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| Salin, Karine; Auer, Sonya K.; Villasevil, Eugenia M.; Anderson, Graeme J.; Cairns, Andrew G.; Mullen, William; Hartley, Richard C.; Metcalfe, Neil B.. |
| In recent years evolutionary ecologists have become increasingly interested in the effects of reactive oxygen species (ROS) on the life-histories of animals. ROS levels have mostly been inferred indirectly due to the limitations of estimating ROS from in vitro methods. However, measuring ROS (hydrogen peroxide, H2O2) content in vivo is now possible using the MitoB probe. Here, we extend and refine the MitoB method to make it suitable for ecological studies of oxidative stress using the brown trout Salmo trutta as model. The MitoB method allows an evaluation of H2O2 levels in living organisms over a timescale from hours to days. The method is flexible with regard to the duration of exposure and initial concentration of the MitoB probe, and there is no... |
| Tipo: Text |
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| Ano: 2017 |
URL: https://archimer.ifremer.fr/doc/00413/52494/53297.pdf |
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| Salin, Karine; Villasevil, Eugenia M.; Auer, Sonya K.; Anderson, Graeme J.; Selman, Colin; Metcalfe, Neil B.; Chinopoulos, Christos. |
| The use of tissue homogenate has greatly aided the study of the functioning of mitochondria. However, the amount of ATP produced per oxygen molecule consumed, that is, the effective P/O ratio, has never been measured directly in tissue homogenate. Here we combine and refine existing methods previously used in permeabilized cells and isolated mitochondria to simultaneously measure mitochondrial ATP production (JATP) and oxygen consumption (JO(2)) in tissue homogenate. A major improvement over existing methods is in the control of ATPases that otherwise interfere with the ATP assay: our modified technique facilitates simultaneous measurement of the rates of uncorrected ATP synthesis and of ATP hydrolysis, thus minimizing the amount of tissue and time needed.... |
| Tipo: Text |
Palavras-chave: ATPase; Fluorescence; Magnesium green; Oxidative phosphorylation; Oxygraph. |
| Ano: 2016 |
URL: https://archimer.ifremer.fr/doc/00414/52500/53300.pdf |
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| Salin, Karine; Villasevil, Eugenia M.; Anderson, Graeme J.; Auer, Sonya K.; Selman, Colin; Hartley, Richard C.; Mullen, William; Chinopoulos, Christos; Metcalfe, Neil B.. |
| Many animals experience periods of food shortage in their natural environment. It has been hypothesised that the metabolic responses of animals to naturally‐occurring periods of food deprivation may have long‐term negative impacts on their subsequent life‐history. In particular, reductions in energy requirements in response to fasting may help preserve limited resources but potentially come at a cost of increased oxidative stress. However, little is known about this trade‐off since studies of energy metabolism are generally conducted separately from those of oxidative stress. Using a novel approach that combines measurements of mitochondrial function with in vivo levels of hydrogen peroxide (H2O2) in brown trout (Salmo trutta), we show here that fasting... |
| Tipo: Text |
Palavras-chave: High-resolution respirometry; In vivo; Liver atrophy; MitoB probe; Mitochondrial respiratory state. |
| Ano: 2018 |
URL: https://archimer.ifremer.fr/doc/00449/56022/57540.pdf |
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| Salin, Karine; Villasevil, Eugenia M.; Anderson, Graeme J.; Selman, Colin; Chinopoulos, Christos; Metcalfe, Neil B.. |
| Mitochondrial efficiency is typically taken to represent an animal’s capacity to convert its resources into ATP. However, the term mitochondrial efficiency, as currently used in the literature, can be calculated as either the respiratory control ratio, RCR (ratio of mitochondrial respiration supporting ATP synthesis to that required to offset the proton leak) or as the amount of ATP generated per unit of oxygen consumed, ATP/O ratio. The question of how flexibility in mitochondrial energy properties (i.e. in rates of respiration to support ATP synthesis and offset proton leak, and in the rate of ATP synthesis) affects these indices of mitochondrial efficiency has tended to be overlooked. Furthermore, little is known of whether the RCR and ATP/O ratio vary... |
| Tipo: Text |
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| Ano: 2018 |
URL: https://archimer.ifremer.fr/doc/00449/56014/57529.pdf |
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