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Koji Kitazawa, Satoshi Kawasaki, Morio Ueno, Hiroshi Tanaka, Katsuhiko Shinomiya, Mina Nakatsukasa, Shigeru Kinoshita; Investigation of protein kinase C alpha in corneal epithelial cells derived from a gelatinous drop-like corneal dystrophy patient. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5506.
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Gelatinous drop-like corneal dystrophy (GDLD) is a rare autosomal recessive disorder caused by biallelic loss-of-function mutations within the tumor-associated calcium signal transducer 2 (TACSTD2) gene, and is characterized by the reduction of tight junction-related proteins and a resultant loss of barrier function. Protein kinase C-alpha (PKC-alpha) is known to be involved in altering the function of tight-junctions. The purpose of this present study was to investigate the involvement of PKC-alpha in GDLD.
Immortalized corneal epithelial cells (CECs), which we previously established to have similar characteristics as in vivo GDLD CECs, were seeded on a culture well and then cultured in growth medium. After the cells had reached 100% confluence, the medium was switched to a differentiation medium and the cells were cultured for an additional 7 days. The expression levels of PKC-alpha mRNA , PKC-alpha protein, and tight junction-related proteins claudin (CLDN)-1 and CLDN7 were then examined using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis.
The expression levels of CLDN1 and CLDN7 proteins in GDLD CECs was significantly lower compared to the normal CECs. The mRNA level of PKC-alpha in the GDLD cells was upregulated compared to the normal cells. In addition, the PKC-alpha protein was found to be highly expressed in the GDLD cells.
The findings of this study show that PKC-alpha may be involved in the tight- junction complex in GDLD.
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