Abstract:
PURPOSE:Primary open angle glaucoma-associated mutations in myocilin (MYOC) cause protein "nonsecretion," rendering secreted MYOC difficult to detect using conventional techniques. This study focused on developing and using an assay that can quickly and easily detect mutant MYOC secretion. METHODS:We fused Gaussia luciferase (eGLuc2) to MYOC variants and expressed the constructs in HEK-293T and NTM-5 cells. Secreted and intracellular levels of MYOC eGLuc2 variants were evaluated by Western blotting and compared to untagged and FLAG-tagged MYOC constructs. Secreted and soluble intracellular MYOC eGLuc2 were measured by a GLuc assay. The secretion of nine additional MYOC mutants was assayed in conditioned media from transfected cells to test the applicability of the assay for monitoring other MYOC variants. RESULTS:Myocilin eGLuc2 behaved similarly to untagged and FLAG-tagged MYOC with respect to secretion, soluble intracellular levels, and in response to drug treatment. The GLuc assay could sensitively detect Y437H MYOC secretion 30 minutes after media change. Gaussia luciferase fused variants followed anticipated trends; nonpathogenic variants (D208E, G244V) were secreted at wild-type (WT) levels, whereas predicted disease-causing variants (C245Y, G246R, E300K, Y437H, I477N) demonstrated substantial secretion defects. Secretion defects caused by the C245Y, G246R, and Y437H mutations were partially rescued by permissive growth temperature. Interestingly, however, this increase in secretion was independent of newly synthesized protein. CONCLUSIONS:Fusion of eGLuc2 to MYOC does not significantly change the behavior of MYOC. This newly developed MYOC reporter system can be used to study engineered MYOC variants and potentially to identify modulators of MYOC secretion and function.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Zadoo S,Nguyen A,Zode G,Hulleman JDdoi
10.1167/iovs.15-18789subject
Has Abstractpub_date
2016-04-01 00:00:00pages
1939-50issue
4eissn
0146-0404issn
1552-5783pii
2516297journal_volume
57pub_type
杂志文章abstract:PURPOSE:Many genome-wide association studies have identified common single nucleotide polymorphisms (SNPs) at the 9p21 glaucoma locus (CDKN2B/CDKN2B-AS1) to be significantly associated with primary open-angle glaucoma (POAG), with association being stronger in normal tension glaucoma (NTG) and advanced glaucoma. We aim...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.16-19401
更新日期:2016-06-01 00:00:00
abstract:PURPOSE:To examine the expression pattern and roles of Sonic hedgehog (Shh) in healing corneal epithelium. METHODS:Immunofluorescent staining and Western blot analysis were used to detect Shh, patched 1 (Ptc 1) receptors, and Gli transcription factors in corneal epithelium of Wistar rats (n = 44) at various intervals ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.04-0001
更新日期:2004-08-01 00:00:00
abstract:PURPOSE:To study relationships between glaucoma-sensitive components identified with frequency-domain analysis of global-flash multifocal electroretinogram (mfERG), regional retinal nerve fiber layer thickness (RNFLT), and local visual field sensitivity (VS). METHODS:Eleven macaque monkeys, including four controls and...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.10-6667
更新日期:2011-06-01 00:00:00
abstract:Purpose:To test the amblyopic suppression at mid to low spatial frequencies when compensating for signal attenuation. Methods:Eight amblyopes with (n = 5) or without (n = 3) strabismus and 10 normal controls with normal or corrected to normal visual acuity (≥20/20) and normal stereopsis (≤40 arcseconds) participated. ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.18-24206
更新日期:2018-08-01 00:00:00
abstract:PURPOSE:Children with cortical or cerebral visual impairment (CVI) often experience photophobia. In a study conducted to test whether this clinical phenomenon affects visual function, the sweep visual evoked potential (VEP) was used to evaluate cortical responses to grating stimuli in two luminance conditions: low and ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1252
更新日期:2006-07-01 00:00:00
abstract:PURPOSE:To characterize the expression patterns of myosin heavy chain (MyHC) isoforms in mouse extraocular muscles (EOMs) during postnatal development. METHODS:MyHC isoform expression in mouse EOMs from postnatal day (P)0 to 3 months was evaluated by quantitative polymerase chair reaction (qPCR) and immunohistochemist...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.10-5937
更新日期:2010-12-01 00:00:00
abstract:PURPOSE:To better understand the cellular contributions to the human multifocal ERG (mfERG), rhesus monkey and human mfERGs were recorded using the same stimulus conditions. The monkey mfERGs were recorded before and after injections of pharmacologic agents known to selectively block activity of particular cells and ci...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:2002-05-01 00:00:00
abstract:PURPOSE:To determine if lens epithelial lines can be established from cryopreserved whole rabbit lenses and from cryopreserved capsule-epithelial preparations (CEPs). METHODS:Lenses or freshly isolated CEPs were cryopreserved and subsequently thawed. Thawed whole lenses were cultured for 48 hours in growth medium and ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1995-02-01 00:00:00
abstract::Ultrastructural cytochemistry was used to analyze the polarized distribution of alkaline phosphatase (AP) on the plasma membranes of normal and regenerating retinal pigment epithelium (RPE) and the ciliary body epithelium in rabbits. The AP activity was concentrated on the basolateral plasma membrane in normal RPE. In...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1991-12-01 00:00:00
abstract:PURPOSE:To identify the underlying molecular defects causing retinal degeneration in seven N-ethyl-N-nitrosourea (ENU) induced mutant alleles of the Pde6b gene and to analyze the timescale of retinal degeneration in these new models of retinitis pigmentosa. METHODS:Conformation sensitive capillary electrophoresis and ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-0254
更新日期:2005-09-01 00:00:00
abstract:PURPOSE:Macrophages are thought to be the first line of defense in many infectious diseases and are present in high numbers in corneas with Acanthamoeba keratitis. Conjunctival macrophage depletion was performed in an animal model of Acanthamoeba infection to determine the importance of macrophages in this disease. ME...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1996-06-01 00:00:00
abstract:PURPOSE:To study Bardet-Biedl syndrome (BBS) in the Tunisian population and determine the presence of triallelism in the eight identified BBS genes. METHODS:DNA samples were collected from 19 consanguineous Tunisian families with BBS. Genome-wide scans were performed with microsatellite markers in 12 families, and two...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1334
更新日期:2006-08-01 00:00:00
abstract:PURPOSE:To evaluate the differential effects of ganglion cell depletion from experimental glaucoma on the relative metabolic activities of neurons in the parvo (P)- and magno (M)-cellular visual pathways of the macaque visual system. METHODS:Monocular experimental glaucoma was induced in monkeys (Macaca mulatta and M....
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:2000-06-01 00:00:00
abstract:PURPOSE:To investigate the relationship between long-term glycemic control and photopic negative response (PhNR) changes in the blue flash ERG in adolescents with type 1 diabetes (T1D) without diabetic retinopathy (DR). METHODS:After light adaptation, ERG responses to 1.60 cd·s/m(2) blue (420 nm) flashes (blue flash E...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-8263
更新日期:2012-02-13 00:00:00
abstract:Purpose:To assess changes in Schlemm's canal (SC), intraocular pressure (IOP), and autonomic nervous system activity in healthy individuals after performing the water-drinking test (WDT). Methods:The SC area (SCAR), trabecular meshwork (TM) thickness, IOP, high frequency (HF) of heart rate variability (HRV), heart rat...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.18-23909
更新日期:2018-07-02 00:00:00
abstract::Critical flicker frequency (CFF) was measured for stimuli varying in chromaticity only and in luminance only for patients with multiple sclerosis (MS) and for matched normal controls. The two CFF measurements showed different underlying linear interdependencies for the two groups, consistent with a greater loss of tem...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1982-08-01 00:00:00
abstract:PURPOSE:To determine whether a novel optical treatment using contact lenses to reduce relative peripheral hyperopia can slow the rate of progress of myopia. METHODS:Chinese children, aged 7 to 14 years, with baseline myopia from sphere -0.75 to -3.50 D and cylinder ≤1.00 D, were fitted with novel contact lenses (n = 4...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,随机对照试验
doi:10.1167/iovs.11-7260
更新日期:2011-12-09 00:00:00
abstract:Purpose:To investigate factors associated with visual field (VF) progression in treated myopic normal-tension glaucoma (NTG) using a novel posterior sclera reconstruction method involving swept-source optical coherence tomography (OCT). Methods:Fifty-six myopic patients on ocular hypotensive therapy with the diagnose ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.19-26794
更新日期:2019-05-01 00:00:00
abstract::Swollen rabbit corneas incubated in vitro with their posterior surfaces blocked with silicone oil maintained fairly constant thickness over an 11 hr. period. Increasing the simulated intraocular pressure from 10 to 30 mm. Hg did not produce stromal thinning. When theophylline was added to stimulate epithelial Cl secre...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1977-10-01 00:00:00
abstract:PURPOSE:To investigate the role of anti-vascular endothelial growth factor (VEGF)-C therapy in corneal graft survival and concomitant suppression of hem- and lymph-angiogenesis. METHODS:Corneal suture model in BALB/c mice was placed and immunohistochemical staining was performed with CD31/PECAM-1 and LYVE-1 to quantif...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-8668
更新日期:2012-03-09 00:00:00
abstract:PURPOSE:Because several polypeptide growth factors are known to influence capillary endothelial cell mitogenesis, the authors investigated the presence of some of these molecules in choroidal neovascular membranes (CNVMs) removed surgically from human subjects with age-related macular degeneration (ARMD). METHODS:The ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1994-07-01 00:00:00
abstract:PURPOSE:Nonsyndromic high myopia, defined by a refractive error greater than -6 diopters (D), is associated with an increased risk of macular choroidal neovascularization (CNV), a vision-threatening complication. The aim of this study was to investigate whether genetic factors associated with age-related macular degene...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-9538
更新日期:2012-07-27 00:00:00
abstract:PURPOSE:Polypoidal choroidal vasculopathy (PCV) is a major cause of serosanguinous maculopathy in Chinese patients with age-related macular degeneration (AMD). Variants in the CFH and HTRA1/LOC387715 genes are strongly associated with AMD in Caucasians and Chinese. Variants in the C2 and BF genes have been found to con...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.07-0860
更新日期:2008-06-01 00:00:00
abstract:Purpose:To determine the effect of voluntary exercise on choroidal neovascularization (CNV) in mice. Methods:Age-matched wild-type C57BL/6J mice were housed in cages equipped with or without running wheels. After four weeks of voluntary running or sedentariness, mice were subjected to laser injury to induce CNV. After...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.61.5.52
更新日期:2020-05-11 00:00:00
abstract::The authors have examined the fate of intravitreally injected liposomes in the aphakic, vitrectomized eye of the rabbit. Liposomes labelled with 125[I]-p-hydroxybenzimidylphosphatidylethanolamine were eliminated rapidly from the intraocular fluid. Nonetheless, a significant fraction of these liposomes were found to bi...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1987-05-01 00:00:00
abstract::Herpetic stromal disease often is treated with combinations of antiviral agents and corticosteroids. The addition of steroids to the antiviral treatment regimen frequently increases the efficacy of therapy in patients; however, many complications may arise as a result of corticosteroid therapy. Using a rabbit model, t...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1991-08-01 00:00:00
abstract:PURPOSE:To determine the profiles of soluble cytokine receptors and cytokines, including mostly their ligands, in the vitreous humor of patients with B-cell vitreoretinal lymphoma (VRL) and uveitis. METHODS:Vitreous samples were collected from immunocompetent patients with VRL (n = 21), uveitis (n = 20), and idiopathi...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.15-17465
更新日期:2015-11-01 00:00:00
abstract:PURPOSE:To evaluate the pharmacokinetics of ciliary neurotrophic factor (CNTF) delivered over a period of up to 2 years by an intraocular encapsulated cell technology (ECT) implant in patients with retinitis pigmentosa (RP) and geographic atrophy (GA). METHODS:Patients from phase 1 RP (CNTF1); phase 2 GA (CNTF2); and ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,多中心研究,随机对照试验
doi:10.1167/iovs.12-9970
更新日期:2012-11-01 00:00:00
abstract:PURPOSE:To evaluate the antilymphangiogenic potential of multi-target tyrosine kinase inhibitor sunitinib in corneal neovascularization (NV). METHODS:Inflammatory corneal NV was induced by thermal cauterization applied in the central cornea of mice, to which sunitinib malate was daily administered by gavage or not. At...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-10856
更新日期:2013-05-03 00:00:00
abstract:PURPOSE:To determine the relationship between changes in contrast sensitivity, if any, after glaucoma therapy and the test-retest reliability of the CSV-1000 contrast sensitivity test. METHODS:Patients with primary open angle glaucoma (N = 16) were retrospectively evaluated to determine changes in visual function, as ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1994-08-01 00:00:00