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Ana Fernandez, Carlos Alonso-Ron, Miguel Coca-Prados, Ignacio Alcalde, Lydia Alvarez, Manuel Chacon, Guilherme Ferrara, Jesus Merayo-Lloves, Carlos Belmonte; Cholecystokinin And Gastrin Expression In Corneal Epithelium. Invest. Ophthalmol. Vis. Sci. 2011;52(14):277.
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To determine whether the regulatory peptides cholecystokinin (CCK) and the CCK-related peptide gastrin are co-expressed by the corneal epithelium.
Total RNA was extracted from healthy whole human corneas and from a human corneal epithelial cell line (SV40-HCE; Araki-Sasaki K. et al., 1995) with TRIzol, purified with Qiagen RNeasy and processed for hybridization using the Illumina BeadChip array platform. Expression of CCK and gastrin in cornea and HCEsv cells was validated by conventional RT-PCR using specific primers. Immunolabeling of CCK and gastrin peptides was carried out in both, paraffin-embedded corneas and in cultured SV40-HCE cells following conventional procedures. Briefly, cultured HCEsv cells and cornea sections were fixed with 4% paraformaldehyde, permeabilized with Triton X-100 and incubated overnight with CCK or gastrin antibodies. Peptides were visualized by indirect immunofluorescence.
Microarray analysis of human corneas and HCEsv cells showed that both CCK and gastrin transcripts are expressed. RT-PCR analysis confirmed these results. Immunolabeling of corneas with CCK and gastrin antibodies indicated that both peptides co-localized along the corneal epithelium cells of the basal layer closest to the Bowman’s membrane. HCEsv cells in culture, also under steady-state conditions, were also immuno-stained for both peptides.
Our data show that corneal epithelial cells, in culture, are able to express CCK present as observed in native tissue. Furthermore, the CCK’s evolutionarily and related peptide gastrin, appears to be also coexpressed with CCK in the cornea epithelium in both in vivo and in vitro. The function of CCK and gastrin in the cornea is at present unknown.
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