Antigenic markers and laminin expression in cultured enteric neural cells

In this study, polyclonal and monoclonal antibodies have been used in conjunction with standard immunohistochemical methods to define markers which can be used to identify and study the main cell types present in the outgrowth area of explant cultures of myenteric plexus from newborn guinea pig. We show that all of the neurons binds antibodies to the glycoprotein Thy-1 and the antibody A2B5 which recognizes gangliosides. All enteric glial cells bind antibodies to the calcium-binding protein S100, and the A2B5 monoclonal antibody and ca. 95% of glia are labeled by the antibody LB1, which recognizes the GD3 ganglioside. Most fibroblasts are labelled by antibodies to Thy-1 and the matrix glycoprotein fibronectin. Thus enteric neurons can be defined serologically as Thy-1+/A2B5+/S100- cells; enteric glia as S100+ cells and fibroblasts as A2B5-/S100-cells. The markers have been used to demonstrate that laminin is made by both enteric glia and fibroblasts. They have also been used to show that ca. 5% of neurons and less than 5% of enteric glia bind the antibody Leu 7 (HNK-1, L2), thus revealing the subpopulations of neurons and glia show differential cell surface expression of the carbohydrate epitope recognized by the antibody. In the accompanying paper, we demonstrate that two of the antibodies (LB1 and Thy-1) can be used to generate purified populations of neurons and glia.