Gene expression and immunolocalisation of a calcium-activated chloride channel during the stratification of cultivated and developing corneal epithelium
Gene expression and immunolocalisation of a calcium-activated chloride channel during the stratification of cultivated and developing corneal epithelium
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DOI：
10.1007/s00441-005-0059-2
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Kinoshita, S
中科院分区：
文献类型：
--
作者：
Connon, CJ;Kawasaki, S;Kinoshita, S
通过荧光免疫组织化学和定量聚合酶链反应在鸡胚角膜和羊膜上扩增离体人角膜干细胞，研究了钙激活氯离子通道（CLCA）及其mRNA在体内和体外复层上皮发育过程中的时空定位。羊膜和早期胚胎鸡角膜上的单层人上皮培养物表达相对较少的人CLCA 2或其鸡同源物。然而，随着两种模型中的上皮细胞成熟和细胞层数的增加，基因表达水平和蛋白质染色强度增加，主要在培养组织和胚胎组织的基底细胞内。这些结果表明，人CLCA2蛋白和mRNA的表达在上皮层化过程中升高，表明该蛋白在自然发育和组织培养过程中在多层角膜上皮的生长中起作用。
The spatial and temporal localisation of a calcium-activated chloride channel (CLCA) and its mRNA was investigated, during the in vivo and in vitro development of stratified epithelia, by fluorescence immunohistochemistry and quantitative polymerase chain reaction in embryonic chicken corneas and the expansion of excised human corneal stem cells on amniotic membrane. Single-layered human epithelial cultures on amniotic membrane and early day embryonic chicken corneas expressed relatively little human CLCA2 or its chicken homologue. However, as the epithelium in both models matured and the number of cell-layers increased, the gene expression level and protein staining intensity increased, primarily within the basal cells of both the cultured and embryonic tissues. These results demonstrate that human CLCA2 protein and mRNA expression are elevated during epithelial stratification, suggesting that this protein plays a role in the growth of multi-layered corneal epithelia during both natural development and tissue cultivation.