Confocal Raman microscopy can quantify advanced glycation end product (AGE) modifications in Bruch's membrane leading to accurate, nondestructive prediction of ocular aging

Josephine V. Glenn, J. Renwick Beattie, Lindsay Barrett, Norma Frizzell, Suzanne R. Thorpe, Mike E. Boulton, John J. McGarvey, Alan W. Stitt

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

The modification of proteins by nonenzymatic glycation leading to accumulation of advanced glycation end products (AGEs) is a well-established phenomenon of aging. In the eyes of elderly patients, these adducts have been observed in retinal pigment epithelium (RPE), particularly within the underlying pentalaminar substrate known as Bruch's membrane. AGEs have also been localized to age-related subcellular deposits (drusen and basal laminar deposits) and are thought to play a pathogenic role in progression of the major sight-threatening condition known as age-related macular degeneration (AMD). The current study has quantified AGEs in Bruch's membrane from postmortem eyes and established age-related correlations. In particular, we investigated the potential of confocal Raman microscopy to identify and quantify AGEs in Bruch's membrane in a nondestructive, analytical fashion. Bruch's membrane and the innermost layers of the underlying choroid (BM-Ch) were dissected from fresh postmortem eye-cups (n=56). AGE adducts were quantified from homogenized tissue using reverse-phase HPLC and GC/MS in combination with immunohistochemistry. For parallel Raman analysis, BM-Ch was flat-mounted on slides and evaluated using a Raman confocal microscope and spectra analyzed by a range of statistical approaches. Quantitative analysis showed that the AGEs pentosidine, carboxymethyllysine (CML), and carboxyethyllysine (CEL) occurred at significantly higher levels in BM-Ch with age (P

Original languageEnglish (US)
Pages (from-to)3542-3552
Number of pages11
JournalFASEB Journal
Volume21
Issue number13
DOIs
StatePublished - Nov 2007
Externally publishedYes

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Bruch Membrane
Advanced Glycosylation End Products
Confocal Microscopy
microscopy
Microscopic examination
Aging of materials
eyes
Membranes
prediction
pentosidine
glycation
Deposits
microscopes
Retinal Pigments
immunohistochemistry
Choroid
Retinal Pigment Epithelium
quantitative analysis
Macular Degeneration
epithelium

Keywords

  • Raman spectroscopy
  • RPE

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Confocal Raman microscopy can quantify advanced glycation end product (AGE) modifications in Bruch's membrane leading to accurate, nondestructive prediction of ocular aging. / Glenn, Josephine V.; Beattie, J. Renwick; Barrett, Lindsay; Frizzell, Norma; Thorpe, Suzanne R.; Boulton, Mike E.; McGarvey, John J.; Stitt, Alan W.

In: FASEB Journal, Vol. 21, No. 13, 11.2007, p. 3542-3552.

Research output: Contribution to journalArticle

Glenn, Josephine V. ; Beattie, J. Renwick ; Barrett, Lindsay ; Frizzell, Norma ; Thorpe, Suzanne R. ; Boulton, Mike E. ; McGarvey, John J. ; Stitt, Alan W. / Confocal Raman microscopy can quantify advanced glycation end product (AGE) modifications in Bruch's membrane leading to accurate, nondestructive prediction of ocular aging. In: FASEB Journal. 2007 ; Vol. 21, No. 13. pp. 3542-3552.
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