Simo P, Bouziges F, Lissitzky J C, Sorokin L, Kedinger M, Simon-Assmann P
INSERM Unité 61, Biologie Cellulaire et Physiopathologie Digestives, Strasbourg, France.
Gastroenterology. 1992 Jun;102(6):1835-45. doi: 10.1016/0016-5085(92)90303-g.
The production of laminin by 14-day fetal rat intestinal endoderm and mesenchyme was investigated. The amount of neosynthesized laminin was measured after purification using affinity chromatography. Chain composition of laminin was analyzed by immunoblotting and immunofluorescence staining. The data show that both embryonic intestinal tissue components synthesize laminin and that A and B1/B2 chains were detected in both endodermal and mesenchymal cells. The cellular source of laminin found at the epithelial basement membrane has been studied by immunocytochemistry in rat/chick or mouse/chick interspecies hybrid intestines taken at various stages of development. Immunodetection of the whole laminin molecule and of the individual A and B1/B2 chains by rodent-specific polyclonal and monoclonal antibodies at the basement membrane level in these hybrid intestines revealed (a) laminin molecules, which originate from both mesenchymal and endodermal cells; (b) deposition of A and B1/B2 chains by endodermal cells, regardless of the stage of growth of the hybrid intestines; and (c) asynchronous deposition of the various chains of laminin into the basement membrane by the mesenchyme. B1/B2 chains are deposited concomitant with contact with the epithelium, whereas A chains appear only later (13 days after grafting). These data reinforce the suggestion from previous studies that cooperation between epithelium and mesenchyme is necessary for the formation of a complete basement membrane in the developing intestine.
研究了14天龄胎鼠肠内胚层和间充质层中层粘连蛋白的产生情况。使用亲和层析法纯化后，测定新合成层粘连蛋白的量。通过免疫印迹和免疫荧光染色分析层粘连蛋白的链组成。数据表明，胚胎肠组织的这两种成分均能合成层粘连蛋白，并且在内胚层细胞和间充质细胞中均检测到A链和B1/B2链。通过免疫细胞化学方法，在处于不同发育阶段的大鼠/鸡或小鼠/鸡种间杂交肠中，研究了上皮基底膜处层粘连蛋白的细胞来源。在这些杂交肠的基底膜水平，用啮齿动物特异性多克隆和单克隆抗体对整个层粘连蛋白分子以及单个A链和B1/B2链进行免疫检测，结果显示：（a）层粘连蛋白分子来源于间充质细胞和内胚层细胞；（b）内胚层细胞均能沉积A链和B1/B2链，与杂交肠的生长阶段无关；（c）间充质层将层粘连蛋白的不同链异步沉积到基底膜中。B1/B2链在与上皮接触时沉积，而A链则在较晚时候（移植后13天）才出现。这些数据进一步支持了先前研究提出的观点，即上皮细胞和间充质细胞之间的合作对于发育中的肠道形成完整的基底膜是必要的。
