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Oxidative Stress in Caenorhabditis elegans: Protective Effects of Spartin.

Truong T, Karlinski ZA, O'Hara C, Cabe M, Kim H, Bakowska JC - PLoS ONE (2015)

Bottom Line: These results suggest an age-dependent decline in motor function in mutant animals.In the behavioral assays, spg-20 mutant animals showed a significant decrease in both crawling speed and thrashing frequency compared with wild-type animals.These data suggest that the protein F57B10.9/SPG-20 might have a protective role against oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pharmacology and Therapeutics, Loyola University Chicago, Maywood, Illinois, United States of America.

ABSTRACT
Troyer syndrome is caused by a mutation in the SPG20 gene, which results in complete loss of expression of the protein spartin. We generated a genetic model of Troyer syndrome in worms to explore the locomotor consequences of a mutation of the Caenorhabditis elegans SPG20 orthologue, F57B10.9, also known as spg-20. Spg-20 mutants showed decreased length, crawling speed, and thrashing frequency, and had a shorter lifespan than wild-type animals. These results suggest an age-dependent decline in motor function in mutant animals. The drug paraquat was used to induce oxidative stress for 4 days in the animals. We measured survival rate and examined locomotion by measuring crawling speed and thrashing frequency. After 4 days of paraquat exposure, 77% of wild-type animals survived, but only 38% of spg-20 mutant animals survived. Conversely, animals overexpressing spg-20 had a survival rate of 95%. We also tested lifespan after a 1 hour exposure to sodium azide. After a 24 hour recovery period, 87% of wild type animals survived, 57% of spg-20 mutant animals survived, and 82% of animals overexpressing spg-20 survived. In the behavioral assays, spg-20 mutant animals showed a significant decrease in both crawling speed and thrashing frequency compared with wild-type animals. Importantly, the locomotor phenotype for both crawling and thrashing was rescued in animals overexpressing spg-20. The animals overexpressing spg-20 had crawling speeds and thrashing frequencies similar to those of wild-type animals. These data suggest that the protein F57B10.9/SPG-20 might have a protective role against oxidative stress.

No MeSH data available.


Related in: MedlinePlus

spg-20 mutant animals are sensitive to oxidative stress.C. elegans transgenic animals expressing GFP were grown on 2 mM paraquat plates for 4 days. The L4 stage was set as Day 0. (A) Percent survival of animals after treatment with 2 mM paraquat. The percentage of animals alive was scored every 24 hours for 4 days. Animals overexpressing spg-20 had a higher survival rate than both wild-type and spg-20 mutant animals (*p<0.0001). Survival data represent at least 100 animals per genotype from four independent trials. Error bars represent SEM; Kaplan-Meier method. (B) Percent survival of animals after treatment with 100 mM sodium azide for 1 hour. The percentage of animals alive was scored after a 24 hour recovery period. Spg-20(tm5514) mutant animals had decreased survival when compared to wild type animals (*p<0.001) and animals overexpressing spartin (*p< 0.001). There was no significant difference found between the survival of wild type animals and animals overexpressing spartin. (C) Average speed of transgenic animals measured each day for 4 days after treatment with 2 mM paraquat. spg-20 mutant animals showed reduced speed, whereas animals overexpressing spg-20 showed similar speed compared with WT animals (*p<0.05). Speed was quantified by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 50. (D) Thrashing frequency (body bends per second) of transgenic animals in M9 liquid on Day 4 of treatment with 2 mM paraquat. Spg-20 mutant animals had a reduced thrashing rate, whereas animals overexpressing spg-20 showed a recovery in thrashing rate compared to WT animals (*p<0.01). Thrashing frequency was measured by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 30.
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pone.0130455.g004: spg-20 mutant animals are sensitive to oxidative stress.C. elegans transgenic animals expressing GFP were grown on 2 mM paraquat plates for 4 days. The L4 stage was set as Day 0. (A) Percent survival of animals after treatment with 2 mM paraquat. The percentage of animals alive was scored every 24 hours for 4 days. Animals overexpressing spg-20 had a higher survival rate than both wild-type and spg-20 mutant animals (*p<0.0001). Survival data represent at least 100 animals per genotype from four independent trials. Error bars represent SEM; Kaplan-Meier method. (B) Percent survival of animals after treatment with 100 mM sodium azide for 1 hour. The percentage of animals alive was scored after a 24 hour recovery period. Spg-20(tm5514) mutant animals had decreased survival when compared to wild type animals (*p<0.001) and animals overexpressing spartin (*p< 0.001). There was no significant difference found between the survival of wild type animals and animals overexpressing spartin. (C) Average speed of transgenic animals measured each day for 4 days after treatment with 2 mM paraquat. spg-20 mutant animals showed reduced speed, whereas animals overexpressing spg-20 showed similar speed compared with WT animals (*p<0.05). Speed was quantified by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 50. (D) Thrashing frequency (body bends per second) of transgenic animals in M9 liquid on Day 4 of treatment with 2 mM paraquat. Spg-20 mutant animals had a reduced thrashing rate, whereas animals overexpressing spg-20 showed a recovery in thrashing rate compared to WT animals (*p<0.01). Thrashing frequency was measured by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 30.

Mentions: To further investigate the protective effects of spg-20 under oxidative stress, we examined three strains of C. elegans; wild-type, spg-20(tm5514) mutants, and animals overexpressing spg-20, all of which expressed green fluorescence protein (GFP). All three of these strains were exposed to 2 mM paraquat for 4 days. We observed that spg-20 mutant animals had a lower survival rate than wild-type animals (p<0.0001), and animals overexpressing spg-20 had a higher survival rate than both wild-type animals (p<0.001) and spg-20 mutant animals (p<0.0001) (Fig 4A). After 4 days of exposure to paraquat, we determined that 77% of wild-type animals survived, 38% of spg-20 mutant animals survived, and 95% of animals overexpressing spg-20 survived (S1 Table). Because animals overexpressing spartin showed significantly longer survival than wild-type animals, we propose that spartin plays an important role in the oxidative stress response.


Oxidative Stress in Caenorhabditis elegans: Protective Effects of Spartin.

Truong T, Karlinski ZA, O'Hara C, Cabe M, Kim H, Bakowska JC - PLoS ONE (2015)

spg-20 mutant animals are sensitive to oxidative stress.C. elegans transgenic animals expressing GFP were grown on 2 mM paraquat plates for 4 days. The L4 stage was set as Day 0. (A) Percent survival of animals after treatment with 2 mM paraquat. The percentage of animals alive was scored every 24 hours for 4 days. Animals overexpressing spg-20 had a higher survival rate than both wild-type and spg-20 mutant animals (*p<0.0001). Survival data represent at least 100 animals per genotype from four independent trials. Error bars represent SEM; Kaplan-Meier method. (B) Percent survival of animals after treatment with 100 mM sodium azide for 1 hour. The percentage of animals alive was scored after a 24 hour recovery period. Spg-20(tm5514) mutant animals had decreased survival when compared to wild type animals (*p<0.001) and animals overexpressing spartin (*p< 0.001). There was no significant difference found between the survival of wild type animals and animals overexpressing spartin. (C) Average speed of transgenic animals measured each day for 4 days after treatment with 2 mM paraquat. spg-20 mutant animals showed reduced speed, whereas animals overexpressing spg-20 showed similar speed compared with WT animals (*p<0.05). Speed was quantified by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 50. (D) Thrashing frequency (body bends per second) of transgenic animals in M9 liquid on Day 4 of treatment with 2 mM paraquat. Spg-20 mutant animals had a reduced thrashing rate, whereas animals overexpressing spg-20 showed a recovery in thrashing rate compared to WT animals (*p<0.01). Thrashing frequency was measured by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 30.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4482654&req=5

pone.0130455.g004: spg-20 mutant animals are sensitive to oxidative stress.C. elegans transgenic animals expressing GFP were grown on 2 mM paraquat plates for 4 days. The L4 stage was set as Day 0. (A) Percent survival of animals after treatment with 2 mM paraquat. The percentage of animals alive was scored every 24 hours for 4 days. Animals overexpressing spg-20 had a higher survival rate than both wild-type and spg-20 mutant animals (*p<0.0001). Survival data represent at least 100 animals per genotype from four independent trials. Error bars represent SEM; Kaplan-Meier method. (B) Percent survival of animals after treatment with 100 mM sodium azide for 1 hour. The percentage of animals alive was scored after a 24 hour recovery period. Spg-20(tm5514) mutant animals had decreased survival when compared to wild type animals (*p<0.001) and animals overexpressing spartin (*p< 0.001). There was no significant difference found between the survival of wild type animals and animals overexpressing spartin. (C) Average speed of transgenic animals measured each day for 4 days after treatment with 2 mM paraquat. spg-20 mutant animals showed reduced speed, whereas animals overexpressing spg-20 showed similar speed compared with WT animals (*p<0.05). Speed was quantified by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 50. (D) Thrashing frequency (body bends per second) of transgenic animals in M9 liquid on Day 4 of treatment with 2 mM paraquat. Spg-20 mutant animals had a reduced thrashing rate, whereas animals overexpressing spg-20 showed a recovery in thrashing rate compared to WT animals (*p<0.01). Thrashing frequency was measured by video analysis, as described in Materials and Methods. Error bars represent SEM. One-way ANOVA with Tukey test; n > 30.
Mentions: To further investigate the protective effects of spg-20 under oxidative stress, we examined three strains of C. elegans; wild-type, spg-20(tm5514) mutants, and animals overexpressing spg-20, all of which expressed green fluorescence protein (GFP). All three of these strains were exposed to 2 mM paraquat for 4 days. We observed that spg-20 mutant animals had a lower survival rate than wild-type animals (p<0.0001), and animals overexpressing spg-20 had a higher survival rate than both wild-type animals (p<0.001) and spg-20 mutant animals (p<0.0001) (Fig 4A). After 4 days of exposure to paraquat, we determined that 77% of wild-type animals survived, 38% of spg-20 mutant animals survived, and 95% of animals overexpressing spg-20 survived (S1 Table). Because animals overexpressing spartin showed significantly longer survival than wild-type animals, we propose that spartin plays an important role in the oxidative stress response.

Bottom Line: These results suggest an age-dependent decline in motor function in mutant animals.In the behavioral assays, spg-20 mutant animals showed a significant decrease in both crawling speed and thrashing frequency compared with wild-type animals.These data suggest that the protein F57B10.9/SPG-20 might have a protective role against oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pharmacology and Therapeutics, Loyola University Chicago, Maywood, Illinois, United States of America.

ABSTRACT
Troyer syndrome is caused by a mutation in the SPG20 gene, which results in complete loss of expression of the protein spartin. We generated a genetic model of Troyer syndrome in worms to explore the locomotor consequences of a mutation of the Caenorhabditis elegans SPG20 orthologue, F57B10.9, also known as spg-20. Spg-20 mutants showed decreased length, crawling speed, and thrashing frequency, and had a shorter lifespan than wild-type animals. These results suggest an age-dependent decline in motor function in mutant animals. The drug paraquat was used to induce oxidative stress for 4 days in the animals. We measured survival rate and examined locomotion by measuring crawling speed and thrashing frequency. After 4 days of paraquat exposure, 77% of wild-type animals survived, but only 38% of spg-20 mutant animals survived. Conversely, animals overexpressing spg-20 had a survival rate of 95%. We also tested lifespan after a 1 hour exposure to sodium azide. After a 24 hour recovery period, 87% of wild type animals survived, 57% of spg-20 mutant animals survived, and 82% of animals overexpressing spg-20 survived. In the behavioral assays, spg-20 mutant animals showed a significant decrease in both crawling speed and thrashing frequency compared with wild-type animals. Importantly, the locomotor phenotype for both crawling and thrashing was rescued in animals overexpressing spg-20. The animals overexpressing spg-20 had crawling speeds and thrashing frequencies similar to those of wild-type animals. These data suggest that the protein F57B10.9/SPG-20 might have a protective role against oxidative stress.

No MeSH data available.


Related in: MedlinePlus