Sinning A R, Lepera R C, Markwald R R
Department of Anatomy and Cellular Biology, Medical College of Wisconsin, Milwaukee 53226.
Dev Biol. 1988 Nov;130(1):167-74. doi: 10.1016/0012-1606(88)90423-x.
Formation of the atrioventricular (AV) mesenchyme is a critical step in early heart development. Endothelial cells are activated and transformed into a mesenchymal population that invades the cell-free myocardial basement membrane. This process can be duplicated in collagen gel culture, where it has been established that myocardium or its secretory products activate the endothelium. The purpose of the present study was to determine when these activated endothelial and/or mesenchymal cells start producing type I collagen in situ. These results were compared to those obtained from a culture model of mesenchyme formation. The production of type I collagen was monitored using a monoclonal antibody (M38) that recognizes the carboxy-terminal propeptide of human type I procollagen. The initial expression of the latter within activated AV endothelial and mesenchymal cells in ovo was 48 hr following activation. Prior to this time, only the myocardium was reactive with M38. AV explants of early hearts on collagen gels revealed staining of activated endothelial and mesenchymal cells with M38 after 48 hr in coculture with myocardial tissue. Explants that were prevented from activating (myocardium removed) never expressed the M38 antigen. Similarly, AV endothelial monolayers grown in the presence of myocardial conditioned medium activated and expressed type I collagen after 48 hr in culture, whereas those grown in standard medium did not. These results establish the initial expression of type I collagen within activated AV endothelium and mesenchyme. In addition, the data suggest that the expression of type I collagen within the AV mesenchyme may be dependent on extrinsic influences that induce the AV endothelium to transform into mesenchyme.
房室间充质的形成是心脏早期发育的关键步骤。内皮细胞被激活并转化为侵入无细胞心肌基底膜的间充质细胞群体。这个过程可以在胶原凝胶培养中重现,在该培养中已证实心肌或其分泌产物可激活内皮细胞。本研究的目的是确定这些被激活的内皮细胞和/或间充质细胞何时开始在原位产生I型胶原蛋白。将这些结果与从间充质形成的培养模型中获得的结果进行比较。使用识别人类I型前胶原羧基末端前肽的单克隆抗体(M38)监测I型胶原蛋白的产生。在卵内被激活的房室内皮细胞和间充质细胞中,后者的初始表达是在激活后48小时。在此之前,只有心肌与M38发生反应。在与心肌组织共培养48小时后,胶原凝胶上早期心脏的房室外植体显示被激活的内皮细胞和间充质细胞被M38染色。被阻止激活的外植体(去除心肌)从未表达M38抗原。同样,在心肌条件培养基存在下生长的房室内皮细胞单层在培养48小时后被激活并表达I型胶原蛋白,而在标准培养基中生长的则没有。这些结果确定了I型胶原蛋白在被激活的房室内皮细胞和间充质细胞中的初始表达。此外,数据表明房室间充质中I型胶原蛋白的表达可能依赖于诱导房室内皮细胞转化为间充质的外在影响。