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Transgenically expressed Parascaris P-glycoprotein-11 can modulate ivermectin susceptibility in Caenorhabditis elegans.

Janssen IJ, Krücken J, Demeler J, von Samson-Himmelstjerna G - Int J Parasitol Drugs Drug Resist (2015)

Bottom Line: This association was recently shown for Parascaris Pgp-11.Ivermectin resistance was correlated with the presence of three pgp-11 single nucleotide polymorphisms and/or increased pgp-11 mRNA levels.This is the first report on the successful functional analysis of a parasitic nematode Pgp in the model organism C. elegans.

View Article: PubMed Central - PubMed

Affiliation: Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.

ABSTRACT
P-glycoproteins (Pgps) are suspected to mediate drug extrusion in nematodes contributing to macrocyclic lactone resistance. This association was recently shown for Parascaris Pgp-11. Ivermectin resistance was correlated with the presence of three pgp-11 single nucleotide polymorphisms and/or increased pgp-11 mRNA levels. In the present study, the ability of Pgp-11 to modulate ivermectin susceptibility was investigated by its expression in a pgp-11-deficient Caenorhabditis elegans strain. Expression of Parascaris pgp-11 in two transgenic lines significantly decreased ivermectin susceptibility in a motility (thrashing) assay conducted in liquid medium. The EC50 values increased by 3.2- and 4.6-fold in the two lines relative to a transgenic control strain. This is the first report on the successful functional analysis of a parasitic nematode Pgp in the model organism C. elegans.

No MeSH data available.


Related in: MedlinePlus

Concentration–response curves to ivermectin of the control and Parascaris pgp-11 transgenic Caenorhabditis elegans in a pgp-11-deficient genetic background (tm0333). After incubation for 18 h in a medium containing various concentrations of ivermectin or only the vehicle DMSO (1%), the motilities of C. elegans worms were recorded for 1 min in liquid medium containing the same drug concentration. Transgenic lines were produced by transformation with pgp-11 expression constructs (Cel-pgp-11::Parascaris-pgp-11(1) (triangle), Cel-pgp-11::Parascaris-pgp-11(2) (square)) or the construct lacking the pgp-11 cDNA (Cel-pgp-11::control (circles)). The motility of single worms was assessed as body bends per minute. The negative control without IVM was set to 10−11 M to allow log10 transformation of the concentrations. Values represent means ± standard error of the mean of at least 12 worms. The bottom and top values for four-parameter logistic regression were constrained to values between 0 and 100%.
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f0010: Concentration–response curves to ivermectin of the control and Parascaris pgp-11 transgenic Caenorhabditis elegans in a pgp-11-deficient genetic background (tm0333). After incubation for 18 h in a medium containing various concentrations of ivermectin or only the vehicle DMSO (1%), the motilities of C. elegans worms were recorded for 1 min in liquid medium containing the same drug concentration. Transgenic lines were produced by transformation with pgp-11 expression constructs (Cel-pgp-11::Parascaris-pgp-11(1) (triangle), Cel-pgp-11::Parascaris-pgp-11(2) (square)) or the construct lacking the pgp-11 cDNA (Cel-pgp-11::control (circles)). The motility of single worms was assessed as body bends per minute. The negative control without IVM was set to 10−11 M to allow log10 transformation of the concentrations. Values represent means ± standard error of the mean of at least 12 worms. The bottom and top values for four-parameter logistic regression were constrained to values between 0 and 100%.

Mentions: A statistically significant increase (P < 0.0001) in the IVM EC50 value was observed in the thrashing assay for both lines injected with the Parascaris pgp-11 expression construct, Cel-pgp-11::Parascaris-pgp-11(1) and Cel-pgp-11::Parascaris-pgp-11(2), relative to the control line Cel-pgp-11::control (Table 1 and Fig. 1). The EC50 values were increased by approximately 4.6- and 3.2-fold in the two expression constructs.


Transgenically expressed Parascaris P-glycoprotein-11 can modulate ivermectin susceptibility in Caenorhabditis elegans.

Janssen IJ, Krücken J, Demeler J, von Samson-Himmelstjerna G - Int J Parasitol Drugs Drug Resist (2015)

Concentration–response curves to ivermectin of the control and Parascaris pgp-11 transgenic Caenorhabditis elegans in a pgp-11-deficient genetic background (tm0333). After incubation for 18 h in a medium containing various concentrations of ivermectin or only the vehicle DMSO (1%), the motilities of C. elegans worms were recorded for 1 min in liquid medium containing the same drug concentration. Transgenic lines were produced by transformation with pgp-11 expression constructs (Cel-pgp-11::Parascaris-pgp-11(1) (triangle), Cel-pgp-11::Parascaris-pgp-11(2) (square)) or the construct lacking the pgp-11 cDNA (Cel-pgp-11::control (circles)). The motility of single worms was assessed as body bends per minute. The negative control without IVM was set to 10−11 M to allow log10 transformation of the concentrations. Values represent means ± standard error of the mean of at least 12 worms. The bottom and top values for four-parameter logistic regression were constrained to values between 0 and 100%.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401813&req=5

f0010: Concentration–response curves to ivermectin of the control and Parascaris pgp-11 transgenic Caenorhabditis elegans in a pgp-11-deficient genetic background (tm0333). After incubation for 18 h in a medium containing various concentrations of ivermectin or only the vehicle DMSO (1%), the motilities of C. elegans worms were recorded for 1 min in liquid medium containing the same drug concentration. Transgenic lines were produced by transformation with pgp-11 expression constructs (Cel-pgp-11::Parascaris-pgp-11(1) (triangle), Cel-pgp-11::Parascaris-pgp-11(2) (square)) or the construct lacking the pgp-11 cDNA (Cel-pgp-11::control (circles)). The motility of single worms was assessed as body bends per minute. The negative control without IVM was set to 10−11 M to allow log10 transformation of the concentrations. Values represent means ± standard error of the mean of at least 12 worms. The bottom and top values for four-parameter logistic regression were constrained to values between 0 and 100%.
Mentions: A statistically significant increase (P < 0.0001) in the IVM EC50 value was observed in the thrashing assay for both lines injected with the Parascaris pgp-11 expression construct, Cel-pgp-11::Parascaris-pgp-11(1) and Cel-pgp-11::Parascaris-pgp-11(2), relative to the control line Cel-pgp-11::control (Table 1 and Fig. 1). The EC50 values were increased by approximately 4.6- and 3.2-fold in the two expression constructs.

Bottom Line: This association was recently shown for Parascaris Pgp-11.Ivermectin resistance was correlated with the presence of three pgp-11 single nucleotide polymorphisms and/or increased pgp-11 mRNA levels.This is the first report on the successful functional analysis of a parasitic nematode Pgp in the model organism C. elegans.

View Article: PubMed Central - PubMed

Affiliation: Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.

ABSTRACT
P-glycoproteins (Pgps) are suspected to mediate drug extrusion in nematodes contributing to macrocyclic lactone resistance. This association was recently shown for Parascaris Pgp-11. Ivermectin resistance was correlated with the presence of three pgp-11 single nucleotide polymorphisms and/or increased pgp-11 mRNA levels. In the present study, the ability of Pgp-11 to modulate ivermectin susceptibility was investigated by its expression in a pgp-11-deficient Caenorhabditis elegans strain. Expression of Parascaris pgp-11 in two transgenic lines significantly decreased ivermectin susceptibility in a motility (thrashing) assay conducted in liquid medium. The EC50 values increased by 3.2- and 4.6-fold in the two lines relative to a transgenic control strain. This is the first report on the successful functional analysis of a parasitic nematode Pgp in the model organism C. elegans.

No MeSH data available.


Related in: MedlinePlus