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GAPDH AND Retinopathy

Xin Liu, Jia'nan Xie, Zaoxia Liu, Qiaoyun Gong, Rui Tian, Guanfang Su
Retinal pigment epithelium (RPE) cell-based gene expression studies performed under hypoxia and/or hyperglycemia show huge potential for modeling cell responses in diabetic retinopathy, retinopathy of prematurity and other retinal diseases. However, normalization of gene expression on RPE cells under those conditions has commonly been done using either GAPDH or β-actin as reference genes without any validation of their expression stability. Therefore, we aimed to establish a suitable set of reference genes for studies on RPE cells cultured under both normal culturing glucose and atmospheric oxygen tension (normoxia, 21%), under a low oxygen tension (hypoxia, 1%), under a high glucose growth medium (25 mmol/l) and under the combination of the two changed conditions above for distinct time points taking together from 24h to 7 days...
April 10, 2016: Gene
Sandra Suarez, Gary W McCollum, Ashwath Jayagopal, John S Penn
Diabetic retinopathy (DR) is a leading cause of blindness worldwide, and its prevalence is growing. Current therapies for DR address only the later stages of the disease, are invasive, and have limited effectiveness. Retinal pericyte death is an early pathologic feature of DR. Although it has been observed in diabetic patients and in animal models of DR, the cause of pericyte death remains unknown. A novel pro-apoptotic pathway initiated by the interaction between glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the E3 ubiquitin ligase, seven in absentia homolog 1 (Siah1), was recently identified in ocular tissues...
November 20, 2015: Journal of Biological Chemistry
Bo Y Baker, Wuxian Shi, Benlian Wang, Krzysztof Palczewski
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the oxidative phosphorylation of d-glyceraldehyde 3-phosphate (G3P) into 1,3-diphosphoglycerate (BGP) in the presence of the NAD cofactor. GAPDH is an important drug target because of its central role in glycolysis, and nonglycolytic processes such as nuclear RNA transport, DNA replication/repair, membrane fusion and cellular apoptosis. Recent studies found that GAPDH participates in the development of diabetic retinopathy and its progression after the cessation of hyperglycemia...
November 2014: Protein Science: a Publication of the Protein Society
Grazyna Adamus, Dongseak Choi, Anitha Raghunath, Jade Schiffman
BACKGROUND: The presence of autoantibodies (AAbs) is the primary serological indicator of autoimmunity. Cancer-associated retinopathy (CAR) is associated with AAbs and different types of cancer. The goal of the study was to examine the profile of serum autoantibodies in women with gynecological cancers with and without paraneoplastic visual manifestation. METHODS: Retrospective studies of a cohort of 46 women with symptoms of CAR and gynecological tumors, including endometrial, cervical, ovarian, and fallopian tubes, 111 women with similar tumors without symptoms of CAR, and 60 age-matched healthy controls...
December 1, 2013: Journal of Clinical & Experimental Ophthalmology
Katrine R Lind, Kelly K Ball, Nancy F Cruz, Gerald A Dienel
Oxidative-nitrosative stress and inflammatory responses are associated with endoplasmic reticulum (ER) stress in diabetic retinopathy, raising the possibility that disturbances in ER protein processing may contribute to CNS dysfunction in diabetics. Upregulation of the unfolded protein response (UPR) is a homeostatic response to accumulation of abnormal proteins in the ER, and the present study tested the hypothesis that the UPR is upregulated in two models for diabetes, cultured astrocytes grown in 25mmol/L glucose for up to 4weeks and brain of streptozotocin (STZ)-treated rats with diabetes for 1-7months...
April 2013: Neurochemistry International
Shereen A El Tarhouny, Khaled M Hadhoud, Magdy M Ebrahem, Nashwa M Al Azizi
In order to assess the potential biochemical markers in the development, diagnosis, and prognosis of diabetic patient with microvascular complication represented with retinopathy, we analyzed the levels of cell-free DNA by two different techniques. The levels of cell-free GAPDH assayed by quantitative PCR were significantly higher in the plasma samples of diabetic patients with and without diabetic retinopathy than in those of the control group; thus, it is a better biomarker than nucleosomes assayed by ELISA in patients with type 2 diabetes for the early detection of development of microvascular complications as retinopathy...
March 2010: Nucleosides, Nucleotides & Nucleic Acids
Sally Madsen-Bouterse, Ghulam Mohammad, Renu A Kowluru
PURPOSE: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been hypothesized as a mediator in the activation of multiple pathways implicated in the pathogenesis of diabetic retinopathy. The objective of this study was to understand the mechanism by which high glucose inactivates GAPDH in retinal microvascular cells. METHODS: Bovine retinal endothelial cells (BRECs), transfected with GAPDH, were incubated in 20 mM glucose. The effect of the overexpression of GAPDH on its activity, apoptosis, and upstream signaling pathways, protein kinase C, and hexosamine pathways was determined...
March 2010: Investigative Ophthalmology & Visual Science
Mamta Kanwar, Renu A Kowluru
OBJECTIVE: Mitochondrial superoxide levels are elevated in the retina in diabetes, and manganese superoxide dismutase overexpression prevents the development of retinopathy. Superoxide inhibits glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which activates major pathways implicated in diabetic complications, including advanced glycation end products (AGEs), protein kinase C, and hexosamine pathway. Our aim is to investigate the role of GAPDH in the development and progression of diabetic retinopathy and to elucidate the mechanism...
January 2009: Diabetes
Ya-Dong Wang, Jin-Dao Wu, Zhong-Li Jiang, Yu-bang Wang, Xue-Hao Wang, Chao Liu, Ming-Qing Tong
PURPOSE: The purpose of this study was to isolate, identify, and analyze diabetes-related protein changes that occur in neural retinas in vivo. METHODS: Total proteins were extracted from neural retinas of normal and 8-weeks diabetic Sprague-Dawley (SD) rats and separated by two-dimensional gel electrophoresis (2-DE). Some protein spots exhibiting statistically significant variations (p < 0.05) were selected randomly and identified by mass spectrometry (MS or MS/MS)...
October 2007: Current Eye Research
Sidney G Shaw, Jane P Boden, Erwin Biecker, Juerg Reichen, Barbara Rothen
Altered activity of retinal endothelin-1 (ET-1) and nitric oxide may play a causal role in the hemodynamic and histopathological changes of diabetic retinopathy. This study evaluated the therapeutic potential of long-term selective blockade of the ET-1(A) receptor (ETRA) to prevent the development of retinopathy in a genetic mouse model of nonobese type 1 diabetes (NOD). Mice with NOD that received subcutaneous implantation of insulin pellets and wild-type control mice were treated for 4 months with the selective ETRA antagonist LU208075 (30 mg/kg/day) via drinking water...
June 2006: Experimental Biology and Medicine
Pál Pacher, Csaba Szabó
Hyperglycemia-induced overproduction of superoxide by mitochondrial electron-transport chain triggers several pathways of injury involved in the pathogenesis of diabetic complications [protein kinase C (PKC), hexosamine and polyol pathway fluxes, advanced glycation end product (AGE) formation] by inhibiting glyceraldehyde- 3-phosphate dehydrogenase (GAPDH) activity. Increased oxidative and nitrosative stress activates the nuclear enzyme, poly(ADP-ribose) polymerase-1 (PARP). PARP activation, on the one hand, depletes its substrate, NAD+, slowing the rate of glycolysis, electron transport, and ATP formation...
November 2005: Antioxidants & Redox Signaling
Levente Kiss, Csaba Szabó
Recent work has demonstrated that hyperglycemia-induced overproduction of superoxide by the mitochondrial electron-transport chain triggers several pathways of injury [(protein kinase C (PKC), hexosamine and polyol pathway fluxes, advanced glycation end product formation (AGE)] involved in the pathogenesis of diabetic complications by inhibiting glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. Increased oxidative and nitrosative stress activates the nuclear enzyme, poly(ADP-ribose) polymerase-1 (PARP)...
March 2005: Memórias do Instituto Oswaldo Cruz
Linda L Kusner, Vijay P Sarthy, Susanne Mohr
PURPOSE: A recent study demonstrated that retinal Müller cells undergo hyperglycemia-induced apoptosis in vitro. Translocation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the cytosol to the nucleus is a critical step in the induction of apoptosis in neuronal cells. R-(-)-deprenyl prevents nuclear translocation of GAPDH and subsequent apoptosis in neuronal cells. In this study, the role of nuclear translocation of GAPDH in hyperglycemia-induced apoptosis in retinal Müller cells and the ability of R-(-)-deprenyl to inhibit the translocation of GAPDH and apoptosis were investigated...
May 2004: Investigative Ophthalmology & Visual Science
Mark E Obrenovich, Vincent M Monnier
Diabetes accelerates the aging process and leads to complications that include blindness, renal failure, nerve damage, stroke, and cardiovascular disease. It has been hypothesized that high plasma glucose concentrations are responsible for increased mitochondrial free radical production and subsequent inactivation of glyceraldehyde phosphate dehydrogenase (GAPDH) in vascular endothelial cells and other cells implicated in these complications. As a result of the decreased ability of GAPDH to process upstream metabolites, three pathways of metabolic damage are activated, which include the advanced glycation end-product formation pathway, the protein kinase C pathway, and the hexosamine pathway...
March 12, 2003: Science of Aging Knowledge Environment: SAGE KE
Steve F Abcouwer, Philip L Marjon, Robyn K Loper, David L Vander Jagt
PURPOSE: Vascular endothelial growth factor (VEGF) plays an important role in initiation of the angiogenesis that leads to proliferative retinopathy. Several environmental conditions and chemical agents that influence the expression of VEGF can also cause endoplasmic reticulum (ER) stress. The hypothesis for the current study was that expression of VEGF is responsive to conditions that cause ER stress, including amino acid deprivation. METHODS: Confluent cultures of a human retinal pigmented epithelial cell line (ARPE-19) were deprived of amino acids or treated with chemical inducers of ER stress...
August 2002: Investigative Ophthalmology & Visual Science
D A Simpson, S Feeney, C Boyle, A W Stitt
PURPOSE: To determine whether continuous monitoring of SYBR Green I fluorescence provides a reliable and flexible method of quantitative RT-PCR. Our aims were (i) to test whether SYBR Green I analysis could quantify a wide range of known VEGF template concentrations, (ii) to apply this method in an experimental model, and (iii) to determine whether 20 existing primer pairs could be used to quantify their cognate mRNAs. METHODS: Real-time quantitative PCR was performed using a LightCycler rapid thermal cycler (Roche)...
October 5, 2000: Molecular Vision
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