Ultrastructural analysis of damage to nuclear fiber cell membranes in advanced age-related cataracts from India.
The primary goal was to characterize the structural alterations that occur at the fiber cell interfaces in nuclei of fully opaque cataracts removed by extracapsular cataract surgery in India. The dark yellow to brunescent nuclei, ages 38-78 years, were probably representative of advanced age-related nuclear cataracts. Thick tissue slices were fixed, en bloc stained and embedded for transmission electron microscopy. Stained thin sections contained well-preserved membranes and junctions, although the complex cellular topology often made it necessary to tilt the grid extensively to visualize the membranes. Damage to the fiber cell membranes was noted in all regions of the nucleus. The most important damage occurred within undulating membrane junctions where the loss of membrane segments was common. These membrane breaks were not sites of fusion as membrane edges were detected and cytoplasm appeared to be in contact with extracellular space, which was enlarged in many regions. Dense deposits of protein-like material were frequently observed within the extracellular space and appeared to be similar to protein in the adjacent cytoplasm. The deposits were often 20-50 nm thick, variable in length and located on specific sites on plasma membranes and between clusters of cells or cell processes. In addition, low density regions were seen within the extracellular space, especially within highly undulating membranes where spaces about 100 nm in diameter were observed. The membrane damage was more extensive and extracellular spaces were larger than in aged transparent donor lenses. Because high and low density regions contribute equally to the fluctuations in refractive index, the changes in density due to the observed damage near membranes are likely to produce significant light scattering based on theoretical analysis. The dimensions of the fluctuations in the range 20-100 nm imply that the scattering is probably similar to that of small particles that would increase high-angle scattering visible in the slit lamp. Such damage to membranes would be expected to contribute to the total opacification of the nucleus as the cataract matures. The main sources of the fluctuations appear to be the degradation of membranes and adjacent cytoplasmic proteins, as well as the redistribution of proteins and fragments.
Costello, MJ; Johnsen, S; Metlapally, S; Gilliland, KO; Ramamurthy, B; Krishna, PV; Balasubramanian, D
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