Western blotting and immunohistochemistry for cultured human and rat dermal fibroblasts and those skin tissues. Finally, whole mount in situ hybridization (WISH) was performed to examine the differences in the expression of the corresponding gene during the developmental stage of mouse embryos.Results: We identified three cDNA clones encoding fibronectin (FN), pregnancy-specific beta 1-glycoprotein 5 and beta-actin, respectively, whose mRNAs were abundantly or specifically expressed by trunk skin fibroblasts. FN and some integrins were further confirmed to be expressed more selectively in human and rat trunk skin fibroblasts, both in terms of the RNA and the protein levels, compared with the fibroblasts derived from plamoplantar skin and oral mucosa. WISH demonstrated that FN was localized around the hair follicles of mouse embryos.Conclusion: FN, one of most potent extracellular matrix molecules, was demonstrated to be spatially transcribed depending on the body sites. The distinct expression of FN was suggestive of the essential commitment in the process of cutaneous development and morphogenesis of appendages originated from hair germ. The paucity of FN in palmoplantar skin and oral mucosa might explain the characteristics of these skin phenotypes.