Kedinger M, Simon-Assmann P, Bouziges F, Arnold C, Alexandre E, Haffen K
INSERM Unité 61, Biologie Cellulaire et Physiopathologie Digestives, Strasbourg, France.
Differentiation. 1990 Apr;43(2):87-97. doi: 10.1111/j.1432-0436.1990.tb00434.x.
Cytodifferentiation of smooth muscle cells has been analyzed immunocytochemically during rat intestinal development and in chimaeric intestines by using monoclonal antibodies reacting specifically with smooth muscle actin species (CGA7 [10] and anti-alpha SM-1 [40]). As development proceeds, the various intestinal muscle layers differentiate in the following order: (1) cells expressing smooth muscle actin appear within the mesenchyme of the 15-day fetal rat intestine, in the circular muscle-forming area, the differentiation of cells in the presumptive longitudinal muscle layer starting with a 48-h delay; (2) smooth muscle fibers appear within the connective tissue core of the villi shortly after birth, in parallel with a progressive formation of the muscularis mucosae, which becomes clear-cut only in the course of the 2nd week after birth; (3) a distinct cell layer in the innermost part of the circular muscle layer arises during the perinatal period. Thereafter, the fluorescence pattern remains unchanged until the adult stage. Chimaeric intestines were constructed by the association of 14-day fetal intestinal epithelium and cultured fetal rat or human skin fibroblasts. These fibroblastic cells did not express actin at the time at which they were associated. The immunocytochemical analysis of smooth muscle actin in the hybrid intestines, which had developed as intracoelomic grafts for 12 days, revealed that the skin fibroblastic cells had been induced by the intestinal epithelial cells to differentiate into smooth muscle cells. Such a result was also obtained with allantoic endoderm. It was not obvious in cocultures of intestinal epithelium with skin fibroblastic cells. However, when intestinal epithelial cells were cocultured with intestinal mesenchymal cells, actin expression was stimulated in the latter cell population.
利用与平滑肌肌动蛋白特异性反应的单克隆抗体(CGA7 [10]和抗α-SM-1 [40]),通过免疫细胞化学方法分析了大鼠肠道发育过程中和嵌合肠道中平滑肌细胞的细胞分化情况。随着发育的进行,肠道各肌层按以下顺序分化:(1)在15天胎鼠肠道的间充质内,在环形肌形成区域出现表达平滑肌肌动蛋白的细胞,推测的纵行肌层细胞的分化开始延迟48小时;(2)出生后不久,绒毛结缔组织核心内出现平滑肌纤维,同时黏膜肌层逐渐形成,仅在出生后第2周才变得清晰;(3)围产期,环形肌层最内层出现明显的细胞层。此后,荧光模式直至成年期保持不变。通过将14天胎鼠肠上皮与培养的胎鼠或人皮肤成纤维细胞联合构建嵌合肠道。这些成纤维细胞在联合时不表达肌动蛋白。对作为体腔内移植物发育12天的杂种肠道中平滑肌肌动蛋白的免疫细胞化学分析表明,皮肤成纤维细胞已被肠上皮细胞诱导分化为平滑肌细胞。尿囊内胚层也得到了这样的结果。在肠上皮与皮肤成纤维细胞的共培养中不明显。然而,当肠上皮细胞与肠间充质细胞共培养时,后者细胞群体中的肌动蛋白表达受到刺激。