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Updated: Oct 9, 2026

RhoC GTPase Activation Assay
Published on: August 22, 2010
Role of the small GTP-binding protein rho in epithelial cell migration in the rabbit cornea
M Nakamura1, T Nagano, T Chikama
1Department of Ophthalmology, Yamaguchi University School of Medicine, Yamaguchi, Japan. Ophthalmic Research Division, Santen Pharmaceutical Co., Ltd., Nara, Japan.
Purpose:
To determine the role of the small guanosine triphosphate (GTP)-binding protein Rho in the migration of corneal epithelial cells.
Methods:
The presence of the Rho target proteins Rho-associated coiled coil-containing protein kinase (ROCK)-1 and ROCK-2 in rabbit cornea was examined by immunohistochemical analysis, and that of the corresponding mRNAs in rabbit corneal epithelial cells was determined by reverse transcription-polymerase chain reaction analysis. The effects of various agents on epithelial cell migration were investigated by measuring the length of the migration path in rabbit corneal blocks in culture.
Results:
Both ROCK-1 and ROCK-2 were detected in the rabbit corneal epithelium at both protein and mRNA levels. The Rho activator lysophosphatidic acid (LPA) stimulated corneal epithelial migration in a dose-dependent manner, whereas exoenzyme C3, a Rho inhibitor, inhibited epithelial migration also in a dose-dependent manner. The stimulatory effect of LPA on corneal epithelial migration was prevented by exoenzyme C3. Both cytochalasin B, an inhibitor of actin filament assembly, and ML-7, an inhibitor of myosin light chain kinase, also prevented LPA stimulation of epithelial migration.
Conclusions:
These results suggest that Rho mediates corneal epithelial migration in response to external stimuli by regulating the organization of the actin cytoskeleton.
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