Marche P, Cassier P, Mathieu H
Cell Tissue Res. 1980;212(1):63-72. doi: 10.1007/BF00234033.
The vitamin D-dependent calcium-binding protein (CaBP) was studied in relation to the age of the cell, in isolated epithelial cell populations removed from rat duodenum. Alkaline phosphatase and thymidine kinase activities were used as markers to characterize differentiated villus cells and undifferentiated (mitotically active) crypt cells, respectively. CaBP distribution along the length of the villus, as established by radioimmunoassay, appears as a gradient increasing from the crypt to the tip of the villus. CaBP concentration in cells is shown to be (i) negatively correlated with the thymidine kinase activity of cells, and (ii) positively correlated with the alkaline phosphatase activity of cells. This indicates that CaBP is absent in crypt cells and appears in differentiated cells with the development of the brush border. Thus CaBP, like alkaline phosphatase, can be considered as an indicator of enterocyte maturation. These data were also confirmed by studying the cellular localization of the protein. In addition both indirect immunofluorescence and immunoperoxidase staining methods reveal that antibody against CaBP decorates the terminal web, but not the microvilli of the brush border of mature absorptive cells. The results suggest that CaBP may act as a modulator of some Ca2+-mediated biochemical processes at the level of the enterocyte brush border.
在从大鼠十二指肠分离出的上皮细胞群体中,研究了维生素D依赖性钙结合蛋白(CaBP)与细胞年龄的关系。分别使用碱性磷酸酶和胸苷激酶活性作为标志物来表征分化的绒毛细胞和未分化(有丝分裂活跃)的隐窝细胞。通过放射免疫测定确定,CaBP沿绒毛长度的分布呈现出从隐窝到绒毛尖端逐渐增加的梯度。细胞中的CaBP浓度显示为:(i)与细胞的胸苷激酶活性呈负相关,(ii)与细胞的碱性磷酸酶活性呈正相关。这表明CaBP在隐窝细胞中不存在,随着刷状缘的发育出现在分化细胞中。因此,CaBP与碱性磷酸酶一样,可被视为肠上皮细胞成熟的指标。通过研究该蛋白的细胞定位也证实了这些数据。此外,间接免疫荧光和免疫过氧化物酶染色方法均显示,抗CaBP抗体可标记成熟吸收细胞刷状缘的终末网,但不标记微绒毛。结果表明,CaBP可能在肠上皮细胞刷状缘水平作为某些Ca2+介导的生化过程的调节剂。