摘要:

A lens epithelial (LE) cell cDNA clone, designatedClone 156, was isolated from a mature rat LE cDNA library by methods of subtractive hybridization. The cDNA sequence ofClone 156was 521 nucleotides in length, excluding the poly(T)-tail, and it encoded an open reading frame of 115 amino acids. The translated protein shared extensive sequence similarities with macrophage migration inhibitory factor (MIF) from mouse lens and human T-cell lymphocytes. Northern blot hybridization showed that rat lens MIF mRNA is about 500 nucleotides in length and that its expression in mature rat lens is relatively low in comparison with that in other rat tissues. The expression of MIF mRNA in LE of normal rats and of rats treated by feeding a diet of 50% (w/w) galactose was studied by quantitative RT-PCR. The results showed that the expression of MIF mRNA in a 20-day galactosemic rat LE increased twelvefold as compared to that found in control LE. From the results of this study and from what we know about the locale of epithelial cell differentiation in mature rat lenses, it is being proposed that the increase in abundance of MIF mRNA in the cataractous rat lens is correlated with the enhanced proliferation of the undifferentiated epithelial cells.

ISSN:0271-3683    

DOI:10.3109/02713689608997404

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

The concentrations of hyaluronan (HA) and chondroitin sulfate (CS) in rabbit tears were determined using high performance liquid chromatography. The tears were taken from both normal eyes and eyes with a corneal epithelial defect. The concentration of CS was 7.61 times higher than that of HA in the tears of normal eyes. When the concentration of CS isomers was compared, the highest concentration was observed in chondroitin 6-sulfate (C6S), followed by chondroitin 4-sulfate (C4S) and chondroitin (COS), in that order. When the corneal epithelium was removed from the left eye using a trephine and microscissors, a significant increase was detected in the concentrations of HA and C6S in tears of the left eye one day after removal, whereas no change was observed in those of the right eye. In addition, a significant positive correlation was observed between the increased amount of HA concentration and the healing rate of the corneal epithelium. These results suggest that the increase of HA concentration in tears is associated with spontaneous corneal epithelial healing.

ISSN:0271-3683    

DOI:10.3109/02713689608997405

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

The effects of natriuretic peptides on cGMP formation and [125I]ANP binding in human trabecular meshwork cells were investigated. CNP at 1μM stimulated cGMP formation∼18–25 fold, with a half maximal effective concentration∼20–30nM. BNP at 1μM stimulated∼7 fold, while ANP stimulated cGMP formation 2-fold at 1μM but had little or no effect at concentrations below 1μM. Displacement binding of [125I]ANP to intact TM cells in the presence of unlabeled ANP indicated a single binding site with a dissociation constant∼0.15nM. c-ANP, which binds specifically to natriuretic peptide C receptors, displaced>95% [125I]ANP binding to surface receptor sites with a half-maximal effective concentration comparable to that of ANP or BNP. c-ANP had no inhibitory effect on CNP stimulation of cGMP formation. The data suggest that human TM cells possess natriuretic peptide B receptors as the primary guanylyl cyclase-containing subtype and C receptors as the numerically predominant subtype of natriuretic peptide receptors.

ISSN:0271-3683    

DOI:10.3109/02713689608997406

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

Visual field test and optic disc evaluation are the standard examination techniques used to detect the onset and progression of glaucoma. This explorative study was performed to assess the temporal correlation between visual field and optic disc changes in eyes with ocular hypertension and well-established glaucoma. Eighty-six hypertensive and 16 glaucomatous eyes were followed up for a period of up to 9 years (average 4.4 yrs) using kinetic and computerized static perimetry and optic disc manual morphometry. Perimetric changes were based on a series of strict criteria and optic disc changes were based as a reduction in the baseline rim area/disc area ratio (R/D) measurement exceeding the 99% confidence interval for intraobserver reproducibility (7.7%). Optic disc changes were found prior to visual field changes in four hypertensive eyes, whereas visual field changes were found prior to disc changes in six glaucomatous eyes (p = 0.042). The results of our explorative study suggest that quantitative optic disc analysis may be more sensitive than visual field examination in detecting early glaucomatous changes, whereas visual field examination may be more sensitive than quantitative optic disc analysis in detecting glaucomatous progressions in eyes with well established glaucoma.

ISSN:0271-3683    

DOI:10.3109/02713689608997407

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

The numbers ofAcanthamoebabinding to new hydrogel contact lenses of different polymer and water content were determined with two quantitative methods, a radiolabeled-cell method and a detaching-fluid method. Numbers of amoebae retained on nonionic lenses increased with increasing water content of the lenses. With both nonionic and ionic lenses numbers of associated amoebae decreased with successive rinsing steps. The retentions of amoebae on unworn hydrogel lenses, in contrast to the irreversible adhesion of bacteria, were tenuous and appeared to be effected mainly by surface tension, surface charge and water content.

ISSN:0271-3683    

DOI:10.3109/02713689608997408

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

Ca2+/calmodulin-and cAMP-dependent protein kinase activities were characterized in two subcellular membrane samples. Membranes from rat lacrimal gland were isolated by differential and density gradient centrifugation into six density windows. The present study focused on membranes from density windows III and V which contain mixtures of apical, Golgi, endosomal, and endoplasmic reticulum membranes in different proportions. Phosphorylation of membrane proteins was measured by incubating the samples in [g-32P]ATP and separating the proteins by discontinuous SDS-PAGE followed by autoradiography. The amount of phosphate incorporated into specific peptide bands was quantified by densitometry. Ca2+/calmodulin-dependent protein kinase phosphorylated a 52,000 MW peptide in membranes from both density windows with a maximal increase from 0.3 to 66μM free Ca2+. Trifluoperazine and promethazine, two inhibitors of Ca2+/calmodulin-dependent protein kinases, inhibited this phosphorylation. cAMP-dependent protein kinase phosphorylated a 22,000 MW peptide and a 91,000 MW peptide which were present in membranes from density window III only. We conclude that a Ca2+/calmodulin-dependent protein kinase activity is present in membranes from both density window III and V whereas a cAMP-dependent protein kinase activity is present only in membranes from density window III.

ISSN:0271-3683    

DOI:10.3109/02713689608997409

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

This study reports on the combined use of an aldehyde fixable, cell viability fluoroprobe, 5-chloromethylfluorescein diacetate (CMFDA), confocal laser scanning microscopy and digital image reconstruction, to produce high resolution images of corneal keratocyte preparationsin situ.The central region of freshly enucleated porcine corneae were removed and stained overnight at 4°C with CMFDA. The tissue was washed, fixed, and frozen for cryosectioning in either a horizontal or antero-posterior orientation. Sections from anterior, central and posterior stroma were examined with a confocal microscope, and the digital images rendered as three-dimensional stereo reconstructions. Fluorescent CMFDA which completely permeated the cell bodies and extremes of the finest ramifying cell processes of all keratocytes provided exceptional high resolution images of the three morphologically distinct cell subpopulations at different levels of the stroma, and enabled improved characterisation of each cell type. Anteriorly was a thin, dense, non-lamellar network of keratocytes subjacent Bowman's membrane. In the central stroma, keratocytes were arranged in layers, the cell bodies had a flattened pyramidal or stellate shape, and the fine cell processes formed extensive distal ramifications. Immediately anterior to Descemet's membrane a small subpopulation of keratocytes with large cell bodies and short branched processes was identified. Extensive and diverse cell-to-cell contacts were orientated in all stromal planes, including ramping cell bridges between keratocyte lamellae in the central stroma. The use of the cell viability dye CMFDA is feasible and valuable for enhancing the visibility of entire keratocyte population in the intact cornea. Diverse multidirectional cell processes and intercellular contacts throughout the keratocyte network suggest a strong capacity for direct communication and cohesion in the maintenance and repair of the stromal matrix. Keratocytes closely related to the epithelium and endothelium have unique morphologies which may relate to specialised functions of these interface cells.

ISSN:0271-3683    

DOI:10.3109/02713689608997410

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

The temporal and spatial expression of vascular endothelial cell growth factor (VEGF) mRNA was studied in normal developing cat retina, and in oxygen induced retinopathy. Unexposed control and oxygen-exposed animals (80 h of 80% oxygen from day 3, n = 16) were studied at 1, 2, 4, and 6 weeks after birth. India ink injected retinal flat mounts were used to study vessel progression, andin situhybridizations using retinal cross sections were used to assess VEGF mRNA accumulation.In controls, as the retina matured, VEGF mRNA hybridization was evident in the ganglion cell layer in a scattered line of distinct cells prior to the ingrowth of vessels, involved the most cells in regions just peripheral to invading vessels and persisted in a fewer positive cells, widely spaced in the vascularized retinas of control, six week animals. In the inner nuclear layer, hybridization initially appeared diffusely and later became localized to a narrow portion of that layer and persisted there. In animals with oxygen induced retinopathy, a substantial increase in hybridization was observed in both the ganglion cell and inner nuclear layers of the avascular retina anterior to the advancing neovascularization. VEGF hybridization decreased abruptly to background levels in both layers at the point were neovascularization met avascular retina. By six weeks, when the neovascularization reached the ora, there was a return of VEGF mRNA in the inner nuclear layer which was similar to normal control expression. A low level of unchanging expression was also observed in the retinal pigment epithelium in both groups at all ages. These results indicate that VEGF mRNA abundance is regulated during retinal vascularization and is increased in relation to oxygen induced neovascularization, suggesting that VEGF may play an important role in both normal retinal vessel development and in the pathophysiology of retinopathy of prematurity.

ISSN:0271-3683    

DOI:10.3109/02713689608997411

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

To define the characteristics of the diurnal variation of intraocular pressure (IOP) in eyes of awake rats, ten male brown Norway rats were entrained to a 12-hour light: 12-hour dark (12L:12D) lighting schedule and were conditioned to IOP measurement with the TonoPen XL tonometer while awake, using only 0.5% proparacaine HC1 anesthesia. The IOP measurements were performed in 4 experiments:Preliminary-IOP was measured at 6-hour intervals in both eyes of each animal, to determine correlation between right and left eyes;Light:Dark-lightingremained the same as in the preliminary experiment, but the measurement schedule was altered so that measurements were obtained at 4-hour intervals in alternating eyes, over two 24-hour light cycles;Dark:Dark-animalswere placed in constant dark (0L:24D) and, after 72 h, measurements were obtained at 4-hour intervals in alternating eyes. Animals were then re-entrained to the previous 12L:12D schedule for 7 days, after which they were returned to constant dark and the experiment was repeated; andDark:Light-animalswere entrained to a reversed light:dark cycle (12D:12L) for 28 days, after which measurements were obtained in the same fashion as in the Light:Dark experiment. Close agreement was found between right-and left-eye IOPs. Animals on a 12L:12D schedule exhibited lowest IOP while the lights were on (19.3±1.9 mm Hg), and highest (31.3±1.3 mm Hg) while the lights were off. Pressure changes anticipated the change from light to dark and dark to light. This pattern persisted in constant dark, and was reversed when the cycle was changed to 12D:12L. Brown Norway rats possess a regular rhythm of IOP that is entrained by the cycle of light and dark, and persistence of this rhythm in constant dark establishes it as a circadian rhythm. Furthermore, our results indicate that reliable and physiologically meaningful IOP measurements can be obtained in awake rats using the TonoPen XL tonometer.

ISSN:0271-3683    

DOI:10.3109/02713689608997412

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor

摘要:

Following thorough removal of adhering aqueous humor, we have succeeded in patch clamping the intact rabbit ciliary epithelium in the cell-attached and inside-out excised-patch modes. Rapidly fluctuating currents (‘chatter activity’) were observed during recordings conducted for as long as 1 h. Chatter activity did not reflect seal instability since interconversion was noted between chatter activity and transitions between stable open and closed states, excision of patches into the bath was associated with substantial shifts in the reversal potential, and chatter activity could be triggered by sustained hyperpolarization, but was insensitive to stretch. The chatter channel was identified as cation-nonselective from the reversal potentials both in the cell-attached and excised-patch modes. The channel's kinetics were similar to those of the cGMP-activated phototransduction channel. The results of PCR amplifications of fragments in cDNA libraries from both human ciliary body and human nonpigmented ciliary epithelial (NPE) cells indicated that human ciliary epithelial cells transcribe message for the retinal phototransduction channel. The possible role of the phototransduction channel in expressing chatter activity was further explored by perfusing preparations with a known activator of that channel (cGMP) and with a known inhibitor (L-cis-diltiazem). Neither agent significantly affected chatter behavior. We conclude that: (1) this is the first demonstration of the feasibility of patch-clamping the intact ciliary epithelium; (2) the NPE cells display chatter activity arising from rapidly fluctuating transitions of a cation-nonselective channel; (3) NPE cells can transcribe message for the cation-nonsclective phototransduction channel; and (4) if the observed chatter activity is from a homologue of the phototransduction channel, the homologue is pharmacologically distinct.

ISSN:0271-3683    

DOI:10.3109/02713689608997413

出版商:Taylor&Francis    

年代:1996

数据来源: Taylor