The hLAMP-1-Positive Particulate Matrix Involved in Cardiac Mesenchyme Formation in the Chick Does Not Include BMP-2

Abstract

Early heart development involves the transformation of endocardial cells in the atrioventricular canal and outflow tract regions into mesenchymal cells, a process called endocardial mesenchymal transformation (EMT). This process is initiated by factors, termed the particulate matrix, that are secreted by the myocardium. The particulate matrix causes a subset of endocardial cells to hypertrophy, lose their cell-cell contacts, form migratory processes, transform into mesenchymal cells, and migrate into the underlying endocardial cushions. The particulate matrix can be extracted using EDTA and the EDTA extract can initiate the EMT process. Earlier reports from our laboratory have shown that the particulate matrix can be detected with the hLAMP-1 antibody in immunostaining and Western blot analysis. In addition, similar proteins have been isolated from the growth media of stage 15-16 chick embryo myocardial cultures (MyoCM). Since other investigators have identified a possible role for bone morphogenetic protein (BMP)-2 during the EMT process in the heart, we asked whether BMP-2 is a part of the chick hLAMP-1-positive particulate matrix. To answer this question, we double stained stage 15-16 chick embryo sections with hLAMP-1 and BMP-2 antibodies. We found that BMP-2 signals do not colocalize with hLAMP-1-stained particles. In addition, using immunoprecipitation-Western blot analysis, we demonstrated no association of BMP-2 with the hLAMP-1-bound fraction of the EDTA extract or MyoCM. Our results indicate that BMP-2 is not a component of the hLAMP-1-positive particulate matrix in the chick.