Use of a potentiometric vital dye to determine the effect of the herbicide bromoxynil octanoate on mitochondrial bioenenergetics in <i>Chlamydomonas reinhardtii</i>

Brickley, M R; Lawrie, Emily; Weise, Veronika; Hawes, Chris; Cobb, A. H.

doi:10.1002/ps.2298

Cited by 9 publications

(12 citation statements)

References 25 publications

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“…That is Cu(II) at intermediate and high concentrations appears to have an ionophore-like effect, which reduces ΔΨm and most likely inhibits respiration. A similar effect (collapse of the mitochondrial membrane potential) was reported in Chlamydomonas reinhardtii exposed to the herbicide bromoxynil octanoate (Brickley et al 2012).…”

Section: Discussionsupporting

confidence: 74%

“…According to the chemiosmotic theory, the passage of electrons through the electron transport chain in the inner mitochondrial membrane generates a transmembrane electrochemical gradient that is required for ATP production by the enzyme ATP synthase (Jastroch et al 2010). Rh123 appears to be a sensitive and useful potentiometric dye for use in the investigation of mitochondrial bioenergetics (Brickley et al 2012). The complete loss of mitochondrial respiratory function due to the treatment of P. subcapitata cells with sodium azide (an electron transport chain inhibitor) led to the collapse of ΔΨm.…”

Section: Discussionmentioning

confidence: 99%

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Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

Machado

Soares

2014

J Appl Phycol

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This work explores the use of fluorescent probes to evaluate the responses of the green alga Pseudokirchneriella subcapitata to the action of three nominal concentrations of Cd(II), Cr(VI), Cu(II) and Zn(II) for a short time (6 h). The toxic effect of the metals on algal cells was monitored using the fluorochromes SYTOX Green (SG, membrane integrity), fluorescein diacetate (FDA, esterase activity) and rhodamine 123 (Rh123, mitochondrial membrane potential). The impact of metals on chlorophyll a (Chl a) autofluorescence was also evaluated. Esterase activity was the most sensitive parameter. At the concentrations studied, all metals induced the loss of esterase activity. SG could be used to effectively detect the loss of membrane integrity in algal cells exposed to 0.32 or 1.3 μmol L −1 Cu(II). Rh123 revealed a decrease in the mitochondrial membrane potential of algal cells exposed to 0.32 and 1.3 μmol L −1 Cu(II), indicating that mitochondrial activity was compromised. Chl a autofluorescence was also affected by the presence of Cr(VI) and Cu(II), suggesting perturbation of photosynthesis. In conclusion, the fluorescence-based approach was useful for detecting the disturbance of specific cellular characteristics. Fluorescent probes are a useful diagnostic tool for the assessment of the impact of toxicants on specific targets of P. subcapitata algal cells.

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Section: Discussionsupporting

confidence: 74%

Section: Discussionmentioning

confidence: 99%

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

Machado

Soares

2014

J Appl Phycol

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“…This fluorescent probe have been used in mitochondrial function studies with a variety of cell types, such as animal [54,55], plant [56], yeast [43] and algal cells [57,58]. In recent years, bioenergetic studies in algal cells carried out with Rh123 were validated using a polarographic oxygen electrode [44].…”

Section: Discussionmentioning

confidence: 99%

“…Rh123 enters freely the cell by simple diffusion through the plasma membrane and is electrophoretically distributed in the mitochondrial matrix in response to mitochondrial membrane potential ( m ) [42,43]. It was shown that Rh123 is a reliable method for mitochondrial function studies with algal cells [44]. In this context, Rh123 was used to investigate the changes in the mitochondrial bioenergetics in response to the presence of heavy metals.…”

Section: Impact Of Heavy Metals On Esterase Activity and Mitochondriamentioning

confidence: 99%

Responses of the alga Pseudokirchneriella subcapitata to long-term exposure to metal stress

Machado

Lopes

Soares

2015

Journal of Hazardous Materials

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The green alga Pseudokirchneriella subcapitata has been widely used in ecological risk assessment, usually based on the impact of the toxicants in the alga growth. However, the physiological causes that lead algal growth inhibition are not completely understood. This work aimed to evaluate the biochemical and structural modifications in P. subcapitata after exposure, for 72 h, to three nominal concentrations of Cd(II), Cr(VI), Cu(II) and Zn(II), corresponding approximately to 72 h-EC10 and 72 h-EC50 values and a high concentration (above 72 h-EC90 values). The incubation of algal cells with the highest concentration of Cd(II), Cr(VI) or Cu(II) resulted in a loss of membrane integrity of ~16, 38 and 55%, respectively. For all metals tested, an inhibition of esterase activity, in a dose-dependent manner, was observed. Reduction of chlorophyll a content, decrease of maximum quantum yield of photosystem II and modification of mitochondrial membrane potential was also verified. In conclusion, the exposure of P. subcapitata to metals resulted in a perturbation of the cell physiological status. Principal component analysis revealed that the impairment of esterase activity combined with the reduction of chlorophyll a content were related with the inhibition of growth caused by a prolonged exposure to the heavy metals.

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“…Mitochondrial area is a good approximation of the area of mitochondrial membrane, and changes in mitochondrial potential can be measured in live cells using potentiometric dye. Previous work 45 has shown that the styrl dye rhodamine 123 (R123) can be used in live cells of phytoplankton to measure mitochondrial potential in real time. For staining, a 10 mg mL −1 stock solution in DMSO was prepared and diluted to a 1μM working solution on the day of measurements.…”

Section: Methodsmentioning

confidence: 99%

Environmental fluctuations accelerate molecular evolution of thermal tolerance in a marine diatom

Schaum

Buckling

Smirnoff

et al. 2017

Preprint

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Diatoms contribute roughly 20% of global primary production, but the factors determining their ability to adapt to global warming are unknown. Here we quantify the capacity for adaptation to warming in the marine diatom Thalassiosira pseudonana. We found that evolutionary rescue under severe warming was slow but adaptation to more realistic scenarios, where temperature increases were moderate or where they fluctuated between benign and severe conditions, was rapid. Adaption to warming was linked to major phenotypic changes in metabolism and elemental composition. Whole genome re-sequencing identified significant genetic divergence both among populations adapted to the different warming regimes and between the evolved and ancestral lineages. Consistent with the phenotypic changes, the most rapidly evolving genes were associated with transcriptional regulation, cellular response to oxidative stress and redox homeostasis. These results demonstrate that evolution of thermal tolerance in marine diatoms can be rapid, particularly in fluctuating environments, and is underpinned by major genomic and phenotypic divergence.

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Use of a potentiometric vital dye to determine the effect of the herbicide bromoxynil octanoate on mitochondrial bioenenergetics in Chlamydomonas reinhardtii

Abstract: R123 is a sensitive potentiometric dye in C. reinhardtii that may find further use in investigations of both mitochondrial bioenergetics in plants and environmental toxicology.

Cited by 9 publications

References 25 publications

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

Responses of the alga Pseudokirchneriella subcapitata to long-term exposure to metal stress

Environmental fluctuations accelerate molecular evolution of thermal tolerance in a marine diatom

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