title: Multiplex analysis of age-related protein and lipid modifications in human Bruch's membrane.
abstract: Aging of the human retina is characterized by progressive pathology, which can lead to vision loss. This progression is believed to involve reactive metabolic intermediates reacting with constituents of Bruch's membrane, significantly altering its physiochemical nature and function. We aimed to replace a myriad of techniques following these changes with one, Raman spectroscopy. We used multiplexed Raman spectroscopy to analyze the age-related changes in 7 proteins, 3 lipids, and 8 advanced glycation/lipoxidation endproducts (AGEs/ALEs) in 63 postmortem human donors. We provided an important database for Raman spectra from a broad range of AGEs and ALEs, each with a characteristic fingerprint. Many of these adducts were shown for the first time in human Bruch's membrane and are significantly associated with aging. The study also introduced the previously unreported up-regulation of heme during aging of Bruch's membrane, which is associated with AGE/ALE formation. Selection of donors ranged from ages 32 to 92 yr. We demonstrated that Raman spectroscopy can identify and quantify age-related changes in a single nondestructive measurement, with potential to measure age-related changes in vivo. We present the first directly recorded evidence of the key role of heme in AGE/ALE formation.
authors.affiliation: Centre for Vision and Vascular Science, School of Medicine and Dentistry, Queen's University Belfast, Belfast, UK.
authors.forename: J Renwick
authors.initials: JR
authors.lastname: Beattie
authors.forename: Anna M
authors.initials: AM
authors.lastname: Pawlak
authors.forename: Michael E
authors.initials: ME
authors.lastname: Boulton
authors.forename: Jianye
authors.initials: J
authors.lastname: Zhang
authors.forename: Vincent M
authors.initials: VM
authors.lastname: Monnier
authors.forename: John J
authors.initials: JJ
authors.lastname: McGarvey
authors.forename: Alan W
authors.initials: AW
authors.lastname: Stitt
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology*; D016570:Bruch Membrane / Q000378:metabolism*; D005136:Eye Proteins / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D050356:Lipid Metabolism / Q000502:physiology*; D008875:Middle Aged; D013059:Spectrum Analysis, Raman
chemical_list: D005136:Eye Proteins; D017127:Glycation End Products, Advanced