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Biomedical subjects

J R Kuszak

Publications and source records attributed to J R Kuszak.

At least 19 recordsLinked to original sources
PubMed2004

Fibre cell organization in crystalline lenses.

This review qualitatively and quantitatively compares the gross shape and size of lenses from different species as a function of their fibre cell organization. Grossly, all vertebrate lenses are asymmetrical, oblate spheroids with size and spheroidicity that varies considerably between species. Correlative LM and SEM analysis of the basic structural element of lenses, the fibre cell, shows that the average equatorial fibre width and thickness is relatively constant between most species. This indicates that inter-species differences in lens size is primarily a function of fibre number. Comparable analysis demonstrates that lens spheroidicity is due at least in part, to differences in the average anterior and posterior fibre end segment thickness in relation to that at the equator. In addition, the above analysis, supplemented by 3D-CAD reconstructions, illustrates how lifelong lens growth produces crescent fibres, that become arranged in age-related, concentric growth shells overlaid in slightly imprecise register. The reason for the non-exact registering of growth shells is that, while the vast majority of fibres are hexagonal in cross-section, a very small minority are pentagonal in cross-section and of inconstant width and thickness. Hexagonal and pentagonal fibres are required because the increase in the circumference of successive growth shells is frequently less than the widths of hexagonal fibres. Thus, lens growth is likely accomplished by a combination of the addition of successive growth shells containing more fibres, as well as by the addition of growth shells containing equal numbers of fibres that are incrementally wider as a function of radial location. Finally, SEM analysis, supplemented by 3D-CAD reconstructions, highlights the fact that the end-to-end arrangement of fibres within growth shells, the suture patterns, is not identical in all vertebrate lenses. This is important because lens optical quality is directly related to lens suture type and a negative influence of sutures on lens optical quality increases with age and as a result of some ocular surgeries (vitrectomy and trabeculectomy). These facts suggest that future research efforts should be directed at determining the factor or factors that influence or direct the differences in fibre shape, size and organization in branched and unbranched suture lenses.

Animals↗
PubMed2002

A quantitative analysis of sutural contributions to variability in back vertex distance and transmittance in rabbit lenses as a function of development, growth, and age.

PURPOSE: To correlate specific parameters of lens structure (anterior and posterior suture branch length and planar area) with variability in back vertex distance (BVD) and scatter in rabbit lenses as a function of development, growth, and age. METHODS: Lenses from juvenile (n = 9), adult (n = 9), and aged (n = 10) New Zealand White rabbits were utilized in this study. After sacrifice, lens suture patterns were photographed using a stereo surgical dissecting microscope. Within 5 min of sacrifice, average BVD, variability in BVD, and scatter were assessed with a Scantox In Vitro Assay System. Laser beams were passed incrementally along anterior and posterior suture planes through right eye (oculus dexter, OD) lenses, and between suture planes through left eye (oculus sinister, OS) lenses. After fixation, lens axial dimensions and suture branch lengths were assessed and used to create scaled, 3-dimensional computer assisted drawings (3-D CADs) depicting gross lens shape and sutural changes throughout life. RESULTS: Whereas average BVD only increased significantly as a function of growth, variability in BVD only increased significantly as a function of aging. However, anterior sutures exerted a greater influence on variability of BVD than posterior sutures throughout growth and aging. This difference is consistent with anterior suture branches being longer, or extending farther peripherally, than posterior sutures. Scatter was essentially unchanged between juvenile and adult lenses but significantly increased in aged lenses. Notably, posterior sutures effected a greater age-related increase in scatter than anterior sutures. This difference was consistent with the formation of numerous small, posterior subbranches and subplanes later in life. Structural analysis also suggested that asymmetric age-related lens compaction had occurred, predominantly affecting posterior lens dimensions. CONCLUSIONS: Lens sutures significantly influence average BVD throughout development and growth, and variability in BVD throughout aging. In addition, even though the rabbit lenses appeared transparent throughout growth and aging, unequal length and area of anterior vs. posterior suture branches and planes respectively, as well as a greater degree of age-related posterior lens compaction, were factors contributing to increased scatter.

Aging↗
PubMed2002

Suppression of post-vitrectomy lens changes in the rabbit by novel benzopyranyl esters and amides.

This study reports for the first time a therapeutic modality for the suppression of posterior subcapsular cataract (PSC) formation in an animal model (rabbit) of vitrectomy. This therapeutic modality may also have the potential to attenuate/prevent the high incidence of loss of vision due to cataract formation in patients that undergo vitrectomy. Unilateral, partial vitrectomy was performed on 2.5 month old Dutch Belted rabbits with vitreous replaced by either commercially available BSS((R)) or BSS PLUS((R)) (n=16). Alternatively, vitreous was replaced with a proprietary, modified BSS PLUS((R)) irrigating solution containing 1.25 microM AL-8417 (n=12), 5.0 microM AL-12615 (n=5) or 5.0 microM AL-17052 (n=9). Age matched, non-operated rabbits were used as controls (n=16). Lenses were analysed by correlative structural (light, scanning electron microscopic and three-dimensional computer-assisted drawings) and optical (low power helium-neon laser scan) quality analysis 6 months following surgery. Results demonstrate that vitreous replacement with an irrigating solution that contains the ester-linked benzopyran, AL-8417, the amide-linked benzopyran pro-drug, AL-17052, or its active metabolite, AL-12615, prevented abnormal post-vitrectomy lens growth, or fiber formation. Focal length variability (FLV) assessments (sharpness of focus) confirmed the beneficial drug effects detected morphologically, with FLV being essentially equal to that of age-matched, non-surgical controls. In contrast, lenses of animals with vitreous replaced solely with BSS((R)) or BSS PLUS((R)) exhibited significantly higher FLV than both age-matched controls and animals that underwent vitrectomy with drug-containing irrigating solutions. The ability of AL-8417, AL-17052 and its active metabolite, AL-12615, to suppress vitrectomy-induced posterior lens fiber changes appears to reside in their unique pharmacological profile, acting as antioxidant, anti-inflammatory and cytostatic agents.

Animals↗
PubMed2001

Structural evidence of human nuclear fiber compaction as a function of ageing and cataractogenesis.

This study was conducted to quantify structural change associated with human nuclear fiber compaction as a function of ageing and nuclear cataract formation. Normal donor lenses in three age ranges, young (15--25 years), middle-aged (36--46 years) and aged (59--81 years) were compared to each other and to age-related nuclear cataracts (55--81 years) surgically removed by extracapsular extraction. Several structural modifications which occurred as a manifestation of fiber compaction were noted. In the fetal nucleus (FN), the average anterior and posterior fiber angles decreased approximately 20% with age. Additionally, there was a reduction in the thickness of both the anterior and posterior segments of fetal fibers with age. On average, the anterior--posterior (A--P) axis in the embryonic nucleus (EN) decreased 33% with age. The average length of EN fibers decreased significantly (37%) as a function of age. This change in EN fiber length was accomplished by effecting compaction folds along fiber length. By comparison, in nuclear cataracts the anterior and posterior angles of FN fibers were about 12% smaller than comparably aged normal lenses. Similarly, the A--P axis and the length of EN fibers were 13% smaller than age-matched normals. Nuclear fiber compaction in early adulthood was significant and may contribute to the lens hardening and loss of accommodative ability symptomatic of presbyopia. 3D-CAD reconstructions of fiber compaction show how the reduction in the spacing of lateral interdigitations along fiber length causes an increase in the fiber membrane complexity along the A--P axis in relation to fiber cytoplasm as light passes through lenses. These results may explain, at least in part, how an increase in large particle scatter occurs as light is transmitted through fiber membranes, resulting in reduced lens optical quality as a function of age. By extrapolation, the significantly increased compaction of nuclear fibers in age-related nuclear cataracts may be a contributing factor for excessive scatter in nuclear opacification.

Adolescent↗
PubMed2001

The effect of aging on glutathione peroxidase-i knockout mice-resistance of the lens to oxidative stress.

Populations of control, C, and glutathione peroxidase-1 (GPx-1) knockout mice, K, were studied over a period of 2 years. No significant difference was observed between the C and K populations with respect to longevity, vitality, weight, lens biochemistry or morphology based on light and electron microscopy. It was concluded that under normal animal room barrier facilities, GPx-1 is not required. Furthermore, C and K lenses placed in organ culture and observed over a 24 hr period were indistinguishable. Organ cultured C lenses degraded medium H(2)O(2)levels at only a slightly greater rate than K lenses and this did not appear to change with age. However, tertiary butyl hydroperoxide (TBHP) was degraded less effectively by K lenses and this deficiency increased with age. No indication of change in redox non-protein SH (equivalent to GSH) status was observed between C and K whole lenses or epithelial cell fractions. With H(2)O(2)stress, the drop in C and K non-protein SH was comparable and there was little change with age. Examination of the impact of photochemical stress with 1.5 microM riboflavin and 4% O(2)upon choline transport indicated considerable damage with both C and K lenses, but little difference between the two populations until 1 or 2 years of age when the K lenses appear more vulnerable. With TBHP, the detrimental effect on the K lenses is greater and is observed earlier than with photochemical stress suggesting that the K lens membrane function is more susceptible to phospholipid hydroperoxide stress than are C lenses. Light and electron microscopy of the oxidative stressed lenses indicates significant damage which was generally somewhat greater in the K lenses. TBHP was a more potent oxidant than photochemically generated oxidants particularly at the anterior pole. The overall results suggest that under normal conditions, at any age, the lens does not require the presence of GPx-1 but depending on the type of oxidative stress, the enzyme may significantly contribute to its defense and this dependency may increase with age.

Aging↗
PubMed2000

The relationship between rabbit lens optical quality and sutural anatomy after vitrectomy.

This study was conducted first, to characterize structural changes in rabbit lenses after vitrectomy; and second, to assess whether such changes correlate with a quantifiable compromise in optical function. Unilateral, partial vitrectomies were performed on 2.5 month old Dutch Belted rabbits (n = 64). Age matched non-operated rabbits (n = 32) were used as controls. Lenses were analysed by correlative structural (light, scanning electron microscopic and three-dimensional computer-assisted drawings) and optical (low power helium-neon laser scan) analysis at 1.5, 3, 6 and 12 months post-surgery (n = 16 lenses from operated animals and n = 8 lenses from non-operated controls at each time point). Results demonstrate that in rabbits lens growth, or fiber formation, is compromised after vitrectomy. From 1.5 to 12 months after surgery, lenses had progressively more crooked posterior line sutures with sub-branches of increasing size and number in successive growth shells. Quantification of lens optical quality specifically along and/or between these atypical suture branches and sub-branches revealed a significant increase in focal length variability (sharpness of focus) after vitrectomy. A peripheral zone of fibers with abnormal posterior ends was produced surrounding the pre-surgical lens mass. This additional zone of aberrant fibers was associated with a quantifiable degradation in lens optics. Studies on the prevention of post-vitrectomy lens changes in this rabbit model may yield useful information applicable to the human condition.

Animals↗
PubMed2000

Noncontact specular microscopy of human lens epithelium.

PURPOSE: To obtain in vivo specular images of human lens epithelial cells (LECs) from persons with or without age-related cataract (ARC); to identify features that describe individual aspects of these complex images; to develop feature scales to quantify the severity of each feature; and to study the association of these features with LEC count, age, Lens Opacity Classification System III (LOCS III) classifications and microscopic features of lens epithelium in ARC. METHODS: One hundred fifty-two individuals underwent ophthalmic examinations and LOCS III cataract classifications. Specular images of lenses were captured using a modified noncontact corneal specular microscope (SML-2; Konan, Hyogo, Japan). Enhanced images were graded in a masked fashion, and the presence or absence and severity of each of four features in the specular image ("columnar organization," "linear furrows," "puffy clouds," and "black holes") was graded on a four-step scale. The generalized linear model with intraclass correlation was used to ascertain the statistical significance of associations between age, sex, LOCS III grade, cell count, and feature grade. Capsulorrhexis specimens from 29 patients were studied with correlative light and electron microscopy. RESULTS: LEC density declined with age and was inversely correlated with the scalar grade for puffy clouds and for the size and number of black holes. The scalar grade for columnar organization was inversely associated with the severity of posterior subcapsular and nuclear cataracts, which was the only feature associated with the LOCS III grade of ARC. No statistically significant associations were found between average cell count and LOCS III grade. CONCLUSIONS: With the use of the corneal specular microscope excellent in vivo specular images of the LECs were obtained, the features in these images that correlated well with microscopic findings were classified, and cell density in vivo was estimated.

Adult↗
PubMed2000

Effects of experimentally induced ametropia on the morphology and optical quality of the avian crystalline lens.

PURPOSE: To examine the effects of refractive error on avian lens morphology and optical quality. METHODS: Hatchling white leghorn chicks were unilaterally goggled for 7 days with either a form-deprivation goggle (n = 12), a -10 D defocus goggle (n = 12), or a +10 D defocus goggle (n = 12) to induce myopia and hyperopia. Optical quality of lenses (focal length and focal length variability) from treated and contralateral control eyes was assessed using a scanning laser apparatus. Lens morphology was examined by light and electron microscopy. RESULTS: Although the induction of refractive errors did not significantly alter lens size, shape, paraxial focal length, or average focal length, average focal length variability increased. Lenses from eyes goggled with form-deprivation and +10 D defocus goggles demonstrated a twofold increase in average focal length variability, when compared with their contralateral controls. The morphology of the lens is not altered by these experimental manipulations. CONCLUSIONS: This study provides evidence that the refractive development of the chick lens is not independent of the refractive development of the ocular globe and that chick lenticular development is influenced by both genetics and visual experience.

Animals↗
PubMed2000

DeltaFosB-induced cataract.

PURPOSE: The objective of this study was to investigate a possible relationship between posterior subcapsular cataract (PSC) formation and expression of the transcription factor DeltaFosB. METHODS: Western blot analysis was performed on bitransgenic NSE-tTA, TetOp-DeltaFosB, and single-transgenic NSE-tTA control mice to determine the pattern of DeltaFosB expression within the eye. Light and scanning electron microscopy and biochemical analyses were also performed. RESULTS: In mice expressing DeltaFosB, cataract developed that initially appeared to be posterior subcapsular and gradually matured to involve the entire lens. The enlarged posterior ends of developing secondary fibers curved away from the visual axis to form an elevated opaque posterior plaque. As a result, posterior suture formation did not occur. At a later time, the attenuated posterior capsule overlying the plaque ruptured and the lens nucleus subluxated into the vitreous. Retinal damage was also observed but only from postnatal day 65, a time when extensive lens degeneration had already occurred. DeltaFosB expression was observed well before the detection of morphologic change in both the lens and the retina. Within the lens, DeltaFosB expression was found in both the epithelium and fibers. The development of cataracts was a direct consequence of DeltaFosB expression and was not due to the disruption of an endogenous gene by transgene integration since cataracts could be prevented by silencing expression of DeltaFosB by feeding bitransgenic animals doxycycline (Dox). Moreover, cataracts were observed in bitransgenic mice derived from two independent TetOp-DeltaFosB founder lines but not in single NSE-tTA transgenic controls. Cataractogenesis was not a consequence of abnormal development, because mice conceived and raised on Dox to prevent expression of DeltaFosB also were subject to formation of PSC when expression of DeltaFosB was turned on in adult animals by removing Dox. Examination of biochemical parameters indicated that the earliest change observed was the disruption of calcium homeostasis with a significant increase in Ca(2+) influx, followed by a gradual but marked decrease in protein content. Significant changes in certain metabolic parameters and protein composition were also observed. CONCLUSIONS: The DeltaFosB-induced cataract in which the major morphologic early event was the disruption of normal posterior fiber formation, may be a good model for PSC. By identifying DeltaFosB-regulated target genes, it should be possible to achieve a better understanding of the molecular mechanisms through which PSC is formed.

Animals↗
PubMed1999

The internalization of posterior subcapsular cataracts (PSCs) in Royal College of Surgeons (RCS) rats. I. Morphological characterization.

PURPOSE: To document lens ultrastructure during and after internalization of posterior subcapsular cataracts (PSCs) in Royal College of Surgeons (RCS) rats, a model for human autosomal retinal degenerative disease. METHODS: RCS rat lenses at 2, 2.5, 3, 4, 6, 9, 12, and 15 months old were enucleated and fixed. For light and transmission electron microscopy (TEM), lenses were embedded in epoxy and sectioned along the visual axis. For scanning electron microscopy, lenses were dissected to expose the posterior fibers in concentric growth shells down to the internalized PSC plaques. RESULTS: Overgrowth of the plaque began between 8 and 9 weeks postnatal and proceeded from the periphery to the posterior pole. This is in contrast to PSC formation which begins centrally and enlarges radially between 4-6 weeks postnatal. Peripheral-to-central overgrowth resulted in the formation of a convexo-concave, disk-shaped suture plane oriented parallel to the capsule. The initial fibers overlying the plaque were extremely flattened at their posterior ends. However, by 3 months postnatal, fiber ultrastructure was relatively normal and displayed only minor morphological irregularities. These temporal and structural changes were used to create 3-dimensional computer assisted-drawing (3D-CAD) reconstructions and animations. TEM examination of plaques revealed scattered fiber defects such as membrane whorls, globular aggregates and intracellular voids in both the internalized plaques and the initial overgrowth. The internalized PSC plaques had comparable morphology in all animals, regardless of age. Specifically, the posterior segments of fibers were enlarged and curved abnormally toward the capsule. CONCLUSIONS: PSC plaques are not internalized and broken down in the classical cell biological sense (i. e. via lysosomal degradation). Rather the plaques retain their structure indefinitely as lens growth proceeds (albeit not entirely normally). This demonstrates that the lens has a restricted ability to respond to growth defects and effect a limited recovery after PSC formation.

Age Factors↗
PubMed1999

The internalization of posterior subcapsular cataracts (PSCs) in Royal College of Surgeons (RCS) rats. II. The inter-relationship of optical quality and structure as a function of age.

PURPOSE: The Royal College of Surgeons (RCS) rat is an animal model for human retinal degenerative disease and posterior subcapsular cataracts (PSCs). The purpose of this study was to correlate the structure and optical quality of RCS lenses with PSCs as a function of their internalization, with normal, non-cataractous, age-matched control lenses. METHODS: Correlative light (LM), scanning electron microscopic (SEM), three-dimensional computer assisted drawings (3D-CADs) and low power helium-neon laser scan analysis were used to examine the structure and function of lenses. RESULTS: The optical properties (average focal length variability; sharpness of focus) of RCS rat lenses are quantitatively compromised by PSCs. Correlative LM and SEM analysis of RCS lenses at various stages of PSC internalization (1.5, 3, 6, 9, 12 and 15 months of age), revealed that the sutures formed by additional fiber growth were progressively more abnormal. During PSC internalization, two to nine small suture branches were formed and arranged in modified line to multiple y configurations rather than the normal three branch y sutures. These temporal changes were also chronicled in animated 3D-CAD videos derived from lens reconstructions based on LM and SEM micrographs from the selected time points stated above. However, laser scan analysis also revealed that as the PSCs of RCS rat lenses were progressively internalized, there was a steady improvement in total sharpness of focus that reached normal levels by 12 months of age. The correlation of laser scan and structural data from specific regions of lenses revealed the following: 1. The abnormal posterior sutures of RCS rats with internalized PSCs effect a greater reduction in optical quality than normal posterior sutures of age-matched controls; 2. However, the resulting abnormal suture plane area was cumulatively similar to that of age-matched controls; 3. Thus, total optical quality was similar between RCS lenses with internalized PSCs and age-matched controls by 12 months of age. CONCLUSIONS: The results of this study show that RCS lenses with internalized PSCs can appear grossly, and indeed optically perform, at levels comparable to aged lenses. These findings are consistent with clinical observations of spontaneous recovery from PSC. The results suggest that human PSCs that occur as a consequence of retinal degenerative disease could also be the result of abnormal posterior suture growth. If this is proven to be the case, such PSCs may have some capacity for repair or recovery thereby obviating their surgical removal.

Aging↗
PubMed1999

Effect of age on the morphology and optical quality of the avian crystalline lens.

The effect of age on the avian lens was examined using White Leghorn chickens of five age groups: hatchling (n =19), 7 day (n = 15), 34 week (n =10), 2 year (n =24), and 5 year (n =25). The chick lens grows steadily up to 34 weeks of age, after which, the rate of growth slows down. During growth, average focal length of the lens becomes longer. However, no significant changes were noted between 2 and 5 year old lenses. An age related increase in average lenticular focal length variability (FLV) was observed, revealing that the optical quality of the lens decreases with age. Scanning electron microscopy demonstrated that: (1) the suture region of the lens becomes more diffuse and less precise with age; (2) the central regions of younger lenses are oblate spheroids, while older lenses become more apple-shaped; (3) individual lens fibres in the young lens are crescent-shaped, while older lens fibres are square-bracket-like in shape; (4) the edges of individual lens fibres become more jagged and irregular with age; and (5) the layering of lens fibres is more disorderly in older lenses, in contrast to the parallel and organized layering of fibres in young lenses.

Aging↗
PubMed1999

Anterior polar cataracts in CS rats: a predictor of mature cataract formation.

PURPOSE: The objective of this study was to characterize the morphology of the anterior opacities formed during recovery from posterior subcapsular cataract (PSC) in Royal College of Surgeons (RCS) rats. METHODS: Lenses from RCS rats at 8 and 12 weeks postnatal (n = 14 and 12, respectively) were examined under a dissecting microscope for the presence of anterior opacities. Lenses with anterior opacities were fixed, embedded in epoxy resin, and sectioned along the optic axis for light microscopy (LM) and transmission electron microscopy (TEM). RESULTS: At eight weeks postnatal, 21.5% of animals (3/14) had anterior cataracts. Light microscopy of 1- to 2-microm-thick sections revealed an anomalous layer of material located at the epithelium-fiber interface, which was identified as a zone of liquefaction by TEM. Epithelial cells had minor structural defects but were not necrotic. Anterior portions of elongating and cortical fibers under the zone of liquefaction were undisrupted, whereas their posterior portions had numerous vacuoles. The anterior opacities were classified as anterior polar cataracts (APCs) based on the location and type of morphologic damage in the affected lenses. At twelve weeks postnatal, 25% of animals (3/12) had APCs that involved prominent vesiculation of the anterior cortex. Ultrastructural examination showed that large vesicles were located between and inside anterior fibers and that most extracellular spaces were abnormally widened. Posteriorly, internalization of the PSC by new fiber growth was disordered and displayed vesiculation and density variations. In the bow region, LM revealed minor structural irregularities that were identified as groups of apparently degenerating fibers by TEM. CONCLUSIONS: APCs in RCS rats are caused by degeneration of elongating fibers in the bow region and subsequent damage in the superficial anterior cortex. The percentage of animals with APCs (25%) was consistent with the percentage of animals in which mature cataracts eventually develop. The morphologic changes, time of onset, and percentage of animals affected suggest that APC is the initial manifestation of mature cataract formation in RCS rats.

Animals↗
PubMed1999

A transgenic animal model of osmotic cataract. Part 1: over-expression of bovine Na+/myo-inositol cotransporter in lens fibers.

PURPOSE: Intracellular osmotic stress is believed to be linked to the advancement of diabetic cataract. Although the accumulation of organic osmolytes (myo-inositol, sorbitol, taurine) is thought to protect the lens by maintaining osmotic homeostasis, the physiologic implication of osmotic imbalance (i.e., hyperosmotic stress caused by intracellular over-accumulation of organic osmolytes) on diabetic cataract formation is not clearly understood. Studies from this laboratory have identified several osmotic compensatory mechanisms thought to afford the lens epithelium, but not the lens fibers, protection from water stress during intervals of osmotic crisis. This model is founded on the supposition that the fibers of the lens are comparatively more susceptible to damage by osmotic insult than is the lens epithelium. To test this premise, several transgenic mouse lines were developed that over-express the bovine sodium/myo-inositol cotransporter (bSMIT) gene in lens fiber cells. METHODS: Of the several transgenic mouse lines generated, two, MLR14 and MLR21, were analyzed in detail. Transgenic mRNA expression was analyzed in adult and embryonic transgenic mice by a coupled reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization on embryonic tissue sections, respectively. Intralenticular myo-inositol content from individual mouse lenses was quantified by anion exchange chromatography and pulsed electrochemical detection. Ocular histology of embryonic day 15.5 (E15.5) embryos from both transgenic (TG) families was analyzed and compared to their respective nontransgenic (NTG) littermates. RESULTS: Both RT-PCR and in situ hybridization determined that transgene expression was higher in line MLR21 than in line MLR14. Consistent with this, intralenticular myo-inositol from MLR21 TG mice was markedly higher compared with NTG littermates or MLR14 TG mice. Histologic analysis of E15.5 MLR21 TG embryos disclosed a marked swelling in the differentiating fibers of the bow region and subcapsular fibers of the central zone, whereas the lens epithelium appeared morphologically normal. The lenticular changes, initiated early during lens development in TG MLR21 embryos, result in severe bilateral nuclear cataracts readily observable in neonates under normal rearing and dietary conditions. In contrast, TG MLR14 pups reared under standard conditions produced no lens opacity. CONCLUSIONS: Lens fiber swelling and related cataractous outgrowth positively correlated to the degree of lens bSMIT gene expression and intralenticular myo-inositol content. The affected (i.e., swollen) lens fibers appeared to be unable to cope with the water stress generated by the transgene-induced over-accumulation of myo-inositol and, as a result of this inability to osmoregulate, suffered osmotic damage due to water influx.

Animals↗
PubMed1998

The structure of posterior subcapsular cataracts in the Royal College of Surgeons (RCS) rats.

The Royal College of Surgeons (RCS) rat is an animal model for human autosomal recessive retinitis pigmentosa. As the retinas of these animals degenerate from two to six weeks after birth, posterior subcapsular cataracts (PSCs) develop, presumably in response to toxic lipid peroxides formed by degenerating rod outer segments. Morphologically, these PSCs are thought to be characterized by a proliferation of dysplastic bladder-like fibers, or Wedl cells, in the meridional region of the lens, that subsequently migrate to, and aggregate at, the posterior pole as the PSC. This report presents the results of correlative scanning (SEM) and transmission (TEM) electron microscopic as well as light microscopic (LM) analysis of the ultrastructure of RCS PSCs. SEM analysis of two, four and six week old lenses (n=6-10 specimens per age group) demonstrated that the PSCs of RCS rats resulted from a growth malformation of the posterior fiber ends from four to six weeks. The PSC is composed of markedly enlarged and irregular posterior fiber ends aberrantly curved away from the polar axis toward the vitreous rather than overlapping and abutting to form suture branches within and between concentric growth shells. LM analysis revealed evidence of progressively more numerous, enlarged, and irregular, ovate cellular profiles at the posterior pole from four to six weeks. However, there was no evidence of Wedl cells either within the meridional row region or along a migratory path from the equator to the posterior pole at any age. TEM analysis confirmed that the size and abnormal shapes of cellular profiles were consistent with SEM analysis and that nuclei were never observed within the plaque. In addition, there was considerable variation in cytoplasmic densities between cells. Also, dense deposits were frequently noted between cells and beneath the capsule. The orientation of posterior fiber end profiles to the posterior capsule was 45, 70 and 90 degrees at respectively two, four and six weeks of age. These results show that RCS PSCs are a consequence of abnormal posterior fiber end growth culminating in a posterior opacity.

Aging↗
PubMed1998

The effect of photochemical stress upon the lenses of normal and glutathione peroxidase-1 knockout mice.

This communication investigates the effect of oxidative stress upon the lenses of young normal and glutathione peroxidase-1 (GSHPx-1) Knockout mice. Both normal and knockout lenses have similar biochemical and morpholigical characteristics and the elimination of GSHPx-1 only decreases slightly the ability of the lens to degrade H2O2. Examination of the effect of a 4 hr photochemical stress on morphological characteristics indicates that there is comparable damage in the normal and knockout lenses in the epithelial and bow regions while the posterior region remains normal. However, at 24 hrs post-insult, the normal lenses appear to recover somewhat in the bow region while the knockout bow and posterior regions have extensive damage. In contrast to the morphological data, the biochemical parameters (14C)choline transport and (3H)thymidine incorporation are affected to a somewhat greater extent in the knockout lenses than in normal lenses. While both of these parameters are further affected in the 24 hr post-insult period, there is no further change in the relative effects upon normal and knockout lenses. Non-protein thiol is affected in a similar manner in both lens types. The effect upon biochemical parameters of tertiary butyl hydroperoxide (TBHP) insult was similar to H2O2 and photochemical stress. The overall conclusion is that young GSHPx-1 knockout lenses handle oxidative stress somewhat less effectively than comparable normal lenses but non-stressed knockout lenses appear normal. These results differ from observations reported by Reddy et al. (1997) under somewhat different conditions.

Animals↗
PubMed1996

Morphology of the normal human lens.

PURPOSE: To provide a quantitative, morphologic description of differentiated lens fiber cells in all regions of aged normal human lenses. METHODS: Transparent normal human lenses (age range, 44 to 71 years) were examined with correlative transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Vibratome sections allowed examination of internal structures, whereas dissected whole lenses revealed surface characteristics. Additionally, image analysis was used to measure cross-sectional areas of fiber cells. RESULTS: Approximate regional dimensions (percentage of diameter and thickness, respectively) were determined for whole lenses: cortex 16%, 17%; adult nucleus 24%, 21%; juvenile nucleus 12%, 9%; fetal nucleus 45%, 49%; and embryonic nucleus 3%, 4%. Cortical cells were irregularly hexagonal, and the average cross-sectional area measured 24 +/- 9 microns2. Adult nuclear cells were flattened with intricate membranous interdigitations and an area of 7 +/- 2 microns2. Juvenile nuclear cells had an area of 14 +/- 5 microns2. Fetal nuclear cells were rounded with an area of 35 +/- 22 microns2. Embryonic nuclear cells also were rounded and had a variable area of 80 +/- 68 microns2. Fiber cell cytoplasm in all lens regions appeared smooth in texture and homogeneous in staining density. CONCLUSIONS: Both TEM and SEM are necessary to obtain a complete description of fiber cells. Cross-sections of fibers give new insights into the lamellar organization of the lens, indicating that each region has characteristic cell shapes and sizes. Furthermore, average dimensions were used to demonstrate that the number of cells and approximate growth rates vary significantly between adjacent regions.

Adult↗