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Impressum
The neurotrophic growth factor Artemin stimulates corneal proliferation, migration and wound healing
Ebner S., You L., Kruse F. E.
Department of Ophthalmology, University of Heidelberg, INF 400, 69120 Heidelberg
Neurotrophins play an important role during migration and wound healing
in the cornea. For further elucidation we investigated the function and
signal transduction of corneal epithelial cells in combination with factors,
which bind to the GDNF-a/ret system and the influence of Artemin, a new
member of the GDNF-family, concerning its role in wound healing and signal
transduction .
Methods: For experiments Primary human and rabbit corneal epithelial
cells as well as SV-40 transformed cells were used. Proliferation was
quantified in a single cell clonal growth assay, while migration was quantified
in a modified Boyden chamber. In-vitro wound healing was evaluated by
time-lapse video-images from scratch wounds. MAP-kinase signaling was
investigated by use of specific antibodies and western-blots.
Results: The number of colonies /60 mm dish was significantly higher
in the presence of 250 ng Artemin than in control (80.5 ± 37.9
vs 36.7 ± 17.6; p < 0.001). The total number of cells/dish increased
significantly due to Artemin (489.25 ± 278.28 vs 218.25 ±
97.85 control; p<0.001) Clonal proliferation (cells/colony) was not
increased (5.78 ± 0.65 vs 5.55 ± 0.65 control). Migration
through 8 µm filters was significantly enhanced by Artemin (59.9
± 10.5 vs 4.3 ± 8.07; p<0.01). Also wound closure was
significantly stimulated by Artemin (38.6 ± 5.64 vs 20.94 ±
4.54; p<0.01). Western blot analysis showed that phosphorylation of
Erk, Raf, MEK and Elk is induced by Artemin.
Conclusion: Our results suggest, that Artemin uses the GDNF/GDNFRa/
ret pathway to regulate proliferation, migration and wound healing in
the human cornea. This enhances the significance of the Map-Kinase pathway
for corneal wound healing. Furthermore topical administration of Artemin
might be usefull to treat wound healing disorders. (Kr 993/12-1)