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Mary Ann Stepp, Gauri Tadvalkar, Victor Perez, Yaohong Tan, Sonali Ghosh, James Zieske, Albert Lee, Vickery Trinkaus-Randall, Ahdeah Pajoohesh-Ganji; Suture placement on the mouse cornea induces K8 and Muc5ac expression within K12+ corneal epithelial cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3219.
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The corneal epithelium is maintained by a stem cell population that is bipotential and can give rise to both corneal epithelial cells and corneal goblet cells which are generated upon injuries close to the limbus (Pajoohesh-Ganji, et al., Stem Cells, 30:2032-2043, 2012). To investigate whether corneal goblet cell progenitors are present in the central mouse cornea, we triggered an immune response by placing sutures in the central cornea and then evaluated the corneal epithelium for the presence of corneal goblet cells.
Adult BALB/c (5) and C57BL6 (4) mouse corneas were sutured as described for corneal transplantation studies studies (Dana and Streilein, IOVS, 37:2485-2494, 1996) and were allowed to respond to the sutures for 14 days. Sutures were classified as low or high risk depending upon the length of the suture thread left in contact with the corneal epithelium. After 14 days, mice were sacrificed, eyes fixed, and corneas processed for whole mount immunofluorescence to localize K12, K8, and Muc5ac within the corneal epithelium. Corneas were stained with the sutures in place.
Data show that sutures induce expression of goblet cell markers including K8 and Muc5ac within K12 positive corneal epithelial cells within 14 days after sutures are placed in the cornea in both BALB/c and C57BL6 mice but the upregulation is more robust in BALB/c compared to C57BL6 mice. Similar upregulation is seen for low and high risk sutures.
The data obtained support the fact that the immune response of the mouse cornea to a suture can induce the conversion of corneal epithelial and goblet progenitor cells located in the central cornea into goblet cells. Since the corneas were not debrided, it is unlikely that the cells that generate the goblet cells migrate from the limbus. We have shown previously that wounding the cornea near the limbus upregulates corneal goblet cell differentiation on the ocular surface. Additional studies are underway to determine whether these events can occur in rat corneas in vitro in organ culture and in human corneal equivalents
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