Exposure to Lysosomotropic Amines and Protease Inhibitors Retard Corneal Endothelial Cell Migration along the Natural Basement Membrane during Wound Repair

• Published in: Experimental cell research 1999-01, Vol.246 (1), p.233-242
• Main Authors: Gordon, Sheldon R., DeMoss, James
• Format: Article
• Language: English
• Subjects: Amines - pharmacology; Animals; Antipain - pharmacology; Basement Membrane - metabolism; Cell Movement - drug effects; Chloroquine - pharmacology; Culture Techniques; Descemet Membrane - metabolism; Endopeptidases - physiology; Endothelium, Corneal - cytology; Endothelium, Corneal - injuries; Endothelium, Corneal - metabolism; Freezing; Leupeptins - pharmacology; Lysosomes - enzymology; Lysosomes - physiology; Metalloendopeptidases - antagonists & inhibitors; Metalloendopeptidases - physiology; Methylamines - pharmacology; Propylamines - pharmacology; Protease Inhibitors - pharmacology; Rats; Rats, Sprague-Dawley; Trypsin Inhibitor, Kunitz Soybean - pharmacology; Wound Healing - drug effects
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1006/excr.1998.4298

Regulation of cell migration along the natural basement membrane during wound repair in the organ culture corneal endothelium was investigated using various lysosomotropic amines and protease inhibitors. Following a circular transcorneal freeze injury, cells within the area die and expose the underlying basement membrane (Descemet's membrane). During normal wound repair, cells traverse this expanse and repopulate the region by approximately 48 h postinjury. During this time, acid phosphatase histochemistry revealed distinct alterations in the lysosomal population of cells that were adjacent to, and migrated into, the wound region. To explore whether relationships may exist between changes in the lysosome population and cell migration, injured endothelia were organ cultured in the presence of either methylamine or chloroquine, two lysosomotropic amines. Methylamine significantly retarded cell translocation (85%) into the injury zone when compared to nontreated controls. In comparison, chloroquine was less effective in restricting injury-induced cell migration and propylamine, also a lysosomotropic amine, had no influence on the repair process. In addition, two serine/thio protease inhibitors, leupeptin and antipain, were both able to impede cell translocation during wound repair by 85 and 52%, respectively, whereas soybean trypsin inhibitor, a serine protease inhibitor, exhibited no inhibitory effect on the repair process. Similarly, incubating injured tissues in either 1,10-phenanthroline or phosphoramidon, both metalloproteinase inhibitors, did not prevent endothelial cell movement nor wound repair. Results indicate that corneal endothelial cell migration along the natural basement membrane is dependent on protease function. Although the precise nature of the proteases involved has yet to be ascertained, results indicate that lysosomal enzymes may have a distinct role in corneal endothelial cell movement along the natural basement membrane during wound repair.