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1 General aspects (0)   1.1 Epidemiology (9)   1.2 Population genetics (2)   1.3 Pathogenesis (3)   1.4 Quality of life (5)   1.5 Glaucomas as cause of blindness (6)   1.6 Prevention and screening (7) 2 Anatomical structures in glaucoma (0)   2.1 Conjunctiva (8)   2.2 Cornea (15)   2.3 Sclera (4)   2.4 Anterior chamber angle (8)   2.5 Meshwork (0)     2.5.1 Trabecular meshwork (7)     2.5.2 Schlemms canal (2)     2.5.3 Scleral outlet channels (0)   2.6 Aqueous humor dynamics (0)     2.6.1 Production (0)     2.6.2 Outflow (0)       2.6.2.1 Trabecular meshwork (1)       2.6.2.2 Uveoscleral (0)     2.6.3 Compostion (3)   2.7 Episcleral veins and venous pressure (1)   2.8 Iris (2)   2.9 Ciliary body (1)   2.10 Lens (0)   2.11 Vitreous body (0)   2.12 Choroid, peripapillary choroid, peripapillary atrophy (5)   2.13 Retina and retinal nerve fibre layer (18)   2.14 Optic disc (19)   2.15 Optic nerve (2)   2.16 Chiasma and retrochiasmal central nervous system (3)   2.17 Stem cells (2)   2.20 Other (0) 3 Laboratory methods (0)   3.1 Microscopy (2)   3.2 Electron microscopy (0)   3.3 Immunohistochemistry (1)   3.4 Molecular genetics (0)     3.4.1 Linkage studies (5)     3.4.2 Gene studies (21)   3.5 Molecular biology incl. SiRNA (13)   3.6 Cellular biology (28)   3.7 Biochemistry (10)   3.8 Pharmacology (11)   3.9 Pathophysiology (12)   3.10 Immunobiology (5)   3.11 Pharmacogenetics (0)   3.12 Proteomics (2)   3.13 In vivo imaging (0)     3.13.1 Laser Scanning (0)       3.13.1.1 Confocal Scanning Laser Ophthalmoscopy (0)       3.13.1.2 Confocal Scanning Laser Polarimetry (0)     3.13.2 Optical Coherence Tomography (0)       3.13.2.1 Anterior Segment (0)       3.13.2.2 Posterior Segment (0)     3.13.3 RGC Imaging (1)     3.13.4 Other (0)   3.20 Other (0) 4 Tissue culture of ocular cells (0) 5 Experimental glaucoma; animal models (0)   5.1 Rodent (18)   5.2 Primates (3)   5.3 Other (9) 6 Clinical examination methods (0)   6.1 Intraocular pressure measurement; factors affecting IOP (1)     6.1.1 Devices, techniques (7)     6.1.2 Fluctuation, circadian rhythms (8)     6.1.3 Factors affecting IOP (10)   6.2 Tonography, aqueous flow measurement (see also 2.6) (0)   6.3 Biomicroscopy (slitlamp) (0)     6.3.1 Anterior segment (1)     6.3.2 Posterior segment (0)   6.4 Gonioscopy (3)   6.5 Ophthalmoscopy (0)   6.6 Visual field examination and other visual function tests (0)     6.6.1 Conventional manual (0)     6.6.2 Automated (10)     6.6.3 Special methods (e.g. color, contrast, SWAP etc.) (3)   6.7 Electro-ophthalmodiagnosis (7)   6.8 Photography (0)     6.8.1 Anterior segment (0)     6.8.2 Posterior segment (3)   6.9 Computerized image analysis (0)     6.9.1 Laser scanning (0)       6.9.1.1 Confocal Scanning Laser Ophthalmoscopy (7)       6.9.1.2 Confocal Scanning Laser Polarimetry (7)     6.9.2 Optical coherence tomography (0)       6.9.2.1 Anterior (12)       6.9.2.2 Posterior (32)     6.9.5 Other (1)   6.10 Fluorescein (ICG) angiography (0)     6.10.1 Anterior segment (0)     6.10.2 Posterior segment (0)   6.11 Bloodflow measurements (11)   6.12 Ultrasonography and ultrasound biomicroscopy (5)   6.13 Provocative tests (3)   6.19 Telemedicine (2)   6.20 Progression (13)   6.30 Other (4) 8 Refractive errors in relation to glaucoma (0)   8.1 Myopia (7)   8.2 Hypermetropia (0)   8.3 Contact lenses (0)   8.4 Refractive surgical procedures (0)   8.5 Other (0) 9 Clinical forms of glaucomas (0)   9.1 Developmental glaucomas (0)     9.1.1 Congenital glaucoma, Buphthalmos (13)     9.1.2 Juvenile glaucoma (8)     9.1.3 Syndromes of Axenfeld, Rieger, Peters, aniridia (2)     9.1.4 Other (0)   9.2 Primary open angle glaucomas (0)     9.2.1 Ocular hypertension (0)     9.2.2 Other risk factors for glaucoma (9)     9.2.3 Open angle glaucoma with elevated IOP (3)     9.2.4 Normal pressure glaucoma (12)   9.3 Primary angle closure glaucomas (0)     9.3.1 Acute primary angle closure glaucoma (pupillary block) (7)     9.3.2 Chronic primary angle closure glaucoma (pupillary block) (1)     9.3.3 Plateau iris syndrome (1)     9.3.4 Primary angle closure suspect (2)     9.3.5 Primary angle closure (10)     9.3.10 Other (0)   9.4 Glaucomas associated with other ocular and systemic disorders (0)     9.4.1 Steroid-induced glaucoma (3)     9.4.2 Glaucomas associated with disorders of the cornea, conjunctiva, sclera (0)       9.4.2.1 Iridocorneal endothelial syndrome (ICE, incl. irisatrophy) (1)       9.4.2.2 Posterior polymorphous dystrophy (0)       9.4.2.3 Fuchs' endothelial dystrophy (0)       9.4.2.5 Other (4)     9.4.3 Glaucomas associated with disorders of the iris and ciliary body (0)       9.4.3.1 Pigmentary glaucoma (2)       9.4.3.5 Other (0)     9.4.4 Glaucomas associated with disorders of the lens (0)       9.4.4.1 Exfoliation syndrome (9)       9.4.4.2 Glaucomas associated with cataracts (2)       9.4.4.3 Glaucomas associated with lens dislocation (2)       9.4.4.5 Other (2)     9.4.5 Glaucomas associated with disorders of the retina, choroid and vitreous (0)       9.4.5.1 Neovascular glaucoma (12)       9.4.5.5 Other (1)     9.4.6 Glaucomas associated with inflammation, uveitis (8)     9.4.7 Glaucomas associated with ocular trauma (1)     9.4.8 Glaucomas associated with intraocular tumors (5)     9.4.9 Glaucomas associated with elevated episcleral venous pressure (0)       9.4.10 Glaucomas associated with hemorrhage (3)     9.4.11 Glaucomas following intraocular surgery (0)       9.4.11.1 Ciliary block (malignant) glaucoma (6)       9.4.11.2 Glaucomas in aphakia and pseudophakia (6)       9.4.11.3 Epithelial, fibrous, and endothelial proliferation (0)       9.4.11.4 Glaucomas associated with corneal surgery (8)       9.4.11.5 Glaucomas associated with vitreoretinal surgery (7)     9.4.15 Glaucoma in relation to systemic disease (18)     9.4.20 Other (0) 10 Differential diagnosis e.g. anterior and posterior ischemic optic neuropathy (4) 11 Medical treatment (0)   11.1 General management, indication (4)   11.2 Cholinergic drugs (0)   11.3 Adrenergic drugs (0)     11.3.1 Epinephrine (0)     11.3.2 Thymoxamine, dapiprazole (0)     11.3.3 Apraclonidine, brimonidine (1)     11.3.4 Betablocker (6)     11.3.5 Other (0)   11.4 Prostaglandins (20)   11.5 Carbonic anhydrase inhibitors (0)     11.5.1 Systemic (3)     11.5.2 Topical (0)   11.6 Osmotic treatment (0)   11.7 Treatment of bloodflow (0)   11.8 Neuroprotection (18)   11.9 Gene therapy (2)   11.13 Combination therapy (0)     11.13.1 Betablocker and pilocarpine (0)     11.13.2 Betablocker and carbon anhydrase inhibitor (3)     11.13.3 Betablocker and brimonidine (2)     11.13.4 Betablocker and prostaglandin (0)     11.13.5 Other (1)   11.14 Investigational drugs; pharmacological experiments (12)   11.15 Other drugs in relation to glaucoma (7)   11.16 Vehicles, delivery systems, pharmacokinetics, formulation (19)   11.17 Cooperation with medical therapy e.g. persistency, compliance, adherence (4)   11.20 Other (1) 12 Surgical treatment (0)   12.1 General management, indication (0)   12.2 Laser iridotomy (8)   12.3 Laser iridoplasty (2)   12.4 Laser trabeculoplasty and other laser treatment of the angle (4)   12.7 Surgical iridectomy (1)   12.8 Filtering surgery (0)     12.8.1 Without tube implant (5)     12.8.2 With tube implant or other drainage devices (13)     12.8.3 Non-perforating (5)     12.8.4 Using laser (0)     12.8.5 Other (0)     12.8.10 Woundhealing antifibrosis (12)     12.8.11 Complications, endophthalmitis (8)   12.9 Trabeculotomy, goniotomy (1)   12.10 Cyclodestruction (5)   12.11 Cyclodialysis (0)   12.12 Cataract extraction (0)     12.12.1 Intracapsular (0)     12.12.2 Extracapsular (0)     12.12.3 Phacoemulsification (3)   12.14 Combined cataract extraction and glaucoma surgery (0)     12.14.1 Intracapsular (0)     12.14.2 Extracapsular (0)     12.14.3 Phacoemulsification (1)   12.15 Capsulotomy (0)   12.16 Vitrectomy (0)   12.17 Anesthesia (1)   12.20 Other (1) 13 Therapeutic prognosis and outcome (0)   13.1 Prognostic factors (2)   13.2 Outcome (0)     13.2.1 IOP (2)     13.2.2 Visual field (0)       13.2.2.1 Progression (0)       13.2.2.2 Improvement (0)     13.2.3 Other (1) 14 Costing studies; pharmacoeconomics (7) 15 Miscellaneous (13)

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Abstract #51820 Published in IGR 14-4

Assessment of mitochondrial damage in retinal cells and tissues using quantitative polymerase chain reaction for mitochondrial DNA damage and extracellular flux assay for mitochondrial respiration activity

Jarrett SG; Rohrer B; Perron NR; Beeson C; Boulton ME
Methods in molecular biology (Clifton, N.J.) 2013; 935: 227-243

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Mitochondrial dysfunction and genomic instability are associated with a number of retinal pathologies including age-related macular degeneration, diabetic retinopathy, and glaucoma. Consequences of mitochondrial dysfunction within cells include elevation of the rate of ROS production due to damage of electron transport chain proteins, mitochondrial DNA (mtDNA) damage, and loss of metabolic capacity. Here we introduce the quantitative polymerase chain reaction assay (QPCR) and extracellular flux assay (XF) as powerful techniques to study mitochondrial behavior. The QPCR technique is a gene-specific assay developed to analyze the DNA damage repair response in mitochondrial and nuclear genomes. QPCR has proved particularly valuable for the measurement of oxidative-induced mtDNA damage and kinetics of mtDNA repair. To assess the functional consequence of mitochondrial oxidative damage, real-time changes in cellular bioenergetics of cell monolayers can be measured with a Seahorse Biosciences XF24 analyzer. The advantages and limitations of these procedures will be discussed and detailed methodologies provided with particular emphasis on retinal oxidative stress.

Department of Molecular and Biomedical Pharmacology, College of Medicine, University of Kentucky, Lexington, KY, USA.

Full article

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Classification:

3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
3.6 Cellular biology (Part of: 3 Laboratory methods)

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