title: Ultrastructural characterization and Fourier analysis of fiber cell cytoplasm in the hyperbaric oxygen treated guinea pig lens opacification model.
abstract: The structural characteristics of differentiated fiber cells in control and hyperbaric oxygen (HBO)-treated guinea pig lenses were examined by transmission electron microscopy (TEM). Emphasis was placed on cell damage, membrane integrity, and cytoplasmic texture. Given the faint gross opacities observed in HBO-treated lenses in previous studies, it was hypothesized that subtle but significant morphological differences due to oxidative damage exist between control and treated animals. Experimental animals received either 70 or 85 treatments with HBO (2.5 atm of 100% O(2) for 2.5 hr, 3 times per week for 5-7 months). All specimens were obtained within 24 hr of death. Freshly cut Vibratome lens sections were fixed and processed for low and high-magnification thin-section TEM analysis. Cytoplasmic texture was analyzed using Fourier and autocorrelation image processing techniques. Low-magnification analysis revealed relatively insignificant differences in general appearance between the fiber cells of the inner fetal and embryonic nuclei in control and HBO-treated guinea pigs. Both groups demonstrated cells of similar morphology with equivalent membrane complexity and homogeneous cytoplasmic texture. Evidence of any major cellular damage or extracellular space debris was not obvious. High-magnification analysis of the cytoplasm of the treated lenses exhibited a mild, yet detectable increase in texture compared with controls and was confirmed by Fourier analysis. Cytoplasmic texture increased in complexity with additional treatments. The absence of major cellular damage in the lenses of HBO-treated animals suggests a less conspicuous source of light scattering. The small changes in cytoplasmic organization observed between treated and control animals may entirely account for the increase in nuclear light scattering observed by slit lamp. The results obtained with this guinea pig/HBO model parallel many of the morphological data associated with human nuclear cataracts. The high-angle scattering observed in the lens of the HBO-treated guinea pig may represent the type of cytoplasmic reorganization that occurs with mild oxidation, effectively making it a valuable model for human lens aging.
authors.affiliation: Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7090, USA.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Freel
authors.forename: Kurt O
authors.initials: KO
authors.lastname: Gilliland
authors.forename: Harold E
authors.initials: HE
authors.lastname: Mekeel
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Giblin
authors.forename: M Joseph
authors.initials: MJ
authors.lastname: Costello
mesh_terms: D000818:Animals; D002386:Cataract / Q000209:etiology / Q000473:pathology*; D002462:Cell Membrane / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure*; D004195:Disease Models, Animal; D005583:Fourier Analysis; D006168:Guinea Pigs; D006931:Hyperbaric Oxygenation; D007908:Lens, Crystalline / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D018384:Oxidative Stress; D012542:Scattering, Radiation