Scleral fibroblast response to experimental glaucoma in mice

Ericka N. Oglesby, Gülgün Tezel, Elizabeth Cone-Kimball, Matthew R. Steinhart, Joan Jefferys, Mary E. Pease, Harry A. Quigley

Research output: Contribution to journalArticle

Abstract

Purpose: To study the detailed cellular and molecular changes in the mouse sclera subjected to experimental glaucoma. Methods: Three strains of mice underwent experimental bead-injection glaucoma and were euthanized at 3 days and 1, 3, and 6 weeks. Scleral protein expression was analyzed with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) using 16O/18O labeling for quantification in 1- and 6-week tissues. Sclera protein samples were also analyzed with immunoblotting with specific antibodies to selected proteins. The proportion of proliferating scleral fibroblasts was quantified with Ki67 and 4’,6-diamidino-2-phenylindole (DAPI) labeling, and selected proteins were studied with immunohistochemistry. Results: Proteomic analysis showed increases in molecules involved in integrin-linked kinase signaling and actin cytoskeleton signaling pathways at 1 and 6 weeks after experimental glaucoma. The peripapillary scleral region had more fibroblasts than equatorial sclera (p=0.001, n=217, multivariable regression models). There was a sixfold increase in proliferating fibroblasts in the experimental glaucoma sclera at 1 week and a threefold rise at 3 and 6 weeks (p=0.0005, univariate regression). Immunoblots confirmed increases for myosin, spectrin, and actinin at 1 week after glaucoma. Thrombospondin-1 (TSP-1), HINT1, vimentin, actinin, and α-smooth muscle actin were increased according to immunohistochemistry. Conclusions: Scleral fibroblasts in experimental mouse glaucoma show increases in actin cytoskeleton and integrin-related signaling, increases in cell division, and features compatible with myofibroblast transition.

Original languageEnglish (US)
Pages (from-to)82-99
Number of pages18
JournalMolecular vision
Volume22
StatePublished - Jan 29 2016

ASJC Scopus subject areas

  • Ophthalmology

Fingerprint Dive into the research topics of 'Scleral fibroblast response to experimental glaucoma in mice'. Together they form a unique fingerprint.

  • Cite this

    Oglesby, E. N., Tezel, G., Cone-Kimball, E., Steinhart, M. R., Jefferys, J., Pease, M. E., & Quigley, H. A. (2016). Scleral fibroblast response to experimental glaucoma in mice. Molecular vision, 22, 82-99.