Schuger L, Skubitz A P, Zhang J, Sorokin L, He L
Department of Pathology and Laboratory Medicine, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
J Cell Biol. 1997 Oct 20;139(2):553-62. doi: 10.1083/jcb.139.2.553.
Laminins, the main components of basement membranes, are heterotrimers consisting of alpha, beta, and gamma polypeptide chains linked together by disulfide bonds. Laminins-1 and -2 are both composed of beta1 and gamma1 chains and differ from each other on their alpha chain, which is alpha1 and alpha2 for laminin-1 and -2, respectively. The present study shows that whereas laminins-1 and -2 are synthesized in the mouse developing lung and in epithelial-mesenchymal cocultures derived from it, epithelial and mesenchymal monocultures lose their ability to synthesize the laminin alpha1 chain. Synthesis of laminin alpha1 chain however returns upon re-establishment of epithelial-mesenchymal contact. Cell-cell contact is critical, since laminin alpha1 chain is not detected in monocultures exposed to coculture-conditioned medium or in epithelial-mesenchymal cocultures in which heterotypic cell-cell contact is prevented by an interposing filter. Immunohistochemical studies on cocultures treated with brefeldin A, an inhibitor of protein secretion, indicated both epithelial and mesenchymal cells synthesize laminin alpha1 chain upon heterotypic cell- cell contact. In a set of functional studies, embryonic lung explants were cultured in the presence of monoclonal antibodies to laminin alpha1, alpha2, and beta/gamma chains. Lung explants exposed to monoclonal antibodies to laminin alpha1 chain exhibited alterations in peribronchial cell shape and decreased smooth muscle development, as indicated by low levels of smooth muscle alpha actin and desmin. Taken together, our studies suggest that laminin alpha1 chain synthesis is regulated by epithelial-mesenchymal interaction and may play a role in airway smooth muscle development.
层粘连蛋白是基底膜的主要成分,是由α、β和γ多肽链通过二硫键连接而成的异源三聚体。层粘连蛋白-1和-2均由β1和γ1链组成,它们的α链不同,层粘连蛋白-1的α链为α1,层粘连蛋白-2的α链为α2。本研究表明,虽然层粘连蛋白-1和-2在小鼠发育中的肺以及从中衍生的上皮-间充质共培养物中合成,但上皮和间充质单培养物失去了合成层粘连蛋白α1链的能力。然而,上皮-间充质接触重新建立后,层粘连蛋白α1链的合成恢复。细胞间接触至关重要,因为在暴露于共培养条件培养基的单培养物中或在上皮-间充质共培养物中(其中异型细胞间接触通过插入滤膜来阻止)未检测到层粘连蛋白α1链。对用布雷菲德菌素A(一种蛋白质分泌抑制剂)处理的共培养物进行的免疫组织化学研究表明,异型细胞间接触时上皮和间充质细胞均合成层粘连蛋白α1链。在一组功能研究中,将胚胎肺外植体在抗层粘连蛋白α1、α2和β/γ链的单克隆抗体存在下培养。暴露于抗层粘连蛋白α1链单克隆抗体的肺外植体表现出支气管周围细胞形状改变和平滑肌发育减少,如平滑肌α肌动蛋白和结蛋白水平较低所示。综上所述,我们的研究表明层粘连蛋白α1链的合成受上皮-间充质相互作用调节,并且可能在气道平滑肌发育中起作用。