Institute of Pathology, RWTH Aachen University, Aachen, Germany.
BMC Gastroenterol. 2010 Mar 7;10:27. doi: 10.1186/1471-230X-10-27.
Maturation of enterocytes along the small intestinal crypt-villus axis is associated with significant changes in gene expression profiles. fls485 coding a putative chaperone protein has been recently suggested as a gene involved in this process. The aim of the present study was to analyze fls485 expression in human small intestinal mucosa.
fls485 expression in purified normal or intestinal mucosa affected with celiac disease was investigated with a molecular approach including qRT-PCR, Western blotting, and expression strategies. Molecular data were corroborated with several in situ techniques and usage of newly synthesized mouse monoclonal antibodies.
fls485 mRNA expression was preferentially found in enterocytes and chromaffine cells of human intestinal mucosa as well as in several cell lines including Rko, Lovo, and CaCo2 cells. Western blot analysis with our new anti-fls485 antibodies revealed at least two fls485 proteins. In a functional CaCo2 model, an increase in fls485 expression was paralleled by cellular maturation stage. Immunohistochemistry demonstrated fls485 as a cytosolic protein with a slightly increasing expression gradient along the crypt-villus axis which was impaired in celiac disease Marsh IIIa-c.
Expression and synthesis of fls485 are found in surface lining epithelia of normal human intestinal mucosa and deriving epithelial cell lines. An interdependence of enterocyte differentiation along the crypt-villus axis and fls485 chaperone activity might be possible.
小肠隐窝-绒毛轴上的肠细胞成熟与基因表达谱的显著变化有关。最近有人提出编码假定伴侣蛋白的 fls485 基因与此过程有关。本研究的目的是分析人小肠黏膜中 fls485 的表达。
采用包括 qRT-PCR、Western blot 和表达策略在内的分子方法研究纯化的正常或受乳糜泻影响的小肠黏膜中 fls485 的表达。分子数据与几种原位技术和新合成的小鼠单克隆抗体的使用结果相吻合。
fls485 mRNA 表达优先存在于人肠黏膜的肠细胞和嗜铬细胞以及包括 Rko、Lovo 和 CaCo2 细胞在内的几种细胞系中。用我们新的抗 fls485 抗体进行 Western blot 分析显示至少有两种 fls485 蛋白。在功能性 CaCo2 模型中，fls485 的表达增加与细胞成熟阶段平行。免疫组织化学显示 fls485 是一种细胞浆蛋白，在隐窝-绒毛轴上的表达呈略有增加的梯度，在乳糜泻 Marsh IIIa-c 中受到损害。
fls485 的表达和合成存在于正常人类小肠黏膜的表面衬里上皮和衍生的上皮细胞系中。沿着隐窝-绒毛轴的肠细胞分化和 fls485 伴侣活性之间可能存在相互依存关系。
