Hauft S M, Kim S H, Schmidt G H, Pease S, Rees S, Harris S, Roth K A, Hansbrough J R, Cohn S M, Ahnen D J
Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110.
J Cell Biol. 1992 May;117(4):825-39. doi: 10.1083/jcb.117.4.825.
The mouse intestinal epithelium represents a unique mammalian system for examining the relationship between cell division, commitment, and differentiation. Proliferation and differentiation are rapid, perpetual, and spatially well-organized processes that occur along the crypt-to-villus axis and involve clearly defined cell lineages derived from a common multipotent stem cell located near the base of each crypt. Nucleotides -1178 to +28 of the rat intestinal fatty acid binding protein gene were used to establish three pedigrees of transgenic mice that expressed SV-40 large T antigen (TAg) in epithelial cells situated in the uppermost portion of small intestinal crypts and in already committed, differentiating enterocytes as they exited these crypts and migrated up the villus. T antigen production was associated with increases in crypt cell proliferation but had no apparent effect on commitment to differentiate along enterocytic, enteroendocrine, or Paneth cell lineages. Single- and multilabel-immunocytochemical studies plus RNA blot hybridization analyses suggested that the differentiation programs of these lineages were similar in transgenic mice and their normal littermates. This included enterocytes which, based on the pattern of [3H]thymidine and 5-bromo-2'-deoxyuridine labeling and proliferating nuclear antigen expression, had reentered the cell cycle during their migration up the villus. The state of cellular differentiation and/or TAg production appeared to affect the nature of the cell cycle; analysis of the ratio of S-phase to M-phase cells (collected by metaphase arrest with vincristine) and of the intensities of labeling of nuclei by [3H]thymidine indicated that the duration of S phase was longer in differentiating, villus-associated enterocytes than in the less well-differentiated crypt epithelial cell population and that there may be a block at the G2/M boundary. Sustained increases in crypt and villus epithelial cell proliferation over a 9-mo period were not associated with the development of gut neoplasms--suggesting that tumorigenesis in the intestine may require that the initiated cell have many of the properties of the gut stem cell including functional anchorage.
小鼠肠道上皮是研究细胞分裂、定向分化和分化之间关系的独特哺乳动物系统。增殖和分化是快速、持续且空间组织良好的过程,沿着隐窝到绒毛轴发生,涉及源自每个隐窝底部附近共同多能干细胞的明确细胞谱系。大鼠肠道脂肪酸结合蛋白基因的核苷酸-1178至+28用于建立三个转基因小鼠谱系,这些小鼠在上部小肠隐窝的上皮细胞以及已定向分化、正在分化的肠细胞从这些隐窝出来并向上迁移到绒毛时表达SV-40大T抗原(TAg)。T抗原的产生与隐窝细胞增殖增加有关,但对沿着肠细胞、肠内分泌细胞或潘氏细胞谱系分化的定向没有明显影响。单标记和多标记免疫细胞化学研究以及RNA印迹杂交分析表明,这些谱系在转基因小鼠及其正常同窝仔鼠中的分化程序相似。这包括肠细胞,根据[3H]胸苷和5-溴-2'-脱氧尿苷标记模式以及增殖细胞核抗原表达,它们在向上迁移到绒毛的过程中重新进入细胞周期。细胞分化状态和/或TAg产生似乎影响细胞周期的性质;对S期与M期细胞的比例(通过长春新碱中期阻滞收集)以及[3H]胸苷对细胞核标记强度的分析表明,分化的、与绒毛相关的肠细胞中S期持续时间比分化程度较低的隐窝上皮细胞群体更长,并且可能在G2/M边界存在阻滞。在9个月期间,隐窝和绒毛上皮细胞增殖持续增加与肠道肿瘤的发生无关——这表明肠道肿瘤发生可能要求起始细胞具有肠道干细胞的许多特性,包括功能性锚定。