Recombinant TIGR/MYOC increases outflow resistance in the human anterior segment

M. P. Fautsch, C. K. Bahler, D. J. Jewison, D. H. Johnson

Research output: Contribution to journalArticlepeer-review

121 Scopus citations

Abstract

PURPOSE. To determine the effect of human recombinant TIGR/myocilin (MYOC) protein on outflow resistance in the human anterior segment. METHODS. A cDNA for MYOC was inserted into a bacterial expression system and purified with nickel ion affinity chromatography. The anterior segments of 12 pairs of human eyes were placed in perfusion organ culture. One eye received an anterior chamber exchange with partially purified recombinant MYOC (25 μg), whereas the other eye received either heat-denatured recombinant MYOC (25 μg), partially purified β-galactosidase (25 or 250 μg), or partially purified control proteins isolated from a null expression lysate (25 μg). Eyes were fixed up to 72 hours after infusion, and immunohistochemistry was performed using anti-MYOC polyclonal antibody. RESULTS. Recombinant MYOC caused an increase in IOP over 12 hours, increasing outflow resistance 94%, whereas the fellow eye infused with null expression sample increased 12% (n = 7; P =0.0005). When compared with recombinant MYOC, neither heat-denatured MYOC, recombinant β-galactosidase, bovine serum albumin, nor fetal calf serum caused an increase in outflow resistance. MYOC IOP remained above baseline levels for 48 to 72 hours. Immunohistochemistry results confirmed the presence of recombinant MYOC in the trabecular meshwork. CONCLUSIONS. Recombinant MYOC increased outflow resistance in human anterior segments, whereas control proteins did not. MYOC may increase outflow resistance by specific interactions within the trabecular meshwork.

Original languageEnglish (US)
Pages (from-to)4163-4168
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume41
Issue number13
StatePublished - Dec 12 2000

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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