Kisalus L L, Herr J C, Little C D
Department of Anatomy and Cell Biology, University of Virginia, Charlottesville 22908.
Anat Rec. 1987 Aug;218(4):402-15. doi: 10.1002/ar.1092180408.
The extracellular matrix (ECM) of first-trimester human decidua was examined with indirect immunofluorescence using affinity-purified antibodies to human collagen types I, III, IV, V, laminin, and fibronectin. In addition, the validity of the classification "mesenchymal-epithelioid" for differentiated decidual cells was addressed using antibodies to the intermediate filament proteins, vimentin, a mesenchymal marker, and keratin, an epithelial marker. Biosynthesis of extracellular matrix components was examined by radiolabeling of decidual explants in culture with 3H-proline, followed by immunoprecipitations of synthesized proteins with collagen type-specific antibodies. Immunofluorescence showed decidual cells are embedded in an extensive network of collagen types I and III, and intracytoplasmic staining suggested synthesis of these collagens by the decidual cells. Collagen type IV and laminin localized in the external lamina which surrounds the differentiated decidual cell, and some fluorescence was evident in the peripheral cytoplasm. Immunoreactive collagen type V was observed in close association with the external lamina and in the peridecidual matrix. Fibronectin localized throughout the decidual ECM and in fibrillar and punctuate patterns in the decidual cell cytoplasm. Differentiated decidual cells retained a "mesenchymal" intermediate filament cytoskeleton containing an abundance of vimentin filaments, but very few, if any, keratin filaments. Collagen types I, III, V, and to a lesser extent, IV, were immunoprecipitated from the medium of decidual explants after 24 hours of culture, demonstrating in vitro synthesis and secretion of these collagens by first trimester human decidua.
利用针对人I、III、IV、V型胶原蛋白、层粘连蛋白和纤连蛋白的亲和纯化抗体，通过间接免疫荧光法检测了孕早期人蜕膜的细胞外基质（ECM）。此外，使用针对中间丝蛋白波形蛋白（一种间充质标志物）和角蛋白（一种上皮标志物）的抗体，探讨了“间充质-上皮样”分类对分化蜕膜细胞的有效性。通过用3H-脯氨酸对培养的蜕膜外植体进行放射性标记，然后用胶原蛋白特异性抗体对合成的蛋白质进行免疫沉淀，检测细胞外基质成分的生物合成。免疫荧光显示蜕膜细胞嵌入I型和III型胶原蛋白的广泛网络中，胞质内染色表明这些胶原蛋白由蜕膜细胞合成。IV型胶原蛋白和层粘连蛋白定位于围绕分化蜕膜细胞的外板中，外周细胞质中可见一些荧光。在与外板紧密相连的位置以及蜕膜周围基质中观察到免疫反应性V型胶原蛋白。纤连蛋白定位于整个蜕膜ECM中，并以纤维状和点状模式存在于蜕膜细胞质中。分化的蜕膜细胞保留了一个“间充质”中间丝细胞骨架，其中含有大量波形蛋白丝，但角蛋白丝极少（如果有的话）。培养24小时后，从蜕膜外植体培养基中免疫沉淀出I、III、V型胶原蛋白，在较小程度上还有IV型胶原蛋白，这表明孕早期人蜕膜在体外合成并分泌这些胶原蛋白。
