Perfusion-cultured bovine anterior segments as an ex vivo model for studying glucocorticoid-induced ocular hypertension and glaucoma

Invest Ophthalmol Vis Sci. 2011 Oct 17;52(11):8068-75. doi: 10.1167/iovs.11-8133.

Abstract

Purpose: To determine whether perfusion-cultured bovine anterior segments would be a suitable model for glaucoma research.

Methods: Fresh bovine eyes were dissected and sealed on a custom-made acrylic dish with an O-ring. Perfusion medium was infused by a syringe pump at a constant infusion rate of 5 μL/min. After intraocular pressure (IOP) was stable, bovine eyes were perfused with medium containing either a vehicle control (0.1% ethanol [ETH]) or dexamethasone (DEX) for up to 7 days. IOP was recorded by a pressure transducer and a computerized system. Perfusion medium was collected for Western immunoblot analysis of myocilin (MYOC).

Results: The morphology of the bovine trabecular meshwork after perfusion culture was similar to that of freshly dissected, nonperfused bovine eyes. Treatment with DEX elevated IOP in some bovine eyes, whereas others showed little change. The authors analyzed the data from 18 ETH-treated control eyes and defined 2.82 mm Hg as the threshold of ocular hypertension (OHT), which equals mean pressure change + 2× SD. Approximately 40% (12/29) of the bovine eyes were DEX responders, which is very close to the DEX-responsive rates observed in human and monkey eyes. Western blot data showed that DEX treatment induced the expression of the DEX-inducible gene MYOC only in the perfusion-cultured anterior segments with DEX-induced OHT.

Conclusions: OHT can be induced by DEX in perfusion-cultured bovine anterior segments. This is a fast, convenient, affordable, and reliable model for studying DEX-induced OHT and the mechanisms of trabecular outflow.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anterior Eye Segment
  • Blotting, Western
  • Cattle
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism
  • Dexamethasone / toxicity*
  • Disease Models, Animal*
  • Eye Proteins / genetics
  • Eye Proteins / metabolism
  • Gene Expression Regulation / drug effects
  • Glaucoma, Open-Angle / chemically induced
  • Glaucoma, Open-Angle / metabolism
  • Glucocorticoids / toxicity*
  • Glycoproteins / genetics
  • Glycoproteins / metabolism
  • Intraocular Pressure / drug effects*
  • Ocular Hypertension / chemically induced*
  • Ocular Hypertension / metabolism
  • Organ Culture Techniques
  • Perfusion
  • Trabecular Meshwork / drug effects
  • Trabecular Meshwork / metabolism

Substances

  • Cytoskeletal Proteins
  • Eye Proteins
  • Glucocorticoids
  • Glycoproteins
  • trabecular meshwork-induced glucocorticoid response protein
  • Dexamethasone