Epinectin, a cell-substratum adhesion promoting molecule, was first isolated from the extracellular matrix of A431 human squamous carcinoma cells. In order to determine the biologic significance of epinectin, we determined the distribution of epinectin in various rat epithelial tissues by indirect immunofluorescence microscopy. Polyclonal antibodies to epinectin stained basal cells and basilar regions of skin, urinary bladder, and vagina. There was predominantly cytoplasmic staining along with amorphous extracellular staining. Strong staining was also noted in sebaceous glands and hair follicles. The immunoreactivity for epinectin in the skin was distinct from that for fibronectin, laminin, and type IV collagen. Antibodies to epinectin also stained subpopulations of neurons in the cerebrum and cerebellum. Epinectin antibodies strongly stained the cytoplasm of some pineal cells and cells of the pars intermedia of the pituitary. The distribution of epinectin suggests a role not only in epithelial cell-substratum adhesion, but in neuronal cell function. Heparan sulfate is known to be involved in the binding of several adhesion promoting molecules to cell surfaces. In order to assess the mechanism of adhesion of epinectin to cells, we measured the binding of 3H-heparin to epinectin. Binding of 3H-heparin was concentration dependent and inhibitable with cold heparin.