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J.-A. Ko, R. Yanai, N. Morishige, T. Nishida; Effects of Semaphorin3A Over-Expressed Corneal Fibroblasts on the Expression of Adherent Junction Proteins in Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4633.
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Semaphorin are a family of glycoproteins that play an important role in repulsive axon guidance during embyogenesis. We have now investigated that effect of Sema3A over-expressed corneal fibroblasts on the expression of junctional proteins in corneal epithelial cells with the use of a coculture system in which the two cell types are separated by a collagen vitrigel membrane.
Sema3A over-expressed human corneal fibroblasts and simian virus 40-transformed human corneal epithelial (HCE) cells were cultured on opposite sides of a collagen vitrigel membrane. Expression of junctional proteins, ZO-1, Occludin, Claudin, E-cadherin, N-cadherin in HCE cells was examined by immunoblot analyses and reverse transcription-polymerase chain reaction (RT-PCR).
Immunoblot and RT-PCR analysis showed that the amounts of E-cadherin and N-cadherin in HCE cells were increased by the presence of Sema3A over-expressed corneal fibroblasts, although no changed in HCE cells by coculture with mock-transfected corneal fibroblasts. And, the amounts of ZO-1, occludin, or claudin, proteins or mRNAs in HCE cells were not affected by the presence of Sema3A over-expressed corneal fibroblasts.
These results suggest that corneal fibroblasts and semaphorin3A released from corneal fibroblasts may play an important role in intercellular communication between HCE cells as well as in maintenance of corneal structure and functions.
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