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.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Malik KJ,Chen CD,Olsen TWdoi
10.1167/iovs.02-1120keywords:
subject
Has Abstractpub_date
2003-06-01 00:00:00pages
2730-5issue
6eissn
0146-0404issn
1552-5783journal_volume
44pub_type
杂志文章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
更新日期:2006-11-01 00:00:00
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
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doi:
更新日期:1997-09-01 00:00:00
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:
更新日期:1992-01-01 00:00:00
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:
更新日期:1992-01-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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doi:10.1167/iovs.17-21942
更新日期:2017-11-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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doi:10.1167/iovs.18-24261
更新日期:2018-06-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2017-02-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2006-06-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2004-04-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
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更新日期:2019-08-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-15920
更新日期:2014-12-23 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2017-05-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.15-16880
更新日期:2015-07-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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doi:10.1167/iovs.10-6588
更新日期:2011-06-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1999-05-01 00:00:00
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
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更新日期:2017-05-01 00:00:00
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更新日期:2009-04-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,评审
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journal_title:Investigative ophthalmology & visual science
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doi:
更新日期:1978-08-01 00:00:00
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
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doi:
更新日期:1985-03-01 00:00:00
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更新日期:2020-04-09 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2016-10-01 00:00:00
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
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更新日期:2014-01-15 00:00:00
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
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更新日期:2014-04-08 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2011-08-22 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2006-10-01 00:00:00
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更新日期:2020-02-07 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:2001-02-01 00:00:00