SEM localization of laminin on the basement membrane of the chick corneal epithelium with immunolatex microspheres

Abstract

Embryonic chick corneal explants were soaked in mild detergent and the anterior corneal epithelium was peeled from its basement membrane, leaving the lamina lucida surface exposed and supported on the subjacent primary stroma. Explants were treated with rabbit anti-laminin IgG, followed by sheep anti-rabbit IgG linked microspheres, and processed for SEM. The lucida surface was heavily decorated with microspheres, whereas controls treated with preimmune rabbit IgG were essentially beadless. Laminin distribution was not regular, appearing denser in some regions than others. However, the connective tissue surface of the basement membrane was never laminin-positive, even after treatment with hyaluronidase. These results suggest the basal lamina of the corneal epithelium is asymmetric, with preferential location of laminin to the lucida surface of the basement membrane.