The Role of Mitochondria, Oxidative Stress, and the Radical-binding Protein A1M in Cultured Porcine Retina
| Autor: | Jesper Bergwik, Oscar Manouchehrian, Bo Åkerström, Martin Cederlund, Ingrid Holmgren Taylor, Linnéa Taylor, Fredrik Ghosh, Karin Arnér |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Survival Swine Oxidative phosphorylation Mitochondrion Biology medicine.disease_cause Real-Time Polymerase Chain Reaction 03 medical and health sciences Cellular and Molecular Neuroscience chemistry.chemical_compound Lactate dehydrogenase Alpha-Globulins medicine In Situ Nick-End Labeling Animals Viability assay Cell damage Cells Cultured Retinal Inner limiting membrane medicine.disease Immunohistochemistry Sensory Systems Cell biology Mitochondria Ophthalmology Disease Models Animal Oxidative Stress 030104 developmental biology medicine.anatomical_structure chemistry sense organs Carrier Proteins Oxidative stress Cell and Molecular Biology |
| Zdroj: | Current Eye Research; 42(6), pp 948-961 (2017) |
| ISSN: | 1460-2202 0271-3683 |
| Popis: | PURPOSE:The purpose of this study was to explore the relationship between oxidative stress, antioxidant defense, mitochondrial structure, and biomechanical tissue support in the isolated porcine retina.METHODS:Full-thickness retinal sheets were isolated from adult porcine eyes. Retinas were cultured for 2 or 48 h using (1) a previously established low-support explant protocol with photoreceptors positioned against the culture membrane (porous polycarbonate) or (2) a high-support procedure developed by our group, apposing the Muller cell endfeet and inner limiting membrane against the membrane. The grafts were analyzed by quantitative polymerase chain reaction (PCR), immunohistochemistry, and transmission electron microscopy (TEM), and culture medium was assayed for the cell damage and oxidative stress markers lactate dehydrogenase and protein carbonyls.RESULTS:In explants cultured with physical support to the inner border, cone photoreceptors were preserved and lactate dehydrogenase levels were reduced, although an initial (2 h), transient, increased oxidative stress was observed. Elevated expression of the antioxidants α1-microglobulin and heme oxygenase-1 was seen in the mitochondria-rich inner segments after 48 h compared to low-support counterparts. Housekeeping gene expression suggested a higher degree of structural integrity of mitochondria in high-support explants, and TEM of inner segments confirmed preservation of a normal mitochondrial morphology.CONCLUSION:Providing retinal explants with inner retinal support leads to mobilization of antioxidant proteins, preservation of mitochondrial function, and increased cell viability. Consequently, the failure of low-support retinal cultures to mobilize an adequate response to the oxidative environment may play a key role in their rapid demise. These findings shed new light on pathological reactions in biomechanically related conditions in vivo. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |