April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Decreased Carbohydrate Metabolism Enzyme Activities in the Glaucomatous Trabecular Meshwork
Author Affiliations & Notes
  • A. K. Junk
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida
    Ophthalmology, Department of Veterans Affairs Medical Center, Miami, Florida
  • M. Goel
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida
  • T. K. Mundorf
    Mundorf Eye Center, Charlotte, North Carolina
  • E. J. Rockwood
    Cole Eye Institute, Cole Eye Institute, Cleveland, Ohio
  • S. K. Bhattacharya
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida
  • Footnotes
    Commercial Relationships  A.K. Junk, None; M. Goel, None; T.K. Mundorf, None; E.J. Rockwood, None; S.K. Bhattacharya, None.
  • Footnotes
    Support  Research to Prevent Blindness
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 3218. doi:
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      A. K. Junk, M. Goel, T. K. Mundorf, E. J. Rockwood, S. K. Bhattacharya; Decreased Carbohydrate Metabolism Enzyme Activities in the Glaucomatous Trabecular Meshwork. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3218.

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

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Abstract

Purpose: : To determine whether activity of carbohydrate metabolism enzymes (Aldolase, Pyruvate Kinase and Isocitrate Dehydrogenase) are altered in the glaucomatous trabecular meshwork (TM) compared to controls.

Methods: : Tissue specimens were obtained from trabeculectomy (n=45 glaucoma; Caucasian, average age 61±8 of both genders) and from cadaver eyes (n= 15 control and n= 5 glaucoma; Caucasian, average age 63±4 of both genders). Protein extracts from TM tissue were prepared in a non-denaturing buffer containing 0.1% genapol. Aldolase activity was measured spectrophotometrically using reaction of 3-phosphoglycerate with hydrazine to form hydrazone that absorbs at 240 nm. Pyruvate Kinase activity was measured by coupling with Lactate Dehydrogenase with NADH and pyruvate. Absorbance was measured at 340 nm. Isocitrate Dehydrogenase activity was measured using reduction of NADP to NADPH resulting in a characteristic absorbance at 340 nm.

Results: : Aldolase, Pyruvate Kinase and Isocitrate Dehydrogenase activities in the glaucomatous TM tissue were found to be reduced 70, 50 and 25 percent, respectively. SDS-PAGE analysis suggests presence of 4-hydorxynonenal modified Isocitrate Dehydrogenase protein in the glaucomatous TM tissue compared to controls.

Conclusions: : Several Krebs cycle enzyme activities are considerably reduced in glaucomatous TM. HNE modified isocitrate dehydrogenase activity is consistent with reduced inactivated form of the protein.

Keywords: trabecular meshwork • metabolism • enzymes/enzyme inhibitors 
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