Knickelbein R G, Aronson P S, Dobbins J W
Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
J Clin Invest. 1988 Dec;82(6):2158-63. doi: 10.1172/JCI113838.
Present evidence suggests that in the small intestine, villus cells are primarily absorptive and crypt cells are primarily secretory. In order to further confirm that there are differences in transport properties between villus and crypt cells, we have separated villus from crypt cells, using calcium chelations techniques, and determined the distribution of Na:H and Cl:HCO3 exchange activity on brush border membrane and basolateral membrane preparations from these two cell populations. Separation of cells was determined utilizing alkaline phosphatase and maltase activity as a marker of villus cells and thymidine kinase activity as a marker of crypt cells. Utilizing these techniques, we were able to sequentially collect cells along the villus-crypt axis. Na-stimulated glucose and alanine uptake in brush border membrane vesicles diminished from the villus to the crypt region in the sequentially collected cells fractions, further suggesting separation of these cells. Brush border and basolateral membranes were then prepared from cells from the villus and crypt areas, utilizing a continuous sucrose gradient. In the villus cells, Na:H exchange activity was found associated with both the brush border and basolateral membrane, whereas, in crypt cells, Na:H exchange activity was only found on the basolateral membrane. Cl:HCO3 exchange activity was found only on the brush border membrane, in both villus and crypt cells. These studies suggest functional heterogeneity in ion transport between villus and crypt cells.
目前的证据表明,在小肠中,绒毛细胞主要负责吸收,隐窝细胞主要负责分泌。为了进一步证实绒毛细胞和隐窝细胞在转运特性上存在差异,我们采用钙螯合技术将绒毛细胞与隐窝细胞分离,并测定了这两种细胞群体的刷状缘膜和基底外侧膜制剂上Na:H和Cl:HCO3交换活性的分布。利用碱性磷酸酶和麦芽糖酶活性作为绒毛细胞的标志物,以及胸苷激酶活性作为隐窝细胞的标志物来确定细胞的分离情况。利用这些技术,我们能够沿着绒毛 - 隐窝轴顺序收集细胞。在顺序收集的细胞组分中,刷状缘膜囊泡中钠刺激的葡萄糖和丙氨酸摄取从绒毛区域到隐窝区域逐渐减少,这进一步表明了这些细胞的分离。然后利用连续蔗糖梯度从绒毛和隐窝区域的细胞制备刷状缘膜和基底外侧膜。在绒毛细胞中,发现Na:H交换活性与刷状缘膜和基底外侧膜均相关,而在隐窝细胞中,Na:H交换活性仅在基底外侧膜上发现。Cl:HCO3交换活性在绒毛细胞和隐窝细胞的刷状缘膜上均有发现。这些研究表明绒毛细胞和隐窝细胞在离子转运方面存在功能异质性。