Selected Publications

Background: Pathological hallmarks of Alzheimer's disease include cerebral β-amyloid (Aβ) deposition, amyloid accumulation, and neuritic plaque formation. We aimed to investigate the hypothesis that molecular pathological findings associated with Alzheimer's disease overlap in the lens and brain.

Methods: We obtained postmortem specimens of eyes and brain from nine individuals with Alzheimer's disease and eight controls without the disorder, and samples of primary aqueous humour from three people without the disorder who were undergoing cataract surgery. Dissected lenses were analysed by slit-lamp stereophotomicroscopy, western blot, tryptic-digest/mass spectrometry electrospray ionisation, and anti-Aβ surface-enhanced laser desorption ionisation (SELDI) mass spectrometry, immunohistochemistry, and immunogold electron microscopy. Aqueous humour was analysed by anti-Aβ SELDI mass spectrometry. We did binding and aggregation studies to investigate Aβ-lens protein interactions.

Findings: We identified Aβ—140 and Aβ1—42 in lenses from people with and without Alzheimer's disease at concentrations comparable with brain, and Aβ1—40 in primary aqueous humour at concentrations comparable with cerebrospinal fluid. Aβ accumulated in lenses from individuals with Alzheimer's disease as electron-dense deposits located exclusively in the cytoplasm of supranuclear/deep cortical lens fibre cells (n=4). We consistently saw equatorial supranuclear cataracts in lenses from people with Alzheimer's disease (n=9) but not in controls (n=8). These supranuclear cataracts colocalised with enhanced Aβ immunoreactivity and birefringent Congo Red staining. Synthetic Aβ bound αB-crystallin, an abundant cytosolic lens protein. Aβ promoted lens protein aggregation that showed protofibrils, birefringent Congo Red staining, and Aβ/αB-crystallin coimmunoreactivity.

Interpretation: Aβ is present in the cytosol of lens fibre cells of people with Alzheimer's disease. Lens Aβ might promote regionally-specific lens protein aggregation, extracerebral amyloid formation, and supranuclear cataracts.

The kynurenine pathway catabolite 3-hydroxykynurenine (3HK) and redox-active metals such as copper and iron are implicated in cataractogenesis. Here we investigate the reaction of kynurenine pathway catabolites with copper and iron, as well as interactions with the major lenticular structural proteins, the α-crystallins. The o-aminophenol kynurenine catabolites 3HK and 3-hydroxyanthranilic acid (3HAA) reduced Cu(II)>Fe(III) to Cu(I) and Fe(II), respectively, whereas quinolinic acid and the nonphenolic kynurenine catabolites kynurenine and anthranilic acid did not reduce either metal. Both 3HK and 3HAA generated superoxide and hydrogen peroxide in a copper-dependent manner. In addition, 3HK and 3HAA fostered copper-dependent R-crystallin cross-linking. 3HK- or 3HAA-modifed R-crystallin showed enhanced redox activity in comparison to unmodified R-crystallin or ascorbate-modified α-crystallin. These data support the possibility that 3HK and 3HAA may be cofactors in the oxidative damage of proteins, such as α-crystallin, through interactions with redox-active metals and especially copper. These findings may have relevance for understanding cataractogenesis and other degenerative conditions in which the kynurenine pathway is activated.