Sergey L. Znoiko

Adhesion of fibronectin, vitronectin, laminin, and collagen type IV to intraocular lens materials in pseudophakic human autopsy eyes. Part 2 : Explanted intraocular lenses

Purpose: To evaluate fibronectin, vitronectin, laminin, and collagen type IV adhesion to poly(methyl methacrylate) (PMMA), silicone, hydrophobic soft acrylate, and hydrogel intraocular lenses (IOLs) in human pseudophakic autopsy eyes. Setting: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. Methods: Thirty‐two autopsy eyes containing PMMA, silicone, soft acrylate, or hydrogel IOLs were assessed. The IOLs were explanted from the capsular bag, and both sides of the IOLs were immunohistochemically stained for fibronectin, vitronectin, laminin, or collagen type IV. The number of cells on the IOL surfaces was counted. The capsular bag from 1 eye containing a soft acrylate IOL was examined for fibronectin and vitronectin. Results: Hydrophobic soft acrylate IOLs had significantly more fibronectin adhering to their surfaces than PMMA (P < .01) or silicone (P < .01) IOLs, as well as more vitronectin. Silicone IOLs had more collagen type IV adhesion than the other IOLs (P < .05–.06). Collective protein adhesion differed significantly between soft acrylate IOLs and PMMA and silicone IOLs, but not between PMMA and silicone IOLs. Conclusions: The greater amount of protein on the hydrophobic soft acrylate (AcrySof®) IOLs seems to support an adhesive mechanism for their attachment to the capsular bag. Fibronectin and vitronectin have functional domains to bind them to lens epithelial cells and the collagenous capsule. This kind of attachment could be a true bioactive bond and may be 1 reason the PCO and neodymium:YAG capsulotomy rates are lower in eyes with a soft acrylate IOL.

Adhesion of fibronectin, vitronectin, laminin, and collagen type IV to intraocular lens materials in pseudophakic human autopsy eyes: Part 1: histological sections

Purpose: To evaluate fibronectin, vitronectin, laminin, and collagen type IV adhesion to poly(methyl methacrylate) (PMMA), silicone, hydrophobic soft acrylate, and hydrogel intraocular lenses (IOLs) in pseudophakic human autopsy eyes. Setting: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. Methods: Thirty‐eight autopsy eyes containing PMMA, silicone, hydrophobic acrylate, or hydrogel IOLs were assessed. Histological sections were prepared from each eye, and immunohistochemical analyses were performed for fibronectin, vitronectin, laminin, and collagen type IV. One hundred fifty‐two specimens were analyzed. Results: A sandwich‐like structure (anterior or posterior capsule/fibronectin/1 cell layer/fibronectin/IOL surface) was seen in 12 of 14 autopsy eyes with soft acrylate IOLs, 3 of 10 with a PMMA IOL (P = .0094), 1 of 10 with a silicone IOL (P = .0022), and 0 of 4 with a hydrogel IOL (P = .0041). The thicker fibrocellular tissue on the inner surface of the anterior or posterior capsule that was in contact with silicone IOLs was lined with collagen type IV. Vitronectin and laminin were not found at the fibrocellular tissue–IOL interface in any specimen. Conclusions: This study seems to confirm the sandwich theory of posterior capsule opacification in eyes with an IOL and suggests that fibronectin may be the major extracellular protein responsible for the attachment of hydrophobic soft acrylate (AcrySof®) IOLs to the capsular bag. This may represent a true bioactive bond between the IOL and lens epithelial cells or between the IOL and the capsular bag and may be one reason the PCO and neodymium:YAG capsulotomy rates are lower in eyes with a soft acrylate IOL.

A Visual Pigment Expressed in Both Rod and Cone Photoreceptors

Rods and cones contain closely related but distinct G protein-coupled receptors, opsins, which have diverged to meet the differing requirements of night and day vision. Here, we provide evidence for an exception to that rule. Results from immunohistochemistry, spectrophotometry, and single-cell RT-PCR demonstrate that, in the tiger salamander, the green rods and blue-sensitive cones contain the same opsin. In contrast, the two cells express distinct G protein transducin alpha subunits: rod alpha transducin in green rods and cone alpha transducin in blue-sensitive cones. The different transducins do not appear to markedly affect photon sensitivity or response kinetics in the green rod and blue-sensitive cone. This suggests that neither the cell topology or the transducin is sufficient to differentiate the rod and the cone response.

Salamander Blue-sensitive Cones Lost During Metamorphosis

The tiger salamander lives in shallow water with bright light in the aquatic phase, and in dim tunnels or caves in the terrestrial phase. In the aquatic phase, there are five types of photoreceptors—two types of rods and three types of cones. Our previous studies showed that the green rods and blue‐sensitive cones contain the same visual pigment and have the same absorbance spectra; however, the green rods have a larger photon‐catch area and thus have higher light sensitivity than the blue‐sensitive cones. Here we show that after metamorphosis, the terrestrial salamander looses the blue‐sensitive cones, while the density of the green rods increases. Moreover, the size of the green rod outer segments is increased in the terrestrial phase, compared to that in the aquatic phase. This switch from the blue‐sensitive cones to the green rods may represent an adaptation to the dim light environment of the terrestrial phase.

Salamander rods and cones contain distinct transducin alpha subunits

The mammalian retina is known to contain two distinct transducins that interact with their respective rod and cone pigments. However, there are no reports of a nonmammalian species having two distinct transducins. In the present study, we report the cloning and cellular localization of two transducin a subunits (G alpha t) from the tiger salamander. Through degenerate polymerase chain reaction (PCR) and subsequent screening of a salamander retina cDNA library, we have identified two forms of G alpha t. When compared to existing sequences in GenBank, the cloned subunits showed high similarity to rod and cone transducins. The salamander G alpha t-1 has 91.2-93.7% amino acid sequence identity to mammalian rod G alpha t subunits and 79.7-80.9% to mammalian cone Gats. The salamander G alpha t-2 has 86.2-87.9% sequence identity to mammalian cone G alpha ts and 78.9-80.9% to mammalian rod G alpha ts at the amino acid level. The G alpha t-1 cDNA encodes 350 amino acids while the G alpha t-2 cDNA encodes 354 residues, which is typical for rod and cone G alpha ts, respectively, and we thus identified the G alpha t- 1 as rod and G alpha t-2 as cone G alpha t. Sequences identified as effector binding sites and GTPase activity regions are highly conserved between the two subunits. Genomic Southern blot analysis showed that rod and cone G alpha t subunits are both encoded by single-copy genes. Northern blot analysis identified retina-specific transcripts of 3.0 kb for rod G alpha t and 2.6 kb for cone G alpha t. Immunohistochemistry in the flat-mounted salamander retina demonstrated that rod G alpha t is localized to rods, predominantly in the outer segments; similarly, cone G alpha t is localized to cone outer segments. The results confirm that the two sequences encode rod and cone transducins and demonstrate that this lower vertebrate contains two distinct transducins that are localized specifically to rod and cone photoreceptors.