April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Role of Nestin and Vimentin in Lacrimal Gland Repair Following Experimentally Induced Inflammation
Author Affiliations & Notes
  • S. you
    General Dentistry, Tufts University School of Dental Medicine, Boston, Massachusetts
  • C. Kublin
    General Dentistry, Tufts University School of Dental Medicine, Boston, Massachusetts
  • D. Zoukhri
    General Dentistry, Tufts University School of Dental Medicine, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  S. you, None; C. Kublin, None; D. Zoukhri, None.
  • Footnotes
    Support  NIH Grant EY12383
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 2957. doi:
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      S. you, C. Kublin, D. Zoukhri; Role of Nestin and Vimentin in Lacrimal Gland Repair Following Experimentally Induced Inflammation. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2957.

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Abstract

Purpose: : It was previously reported that the amount of nestin (a type VI intermediate filament protein and a marker of stem cells) increased during lacrimal gland repair. The purpose of the present studies was to determine whether the amount of vimentin, another intermediate filament (type III) protein, changes during lacrimal gland repair.

Methods: : Injury was induced by direct injection of recombinant human interleukin-1 (IL-1) α (1 µg in 2 µl) into the exorbital lacrimal glands of anesthetized female BALB/c mice. Lacrimal glands from saline (vehicle for IL-1) injected and non-injected animals served as controls. Animals were sacrificed at 1, 2, 3, 4, 5, 6 and 7 days after IL-1 injection. The lacrimal glands were excised and processed for histopathology, immunohistochemistry or Western blotting. Lacrimal glands from 2.5-day IL-1 treated animals were used for explant tissue culture to isolate and propagate mesenchymal stem cells. For immunohistochemistry studies, cells in passage 2-3 were grown on 8-well chamber slides in complete Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Results: : Lacrimal gland repair occurred between 2 and 3 days post-IL-1 injection and was complete by 7 days. Compared to control, the amount of vimentin protein in 2-day injected lacrimal glands was significantly increased by 5.5-fold. Vimentin levels remained high until around day 5 and decreased to control levels by day 7 after IL-1 injection. Immunohistochemistry studies on tissue sections showed that compared to the control, the number of vimentin+ cells increased in glands removed from 2- and 3-day injected animals. Double-labeling experiments revealed three patterns of staining: vimentin+ cells, nestin+ cells, and vimentin+/nestin+ double positive cells. Similar to nestin, some vimentin+ cells were also positive for α-smooth muscle actin (a marker of myoepithelial cells). Cultured mesenchymal stem cells prepared from IL-1 injected lacrimal glands, retained immunoreactivity for nestin, vimentin and α-smooth muscle actin.

Conclusions: : We concluded from these studies that, similar to nestin, vimentin protein is upregulated during lacrimal gland repair after experimentally induced inflammation. We hypothesize that re-expression of nestin and vimentin in mesenchymal stem cells is necessary for their migration and repair of the lacrimal gland.

Keywords: cornea: tears/tear film/dry eye • lacrimal gland • inflammation 
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