| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
From the Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, Florida.
PURPOSE. To determine whether the cellular distribution of cell adhesion–associated protein, pinin, is altered during corneal epithelial migration in response to debridement wounding and to determine the effect of overexpression of pinin in cultured epithelial cells.
METHODS. Corneas from guinea pig and embryonic (day 17) chickens were excised, wounded, and placed on organ-culture rafts. At time points from 0 to 24 hours, corneas were cryosectioned and subsequently analyzed by immunofluorescence or immunoelectron microscopy for the presence and distribution of pinin. Cultured epithelial cell line MDCK (Madin Darby canine kidney) confluent monolayers were wounded by scraping and examined by immunofluorescence for pinin and desmoplakin. MDCK cells were transfected with full-length pinin cDNA. After selection in Geneticin, clones of pinin-transfected cells were isolated. Monolayers of transfected cells were scrape-wounded and assayed for their ability to migrate.
RESULTS. Within 2 hours after wounding, although morphologically identifiable desmosomes were present on migrating epithelial cells, the association of pinin to desmosomes was greatly reduced. Finally, after completion of wound closure, pinin returned to the corneal epithelial desmosome. Wounding of confluent epithelial monolayers (MDCK) in vitro demonstrated a very similar change in the distribution of pinin, whereas desmoplakin remained cell boundary–associated. Transfection of pinin into cultured epithelial cells resulted in an overexpression of pinin. Clones of cells expressing high levels of pinin exhibited marked reduction in their ability to migrate after wounding.
CONCLUSIONS. Pinin is involved in corneal epithelium migration. The localization of pinin at or near the desmosome is correlated with the epithelial quiescence. The loss of pinin from the cell boundary correlates with the transition from quiescence to actively migrating. Overexpressing pinin in cultured epithelial cells affects epithelial homeostasis and, in turn, drives the epithelial cells to a hyperstable epithelial adhesive state and inhibits the transition from quiescence to migratory.
This article has been cited by other articles:
|
|
 |
|
  R. Alpatov, Y. Shi, G. C. Munguba, B. Moghimi, J.-H. Joo, J. Bungert, and S. P. Sugrue Corepressor CtBP and Nuclear Speckle Protein Pnn/DRS Differentially Modulate Transcription and Splicing of the E-Cadherin Gene Mol. Cell. Biol., March 1, 2008; 28(5): 1584 - 1595. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Alpatov, G. C. Munguba, P. Caton, J. H. Joo, Y. Shi, Y. Shi, M. E. Hunt, and S. P. Sugrue Nuclear Speckle-Associated Protein Pnn/DRS Binds to the Transcriptional Corepressor CtBP and Relieves CtBP- Mediated Repression of the E-Cadherin Gene Mol. Cell. Biol., December 1, 2004; 24(23): 10223 - 10235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Zimowska, J. Shi, G. Munguba, M. R. Jackson, R. Alpatov, M. N. Simmons, Y. Shi, and S. P. Sugrue Pinin/DRS/memA Interacts with SRp75, SRm300 and SRrp130 in Corneal Epithelial Cells Invest. Ophthalmol. Vis. Sci., November 1, 2003; 44(11): 4715 - 4723. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Shi and S. P. Sugrue Dissection of Protein Linkage between Keratins and Pinin, a Protein with Dual Location at Desmosome-Intermediate Filament Complex and in the Nucleus J. Biol. Chem., May 12, 2000; 275(20): 14910 - 14915. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
