Purchase this article with an account.
SR Gordon; Microfilament Disruption Initiates Mitosis in the Corneal Endothelium . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3185.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine whether microfilaments play a role in regulating mitosis in the corneal endothelium. Methods: Non-injured rat corneas were organ cultured in BME medium containing 10% serum supplemented with 0.25µg/ml cytochalasin B (CB) for 48 h to disrupt actin microfilaments. In other experiments the medium was additionally supplemented with either insulin (50µg/ml) or IGF-2 (100ng/ml). To capture mitotic figures, colchicine (final concentration 10-8M) was added to the medium after 24 h post explantation. In tissues not exposed to colchicine, cell density was determined by averaging the cell counts of several fields along the x-axis of flat mount preparations and extrapolating the average value to cells/mm2. Microfilament integrity was assessed by fluorescence and confocal microscopy using TRITC-phalloidin staining. In some instances nuclei were counterstained with DAPI. Results: Endothelia incubated without CB showed little or no mitosis, whereas, those exposed to the drug showed a small amount of proliferation. Supplementing the media with either insulin or IGF-2 substantually increased the number of division figures relative to controls. Cell density determinations indicated that both insulin and IGF-2 demonstrated a propensity to augment endothelial cell numbers. Fluorescence and confocal microscopy revealed that while the cellular circumferential microfilament bands appeared relatively unaffected by CB, the cytoplasmic actin cytoskeleton seemed to be completely disrupted. Conclusion: These results suggest the possibility that the actin cytoskeleton may be involved in the mitotic regulation of the corneal endothelium.
This PDF is available to Subscribers Only