Stability of RNA from the retina and retinal pigment epithelium in a porcine model simulating human eye bank conditions.

Abstract:

PURPOSE:To assess RNA stability after death in a porcine model to simulate current human eye bank techniques. METHODS:Eye bank time interval data were collected from 191 donor specimens: death to refrigeration, enucleation, and tissue processing. A control porcine eye was enucleated, retina and RPE isolated, and specimens frozen (-80 degrees C). Fourteen porcine eyes remained at room temperature for 2 hours and then cooled to 4 degrees C. Retina and RPE were isolated and frozen (-80 degrees C) at 5, 12, 24, 29, 36, 48, and 72 hours. Four globes remained in a moist chamber, five whole and five sectioned globes were immersed in RNAlater (Ambion, Austin, TX) at 5, 12, 24, or 48 hours. RNA was isolated. The 28S and 18S rRNA peaks were analyzed by electrophoresis. RT-PCR was performed on each sample. Messenger RNA for GAPDH, beta-actin, mouse rhodopsin from retina (mRHO), and RPE-65 (from RPE) were analyzed with gel electrophoresis. RESULTS:The average time from death to refrigeration was 4.2 hours, to enucleation 6.4 hours, and to tissue processing 10.7 hours. RT-PCR gel electrophoresis patterns from retinal tissue had bands of similar intensity at each interval from beta-actin, GAPDH, and RHO. Band patterns from RPE demonstrated decay of the RT-PCR gene products after 5 hours. This decay was delayed by at least 24 hours with the use of RNAlater. The 28S rRNA decay was similar for retina and RPE. CONCLUSIONS:Retinal tissue RNA can be analyzed within the time constraints of current eye bank tissue processing, whereas analysis of RPE necessitates either rapid processing or use of RNAlater. These results should aid in future studies in which eye bank tissue is used for RNA analysis.

authors

Malik KJ,Chen CD,Olsen TW

doi

10.1167/iovs.02-1120

keywords:

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

2730-5

issue

6

eissn

0146-0404

issn

1552-5783

journal_volume

44

pub_type

杂志文章
  • Triamcinolone acetonide inhibits IL-6- and VEGF-induced angiogenesis downstream of the IL-6 and VEGF receptors.

    abstract:PURPOSE:To test whether triamcinolone acetonide (TA) inhibits angiogenesis induced by IL-6 or VEGF and whether this inhibition is through antagonism of the IL-6 or the VEGF receptor 2. METHODS:A rat cornea micropocket assay was used to initiate IL-6- and VEGF-mediated angiogenesis. The ability of TA or neutralizing VE...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.05-1651

    authors: Ebrahem Q,Minamoto A,Hoppe G,Anand-Apte B,Sears JE

    更新日期:2006-11-01 00:00:00

  • Cell attachment to, and contraction of, the retina in vitro.

    abstract:PURPOSE:To examine the behavior of fibroblasts and retinal pigment epithelial cells after attachment to the retinal surface in vitro to elucidate the pathobiology of the early stages of epiretinal membrane formation. METHODS:Human retinal pigment epithelial (HRPE) cells and bovine Tenon's capsule fibroblasts (BTFs) we...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Allamby D,Foreman D,Carrington L,McLeod D,Boulton M

    更新日期:1997-09-01 00:00:00

  • Redistribution and reduction of interphotoreceptor retinoid-binding protein during ocular coronavirus infection.

    abstract::Inoculation of the neurotropic coronavirus mouse hepatitis virus strain JHM intravitreally or into the anterior chamber causes acute infection of the retinal pigment epithelium (RPE) and neural retina. Weeks later, many retinas have foci of moderate to severe atrophy. The effect of coronavirus infection (after intravi...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Robbins SG,Wiggert B,Kutty G,Chader GJ,Detrick B,Hooks JJ

    更新日期:1992-01-01 00:00:00

  • Aging of the human retina. Differential loss of neurons and retinal pigment epithelial cells.

    abstract::The impact of aging on cell loss in the human retina was examined in foveal and temporal equatorial regions in eyes from 35 donors with ages spanning a 78-yr period from the second to the ninth decade of life. Equatorial cones and retinal pigment epithelial cells (RPE) decreased at uniform rates from the second to the...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Gao H,Hollyfield JG

    更新日期:1992-01-01 00:00:00

  • Intraocular Pressure and Glaucomatous Optic Neuropathy in High Myopia.

    abstract:Purpose:To examine the association between intraocular pressure (IOP) and the prevalence of glaucomatous optic neuropathy (GON) in high myopia. Methods:The hospital-based observational study consisted of patients treated in the Tokyo High Myopia Clinics and for whom fundus photographs and IOP readings were available. ...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.17-21942

    authors: Jonas JB,Nagaoka N,Fang YX,Weber P,Ohno-Matsui K

    更新日期:2017-11-01 00:00:00

  • IGF-1 Regulates the Extracellular Level of Active MMP-2 and Promotes Müller Glial Cell Motility.

    abstract:PURPOSE:In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like gr...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.15-17496

    authors: Lorenc VE,Jaldín-Fincati JR,Luna JD,Chiabrando GA,Sánchez MC

    更新日期:2015-10-01 00:00:00

  • The aging Meibomian gland: an in vivo confocal study.

    abstract:PURPOSE:To evaluate age-related Meibomian gland (MG) changes by in vivo laser scanning confocal microscopy (LSCM). METHODS:Asymptomatic healthy subjects (n=100, age range 20-83 years) with an Ocular Surface Disease Index score of less than 13 were consecutively enrolled. Two additional groups, one composed of subjects...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.13-11914

    authors: Villani E,Canton V,Magnani F,Viola F,Nucci P,Ratiglia R

    更新日期:2013-07-16 00:00:00

  • A Novel Tree Shrew (Tupaia belangeri) Model of Glaucoma.

    abstract:Purpose:Primates and rodents are used widely as animal models of glaucoma, but each has significant limitations. Researchers need additional animal models that closely resemble the relevant anatomy and pathologic features of the human disease to more quickly advance research. We validate a novel glaucoma animal model i...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.18-24261

    authors: Samuels BC,Siegwart JT,Zhan W,Hethcox L,Chimento M,Whitley R,Downs JC,Girkin CA

    更新日期:2018-06-01 00:00:00

  • Senescent Changes and Topography of the Dark-Adapted Multifocal Electroretinogram.

    abstract:Purpose:To investigate the topographic changes of the dark-adapted multifocal electroretinogram (mfERG) across adulthood in the central retina and compare the topography between macular versus extramacular, nasal versus temporal, and inferior versus superior retinal areas. Methods:Sixty-five subjects (18-88 years) rec...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.16-20953

    authors: Panorgias A,Tillman M,Sutter EE,Moshiri A,Gerth-Kahlert C,Werner JS

    更新日期:2017-02-01 00:00:00

  • Treatment of autoimmune anterior uveitis with recombinant TCR ligands.

    abstract:PURPOSE:To determine protective properties of recombinant TCR ligands (RTLs) as a new treatment for experimental autoimmune anterior uveitis (AU). RTLs comprise the rat RT1.B beta1alpha1 domains, linked either to the guinea pig MBP69-89 peptide (RTL201), to the corresponding rat MBP69-89 peptide (RTL200), or to the car...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.05-1242

    authors: Adamus G,Burrows GG,Vandenbark AA,Offner H

    更新日期:2006-06-01 00:00:00

  • Characterization of a bradykinin-hydrolyzing protease from the bovine lens.

    abstract:PURPOSE:To isolate and characterize bovine lens endopeptidase activity that cleaves the Phe-Ser bond in peptide substrates. METHODS:The protease activity in young bovine lens homogenate was measured using the Mca-(Ala(7),Lys(Dnp)(9))-bradykinin substrate. Degradation of bradykinin and other unlabeled peptide substrate...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.03-0769

    authors: Chaerkady R,Sharma KK

    更新日期:2004-04-01 00:00:00

  • Diabetic Optic Neuropathy and Its Risk Factors in Chinese Patients With Diabetic Retinopathy.

    abstract:Purpose:To investigate diabetic optic neuropathy (DON) prevalence and risk factors in Chinese diabetic retinopathy (DR) patients. Methods:This retrospective study included 1067 eyes (550 patients) that underwent ocular imaging. The diabetes duration, systolic blood pressure (SBP), hemoglobin A1c (HbA1c), and high-dens...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.19-26825

    authors: Hua R,Qu L,Ma B,Yang P,Sun H,Liu L

    更新日期:2019-08-01 00:00:00

  • A DNA dot hybridization model for assessment of bacterial bioburden in orthokeratology lens storage cases.

    abstract:PURPOSE:The aim of this study was to evaluate a DNA dot hybridization assay (DHA) for assessing bacterial bioburden in orthokeratology lens (OK) storage cases. METHODS:Forty-one OK wearers participated in this study. The dot hybridization assay was used to assess the bacterial bioburden of OK after removal and 6-hour ...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.14-15920

    authors: Kuo MT,Chien CC,Lo J,Hsiao CC,Tseng SL,Lai YH,Fang PC,Chang TC

    更新日期:2014-12-23 00:00:00

  • Biomarkers and Surrogate Endpoints in Uveitis: The Impact of Quantitative Imaging.

    abstract:Purpose:Uveitis is a major cause of sight loss across the world. The reliable assessment of intraocular inflammation in uveitis ('disease activity') is essential in order to score disease severity and response to treatment. In this review, we describe how 'quantitative imaging', the approach of using automated analysis...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章,评审

    doi:10.1167/iovs.17-21788

    authors: Denniston AK,Keane PA,Srivastava SK

    更新日期:2017-05-01 00:00:00

  • Microsaccades and Prediction of a Motor Act Outcome in a Dynamic Sport Situation.

    abstract:PURPOSE:Microsaccades could indicate the place where our mind is unconsciously focusing, although our gaze is directed elsewhere. Many studies report the importance of microsaccades in visual scene perception, but none of them has addressed their relationship with the perception of a dynamic action and the prediction o...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.15-16880

    authors: Piras A,Raffi M,Lanzoni IM,Persiani M,Squatrito S

    更新日期:2015-07-01 00:00:00

  • Early multifocal electroretinogram findings during intravitreal ranibizumab treatment for neovascular age-related macular degeneration.

    abstract:PURPOSE:To evaluate changes in the multifocal electroretinogram (mfERG) in patients with neovascular age-related macular degeneration (nAMD) undergoing ranibizumab treatment. METHODS:This was an observational, longitudinal, prospective study. Treatment-naive patients with nAMD who met the inclusion and exclusion crite...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.10-6588

    authors: Campa C,Hagan R,Sahni JN,Brown MC,Beare NA,Heimann H,Harding SP

    更新日期:2011-06-01 00:00:00

  • Corneal wound healing in tenascin knockout mouse.

    abstract:PURPOSE:Tenascin (TN) is a large hexameric extracellular matrix glycoprotein that is expressed in developing organs and tumors. It has also been reported that TN is expressed in the embryonic cornea and during corneal wound healing. However, the role of TN in the cornea is not fully known. In this study, the role of TN...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Matsuda A,Yoshiki A,Tagawa Y,Matsuda H,Kusakabe M

    更新日期:1999-05-01 00:00:00

  • Downregulation of Lysyl Oxidase Protects Retinal Endothelial Cells From High Glucose-Induced Apoptosis.

    abstract:Purpose:To investigate the effect of reducing high glucose (HG)-induced lysyl oxidase (LOX) overexpression and increased activity on retinal endothelial cell apoptosis. Methods:Rat retinal endothelial cells (RRECs) were grown in normal (N) or HG (30 mM glucose) medium for 7 days. In parallel, RRECs were grown in HG me...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.16-21340

    authors: Kim D,Mecham RP,Trackman PC,Roy S

    更新日期:2017-05-01 00:00:00

  • Abundant expression of ponsin, a focal adhesion protein, in lens and downregulation of its expression by impaired cytoskeletal signaling.

    abstract:PURPOSE:This study was undertaken to improve understanding of the defective lens developmental changes induced by the transgenic overexpression of the Rho GDP dissociation inhibitor RhoGDIalpha. The study was focused on a single differentially expressed gene encoding ponsin, a cell adhesion interacting signaling adapto...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.08-2909

    authors: Rao PV,Maddala R

    更新日期:2009-04-01 00:00:00

  • The Corneal Basement Membranes and Stromal Fibrosis.

    abstract:Purpose:The purpose of this review was to provide detailed insights into the pathophysiology of myofibroblast-mediated fibrosis (scarring or late haze) after corneal injury, surgery, or infection. Method:Literature review. Results:The epithelium and epithelial basement membrane (EBM) and/or endothelium and Descemet's...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章,评审

    doi:10.1167/iovs.18-24428

    authors: Medeiros CS,Marino GK,Santhiago MR,Wilson SE

    更新日期:2018-08-01 00:00:00

  • Congenital stationary night blindness: an animal model.

    abstract::Electroretinographic studies of myctalopic Appaloosa horses demonstrated photopic and scotopic abnormalities similar to those in humans with congenital stationary night blindness (CSNB) of the Schubert-Bornschein type. The phototopic abnormalities consisted of reduced b-wave amplitudes and slower than normal b-wave im...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Witzel DA,Smith EL,Wilson RD,Aguirre GD

    更新日期:1978-08-01 00:00:00

  • Spatial contrast transfer functions of the pattern-evoked electroretinogram.

    abstract::Pattern ERGs in response to pattern-reversal stimuli varying in spatial frequency (range: 0.2-9.03 cycles/deg) and contrast (range: 0.03-0.93) were recorded from two normal male subjects. At most spatial frequencies the response amplitude increased linearly up to the highest contrast without saturation. From the ampli...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Korth M,Rix R,Sembritzki O

    更新日期:1985-03-01 00:00:00

  • Endothelial Yes-Associated Protein 1 Promotes Astrocyte Proliferation and Maturation via Cytoplasmic Leukemia Inhibitory Factor Secretion in Oxygen-Induced Retinopathy.

    abstract:Purpose:Purpose The role of endothelial Yes-associated protein 1 (YAP) in the pathogenesis of retinal angiogenesis and the astrocyte network in the mouse oxygen-induced retinopathy (OIR) model is unknown. Methods:For in vivo studies, OIR was induced in conditional endothelial YAP knockout mice and their wild-type litt...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.61.4.1

    authors: Ai LQ,Zhu JY,Chen X,Li X,Luo LL,Hu QM,Lin S,Ye J

    更新日期:2020-04-09 00:00:00

  • Molecular Mechanisms Mediating Antiangiogenic Action of the Urokinase Receptor-Derived Peptide UPARANT in Human Retinal Endothelial Cells.

    abstract:Purpose:To investigate the molecular mechanisms of the antiangiogenic activity of UPARANT, an antagonist of the urokinase-type plasminogen activator receptor (uPAR), on primary human retinal endothelial cells (HREC) as a model of in vitro angiogenesis. Methods:The antiangiogenic activity of UPARANT was evaluated on en...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.16-19909

    authors: Motta C,Lupo G,Rusciano D,Olivieri M,Lista L,De Rosa M,Pavone V,Anfuso CD

    更新日期:2016-10-01 00:00:00

  • Visually impaired drivers who use bioptic telescopes: self-assessed driving skills and agreement with on-road driving evaluation.

    abstract:PURPOSE:To compare self-assessed driving habits and skills of licensed drivers with central visual loss who use bioptic telescopes to those of age-matched normally sighted drivers, and to examine the association between bioptic drivers' impressions of the quality of their driving and ratings by a "backseat" evaluator. ...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.13-13520

    authors: Owsley C,McGwin G Jr,Elgin J,Wood JM

    更新日期:2014-01-15 00:00:00

  • Aberrant cell and basement membrane architecture contribute to sidestream smoke-induced choroidal endothelial dysfunction.

    abstract:PURPOSE:Environmental tobacco smoke (ETS) is widely regarded as a major modifiable risk factor for age-related macular degeneration (AMD). Yet, precisely how it exerts its pathologic effects is poorly understood. Since early-stage AMD is characterized by choroidal capillary loss, this study examined the effect of sides...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.13-13659

    authors: Yang X,Scott HA,Ardekani S,Williams M,Talbot P,Ghosh K

    更新日期:2014-04-08 00:00:00

  • Inhibitory effect of aminoimidazole carboxamide ribonucleotide (AICAR) on endotoxin-induced uveitis in rats.

    abstract::PURPOSE. To investigate the anti-inflammatory effect of aminoimidazole carboxamide ribonucleotide (AICAR), an analog of adenosine monophosphate (AMP), in endotoxin-induced uveitis (EIU). METHODS. EIU was induced by subcutaneous injection of lipopolysaccharide (LPS) (200 μg) in Lewis rats. AICAR (50 mg/kg, intraperiton...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.11-7331

    authors: Suzuki J,Manola A,Murakami Y,Morizane Y,Takeuchi K,Kayama M,Miller JW,Sobrin L,Vavvas DG

    更新日期:2011-08-22 00:00:00

  • Role of swarming migration in the pathogenesis of bacillus endophthalmitis.

    abstract:PURPOSE:Bacillus cereus causes one of the most rapidly blinding forms of bacterial endophthalmitis. Migration of B. cereus throughout the eye during endophthalmitis is a unique aspect of this disease that may contribute to intraocular virulence. This study was conducted to analyze the contribution of swarming and intra...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.06-0301

    authors: Callegan MC,Novosad BD,Ramirez R,Ghelardi E,Senesi S

    更新日期:2006-10-01 00:00:00

  • Electroretinographic Abnormalities and Sex Differences Detected with Mesopic Adaptation in a Mouse Model of Schizophrenia: A and B Wave Analysis.

    abstract:Purpose:Mesopic flash electroretinography (fERG) as a tool to identify N-methyl-d-aspartate receptor (NMDAR) hypofunction in subjects with schizophrenia shows great potential. We report the first fERG study in a genetic mouse model of schizophrenia characterized by NMDAR hypofunction from gene silencing of serine racem...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:10.1167/iovs.61.2.16

    authors: Torres Jimenez N,Lines JW,Kueppers RB,Kofuji P,Wei H,Rankila A,Coyle JT,Miller RF,McLoon LK

    更新日期:2020-02-07 00:00:00

  • Group II PLA(2) content of tears in normal subjects.

    abstract:PURPOSE:To determine the concentration of group II phospholipase (PL) A(2), an antimicrobial molecule, in tears of normal subjects in different age and sex groups. METHODS:PLA(2) content of tears was measured in 122 healthy volunteers with ages ranging from 20 to 89 years (mean, 49.5 years) by a time-resolved fluoroim...

    journal_title:Investigative ophthalmology & visual science

    pub_type: 杂志文章

    doi:

    authors: Saari KM,Aho V,Paavilainen V,Nevalainen TJ

    更新日期:2001-02-01 00:00:00