Functional and energetic characterization of P-gp-mediated doxorubicin transport in rainbow trout (Oncorhynchus mykiss) hepatocytes

Hildebrand, Jennifer; Bains, Onkar S.; Lee, Denny S.H.; Kennedy, Christopher J.

doi:10.1016/j.cbpc.2008.07.003

Cited by 16 publications

(9 citation statements)

References 90 publications

(97 reference statements)

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“…Several studies on freshly isolated suspensions of fish hepatocytes also reported high costs associated with transporter activity. These studies showed an 18-25% increase in respiration in the presence of the transporter substrate rhodamine 123 (R123) (Bains and Kennedy, 2005), and a similar percent decrease in steady-state concentrations of ATP in the presence of 125μmoll −1 doxorubicin (Hildebrand et al, 2009). However, Bow et al have reported that freshly isolated hepatocytes exhibit rapid internalization of membrane-bound ABCB1 and ABCC2 transporter proteins, causing the authors to conclude that suspended hepatocytes 'give an inaccurate assessment of true apical elimination' (Bow et al, 2008), and thus also of transporter costs.…”

Section: Discussionmentioning

confidence: 90%

“…Initial attempts to measure costs of transporter activity used drugresistant cell lines or proteoliposomes, and indicated high costs associated with efflux transporter activity (Bains and Kennedy, 2005;Broxterman et al, 1988;Broxterman et al, 1989;Hildebrand et al, 2009). However, the actual cost of this defense has not previously been determined in a naturally occurring system under environmentally relevant conditions.…”

Section: Introductionmentioning

confidence: 99%

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Cost, effectiveness and environmental relevance of multidrug transporters in sea urchin embryos

Cole

Hamdoun

Epel

2013

Journal of Experimental Biology

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SUMMARY ATP-binding cassette transporters protect cells via efflux of xenobiotics and endogenous byproducts of detoxification. While the cost of this ATP-dependent extrusion is known at the molecular level, i.e. the ATP used for each efflux event, the overall cost to a cell or organism of operating this defense is unclear, especially as the cost of efflux changes depending on environmental conditions. During prolonged exposure to xenobiotics, multidrug transporter activity could be costly and ineffective because effluxed substrate molecules are not modified in the process and could thus undergo repeated cycles of efflux and re-entry. Here we use embryos of the purple sea urchin, Strongylocentrotus purpuratus, as a model to determine transport costs and benefits under environmentally relevant xenobiotic concentrations. Strikingly, our results show that efflux transporter activity costs less than 0.2% of total ATP usage, as a proportion of oxygen consumption. The benefits of transport, defined as the reduction in substrate accumulation due to transporter activity, depended largely, but not entirely, on the rate of passive flux of each substrate across the plasma membrane. One of the substrates tested exhibited rapid membrane permeation coupled with high rates of efflux, thus inducing rapid and futile cycles of efflux followed by re-entry of the substrate. This combination significantly reduced transporter effectiveness as a defense and increased costs even at relatively low substrate concentrations. Despite these effects with certain substrates, our results show that efflux transporters are a remarkably effective and low-cost first line of defense against exposure to environmentally relevant concentrations of xenobiotics.

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Section: Discussionmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Cost, effectiveness and environmental relevance of multidrug transporters in sea urchin embryos

Cole

Hamdoun

Epel

2013

Journal of Experimental Biology

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“…The cultures were collected by centrifugation (10,000 rpm) for 6 min at room temperature. Phosphate in culture supernatant was estimated using "QuantiChrom TM Phosphate Assay Kit (DIPI-500)" (Hildebrand et al, 2009). …”

Section: Quantification Of Soluble Phosphate and Ph In Liquid Mediamentioning

confidence: 99%

Biocontrol of Fusarium spp., Causal Agents of Damping-off in Cotton Plants by Native Bacillus subtilis Isolated from Prosopis juliflora

Abdelmoteleb

Troncoso‐Rojas

Tzintzun-Camacho

et al. 2017

IJAB

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The present study was conducted to evaluate the application of native phosphate solubilizing Bacillus sp., isolated from Prosopis juliflora rhizosphere in the biocontrol of three different Fusarium species. The native Bacillus sp., was further confirmed as B. subtilis using the Polymerase Chain Reaction (PCR). The results showed that the phosphate solubilizing ability of B. subtilis strain ALICA was tested in liquid medium with 0.5% Ca3(PO4)2, and maximum of the value of soluble phosphate was 202.02 µg/mL after 3 days of incubation. On the other hand, the growth inhibition of Fusarium solani, F. equiseti and F. oxysporum were more evident in cell-free medium filtrates with respect to dual medium with a range of percentage inhibition from 51, 66 and 47% after 5 days, respectively. Our result of the DNA-PCR amplification of the lipopeptide genes from B. subtilis strain ALICA, showed 94% identity with subtilosin and subtilisin of different Bacillus spp. This result turns B. subtilis strain ALICA into a potential alternative for the biological control of Fusarium species present in the soil of transgenic insect-resistant cotton plants in Mexico.

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“…Primary hepatocytes obtained from trout, like those from mammals, largely maintain their epithelial phenotype, including functional glucose and lipid metabolism (Polakof et al, 2011), and the activities of phase I and II biotransformation enzymes (Segner and Cravedi, 2001). These cells possess functional membrane transporters, and have been studied using well-known transporter substrates and inhibitors (Sturm et al, 2001;Bains and Kennedy, 2005;Zaja et al, 2008;Hildebrand et al, 2009). Additional studies using primary trout hepatocytes have examined the effects of carcinogens (Bailey et al, 1987), hormones (Bailey et al, 1987;Sathiyaa et al, 2001), naturally derived toxins (Boaru et al, 2006), pharmaceuticals (Ellesat et al, 2010;Ings et al, 2012), nanoparticles (Massarsky et al, 2014), and a variety of other xenobiotic compounds or mixtures (Gagné et al, 2013;Lacaze et al, 2014;Sovadinová et al, 2014).…”

Section: Background Informationmentioning

confidence: 99%

Determination of Metabolic Stability Using Cryopreserved Hepatocytes from Rainbow Trout (Oncorhynchus mykiss)

et al. 2015

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Trout provide a relatively easy source of hepatocytes that can be cryopreserved and used for a range of applications including toxicity testing and determination of intrinsic clearance. Standard protocols for isolating, cryopreserving, and thawing rainbow trout hepatocytes are described, along with procedures for using fresh or cryopreserved hepatocytes to assess metabolic stability of xenobiotics in fish by means of a substrate depletion approach. Variations on these methods, troubleshooting tips, and directions for use of extrapolation factors to express results in terms of in vivo intrinsic clearance are included. These protocols have been developed for rainbow trout, but can be adapted to other fish species with appropriate considerations.

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Functional and energetic characterization of P-gp-mediated doxorubicin transport in rainbow trout (Oncorhynchus mykiss) hepatocytes

Cited by 16 publications

References 90 publications

Cost, effectiveness and environmental relevance of multidrug transporters in sea urchin embryos

Cost, effectiveness and environmental relevance of multidrug transporters in sea urchin embryos

Biocontrol of Fusarium spp., Causal Agents of Damping-off in Cotton Plants by Native Bacillus subtilis Isolated from Prosopis juliflora

Determination of Metabolic Stability Using Cryopreserved Hepatocytes from Rainbow Trout (Oncorhynchus mykiss)

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