Caenorhabditis elegans Battling Starvation Stress: Low Levels of Ethanol Prolong Lifespan in L1 Larvae
Publication Date
January 18, 2012
Journal
PLOS ONE
Authors
Paola V. Castro, Shilpi Khare, Brian D. Young & Steven G. Clarke
Volume
7
Issue
1
Pages
e29984
DOI
https://dx.plos.org/10.1371/journal.pone.0029984
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0029984
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/22279556
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3261173
Europe PMC
http://europepmc.org/abstract/MED/22279556
Web of Science
000299771900034
Scopus
84855934672
Mendeley
http://www.mendeley.com/research/caenorhabditis-elegans-battling-starvation-stress-low-levels-ethanol-prolong-lifespan-l1-larvae
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Mendeley 6325 Apr 20:18 UTC

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Scopus 1720 May 07:24 UTC

Scopus | Further Information

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Figshare25 Dec 12:22 UTC

Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/352111", "https://ndownloader.figshare.com/files/352120", "https://ndownloader.figshare.com/files/352135", "https://ndownloader.figshare.com/files/352150", "https://ndownloader.figshare.com/files/352171", "https://ndownloader.figshare.com/files/352192", "https://ndownloader.figshare.com/files/352212", "https://ndownloader.figshare.com/files/352228", "https://ndownloader.figshare.com/files/352239"], "description"=>"<div><p>The nematode <em>Caenorhabditis elegans</em> arrests development at the first larval stage if food is not present upon hatching. Larvae in this stage provide an excellent model for studying stress responses during development. We found that supplementing starved larvae with ethanol markedly extends their lifespan within this L1 diapause. The effects of ethanol-induced lifespan extension can be observed when the ethanol is added to the medium at any time between 0 and 10 days after hatching. The lowest ethanol concentration that extended lifespan was 1 mM (0.005%); higher concentrations to 68 mM (0.4%) did not result in increased survival. In spite of their extended survival, larvae did not progress to the L2 stage. Supplementing starved cultures with n-propanol and n-butanol also extended lifespan, but methanol and isopropanol had no measurable effect. Mass spectrometry analysis of nematode fatty acids and amino acids revealed that L1 larvae can incorporate atoms from ethanol into both types of molecules. Based on these data, we suggest that ethanol supplementation may extend the lifespan of L1 larvae by either serving as a carbon and energy source and/or by inducing a stress response.</p> </div>", "links"=>[], "tags"=>["battling", "starvation", "levels", "ethanol", "prolong", "lifespan", "l1", "larvae"], "article_id"=>129601, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.s001", "https://dx.doi.org/10.1371/journal.pone.0029984.s002", "https://dx.doi.org/10.1371/journal.pone.0029984.s003", "https://dx.doi.org/10.1371/journal.pone.0029984.s004", "https://dx.doi.org/10.1371/journal.pone.0029984.s005", "https://dx.doi.org/10.1371/journal.pone.0029984.s006", "https://dx.doi.org/10.1371/journal.pone.0029984.s007", "https://dx.doi.org/10.1371/journal.pone.0029984.s008", "https://dx.doi.org/10.1371/journal.pone.0029984.s009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Caenorhabditis_elegans_Battling_Starvation_Stress_Low_Levels_of_Ethanol_Prolong_Lifespan_in_L1_Larvae/129601", "title"=>"<em>Caenorhabditis elegans</em> Battling Starvation Stress: Low Levels of Ethanol Prolong Lifespan in L1 Larvae", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-01-18 02:40:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/690164"], "description"=>"<p>Following hypochlorite treatment of wild-type N2 (black line) or <i>pcm-1</i> mutant (red line) gravid adults, eggs were isolated and incubated in M9 medium for 24 h at 20°C on a rotary shaker at 160 rpm as described in the “<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#s2\" target=\"_blank\">Methods</a>” section. The resulting starved L1 larvae (300 N2 larvae/ml or 1000 <i>pcm-1</i> larvae/ml) were then incubated at 20°C at 160 rpm in either 70 ml of M9 medium (panel A) or 70 ml of M9 medium supplemented with 17 mM ethanol (0.1%; panel B) or 17 mM ethanol and 13 µM cholesterol (5 µg/ml; panel C) and survival was scored every 48 hours until all larvae were dead. Using the Mantel-Cox logrank test, a p-value of 0.03 was found for the significance of the difference between the <i>pcm-1</i> and N2 strains in panel B and a p-value of <0.0001 for these strains in panel C.</p>", "links"=>[], "tags"=>["concentrations", "ethanol", "lifespan", "arrested", "wild-type", "mutant", "l1"], "article_id"=>360643, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Low_concentrations_of_ethanol_extend_lifespan_of_arrested_wild_type_N2_and_pcm_1_mutant_L1_larvae_/360643", "title"=>"Low concentrations of ethanol extend lifespan of arrested wild-type (N2) and <i>pcm-1</i> mutant L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:10:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/690268"], "description"=>"<p>L1 larvae (70,000) were isolated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a> and were incubated at 20°C in either M9 medium only or M9 medium supplemented with 4 mM, 17 mM, or 68 mM ethanol (panels A and B; legend for both in panel B) and 0.001, 0.01, 0.1, or 1 mM ethanol (panel C). Survival was scored every 48 hour until all larvae were dead. Mantel-Cox logrank analyses gave a p-value of <0.0001 for the difference between the control (no ethanol) and all concentrations of ethanol at 1 mM or above. The survival at 68 mM ethanol is significantly different from that at 17 mM ethanol in panel A (p = 0.0001) and in panel B (p = 0.001).</p>", "links"=>[], "tags"=>["concentrations", "ethanol", "lifespan", "arrested", "wild-type", "l1"], "article_id"=>360756, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Small_concentrations_of_ethanol_1_mM_8211_68_mM_extend_the_lifespan_of_arrested_wild_type_L1_larvae_/360756", "title"=>"Small concentrations of ethanol (1 mM–68 mM) extend the lifespan of arrested wild-type L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:12:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/690411"], "description"=>"<p>Wild-type L1 larvae (70,000) were isolated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a> and were transferred to 70 ml of M9 medium supplemented with ethanol (4 mM) after 1, 5, 7, 10 and 13 days of incubation under starvation conditions in M9 medium at 20°C and 160 rpm. Survival was scored every 48 hours until all larvae were dead. Mantel-Cox logrank analyses indicated that the control was significantly different from the 1, 5, 7, and 10 day samples (p<0.0001) and the 13 day sample (p = 0.01).</p>", "links"=>[], "tags"=>["m9", "10", "days", "post-hatching", "extends", "lifespan", "arrested", "wild-type", "l1"], "article_id"=>360890, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ethanol_added_to_M9_medium_up_to_10_days_post_hatching_extends_lifespan_of_arrested_wild_type_L1_larvae_/360890", "title"=>"Ethanol added to M9 medium up to 10 days post-hatching extends lifespan of arrested wild-type L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:14:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/690490"], "description"=>"<p>L1 larvae (70,000) were isolated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a> and were incubated at 20°C at 160 rpm in 70 ml of M9 medium supplemented with 4 mM, 17 mM, and 68 mM ethanol (panels A, B, and C, respectively). The concentration of ethanol in the medium with L1 larvae (green curve) and a control medium without larvae (orange) was measured over time as described in the “<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#s2\" target=\"_blank\">Methods</a>” section. Measurements were taken until all larvae were dead. The error bars correspond to the standard deviation of triplicate assays. We note the small differences between the nominal concentrations of ethanol added and the initial concentrations detected - the origin of this difference is not clear.</p>", "links"=>[], "tags"=>["ethanol", "incubated", "m9", "arrested", "wild-type", "l1"], "article_id"=>360971, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Concentrations_of_ethanol_in_incubated_M9_medium_and_in_M9_medium_with_arrested_wild_type_L1_larvae_/360971", "title"=>"Concentrations of ethanol in incubated M9 medium and in M9 medium with arrested wild-type L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:16:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/690600"], "description"=>"<p>Wild-type (N2) and <i>eat-2(ad1116)</i> mutant L1 larvae (70,000) were prepared as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a>, and were transferred to 70 ml of M9 medium only (wild-type (black line), <i>eat-2</i> (gray line)) or 70 ml of M9 medium supplemented with 4 mM ethanol (wild-type (red line), <i>eat-2</i> (blue line)) and survival at 20°C was scored every 48 hours until all larvae were dead. Mantel-Cox logrank analyses showed that lifespan was significantly different in the wild-type and <i>eat-2</i> mutants in the presence of 4 mM ethanol (p<0.0001).</p>", "links"=>[], "tags"=>["pharyngeal", "pumping", "machinery", "ethanol-mediated", "lifespan", "arrested", "wild-type", "l1"], "article_id"=>361075, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Intact_pharyngeal_pumping_machinery_is_necessary_for_full_ethanol_mediated_lifespan_extension_of_arrested_wild_type_L1_larvae_/361075", "title"=>"Intact pharyngeal pumping machinery is necessary for full ethanol-mediated lifespan extension of arrested wild-type L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:17:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/690682"], "description"=>"<p>Survival was analyzed for wild-type L1 larvae (70,000) incubated at 20°C and 160 rpm as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a> in either 70 ml of M9 medium only, or 70 ml of M9 medium supplemented with 4 mM methanol, n-propanol, or n-butanol (panel A), or M9 medium only, or M9 medium supplemented with 4 mM ethanol, methanol, n-propanol, n-butanol, isopropanol, or isobutanol (panel B). Survival was scored every 48 hours until all nematodes were dead. Mantel-Cox logrank analyses demonstrated significance (p<0.0001) for the lifespan difference of all alcohols versus controls except for methanol (p = 0.58 in panel A and p = 0.28 in panel B) and isopropanol (p = 0.43).</p>", "links"=>[], "tags"=>["containing", "carbons", "lifespan", "arrested", "wild-type", "l1"], "article_id"=>361159, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Alcohols_containing_3_and_4_carbons_can_extend_the_lifespan_of_arrested_wild_type_L1_larvae_/361159", "title"=>"Alcohols containing 3 and 4 carbons can extend the lifespan of arrested wild-type L1 larvae.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:19:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/690795"], "description"=>"<p>Starved L1 larvae (100,000) were incubated in 100 ml of M9 only or 100 ml of M9 supplemented with 1 mM deuterated ethanol (C<sub>2</sub>D<sub>5</sub>OD) medium, under conditions described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a>, and collected at six different time points. For each time point L1 larvae (100,000) were collected and total fatty acids analyzed after saponification and conversion to fatty acid methyl esters as described in the “<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#s2\" target=\"_blank\">Methods</a>” section. Fatty acid methyl esters were fractionated and analyzed by GC-MS to look for the incorporation of deuterium atoms from the deuterated ethanol. Panel A shows the chromatogram for L1 larvae one day after hatching and before fully-deuterated ethanol was added. Panels C, E, G, I and K show the lipid profile for L1 larvae incubated in M9 medium for 1, 5, 7, 10 and 13 days, respectively. Panels D, F, H, J and L show the lipid profile for L1 larvae incubated in deuterated ethanol for 1, 5, 7, 10 and 13 days, respectively. Red arrows indicate the peaks containing deuterated fatty acids.</p>", "links"=>[], "tags"=>["arrested", "wild-type", "l1", "larvae", "incubated", "m9", "deuterated", "ethanol"], "article_id"=>361270, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fatty_acid_profile_of_arrested_wild_type_L1_larvae_incubated_in_M9_or_deuterated_ethanol_over_time_/361270", "title"=>"Fatty acid profile of arrested wild-type L1 larvae incubated in M9 or deuterated ethanol over time.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:21:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/690938"], "description"=>"<p>(panel A) A portion of the GC-MS chromatogram showing the peak of stearic acid methyl ester obtained form starved L1 larvae incubated for 7 days in 1 mM non-deuterated ethanol. (panel B) A similar portion of the GC-MS chromatogram showing deuterated (left) and non-deuterated (right) peaks of stearic acid for L1 larvae incubated for 7 days in deuterated ethanol. Deuterated fatty acids have shorter retention times and elute in a wide peak immediately before non-deuterated fatty acids. Panels D–G show the peak of deuterated stearic acid broken down into four quartiles as shown in panel B. Stearic acid methyl esters containing more deuterium atoms elute before stearic acid methyl esters containing fewer deuterium atoms. (panel D) Stearic acid in quartile I contains 9–10 additional mass units corresponding to 9–10 deuterium atoms. (panel E) Stearic acid in quartile II contains 7–9 additional mass units corresponding to 7–9 deuterium atoms. (panel F) Stearic acid in quartile III contains 5–8 additional mass units corresponding to 5–8 deuterium atoms. (panel G) Stearic acid in quartile IV contains 4–6 additional mass units corresponding to 4–6 deuterium atoms. The non-deuterated peak shows additional mass due to natural abundance of <sup>13</sup>C (panel C).</p>", "links"=>[], "tags"=>["wild-type", "l1", "larvae", "deuterium", "deuterated", "ethanol", "fatty"], "article_id"=>361406, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Arrested_wild_type_L1_larvae_incorporate_deuterium_from_deuterated_ethanol_into_fatty_acids_/361406", "title"=>"Arrested wild-type L1 larvae incorporate deuterium from deuterated ethanol into fatty acids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:23:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/691014"], "description"=>"<p>The isotopic distribution of stearic acid methyl ester and palmitic acid methyl ester from L1 larvae incubated for 7 days in 1 mM fully-deuterated ethanol is shown for the non-deuterated “sharp” peak (panels A and C) and for the earlier-eluting broad deuterated peak (panels B and D) (compare with <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g008\" target=\"_blank\">Figure 8</a>, panel B). Isotopic analysis of the broad peaks shows deuterium incorporation into stearic acid methyl ester that results in addition of 4–10 mass units (panel B) not seen in non-deuterated peak (panel A). Similarly, the earlier-eluting peak for palmitic acid methyl ester shows deuterium incorporation that results in addition of 4–7 mass units (panel D) not seen in the non-deuterated material (panel C).</p>", "links"=>[], "tags"=>["wild-type", "l1", "larvae", "ethanol", "stearic", "palmitic"], "article_id"=>361485, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Arrested_wild_type_L1_larvae_incorporate_medium_ethanol_into_stearic_and_palmitic_acids_/361485", "title"=>"Arrested wild-type L1 larvae incorporate medium ethanol into stearic and palmitic acids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:24:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/691117"], "description"=>"<p>L1 larvae were isolated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#pone-0029984-g001\" target=\"_blank\">Figure 1</a>. For each time point (T = 0, 3 and 7 days) two 100-ml cultures containing 1000 starved L1 larvae/ml were prepared in triplicate. The zero day time point consisted of starved L1 larvae one day after hatching; the 3 day and 7 day time points consisted of larvae incubated with 1 mM fully-deuterated ethanol. Protein was isolated and hydrolyzed and amino acids were derivatized and analyzed using GC-MS for deuterium incorporation as described in the “<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029984#s2\" target=\"_blank\">Methods</a>” section. The M-15 and M-117 ions for glutamate and proline were analyzed and data are shown as the average of 3 separate trials analyzed twice by GC-MS. Asterisks indicate a p-value from Student's T-test of less than 0.05 when the 3-day and 7-day peaks were compared to the 0-day controls.</p>", "links"=>[], "tags"=>["wild-type", "l1", "larvae", "ethanol", "protein-bound", "glutamate"], "article_id"=>361589, "categories"=>["Molecular Biology", "Biochemistry", "Developmental Biology"], "users"=>["Paola V. Castro", "Shilpi Khare", "Brian D. Young", "Steven G. Clarke"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029984.g010"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Arrested_wild_type_L1_larvae_incorporate_medium_ethanol_into_protein_bound_glutamate_and_proline_/361589", "title"=>"Arrested wild-type L1 larvae incorporate medium ethanol into protein-bound glutamate and proline.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-18 00:26:29"}
PMC Usage Stats 168917 May 14:49 UTC

PMC Usage Stats | Further Information

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Relative Metric 17865626 Apr 18:24 UTC

Relative Metric

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