L M Hjelmeland's research while affiliated with University of California, Davis and other places
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Publications (98)
Purpose
. To identify retinal pigment epithelium (RPE)/choroid genes and their relevant expression pathways affected by intravitreal injections of dexamethasone and triamcinolone acetonide in mice at clinically relevant time points for patient care.
Methods
. Differential gene expression of over 34,000 well-characterized mouse genes in the RPE/chor...
Supplemental material contains the complete list of the differentially expressed genes identified by Volcano plot at -1.5≥ FC ≥1.5; p≤0.05 (Tables S1-S6), and ANOVA at p≤0.01 and p≤0.05 (Tables S7 and S8).
Previously, two cytosolic antioxidant enzymes, Glutathione S-transferase Mu 1 (GSTM1) and Mu 5 (GSTM5), were reduced in retinas with age-related macular degeneration (AMD). This study compared genomic copy number variations (gCNV) of these two antioxidant enzymes in AMD versus controls.
Genomic copy number (gCN) assays were performed using Taqman G...
To determine the localization of complement factor H (Cfh) mRNA and its protein in the mouse outer retina.
Quantitative real-time PCR (qPCR) was used to determine the expression of Cfh and Cfh-related (Cfhr) transcripts in the RPE/choroid. In situ hybridization (ISH) was performed using the novel RNAscope 2.0 FFPE assay to localize the expression o...
MicroRNA-34a (miR-34a) has been implicated in neurodegeneration. MiR-34a belongs to a signaling network involving p53 and Sirt-1. This network responds to DNA damage with further downstream signals that induce senescence or apoptosis. Our goal was to measure the expression level of miR-34a in the mouse retina and RPE as a function of age.
The age-d...
To identify retinal genes and their relevant expression pathways affected by intravitreal injections of dexamethasone (Dex) and triamcinolone acetonide (TAA) in mice at clinically relevant time points for patient care.
Differential gene expressions of over 34,000 well-characterized mouse genes, in the retinas of 6-week-old C57BL/6J mice, were analy...
Missing heritability in the study of complex disease has been called dark matter. This review enlarges the scope of this term to include epigenetics and stochasticity, nongenetic sources of risk in complex diseases such as age-related macular degeneration.
Quantifying phenotypic variation at the level of protein expression (variegation) within populations of retinal pigment epithelium (RPE) cells may be important in the study of pathologies associated with this variation. The lack of quantitative methods for examining single cells, however, and the variable presence of pigment and/or lipofuscin compl...
Hyperoxia-related retinal degeneration (HRRD) is a model system in the mouse in which elevated oxygen levels are used to induce retinal degeneration. The hypothesis for the present study was that strain differences in HRRD susceptibility are due to allelic variants of one or more genes in the mouse genome whose human orthologues should be important...
Cystatin C is the major inhibitor of the cysteine cathepsins. Polymorphisms in the cystatin C gene have recently been associated with the risk of developing Age-related Macular Degeneration (AMD). Oxidative stress is also thought to play a key role in the pathogenesis of AMD. We surveyed the retinal pigment epithelium (RPE) and choroid of the C57BL...
To determine the transcriptional proximity of retinal pigment epithelium (RPE) cells grown under different culture conditions and native RPE.
ARPE-19 cells were grown under five conditions in 10% CO(2): "subconfluent" in DMEM/F12+10% FBS, "confluent" in serum and serum withdrawn, and "differentiated" for 2.5 months in serum and serum withdrawn medi...
The retinal pigment epithelium (RPE) is a monolayer of cells that appear phenotypically regular, but which exhibit striking cell-cell variability in content of melanin and lipofuscin granules, and in expression of many proteins. This naturally occurring cell heterogeneity likely arises by normal mechanisms regulating gene expression during developm...
We recently used microarray and reverse transcriptase PCR (RT-PCR) analysis to show an upregulation of cathepsin S (CatS) and glutathione peroxidase 3 (GPX3) in the aging mouse RPE/choroid. To evaluate the mRNA distribution and levels in the RPE and choroid, in situ hybridizations were performed.
Eye sections from 2-month-old and 24-month-old C57BL...
Low-dose D-galactose treatment in mice induces accelerated aging due to advanced glycation endproduct (AGEs) formation. The purpose of this study was to identify ultrastructural aging in the retinal pigment epithelium (RPE)-Bruch's membrane-choriocapillaris.
Five-month-old C57Bl6 mice were injected daily with D-galactose or control buffer for 8 wee...
cDNA libraries from the mouse retina have recently been reported, but no well characterized library from the retinal pigment epithelium (RPE) or choroid of the mouse has yet appeared in the literature. To complement these libraries and to provide the first mouse RPE/choroid library, we used freshly dissected tissue from adult C57BL/6J mice to const...
To investigate the hypothesis that there are topographic and age-related changes in the expression of heme oxygenase (HO)-1 and catalase in the RPE.
Cryosections of the macula and periphery of human eyes (n = 18; aged 27-87 years) were subjected to a high-sensitivity digoxigenin (DIG)-labeled cRNA in situ hybridization protocol to determine the exp...
To evaluate the age-related changes in gene expression occurring in the complex of retinal pigmented epithelium, Bruch's membrane, and choroid (RPE/choroid), we examined the gene expression profiles of young adult (2 mo) and old (24 mo) male C57BL/6 mice. cDNA probe sets from individual animals were synthesized using total RNA isolated from the RPE...
Oxidative stress has been studied in the retinal pigmented epithelium (RPE) in vitro but not in vivo. Our purpose, therefore, was to develop an in vivo model of acute oxidative stress in the C57BL/6J mouse. Mice were exposed to > or = 98% oxygen for 0, 2, or 6 h, and amplified total RNA from the RPE/choroid was applied to microarrays examining abou...
To evaluate the expression and subcellular distribution of proton-coupled monocarboxylate transporters (MCTs) in human RPE in vivo and determine whether ARPE-19 cells retain the ability to express and differentially polarize these transporters.
Total RNA was prepared from human donor eyes and from ARPE-19 cell cultures. Expression of MCT transcript...
The expression of splice variants of FGF receptors, which differ in the third Ig domain, was investigated in retinal pigment epithelium (RPE) cells in vitro and in vivo. This region of the protein determines ligand-binding specificity. Additionally, the expression of potential ligands for these receptors was investigated. Expression of FGF receptor...
Oxidative stress plays a key role in aging diseases of the posterior pole of the eye such as age-related macular degeneration. The oxidative stress response of in vitro RPE cells has been studied for a small number of genes. However, a comprehensive transcriptional response has yet to be elucidated. The purpose of this study was to determine if the...
To determine if mild hyperoxia induces senescence in retinal pigment epithelium (RPE) cells in vitro.
RPE340 cells and WI38 cells were grown in 20% oxygen and 40% oxygen until proliferative exhaustion. A combined senescence associated beta galactosidase (SABG) and 5-bromo-2'-deoxyuridine (BrdU) double labelling technique was performed at different...
To investigate how the differentiation of ARPE-19 cells affects the relative expression of the FGFR genes in response to oxidative stress.
After differentiation in vitro, APRE-19 cells were treated with t-butyl hydroperoxide (tBH) or hydrogen peroxide (H2O2) to induce oxidative stress. Viability and reactive oxygen intermediate (ROI) production wer...
To determine an extensive mRNA phenotype of the established RPE cell line ARPE-19 when grown on a matrix modified by advanced glycation end products (AGEs).
Growth Factor Reduced Matrigel (Collaborative Biomedical Products, Bedford, MA) was nonenzymatically glycated with glycolaldehyde. ARPE-19 cells were seeded on both AGE-Matrigel and Matrigel an...
To demonstrate that chronic hyperoxia induces single-stranded breaks in chromosomal telomeres as a measure of oxidative DNA damage in cultured RPE cells.
RPE340 cells were cultured in 40% and 20% (control) O(2). DNA damage was assessed by mean terminal restriction fragment (TRF) length, and the S1 nuclease assay was used to determine the frequency...
The purpose of this study is to demonstrate the effect of culture density on the steady state mRNA levels of fibroblast growth factor-2 (FGF-2) when retinal pigment epithelial (RPE) cells are subjected to oxidative stress in vitro.
Subconfluent and confluent cultures of the established RPE cell line ARPE-19, were treated with increasing concentrati...
Previous studies have shown that insulin-like growth factor-binding protein (IGFBP)-2 is markedly upregulated in senescent RPE cells in vitro, and might therefore be a marker of senescent cells in vivo. This study was conducted to determine whether IGFBP-2 expression in human RPE cells from the macula and periphery varies with age in vivo.
Paraform...
To establish a model of mild and chronic oxidative stress using hyperoxia for retinal pigment epithelial (RPE) cells in vitro.
RPE340 cells and WI38 lung fibroblasts were grown in normal oxygen (20% O2) and hyperoxia (40% O2 or 60% O2). After cell viability was examined, the levels of reactive oxygen intermediates (ROI) by flow cytometry and heme o...
Cryopreservation can alter cellular function under certain conditions. In this report, we demonstrate the induction of cellular senescence after cells have been cryopreserved using a standard protocol. A retinal pigment epithelial cell line frozen at a specific freezing rate and subsequently thawed showed severely impaired proliferation compared to...
Our goals were to produce a functional recombinant RPE retinal G protein-coupled receptor (RGR) opsin for biochemical studies and to test the efficiency of a lentiviral vector for transgene expression of human RGR.
A human RGR cDNA was cloned into a replication-defective lentiviral vector, and recombinant hRGR-Lentivirus was prepared for transducti...
The expression and alternative splicing of the four FGF receptor (FGFR) mRNAs are regulated in a developmental- and tissue-specific fashion. Capability of differentiation in vitro of the retinal pigment epithelial cell line ARPE-19 has been previously demonstrated. In this study, the hypothesis that FGF receptor gene expression and the alternative...
To explore the changes in expression of a set of genes in a single retinal pigment epithelial (RPE) cell line and two fibroblast cell lines as controls under culture conditions previously used for the analysis of senescent gene expression.
A single human RPE cell line, which had previously been characterized using known markers of senescence, and t...
Senescence of human cells has largely been studied as an in vitro phenomenon resulting from replicative exhaustion. The literature contains many studies of retinal pigment epithelium (RPE) cells which document replicative senescence. Several studies by Burke and others illustrate the relationship between donor age and replicative lifespan, the rela...
Preretinal neovascular formations called sea fans develop at the border of non-perfused peripheral retina in sickle cell retinopathy. Angiogenic factors which could contribute to their development, however, have not been examined previously. The objective of this study was to determine immunohistochemically if vascular endothelial growth factor (VE...
To develop an antibody that recognizes a variety of advanced glycation endproduct (AGE) epitopes.
Glycolaldehyde was used to modify bovine serum albumin and HPLC analysis was used to measure pentosidine formation as an indicator of AGE formation. A polyclonal anti-AGE antibody was synthesized by injecting glycolaldehyde-incubated keyhole limpet hem...
To develop a senescence-associated beta-galactosidase histochemistry and bleaching protocol for the primate posterior pole.
Rhesus monkey eyes of different ages were enucleated after death, fixed in 4% paraformaldehyde for up to 16 hours, and cryoprotected using a graded sucrose infiltration technique. Ten-micrometer tissue sections were treated wi...
PURPOSE. To determine whether there is an age-related increase of pentosidine in human Bruch's membranes and to localize pentosidine and carboxymethyllysine (CML), two well-characterized, advanced glycation end products (AGEs) in aged human Bruch's membranes and choroid in vivo. METHODS. Human Bruch's membrane samples were isolated from the retinal...
To determine whether there is an age-related increase of pentosidine in human Bruch's membranes and to localize pentosidine and carboxymethyllysine (CML), two well-characterized, advanced glycation end products (AGEs) in aged human Bruch's membranes and choroid in vivo.
Human Bruch's membrane samples were isolated from the retinal pigment epitheliu...
Senescence of human cells has largely been studied as an in vitro phenomenon resulting from replicative exhaustion. The literature contains many studies of retinal pigment epithelium (RPE) cells which document replicative senescence. Sev- eral studies by Burke and others illustrate the relationship between donor age and replicative lifespan, the re...
To investigate the relation of senescence-related beta-galactosidase activity and telomere shortening to replicative senescence in cultured human retinal pigment epithelial (RPE) cells.
A human RPE cell line was serially passaged until 80% of cells were nondividing in a 72-hour 5-bromo-2'-deoxyuridine (BrdU) labeling study. Early- and late-passage...
To determine the polarity of fibroblast growth factor 5 (FGF5) secretions from retinal pigment epithelium (RPE) cells and to examine the viability and utility of the ARPE-19 cell line as a model for the study of RPE polarity.
Influenza infection and adenovirus-mediated gene transfer were used to deliver and express genes encoding influenza hemagglu...
Differentiated retinal pigment epithelial (RPE) cells in vivo express basal levels of FGF-5, a secreted member of the FGF gene family. RPE cells proliferate in response to pathological events, resulting in a transient increase in FGF-5 gene expression. The goal of this study is to identify cis-acting sequences in the FGF-5 gene promoter which upreg...
This study aimed to histologically localize indocyanine green (ICG) dye in the geriatric primate and human eye and to correlate these findings with clinical ICG angiography.
The study design was a clinicopathologic correlation.
Six eyes of three geriatric monkeys (Maccaca mulatta) with macular drusen, 19 to 29 years of age, housed at the California...
Advanced glycation endproducts have been implicated in a number of diabetic and aging changes. Some of these effects occur in part through induction of cytokines such as platelet-derived growth factor (PDGF), which is expressed by the retinal pigment epithelium (RPE). In this study, cultures of RPE were evaluated for PDGF expression after treatment...
To determine by immunocytochemical analysis of epiretinal membranes whether vascular endothelial growth factor and the fibroblast growth factor FGF-5 are present in patients with proliferative diabetic retinopathy or proliferative vitreoretinopathy.
Human surgical specimens of epiretinal membranes were obtained from 11 eyes with proliferative diabe...
To demonstrate the expression of fibroblast growth factor-5 (FGF-5) by bovine choroidal microvascular endothelial (BCME) cells and to investigate its possible role as an autocrine mitogen in these cells.
Expression of FGF-5 by BCME cells was studied by a combination of Northern and Western blot analyses. Total RNA was isolated from BCME cultures at...
The fibroblast growth factors (FGFs) are a family of 9 heparin binding proteins which have been proposed to play key roles in angiogenesis. Basic FGF (bFGF), acidic FGF (aFGF) and FGF5 have previously been demonstrated to be expressed in the normal retina and RPE. In this study, the expression of FGF5 was explored in choroidal neovascular membranes...
Apolipoprotein E plays a key role in lipoprotein metabolism and is believed to be an important protein in other biologic functions such as wound healing processes of nerve tissue. Because the neurosensory retina is rich in lipids, we analyzed the subretinal fluid from patients with acute and chronic retinal detachments to determine whether apolipop...
FGF-5 is a secreted member of the fibroblast growth factor (FGF) gene family which has been shown to be expressed at basal levels in the retinal pigmented epithelium (RPE) in rive. When RPE cells are stimulated to divide in rive, or cultured in vitro, FGF5 gene expression is dramatically increased. The goal of this project is to identify cis-acting...
Purpose. FGF-5 is a secreted member of the FGF gene family which has been shown to be expressed at basal levels in differentiated cells of the CNS. The goal of this project is to identify cu-acting sequences in the FGF-5 5' flanking region important in the differential regulation of FGF-5 gene expression in proliferating vs differentiated human ARP...
Human recombinant tissue plasminogen activator (rt-PA) fibrinolysis of subretinal haemorrhage with concomitant removal has been shown to reverse the natural history of photoreceptor degeneration in experimental subretinal haemorrhages if evacuated within 7 days. The aim of the study was to determine whether fibrinolysis of subretinal haemorrhage wi...
Mutant cDNAs coding for dominant-negative forms of the fibroblast growth factor receptors 1 (FGFR-1) and 2 (FGFR-2) that lack tyrosine kinase activity were ligated to a 2.2 kb DNA fragment containing the bovine rhodopsin promoter and used to generate transgenic mice. Six independent lines were generated with the FGFR-1 construct, and five were gene...
The retinal pigment epithelium (RPE) plays a critical role in the development and maintenance of adjacent photoreceptors in the vertebrate retina. This study describes the development and characterization of ARPE-19, a spontaneously arising human RPE cell line with normal karyology which forms polarized epithelial monolayers on porous filter suppor...
Purpose. Proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR) remain the two most common intraocular proliferative diseases with devastating outcome. Growth factors are thought to play an important role in the pathogenesis of these diseases. Apart the better known Fibroblast Growth Factor-1 and -2 (FGF-1 and -2), littl...
Purpose. Previously we have shown that bovine choroidal microvascular endothelial cells exhibit in vitro expression of FGF-5 at the mRNA and protein levels. The purpose of this study was to investigate the possible biological roles of FGF-5 for cell types involved in the process of choroidal neovascularization. Methods. The mitogenic activity of co...
Purpose. FGF-5 is a secreted member of the FGF gene family which has been shown to be expressed in the outer retina where it may be involved in trophic support of photoreceptors or pathological neovascularization. The goal of this project is identify cis-acting sequences in the FGF-5 5′ flanking region important in the regulation of cell type-speci...
Purpose. To determine if the human RPE cell line, ARPE-19 (Dunn et al., IOVS 36:S764), exhibits polarized structural and functional features. Methods. ARPE-19 cells were plated on permeable filter supports and examined by scanning and transmission electron microscopy. The distribution of several in vivo markers of polarity was examined using laser...
Research into the mechanisms underlying the development of age-related macular degeneration (AMD), the leading cause of visual loss in the United States and Europe in people over 60 years old, has been limited in part by the lack of animal models for this disease. In the current study, we examined 62 elderly (> or = 20 years old) rhesus macaques (M...
To assess the optimum probe design and treatment parameters for transscleral diode laser retinopexy.
Transscleral diode photocoagulation was performed on the eyes of Dutch-belted rabbits using three different transscleral probes: a straight, 400-microns diameter probe; a prism-tipped 400-microns diameter probe; and a prism-tipped 900-microns diamet...
Age-related macular degeneration (ARMD) may result from events initiated by reactive oxygen species. Blood samples from 18 patients with ARMD and 18 similarly aged controls were analysed for activities of important antioxidants. Blood glutathione reductase activity was lower in patients with ARMD compared with controls (p = 0.035). The activities o...
To determine the distribution of mRNA transcribed from the FGF-5 gene and the distribution of FGF-5 protein in the normal adult retina of the rhesus macaque.
Freshly enucleated globes from rhesus macaque were prepared for Northern blot analysis, in situ hybridization, and immunohistochemical studies. A 350 base pair sequence from the human FGF-5 wa...
It has previously been shown that cell density has a profound effect on the expression of the basic fibroblast growth factor (bFGF) gene by human retinal pigment epithelial (RPE) cells in vitro. This study was designed to examine cellular mechanisms which may be responsible for the density-dependent expression of bFGF. Examination of RPE labeling i...
The MAS proto-oncogene codes for a seven transmembrane protein which has been previously localized to specific regions of the hippocampus and cerebral cortex in the rat central nervous system. Because MAS has biological properties related to the growth and differentiation of cells of neuroectodermal origin, we investigated the distribution of MAS e...
To better determine the surgical window for removing experimental subretinal hemorrhages in the cat model and to compare the histopathologic effect of such removal with the natural history of untreated subretinal hemorrhages.
Twenty-three large experimental subretinal hemorrhages were created with a neodymium:YAG laser focused through a performed r...
Previous studies have reported the expression of acidic fibroblast growth factor (aFGF) by rat, bovine, and human retinal pigment epithelium (RPE) in vivo. To critically examine the expression of aFGF by RPE cells, we studied the density dependence of steady-state levels of mRNA and protein expression in vitro. Northern blot analysis demonstrated 5...
To examine the expression of bFGF in the developing bovine retina.
Fetal bovine eyes at 90, 120, 150, and 180 days gestational age, as well as adult bovine eyes, were immunohistochemically stained for the presence of bFGF. Detailed characterization of the anti-bFGF antibodies by immunoblot and Western blot analysis against pure FGF gene family stan...
In vitro cultures of human retinal pigment epithelial (RPE) cells were used to study the regulation of basic fibroblast growth factor (bFGF) gene expression. Four transcripts of 7.0, 3.7, 2.2, and 1.2 kB are produced from the bFGF gene. Increasing cell density has a profound effect on the expression of the 7.0 kB transcript relative to the 3.7 kB t...
Members of the fibroblast growth factor (FGF) gene family have been proposed to play critical roles in the biology of the outer retina. In this study, in vitro cultures of human retinal pigment epithelial (RPE) cells were surveyed for the expression of FGF gene family members. Northern analysis provided evidence for the expression of the previously...
Basic fibroblast growth factor (bFGF) delays photoreceptor degeneration when injected intraocularly in Royal College of Surgeons (RCS) rats with inherited retinal dystrophy. In the present study, we have determined the localization of endogenous bFGF in retinas of normal and RCS rats during the normal developmental period (postnatal days 0-20) and...
The purpose of this study is to compare photocoagulation with the argon green, krypton red, and diode infrared laser indirect ophthalmoscopes in an experimental setting.
Photocoagulation was performed with each of the laser indirect ophthalmoscopes in a grid pattern within one sector of the same eye of 14 Dutch-belted rabbits. Treatment was perform...
Basic fibroblast growth factor is a polypeptide mitogen with potential biological roles in angiogenesis, differentiation, and the survival of neurons. To study the expression and subcellular distribution of basic fibroblast growth factor in human retinal pigment epithelium in vitro, affinity-purified antipeptide antibodies were generated against a...
The project was carried out in response to the increasing incidence of eye injuries, particularly of the retina and choroid, as a result of accidental and other exposures to medium to high energy lasers in the military environment. This research was directed at developing techniques of surgical intervention in an attempt to prevent visual loss whic...
Neovascularization of the iris (NVI) is one of the most frequently studied intraocular vascular proliferations in animal models. Ectropion uveae has not been a consistent finding in these studies. In this study, a surgical model of ectropion uveae and iris neovascularization was developed that involved lensectomy, vitrectomy, bipolar cautery and tr...
The avian gene 9E3/CEF4 belongs to a group of genes whose products are highly conserved and are homologous to inflammatory mediators. These genes, sometimes referred to as the gro family, are also expressed upon wounding or serum-stimulation of quiescent cells, suggesting that they may be important in aspects of growth and/or wound healing. We have...
Using in situ hybridization and platelet-derived growth factor (PDGF) cDNA probes labeled with horseradish peroxidase, PDGF-A and -B (c-cis proto-oncogene) mRNA transcripts were identified and localized in proliferating cultures. A human retinal pigment epithelial (RPE) cell line and a glial cell line were treated with either transforming growth fa...
Subretinal hemorrhages are associated with progressive degeneration of the outer retina and a corresponding poor visual prognosis. Mixed results have been reported in previous attempts to remove such subretinal hemorrhages. We developed an ultramicrosurgical system that used the control of a stereotactic micromanipulator to direct a micropipette ti...
The feasibility of gamma (gamma)-interferon injection for the treatment of ocular fibrotic conditions was studied in rabbits using recombinant rabbit gamma-interferon and the cell-injection model of tractional detachment. A toxicity study revealed that intravitreal injection of greater than 10(4) units gamma-interferon consistently produced panuvei...
To explore the feasibility of using human recombinant tissue-type plasminogen activator in the management of subretinal hemorrhage, we examined the toxic effects of a commercial preparation of human recombinant tissue-type plasminogen activator in the subretinal space of the holangiotic retina of the cat. Various concentrations of human recombinant...
Basic fibroblast growth factor (FGF) is a potent endothelial cell mitogen that has been proposed to play a role in proliferative diabetic retinopathy and other neovascular processes. Our understanding of the in vivo role of basic FGF in the pathogenesis of these disorders is limited. We studied the immunolocalization of basic FGF in 16 clinical cas...
Subretinal blood within the macula may cause visual loss in a number of macular diseases. The clinical and histopathologic effects of experimental subretinal hemorrhage were evaluated in the cat. Subretinal hemorrhages were produced by creating a focal neurosensory retinal detachment with micropipette techniques, then inserting a needle tip transsc...
Glial cells of the human retina participate in various pathologic processes characterized by cell migration, proliferation, and extracellular matrix production. To study these events in vitro, a procedure was developed to obtain primary cultures of human retinal glial cells. The cultures resulting from the processing of 130 globes contained cells w...
Retinal pigment epithelial (RPE) cells, retinal glial, and fibroblasts, three cell types believed to play a role in the pathogenesis of epiretinal membrane formation, were maintained in vitreous culture to determine the influence of vitreous on their ultrastructure and expression of cytokeratin, glial fibrillary acidic protein (GFAP), vimentin, and...
Introduction: We have demonstrated that Glucose-6-phosphatase (G6Pase) activity is localized to the endoplasmic reticulum and nuclear envelope of Mueller glia in the normal and pathological guinea pig retina. Using a combination of this cytochemical technique and high voltage electron microscopy, the distribution of nuclear pores could be clearly o...
This chapter describes the different circumstances under which detergent removal may be advisable. It also discusses the detergent properties, which may influence the ease with which detergents may be removed. During the course of the purification of a membrane protein, it may be necessary to remove excess detergent at three distinct points in the...
This chapter describes the essential criteria on which a useful choice of detergents may be made. It also discusses the considerations for the selection of solubilization conditions. For an appropriate choice of a detergent for the solubilization of membrane proteins, it is necessary to consider several of their physical properties. In addition to...
Human retinal pigment epithelial cells at confluence were used to condition serum-free Dulbecco's modified Eagle's medium. Conditioned media were exhaustively dialyzed against 0.5 N acetic acid, lyophilized, and subjected to Western blot analysis, using as primary antibody an IgG fraction prepared from goat antiserum directed against human platelet...