Abstract：
:The suitability of the rat derived SV-40T immortalized RPE-J cell line for identifying proteome changes associated with RPE differentiation was evaluated by surveying changes in protein expression levels. Rat RPE-J cells were induced to undergo differentiation in culture by growth at the nonpermissive temperature of 40 degrees C in the presence of retinoic acid. Total proteins were extracted from cells grown under proliferating or differentiating conditions and separated by 1D and 2D gel electrophoresis. Gel spots were excised, digested in situ with trypsin, and analysed by mass spectrometry to identify proteins. Computer assisted image analysis was used to align gel patterns and quantify spot intensities. Neither proliferating nor differentiating RPE-J cell cultures exhibited detectable levels of cellular retinaldehyde-binding protein, RPE65, 11- cis -retinol dehydrogenase or lecithin retinol acyl transferase, suggesting that RPE-J cells are not appropriate for visual cycle studies. About 18% of the 61 identified proteins appear to change expression levels with the cell growth conditions. Seven proteins appeared to be up-regulated and four proteins down-regulated when the cells were changed from proliferating to differentiating culture conditions. The majority of the apparent changes in protein expression levels were associated with stress response genes. Significant changes in the apparent mass and charge properties of proteins were also observed and for select proteins, the modifications appeared to be correlated with cell growth conditions. The results demonstrate that proteome differences in RPE-J cells associated with growth conditions can be identified and support the suitability of RPE-J cells for more targeted and/or more global proteome analysis of RPE differentiation.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
West KA,Yan L,Miyagi M,Crabb JS,Marmorstein AD,Marmorstein L,Crabb JWdoi
10.1006/exer.2001.1058keywords:
subject
Has Abstractpub_date
2001-10-01 00:00:00pages
479-91issue
4eissn
0014-4835issn
1096-0007pii
S0014-4835(01)91058-9journal_volume
73pub_type
杂志文章abstract：:Human, monkey, and bovine retinal pigment epithelial (RPE) cells exhibit an M-type K+ current, which in many other cell types is mediated by channels composed of KCNQ α-subunits and KCNE auxiliary subunits. Recently, we demonstrated the expression of KCNQ1, KCNQ4, and KCNQ5 in the monkey RPE. Here, we investigated the...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：
更新日期：2013-11-01 00:00:00
abstract：:Although the presence of 'cartilage-specific' collagens in the eye has been documented earlier, very little is known about their synthesis rates during ocular development, growth and aging. The purpose of the present study was to follow changes in the mRNA levels and distribution of key components of the extracellular...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.2000.0972
更新日期：2001-04-01 00:00:00
abstract：:Neovascular age-related macular degeneration (AMD) is a leading cause of vision loss worldwide. Although intravitreal injection of anti-VEGF antibodies and VEGF Trap have significant clinical benefits, the complications of intravitreal injection, drug resistance and patient compliance still need to be concerned. In th...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2017.12.009
更新日期：2018-03-01 00:00:00
abstract：:A duplex fluorescence assay to assess the viability of cells cultured in multi-well plates is described, which can be carried out in the original culture plate using a plate reader, without exchanges of culture or assay medium, or transfer of cells or cell supernatant. The method uses freshly prepared reagents and doe...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2017.05.011
更新日期：2017-08-01 00:00:00
abstract：:The aim of this study was to develop consistently focal elevated choroidal masses of human choroidal melanoma in immunosuppressed rabbits and to correlate the visualization of prognostically significant microcirculation patterns from confocal indocyanine green angiography with histologic microcirculation patterns. A h...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1998.0650
更新日期：1999-06-01 00:00:00
abstract：:cDNA clones encoding S-antigen (arrestin) were isolated from the expression library constructed from porcine retina and sequenced. The 1490 base pair fragment contained a 1215 base pair open reading frame. From the nucleotide sequence, an amino acid sequence consisting of 405 residues was deduced and a molecular mass ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1996.0036
更新日期：1996-03-01 00:00:00
abstract：:Evidence is presented that alpha-crystallin isolated at 37 degrees C exists as a species, alpha m, which has a minimum molecular weight of about 320000 and a sedimentation coefficient of about 12 S. The amino acid composition, subunit distribution, near- and far-UV CD spectra and immunochemical properties were identic...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(83)90173-2
更新日期：1983-10-01 00:00:00
abstract：:The purpose of this research was to study the early histologic changes occurring in the lens during the formation of selenite overdose cataract. Fourteen-day-old rat pups received a single injection of 2.25 mg Se (kg body wt)-1. Lenses at each of three stages of cataract formation were observed biomicroscopically, and...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(85)90077-6
更新日期：1985-04-01 00:00:00
abstract：:Tear samples were collected from 46 healthy volunteers evenly distributed according to sex and age (mean age 43.5 years). Samples were denatured in a Tris-HCl sample buffer containing 2-mercaptoethanol and SDS, and applied to a gradient SDS-polyacrylamide gel for electrophoresis. The proteinaceous material was transfe...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(91)90160-g
更新日期：1991-10-01 00:00:00
abstract：:The autonomic innervation in the anterior chamber (AC) structures might play an efferent role in neural intraocular pressure (IOP) regulation, the center of which is thought to be located in the hypothalamus. In this study, we identified the efferent pathway from the hypothalamus to the autonomic innervation in the AC...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2020.108367
更新日期：2021-01-01 00:00:00
abstract：:Our previous gene expression analysis suggested that conjunctival epithelial cells of Sjögren's syndrome (SS) are inclined to hyper-proliferation and keratinisation status. The goal of this study is to elucidate whether such pathological situations really exist in the conjunctival epithelium of SS. Also, involvement o...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2005.07.009
更新日期：2006-03-01 00:00:00
abstract：:Our study aimed to investigate metabolites alterations in the blood plasma of central serous chorioretinopathy (CSC) patients and to identify the key biomarkers to increase the understanding of the mechanism of CSC at the molecular level. Quantitative and targeted metabolomics using liquid chromatography tandem-mass s...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2020.108401
更新日期：2020-12-14 00:00:00
abstract：:We aim to determine whether lymphatic drainage from the eye changes with age. Using quantitative photoacoustic tomography, groups of young and older mice were studied in the live state. 10 CD-1 mice of 2-3 months (5M/5F) were studied in addition to 13 older mice of 12-13 months (6M/7F). In each of 23 mice, near-infrar...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2020.108029
更新日期：2020-05-01 00:00:00
abstract：:This study sought to pharmacologically characterize bradykinin receptors on SV40-immortalized human trabecular meshwork (HTM3) cells. Phosphoinositide (PI) turnover studies were conducted using [3H]myo-inositol-labeled HTM3 cells and anion exchange chromatography to quantify [3H]inositol phosphates generated in respon...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1996.0157
更新日期：1996-12-01 00:00:00
abstract：:Utilizing a human beta-actin promoter, a catalase cDNA expression vector was constructed. This construct was used to transfect two immortal cell lines, mouse alpha TN4-1 and rabbit N/N 1003A. The catalase activity was increased about 3.4 fold in the alpha TN4-1 cells and 38 fold in the N/N 1003A cells. Some changes in...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1998.0560
更新日期：1998-12-01 00:00:00
abstract：:Oxidative stress and inflammation are important pathological mechanisms in many neurodegenerative diseases, including age-related macular degeneration (AMD). The very low-density lipoprotein receptor knockout mouse (Vldlr-/-) has been identified as a model for AMD and in particular for retinal angiomatous proliferatio...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2013.08.003
更新日期：2013-11-01 00:00:00
abstract：:The mechanisms by which sulfhydryl (SH-) binding reagents modulate the light-sensitive conductance of retinal rods were investigated by current recording from single rods, by patch clamp recording from the plasma membrane of the rod outer segment (ROS), and by biochemical study of their effects on the light-induced hy...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(90)90176-u
更新日期：1990-07-01 00:00:00
abstract：:By methods of subtraction-hybridization of lambda ZAP cDNA libraries, prepared from 4-week-old rat lens epithelial cells (capsule) and lens fiber cells (decapsulated lens), we have isolated a specific cDNA clone whose target mRNA is about 600 b long. Northern blot hybridization analysis data showed that the target mRN...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(05)80009-0
更新日期：1995-06-01 00:00:00
abstract：:The rd (retinal degeneration) chick possesses an autosomal recessive mutation which results in behavioral and electrophysiological blindness at hatch. Using the technique of two-dimensional gel electrophoresis, we have identified two groups of proteins whose expression in the retina/pigment epithelium/choroid (RET/PE/...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/s0014-4835(89)80061-2
更新日期：1989-10-01 00:00:00
abstract：:Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects of this humanized ...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2014.08.005
更新日期：2014-10-01 00:00:00
abstract：:Lens epithelial cells undergo a dramatic transformation during the process of differentiation into elongated fiber cells. The membrane-associated actin cytoskeleton is likely to play a critical role in the stabilization and maintenance of the highly elongated fiber cell shape. Tropomodulin is a tropomyosin-binding pro...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1996.0111
更新日期：1996-08-01 00:00:00
abstract：:Human retinal pigmented epithelial (RPE) cells cultured on plastic, unlike RPE in situ, often fail to density arrest, do not produce melanin, and do not express mRNA for cellular retinaldehyde binding protein (CRALBP). When human RPE are cultured on the extracellular matrix produced by bovine corneal endothelial cells...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1993.1158
更新日期：1993-11-01 00:00:00
abstract：:The results reported here clearly document changes in the properties and the organization of fiber-cell membrane lipids that occur with age, based on electron paramagnetic resonance (EPR) analysis of lens membranes of clear lenses from donors of age groups from 0 to 20, 21 to 40, and 61 to 80 years. The physical prope...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2015.01.018
更新日期：2015-03-01 00:00:00
abstract：:The purpose of these experiments was to develop a hypothesis to explain activation of m-calpain in cataractogenesis observed in rodents. The in vitro model used to study m-calpain activation was to correlate breakdown of the 'reporter' protein alpha-crystallin with the appearance of activated m-calpain using protein s...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1996.0234
更新日期：1997-04-01 00:00:00
abstract：:In chickens, transient changes in choroidal thickness are found in conditions in which the eye is slowing its growth in response to visual episodes that prevent excessive elongation. To test the hypothesis that the choroidal and ocular growth responses are linked, we used a variety of "brief daily" stimuli known to am...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2007.01.017
更新日期：2007-05-01 00:00:00
abstract：:The P23H-1 rat strain carries a rhodopsin mutation frequently found in retinitis pigmentosa patients. We investigated the progressive degeneration of the inner retina in this strain, focussing on retinal ganglion cells (RGCs) fate. Our data show that photoreceptor death commences in the ventral retina, spreading to th...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2010.10.003
更新日期：2010-12-01 00:00:00
abstract：:In order to determine whether neuronal cell death in the retina accompanies senescence, retinas from pigmented ACI rats of various ages were analysed for changes in dimensions, cell densities, DNA content, and rhodopsin levels. Between the ages of 4 and 30 months, there were significant declines in the densities of nu...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/0014-4835(86)90022-9
更新日期：1986-04-01 00:00:00
abstract：:Ciliary ganglionectomy inhibits the development of myopia in chicks (Schmid et al., 1999), but has no effect on the compensatory responses to spectacle lenses (Schmid and Wildsoet, 1996). This study was done to assess the potential influence of the other parasympathetic input to the choroid, the pterygopalatine gangli...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2012.07.002
更新日期：2012-09-01 00:00:00
abstract：:The conjunctival associated lymphoid tissue is considered to be an integral part of the mucosal immune system. Under normal circumstances immune mechanisms in mucosal associated lymphoid tissue of the gut and bronchus can selectively suppress, rather than enhance, immune responsiveness to encountered antigens, inducin...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1006/exer.1997.0297
更新日期：1997-06-01 00:00:00
abstract：:Recently we determined that the Transient Receptor Potential Vanilloid 4 ion channel (TRPV4) has a crucial signaling role in a pathway that regulates various aspects of lens epithelium function. Here, we report on a different TRPV channel, TRPV1, in porcine lens. The presence of TRPV1 in the lens was evident from RT-P...
journal_title：Experimental eye research
pub_type： 杂志文章
doi：10.1016/j.exer.2018.04.006
更新日期：2018-07-01 00:00:00
