June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Latent Transforming Growth Factor-β Binding Protein 1 expression in the corneal stroma
Author Affiliations & Notes
  • Edgar M Espana
    USF Health Morsani College of Medicine, Tampa, Florida, United States
  • Sarah Salvatori
    USF Health Morsani College of Medicine, Tampa, Florida, United States
  • Mei Sun
    USF Health Morsani College of Medicine, Tampa, Florida, United States
  • Devon Cogswell
    USF Health Morsani College of Medicine, Tampa, Florida, United States
  • Footnotes
    Commercial Relationships   Edgar Espana GSK, Code C (Consultant/Contractor); Sarah Salvatori None; Mei Sun None; Devon Cogswell None
  • Footnotes
    Support  NIH/NEI grant EY029395
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2642. doi:
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    • Get Citation

      Edgar M Espana, Sarah Salvatori, Mei Sun, Devon Cogswell; Latent Transforming Growth Factor-β Binding Protein 1 expression in the corneal stroma. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2642.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Latent Transforming Growth Factor-β Binding Protein 1 (LTBP1) is poorly characterized in the cornea, and our knowledge of its function is mostly unknown. The aim of the current study was to elucidate the expression and role(s) of LTBP1 in regulating corneal stromal structure and function during homeostasis and following injury. The biological importance of LTBP1 radicates in its property to maintain Transforming Growth Factor (TGF) - β latent.

Methods : Analysis of LTBP1 expression, temporal and spatial, was performed at different postnatal days (P) in wild-type C57BL/6 mouse corneal stromas and after injury. Ltbp1 mRNA expression as well as LTBP1 protein content and locatization with stromal maturation were studied. Total (latent + active) TGF- β was analyzed in vitro using transformed mink lung cells transfected with luciferase cDNA driven by PAI-1 promoter. To test the amount of total TGF-β stored in the stromal matrix at different ages, we measured exactly similar aliquots of stromal extract at different ages.

Results : Ltbp1 mRNA stromal expression, obtained after enymatic removal of corneal epithelium, was present at P4 but decreased with corneal maturation at P10, P30 and P140. There was a signficant decrease in Ltbp1 mRNA expression with corneal maturation. In contrast, Wes protein analysis showed increased deposition of LTBP1 with corneal maturation. Highest levels were noted at P140, the oldest age in which protein expression was investigated. Immunofluorescent microscopy showed LTBP1expresion in all ages studied with expression of LTBP1 localized to the extracellular matrix in the entire stromal thickness. Ltbp1 mRNA was significantly upregulated after injury and LTBP1 expression up-regulated following two different injury models: stromal abrasion and full thickness laceration. Besides, we investigated whether increased LTBP1 expression with corneal maturation correlated with increased TGF- β activity since it is well established that LTBP1 is the main regulator of latent TGF- β. We found a statistically significant increase in total TGF- β stromal corneal with corneal maturation, P30 vs P150, t-test, unpaired, p=0.04.

Conclusions : This study indicates that LTBP1 plays a regulatory role in corneal development and in the function of the adult cornea. LTBP1 deposition and total TGF-β storage increase during stromal maturtion. The expression of LTBP1 is recapitulated during wound healing.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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