Endostatin overexpression specifically in the lens and skin leads to cataract and ultrastructural alterations in basement membranes.
Endostatin, a proteolytic fragment of type XVIII collagen, has been shown to inhibit angiogenesis, tumor growth, and endothelial cell proliferation and migration. We analyzed its functions in vivo by generating transgenic mice in which it was overexpressed in the skin and lens capsule under the keratin K14 promoter. Opacity of the lens occurred at 4 months of age in the mouse line J4, with the highest level of endostatin expression. The lens epithelial cells appeared to lose contact with the capsule and began to vacuolize. In 1-year-old mice the lens epithelial cell layer had entirely degenerated, and instead, large plaques of spindle-shaped cells had formed in the anterior region of the lens. Moreover, a widening of the epidermal basement membrane (BM) zone of the skin was observed in electron microscopy. The epidermal BM was conspicuously altered in the J4 mice with high transgene expression, including clear broadening and occurrence of pearl-like protrusions in some areas, whereas the BM was more even in appearance but consistently broadened in the mouse line G20 with moderate transgene expression. In both lines the BM was continuous. Measurements indicated that the lamina densa was 78.54 +/- 53.10 nm in line J4, the large variation reflecting the protrusions of the lamina densa, and 44.24 +/- 11.52 nm in line G20, compared with 33.74 +/- 9.96 nm in wild-type adult mice. Immunoelectron microscopy of wild-type mouse skin type XVIII collagen showed a polarized orientation in the BMs, with the C-terminal endostatin region localized in the lamina densa and the N terminus in average approximately 40 nm more on the dermal side. Type XVIII collagen was dispersed in the transgenic skin, suggesting that the transgene-derived endostatin fragment displaces the full-length collagen XVIII. This may impair the anchoring of the lamina densa to the dermis and thereby lead to loosening of the BMs, resembling the previously observed situation in collagen XVIII-null mice.
Elamaa
Harri
H
Department of Medical Biochemistry and Molecular Biology, Biocenter Oulu, University of Oulu, Aapistie 7, FI 90220, Oulu, Finland.
Sormunen
Raija
R
Rehn
Marko
M
Soininen
Raija
R
Pihlajaniemi
Taina
T
D020533:Angiogenesis Inhibitors; D000818:Animals; D001485:Basement Membrane / Q000473:pathology* / Q000648:ultrastructure; D002386:Cataract / Q000473:pathology*; D017931:DNA Primers; D043169:Endostatins / Q000502:physiology*; D006801:Humans; D007908:Lens, Crystalline / Q000473:pathology* / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D016253:Microscopy, Immunoelectron; D016133:Polymerase Chain Reaction; D012867:Skin / Q000473:pathology* / Q000648:ultrastructure
D020533:Angiogenesis Inhibitors; D017931:DNA Primers; D043169:Endostatins