Age-related changes in human macular Bruch's membrane as seen by quick-freeze/deep-etch

Abstract

Lipid-containing inclusions have been observed in human Bruch's membrane (BrM) and are postulated to be associated with age-related maculopathy (ARM), a major cause of legal blindness in developed countries. The dehydration associated with specimen preparation for thin-section transmission electron microscopy causes loss of these inclusions. Better preservation of the ultrastructure of the inclusions and tissue is achieved by using a quick-freeze/deep-etch preparation. We use this technique to examine normal human macular BrM in order to characterize the deposition of the lipid-rich inclusions and their age-related accumulation within different layers of the tissue. We find that various inclusions mentioned in other studies can be formed by combinations of three basic structures: lipoprotein-like particles (LLPs), small granules (SGs) and membrane-like structures. These inclusions are associated with collagen and elastic fibrils by fine filaments. In younger eyes, these inclusions are found mostly in the elastic (EL) and outer collageneous layer (OCL) and occupy a small fraction of the interfibrillar spacing. As age increases, LLPs and SGs gradually fill the interfibrillar spacing of the EL and inner collageneous layer (ICL) of the tissue, and later form a new sublayer, the lipid wall, within the boundary region between the basal lamina of retinal pigment epithelium (RPE) and ICL. Because the formation of the lipid wall only occurs after these inclusions fill the ICL, and it seems unlikely that the LLPs can pass through the packed layer, this result suggests a possible RPE origin of the LLPs that make up the lipid wall.

Effect of aging and lifestyle on photoreceptors and retinal pigment epithelium: cross-sectional study in a healthy Danish population

“…Histological studies find increased lipid and lipoprotein accumulation with age, which seems to be particularly abundant in individuals aged > 60 years [41]. Using electron microscopy, lipoproteins in aged individuals are seen as spread out in Bruch’s membrane as small particles with a diameter of 60–100 nm [41–43]. In an experimental mouse model, exposure to cigarette smoke thickened Bruch’s membrane due to accumulation of deposits within the membrane [44].…”

Apolipoprotein B-containing lipoproteins in retinal aging and age-related macular degeneration

“…Used to examine BrM, QFDE revealed solid particles accumulating with age that also exhibited a shell and core structure ( Fig. 5A ) ( 92,93 ). Particles typically varied in size from 60-100 nm but could be as large as 300 nm.…”

Comparison of Morphology of Human Macular and Peripheral Bruch's Membrane in Older Eyes

“…6,7 Along with LLPs, we saw an accumulation in BrM of small granules, roughly 10 nm in diameter. 7 We do not know the identity of these particles. However, they look similar to particles seen in the basal lamina, particularly the lamina densa (Figs.…”