Interclass small leucine-rich repeat proteoglycan interactions regulate collagen fibrillogenesis and corneal stromal assembly

Authors

Shoujun Chen, Marian F. Young, Shukti Chakravartic, David E. Birk

Abstract

The corneal stroma is enriched in small leucine-rich proteoglycans (SLRPs), including both class I (decorin and biglycan) and class II (lumican, keratocan and fibromodulin). Transparency is dependent on the assembly and maintenance of a hierarchical stromal organization and SLRPs are critical regulatory molecules. We hypothesize that cooperative interclass SLRP interactions are involved in the regulation of stromal matrix assembly. We test this hypothesis using a compound Bgn−/o/Lum−/− mouse model and single Lum−/− or Bgn−/o mouse models and wild type controls. SLRP expression was investigated using immuno-localization and immuno-blots. Structural relationships were defined using ultrastructural and morphometric approaches while transparency was analyzed using in vivo confocal microscopy. The compound Bgn−/o/Lum−/− corneas demonstrated gross opacity that was not seen in the Bgn−/o or wild type corneas and greater than that in the Lum−/− mice. The Bgn−/o/Lum−/− corneas exhibited significantly increased opacity throughout the stroma compared to posterior opacity in the Lum−/− and no opacity in Bgn−/o or wild type corneas. In the Bgn−/o/Lum−/− corneas there were abnormal lamellar and fibril structures consistent with the functional deficit in transparency. Lamellar structure was disrupted across the stroma with disorganized fibrils, and altered fibril packing. In addition, fibrils had larger and more heterogeneous diameters with an abnormal structure consistent with abnormal fibril growth. This was not observed in the Bgn−/o or wild type corneas and was restricted to the posterior stroma in Lum−/− mice. The data demonstrate synergistic interclass regulatory interactions between lumican and biglycan. These interactions are involved in regulating both lamellar structure as well as collagen fibrillogenesis and therefore, corneal transparency.