Abstract:
:PAX6-related Aniridia is a sight-threatening disease involving progression of secondary glaucoma and aniridia related keratopathy (ARK). Change or loss of limbal epithelial progenitors causes epithelial surface defects. We analyzed the effect of PAX6 on mRNA expression changes with a two-step approach, as follows. First, we sequenced mRNA from limbal epithelial cells isolated from controls and aniridia patients. Second, we confirmed the bioinformatics and literature-based result list for a small interfering RNA (siRNA)-based primary aniridia cell model (PAX6 knockdown). With this approach, we expected that the genes directly influenced by PAX6 would be distinguishable from those affected secondarily by the ARK disease state. Therefore, epithelial cells were isolated from the limbus region of two patients with aniridia and cultured in keratinocyte serum-free medium. Normal control cells were obtained from the limbus region of corneal donors. For the siRNA-based aniridia cell model, cells were transfected with Lipofectamine and 5 nM siRNA against PAX6 or control treatment. All cells were lysed to yield DNA, RNA, and protein. Reduction of PAX6 protein was assessed by western blot. Aniridia and control Poly-A-enriched RNA libraries were subjected to next-generation sequencing. The differential analysis was a combination of quantification with RSEM and differential tests with edgeR. Gene lists were filtered by comparison to NCBI GEO datasets, annotated with DAVID, and manually annotated using a literature search. Based on the resulting filtered gene list, qPCR primers were purchased, and candidate genes (TP63, ABCG2, ADH7, ALDH1A1, PITX1, DKK1, DSG1, KRT12, KRT3, KRT13, SPINK6, SPINK7, CTSV, SERPINB1) were verified by qPCR on the siRNA-based aniridia cell model. We identified genes that might be regulated by PAX6 and showed that SPINK7 mRNA, which codes for a protease inhibitor, is downregulated in patients as well as in our primary aniridia cell model. ALDH1A1 and AHD7 mRNA levels were reduced in limbal epithelial cells of aniridia patients, and both transcripts were downregulated by PAX6 knockdown in our cell model. This siRNA-based aniridia cell model is a valuable tool for confirming identified PAX6-affected genes that might promote ARK pathogenesis. The model recapitulated expression changes for SPINK7, ADH7, and ALDH1A1 that were also observed in patient samples. These results provide evidence that PAX6 might drive corneal epithelial differentiation by direct or indirect control of retinoic acid signaling processes through ADH7 and ALDH1A1.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
Latta L,Nordström K,Stachon T,Langenbucher A,Fries FN,Szentmáry N,Seitz B,Käsmann-Kellner Bdoi
10.1016/j.exer.2018.10.002subject
Has Abstractpub_date
2019-02-01 00:00:00pages
8-17eissn
0014-4835issn
1096-0007pii
S0014-4835(18)30359-2journal_volume
179pub_type
杂志文章abstract::Mounting evidence suggests neuroinflammation is a key process in glaucoma, yet the precise roles are not known. Understanding these complex processes, which may also be a key in other common neurodegenerations such as Alzheimer's disease, will lead to targeted therapeutics for a disease that affects as many as 80 mill...
journal_title:Experimental eye research
pub_type: 杂志文章,评审
doi:10.1016/j.exer.2017.02.014
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abstract::The purpose of this study was to investigate the impact of different diffusion times of mitomycin-C (MMC) on the intrascleral concentration vs depth profile of MMC in an experimental model. Scleral quadrants of eight human donor eyes were exposed to sponges soaked with MMC for an application time of 1 min. After irrig...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.2000.0901
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journal_title:Experimental eye research
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doi:10.1016/s0014-4835(03)00211-2
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abstract::The redox homeostasis is controlled by several enzyme systems. Sulfhydryl groups in lens proteins are very sensitive to oxidative stress and can easily conjugate with nonprotein thiols (S-thiolation) to form protein-thiol mixed disulfides. We have observed an elevation of protein S-S-glutathione (PSSG) and protein-S-S...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1997.0464
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abstract::For in vivo mouse retinal imaging, especially with Adaptive Optics instruments, application of a contact lens is desirable, as it allows maintenance of cornea hydration and helps to prevent cataract formation during lengthy imaging sessions. However, since the refractive elements of the eye (cornea and lens) serve as ...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2018.03.027
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abstract::Previous histochemical and biochemical studies have documented the presence of carbohydrate-containing molecules in the retinal interphotoreceptor matrix (IPM). The lectin peanut agglutinin (PNA), which preferentially binds galactose-containing carbohydrates, especially galactose-galactosamine linkages, selectively la...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/s0014-4835(87)80163-x
更新日期:1987-04-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(88)90058-9
更新日期:1988-09-01 00:00:00
abstract::The binding of monoclonal and polyclonal antibodies to glial fibrillary acidic protein (GFAP) antigenic sites in the rabbit retina was shown to be sensitive to aldehyde fixation. In chemically unfixed retina, the polyclonal anti-GFAP labeled Müller cells, astrocytes, and unidentified profiles in the outer plexiform la...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(90)90139-l
更新日期:1990-04-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.2000.0843
更新日期:2000-06-01 00:00:00
abstract::Basement membranes isolated from the entire neural retinae of normal cows consist of a heterogeneous mixture of polypeptides resolvable as two major populations, one collagenous and the other non-collagenous. The non-collagenous population consists of disulfide-bonded aggregates of which the major fraction is soluble ...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(83)90010-6
更新日期:1983-02-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(86)90025-4
更新日期:1986-04-01 00:00:00
abstract::The purpose of this study was to investigate the expression of the EGF family of growth factors and EGF receptor subtypes (ErbB1-4) present in lacrimal gland and determine the effects of these growth factors on different functions of rat lacrimal gland. RT-PCR was used to detect mRNA expression in the lacrimal gland o...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2004.10.006
更新日期:2005-03-01 00:00:00
abstract::Anandamide (arachidonylethanolamide) is an endogenous ligand for cannabinoid receptors, and exerts various cannabimimetic activities. Since cannabinoids and anandamide were pharmacologically active with the eye, we examined metabolism of anandamide in a variety of porcine ocular tissues. In the presence of ethanolamin...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1996.0265
更新日期:1997-05-01 00:00:00
abstract::Mutations in the optic atrophy type 1 (OPA1) gene give rise to human autosomal dominant optic atrophy. The purpose of this study is to investigate OPA1 protein expression in the human retina and optic nerve. A rabbit polyclonal antiserum was generated using a fusion protein covering amino acids 647 to 808 of the human...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2006.06.004
更新日期:2006-11-01 00:00:00
abstract::The vitreous is a clear, gel-like mass in the posterior cavity of the eye. Its major components are type II collagen, hyaluronic acid, and water. A small amount of proteoglycan is also present. All of these molecules are found in cartilage as well. We have now identified a non-collagenous protein in bovine and human v...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/s0014-4835(86)80074-4
更新日期:1986-09-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2018.04.013
更新日期:2018-08-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1997.0331
更新日期:1997-09-01 00:00:00
abstract::(O-acyl) ω-hydroxy fatty acids (OAHFAs) are a recently found group of polar lipids in meibum. Since these lipids can potentially serve as a surfactant in the tear film lipid layer, the surface properties of a molecule of this lipid class was investigated and compared with a structurally related wax ester and a fatty a...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2013.06.016
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journal_title:Experimental eye research
pub_type: 杂志文章
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journal_title:Experimental eye research
pub_type: 杂志文章
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journal_title:Experimental eye research
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abstract::The ability of diinosine polyphosphates, diinosine triphosphate (Ip(3)I), diinosine tetraphosphate (Ip(4)I) and diinosine pentaphosphate (Ip(5)I) to modify intraocular pressure in normotensive New Zealand white rabbits was tested. Ip(5)I produced increase in intraocular pressure, while Ip(3)I and Ip(4)I produced a dec...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2012.05.011
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abstract::The purpose of this study was to examine the localization and relative levels of vascular endothelial growth factor (VEGF; an angiogenic factor) and pigment epithelium-derived factor (PEDF; an antiangiogenic factor) in aged human choroid and to determine if the localization or their relative levels changed in age-rela...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2005.05.007
更新日期:2006-01-01 00:00:00
abstract::Macular corneal dystrophy (MCD) is an autosomal recessive disease characterized by abnormal deposition of glycosaminoglycans in corneal stroma, keratocytes, Descemet's membrane and corneal endothelium. According to the presence and distribution of sulfated keratan sulfate (KS)-epitopes in serum and cornea (using mAb 5...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.2001.1080
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abstract::The protein phosphatase 2B inhibitor, FK506, is an immunomodulatory polypeptide that has neuroprotective properties, the mechanisms of which have not been elucidated. A possible mechanism may be phosphorylation-mediated regulation of glutamate transporter activity. In the present study, we investigated the effect of F...
journal_title:Experimental eye research
pub_type: 信件
doi:10.1016/j.exer.2010.04.011
更新日期:2010-08-01 00:00:00
abstract::The purpose of this study was to determine the aqueous levels and pharmacokinetics of topical fluconazole 0.2% upon single and multiple drop applications. Forty-nine patients undergoing cataract surgery were given topical fluconazole 0.2%. They either received single drop or a loading dose of 1 drop per 5 min for 20 m...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.2000.0936
更新日期:2001-02-01 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::Although a range of changes in anterior segment structures have been documented to occur during accommodation, the quantification of changes in the structure of the anterior sclera during the accommodation process in human subjects has yet to be examined. This study therefore aimed to investigate the presence of short...
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pub_type: 杂志文章
doi:10.1016/j.exer.2018.07.023
更新日期:2018-12-01 00:00:00
abstract::Fibroblast growth factors (FGF) are multipotent cytokines with demonstrated mitogenic, neurotrophic and angiogenic properties. There is evidence that they have multiple functions during and after development of the vertebrate eye. Amongst these, the role of FGF receptor mediated signaling in the retinal pigmented epit...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.2000.0892
更新日期:2000-10-01 00:00:00
abstract::Owing to a rapidly aging population, vision impairment caused by age-related cataract has become very common. Age-related cataract has also become one of the principal causes of blindness, and apoptosis of lens epithelial cells contributes to non-congenital cataract development. Previous studies have reported that mic...
journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2016.04.018
更新日期:2016-06-01 00:00:00