Myocilin levels in primary open-angle glaucoma and pseudoexfoliation glaucoma human aqueous humor

Kyle G. Howell, Anne M. Vrabel, Uttio Roy Chowdhury, William Daniel Stamer, Michael P Fautsch

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

PURPOSE: To determine the concentration of myocilin in primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PEXG) aqueous humor. METHODS: Aqueous humor was collected during surgery from patients with POAG, PEXG, and elective cataract removal (control). Volume-equivalent aqueous samples were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gradient gels. Quantification of myocilin levels was performed using Western blots probed with 2 independent N-terminal polyclonal anti-myocilin antibodies (AB1 and AB2) followed by densitometry. Myocilin levels in aqueous humor were quantified by plotting the densitometry readings of the aqueous samples against a recombinant myocilin standard curve. Total protein concentration was determined by Bradford protein assay. Transforming growth factor β 2 levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Myocilin levels are significantly elevated in human POAG aqueous humor when compared with control aqueous humor (AB1: 0.66±0.53 ng/μL vs. 0.23±0.20 ng/μL, P<0.001; AB2: 0.98±0.59 ng/μL vs. 0.65±0.5 ng/μL, P<0.03; mean±SD). Myocilin makes up a larger percent of the total protein in POAG aqueous humor compared with control aqueous (AB1: 0.26±0.20% vs. 0.10±0.20%, P<0.001; AB2: 0.43±0.32% vs. 0.28±0.18%, P<0.05). In contrast to POAG, myocilin levels were not elevated in PEXG aqueous humor when compared with control aqueous humor. No correlation between myocilin and transforming growth factor β 2 levels was observed. CONCLUSIONS: Myocilin is elevated in POAG, but not in PEXG aqueous humor.

Original languageEnglish (US)
Pages (from-to)569-575
Number of pages7
JournalJournal of Glaucoma
Volume19
Issue number9
DOIs
StatePublished - Dec 2010

Fingerprint

Aqueous Humor
Glaucoma
Densitometry
Transforming Growth Factors
trabecular meshwork-induced glucocorticoid response protein
Primary Open Angle Glaucoma
Proteins
Sodium Dodecyl Sulfate
Cataract
Reading
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Western Blotting
Gels
Enzyme-Linked Immunosorbent Assay

Keywords

  • aqueous humor
  • glaucoma
  • myocilin
  • PEXG
  • POAG
  • TGFb2
  • TIGR

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Myocilin levels in primary open-angle glaucoma and pseudoexfoliation glaucoma human aqueous humor. / Howell, Kyle G.; Vrabel, Anne M.; Chowdhury, Uttio Roy; Stamer, William Daniel; Fautsch, Michael P.

In: Journal of Glaucoma, Vol. 19, No. 9, 12.2010, p. 569-575.

Research output: Contribution to journalArticle

Howell, Kyle G. ; Vrabel, Anne M. ; Chowdhury, Uttio Roy ; Stamer, William Daniel ; Fautsch, Michael P. / Myocilin levels in primary open-angle glaucoma and pseudoexfoliation glaucoma human aqueous humor. In: Journal of Glaucoma. 2010 ; Vol. 19, No. 9. pp. 569-575.
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abstract = "PURPOSE: To determine the concentration of myocilin in primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PEXG) aqueous humor. METHODS: Aqueous humor was collected during surgery from patients with POAG, PEXG, and elective cataract removal (control). Volume-equivalent aqueous samples were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gradient gels. Quantification of myocilin levels was performed using Western blots probed with 2 independent N-terminal polyclonal anti-myocilin antibodies (AB1 and AB2) followed by densitometry. Myocilin levels in aqueous humor were quantified by plotting the densitometry readings of the aqueous samples against a recombinant myocilin standard curve. Total protein concentration was determined by Bradford protein assay. Transforming growth factor β 2 levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Myocilin levels are significantly elevated in human POAG aqueous humor when compared with control aqueous humor (AB1: 0.66±0.53 ng/μL vs. 0.23±0.20 ng/μL, P<0.001; AB2: 0.98±0.59 ng/μL vs. 0.65±0.5 ng/μL, P<0.03; mean±SD). Myocilin makes up a larger percent of the total protein in POAG aqueous humor compared with control aqueous (AB1: 0.26±0.20{\%} vs. 0.10±0.20{\%}, P<0.001; AB2: 0.43±0.32{\%} vs. 0.28±0.18{\%}, P<0.05). In contrast to POAG, myocilin levels were not elevated in PEXG aqueous humor when compared with control aqueous humor. No correlation between myocilin and transforming growth factor β 2 levels was observed. CONCLUSIONS: Myocilin is elevated in POAG, but not in PEXG aqueous humor.",
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AU - Howell, Kyle G.

AU - Vrabel, Anne M.

AU - Chowdhury, Uttio Roy

AU - Stamer, William Daniel

AU - Fautsch, Michael P

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N2 - PURPOSE: To determine the concentration of myocilin in primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PEXG) aqueous humor. METHODS: Aqueous humor was collected during surgery from patients with POAG, PEXG, and elective cataract removal (control). Volume-equivalent aqueous samples were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gradient gels. Quantification of myocilin levels was performed using Western blots probed with 2 independent N-terminal polyclonal anti-myocilin antibodies (AB1 and AB2) followed by densitometry. Myocilin levels in aqueous humor were quantified by plotting the densitometry readings of the aqueous samples against a recombinant myocilin standard curve. Total protein concentration was determined by Bradford protein assay. Transforming growth factor β 2 levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Myocilin levels are significantly elevated in human POAG aqueous humor when compared with control aqueous humor (AB1: 0.66±0.53 ng/μL vs. 0.23±0.20 ng/μL, P<0.001; AB2: 0.98±0.59 ng/μL vs. 0.65±0.5 ng/μL, P<0.03; mean±SD). Myocilin makes up a larger percent of the total protein in POAG aqueous humor compared with control aqueous (AB1: 0.26±0.20% vs. 0.10±0.20%, P<0.001; AB2: 0.43±0.32% vs. 0.28±0.18%, P<0.05). In contrast to POAG, myocilin levels were not elevated in PEXG aqueous humor when compared with control aqueous humor. No correlation between myocilin and transforming growth factor β 2 levels was observed. CONCLUSIONS: Myocilin is elevated in POAG, but not in PEXG aqueous humor.

AB - PURPOSE: To determine the concentration of myocilin in primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PEXG) aqueous humor. METHODS: Aqueous humor was collected during surgery from patients with POAG, PEXG, and elective cataract removal (control). Volume-equivalent aqueous samples were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gradient gels. Quantification of myocilin levels was performed using Western blots probed with 2 independent N-terminal polyclonal anti-myocilin antibodies (AB1 and AB2) followed by densitometry. Myocilin levels in aqueous humor were quantified by plotting the densitometry readings of the aqueous samples against a recombinant myocilin standard curve. Total protein concentration was determined by Bradford protein assay. Transforming growth factor β 2 levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Myocilin levels are significantly elevated in human POAG aqueous humor when compared with control aqueous humor (AB1: 0.66±0.53 ng/μL vs. 0.23±0.20 ng/μL, P<0.001; AB2: 0.98±0.59 ng/μL vs. 0.65±0.5 ng/μL, P<0.03; mean±SD). Myocilin makes up a larger percent of the total protein in POAG aqueous humor compared with control aqueous (AB1: 0.26±0.20% vs. 0.10±0.20%, P<0.001; AB2: 0.43±0.32% vs. 0.28±0.18%, P<0.05). In contrast to POAG, myocilin levels were not elevated in PEXG aqueous humor when compared with control aqueous humor. No correlation between myocilin and transforming growth factor β 2 levels was observed. CONCLUSIONS: Myocilin is elevated in POAG, but not in PEXG aqueous humor.

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KW - glaucoma

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KW - POAG

KW - TGFb2

KW - TIGR

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