Effect of hevin deletion in mice and characterization in trabecular meshwork

Invest Ophthalmol Vis Sci. 2011 Apr 6;52(5):2187-93. doi: 10.1167/iovs.10-5428.

Abstract

Purpose: Hevin is a matricellular protein and the result of a gene duplication of SPARC. SPARC-null mice have lower intraocular pressure (IOP). The function of hevin in trabecular meshwork (TM) is unknown. The authors hypothesized that hevin is expressed in TM and has a functional consequence on IOP.

Methods: Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting were performed to identify transcription and protein expression in TM and cultured TM cells. Toluidine blue stain was performed to compare anterior segments in wild-type (WT) and hevin-null mice. Confocal microscopy localized the structural distribution of hevin in human TM and hevin/SPARC in mouse anterior segments. IOP was measured in WT (C57BL6 × 129SvJ) and hevin-null mice using both rebound tonometry and cannulation tonometry. Central corneal thickness (CCT) was measured by ocular coherence tomography. Cultured TM cells were treated with TGF-β2 because TGF-β2 is associated with primary open-angle glaucoma.

Results: Hevin mRNA and protein were expressed in TM tissues but not in cultured TM cells. No structural differences were observed in anterior segments of WT and hevin-null mice. IOP between hevin-null (n = 46) and WT (n = 44) mice was equivalent (15.3 ± 1.92 mm Hg and 15.9 ± 2.01 mm Hg, respectively; P = 0.15). CCT was similar between hevin-null and WT mice (107.95 ± 5.06 μm and 106.76 ± 3.46 μm, respectively; P = 0.11). TGF-β2 did not induce hevin, whereas SPARC expression was induced in a dose-dependent manner in human TM cell cultures.

Conclusions: Hevin does not appear to be critical to regulating IOP. Hevin is expressed in TM but, in contrast to SPARC, does not appear to be regulated by TGF-β2.

Publication types

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

MeSH terms

  • Adult
  • Animals
  • Calcium-Binding Proteins / genetics*
  • Cells, Cultured
  • Extracellular Matrix Proteins / genetics*
  • Gene Deletion*
  • Gene Expression Regulation / physiology*
  • Humans
  • Immunoblotting
  • Intraocular Pressure / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Confocal
  • Middle Aged
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tomography, Optical Coherence
  • Tonometry, Ocular
  • Trabecular Meshwork / drug effects
  • Trabecular Meshwork / metabolism*
  • Transforming Growth Factor beta2 / pharmacology

Substances

  • Calcium-Binding Proteins
  • Extracellular Matrix Proteins
  • RNA, Messenger
  • Sparcl1 protein, mouse
  • Transforming Growth Factor beta2