Stromal assemblies containing collagen types IV and VI and fibronectin in the developing embryonic avian cornea

The morphogenesis of type IV collagen-containing structures in the stromal matrix of the developing avian cornea was investigated using immunofluorescence and immunoelectron microscopic histochemistry. Two forms of type IV collagen-containing structures were seen; these differed in their probable origin, structure, molecular composition, and developmental fate. The major form of stromal type IV collagen-containing material, termed "strings," was observed only after swelling of the primary stroma and the onset of mesenchymal invasion. These strings are presumed to be products of the stromal cells. In immunofluorescence histochemistry they appeared as linear segments of type IV collagen-specific immunoreactivity. In immunoelectron microscopy, they appeared initially as electron-dense sausages of variable length and orientation. They frequently were associated with cell surfaces and, in fortuitous sections, appeared to connect adjacent cells. The strings also contained type VI collagen and fibronectin, but very little, if any, of the basement membrane components laminin and heparin sulfate proteoglycan (HSPG). As the stroma continued to expand in thickness, more of these structures were observed in a radial orientation, becoming quite long and less tortuous. Later in development, as stromal condensation proceeded, they disappeared. We suggest that the strings function to stabilize the stromal matrix, and perhaps to limit the rate and/or extent of stromal expansion, during a phase of rapid swelling and matrix deposition. The other form of type IV collagen-containing stromal material appeared as irregularly shaped plaques of basement membrane-like material identical to those previously described in mature corneas. These are likely derived from the corneal endothelial cells. They contained other basement membrane-associated components (laminin, HSPG) and fibronectin, but not type VI collagen. This material persists in mature corneas as sparse irregular stromal plaques and as matrix in the interface between Descemet's membrane and the corneal stroma.