Ultrastructural characterization and Fourier analysis of fiber cell cytoplasm in the hyperbaric oxygen treated guinea pig lens opacification model

Freel, Christopher D.; Gilliland, Kurt O.; Mekeel, Harold E.; Giblin, Frank J.; Costello, M. Joseph

doi:10.1016/s0014-4835(03)00004-6

Cited by 32 publications

(21 citation statements)

References 23 publications

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“…Biochemical evidence indicates that highmolecular-weight aggregates exist (Jedziniak et al, 1973;Jedziniak et al, 1975;Spector and Roy, 1978;Zigler, 1994) and that they are potential sources of scattering in the cytoplasm. However, as noted above, they were not found as distinct particles in the current or previous ultrastructural studies (Costello et al, 1992;Freel et al, 2002;Freel et al, 2003;Costello and Johnsen, 2004). The filters used in the calculations ensured that aggregates in the size range most likely encountered in the cytoplasm (20-200 nm) were analyzed.…”

Section: Discussioncontrasting

confidence: 46%

“…Fourier analysis provides us with a powerful tool to analyze variations not readily visualized by eye and has been used successfully in earlier studies to characterize the fluctuations seen in the fiber cell cytoplasm (Freel et al, 2002;Freel et al, 2003;Costello and Johnsen, 2004;Marsili et al, 2004). The preservation of a variety of textures from human and animal lenses suggests that the chemical fixation procedures have stabilized the native protein distributions.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of nuclear fiber cell cytoplasmic texture in advanced cataractous lenses from Indian subjects using Debye–Bueche theory

Metlapally

Costello

Gilliland

et al. 2008

Experimental Eye Research

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Alterations in ultrastructural features of the lens fiber cells lead to scattering and opacity typical of cataracts. The organelle-free cytoplasm of the lens nuclear fiber cell is one such component that contains vital information about the packing and organization of crystallins critical to lens transparency. The current work has extended analysis of the cytoplasmic texture to transparent and advanced cataractous lenses from India and related the extent of texturing to the nuclear scattering observed using the Debye-Bueche theory for inhomogeneous materials. Advanced age-related nuclear cataracts (age-range 38-75 years) and transparent lenses (age-range 48-78 years) were obtained following extracapsular cataract removal or from the eye bank, at the L. V. Prasad Eye Institute. Lens nuclei were Vibratome-sectioned, fixed and prepared for transmission electron microscopy using established techniques. Electron micrographs of the unstained thin sections of the cytoplasm were acquired at 6500X and percent scattering for wavelengths 400-700 nm was calculated using the Debye-Bueche theory. Electron micrographs from comparable areas in an oxidative-damage sensitive (OXYS) rat model and normal rat lenses preserved from an earlier study were used, as they have extremely textured and smooth cytoplasms, respectively. The Debye-Bueche theoretical approach produces plots that vary smoothly with wavelength and are sensitive to spatial fluctuations in density. The central lens fiber cells from advanced cataractous lenses from India and the OXYS rat, representing opaque lens nuclei, produced the greatest texture and scattering. The transparent human lenses from India had a smoother texture and less predicted scattering, similar to early cataracts from previous studies. The normal rat lens had a homogeneous cytoplasm and little scattering. The data indicate that this method allowed easy comparison of small variations in cytoplasmic texture and robustly detected differences between transparent and advanced cataractous human lenses. This may relate directly to the proportion of opacification contributed by the packing of crystallins. The percent scattering calculated using this method may thus be used to generate a range of curves with which to compare and quantify the relative contribution of the packing of crystallins to the loss of transparency and scattering observed.

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Section: Discussioncontrasting

confidence: 46%

Section: Discussionmentioning

confidence: 99%

Analysis of nuclear fiber cell cytoplasmic texture in advanced cataractous lenses from Indian subjects using Debye–Bueche theory

Metlapally

Costello

Gilliland

et al. 2008

Experimental Eye Research

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“…Sections were cut with a vibrotome and ultramicrotome, and staining was performed as described previously. 25 Specimens (70 nm thick sections) were examined using the FEI Philips Tecnai 12 TEM (FEI Company, Hillsboro, OR, USA) at 80 kV. Images were captured using the Gatan 794 digital camera and Digital Micrograph software.…”

Section: Histological Evaluationmentioning

confidence: 99%

Impaired cytoskeletal organization and membrane integrity in lens fibers of a Rho GTPase functional knockout transgenic mouse

Maddala

Pingping

Costello

et al. 2004

Laboratory Investigation

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To investigate the effects of Rho GTPase inactivation on lens fiber cell cytoskeletal and morphological integrity, a transgenic mouse model expressing C3-exoenzyme (a bacterial toxin) in a lens-specific manner was utilized. Cryosections of whole eyes from C3 transgenic mice and littermate controls were stained for F-actin with rhodamine-phalloidin or immunostained for b-catenin, aquaporin-0 or connexin-50, and confocal images were recorded. Lens fiber cell morphology was examined at both light and electron microscopic levels. To investigate the influence of Rho GTPase inactivation on the profiles of gene expression, cDNA libraries generated from transgenic and littermate control mouse lenses were screened by cDNA microarray analysis. In contrast to the wild-type lens, fiber cells of the transgenic lens were grossly swollen and disorganized, with abnormal membrane architecture. Staining of F-actin, b-catenin, aquaporin-0 and connexin-50 was reduced dramatically in the C3 transgenic lens as compared to controls. Western blot analysis and cDNA microarray analysis did not reveal any noticeable decreases in actin, b-catenin and aquaporin-0 protein levels or expression in C3 transgenic lenses, indicating that altered cytoskeletal organization in response to Rho GTPase inactivation might underlie the noted changes in staining for these proteins. Additionally, cDNA microarray analysis of C3 lens revealed altered expression (at least two-fold, compared to littermate controls) of 44 genes. These include genes encoding extracellular matrix and basement membrane proteins, cell survival and apoptotic pathways, and ion and protein transport. These data indicate that disruption of Rho GTPase function in the developing mouse lens results in abnormal cytoskeletal organization, fiber cell interactions, impaired lens fiber cell morphology and altered gene expression of cellular proteins involved in diverse functions. This work reveals that the morphological and cytoskeletal abnormalities triggered upon Rho GTPase inactivation in lens could be one of the important insults associated with cataract formation in C3 transgenic mouse lens.

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“…The marked pathological changes of the lens were not found in patients with age-related cortical cataract [22]. Important pathogenetic differences in age-related cortical and nuclear cataract were repeatedly confirmed by experimental models cataractogenesis [16,23].…”

Section: General Clinical and Morphological Changes In The Capsule Cementioning

confidence: 90%

New Fluorescent-Histochemical and Immuno-Histochemical Aspects of Secondary Cataract Pathogenesis in Humans

Korsakova¹

2017

IJI

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Secondary cataract is the most common postoperative complication, which causes repeated significant reduced visual acuity, even after a successful operation on the lens. Due to significant clinical heterogeneity of secondary cataract of a man a comparative morphological and functional study in the formation of its fibrous and proliferative forms is especially important for identification of pathogenesis foundations of this disease. The used morphological, histochemical and clinical techniques in the conducted study have provided some evidence of the important role of trophic nervous influences on the type formation of secondary cataract in humans. It is important to note that the revealed similarities in changes of bioamine status, immunoreactivity of the epithelium and fibers of the human crystalline lens in different types of age-related cataract (cortical and nuclear) and its postoperative complications (proliferative and fibrous types of secondary cataract, respectively) can serve as an important proof of their common pathogenesis and, consequently, the type of age-related cataract can be offered as an accessible marker for predicting the risk of postoperative secondary cataract of certain type and its early prevention. The results allow for the first time to put forward a new concept of the secondary cataract pathogenesis. The presented in this chapter concept emphasizes the importance of the fundamental problems of medicine -maintaining the stability of tissue differentiation of a living organism in a variety of physiological and pathological conditions, under the influence of neurotrophic factors.

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Ultrastructural characterization and Fourier analysis of fiber cell cytoplasm in the hyperbaric oxygen treated guinea pig lens opacification model

Cited by 32 publications

References 23 publications

Analysis of nuclear fiber cell cytoplasmic texture in advanced cataractous lenses from Indian subjects using Debye–Bueche theory

Analysis of nuclear fiber cell cytoplasmic texture in advanced cataractous lenses from Indian subjects using Debye–Bueche theory

Impaired cytoskeletal organization and membrane integrity in lens fibers of a Rho GTPase functional knockout transgenic mouse

New Fluorescent-Histochemical and Immuno-Histochemical Aspects of Secondary Cataract Pathogenesis in Humans

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