Insulin-like peptides and the mTOR-TFEB pathway protect Caenorhabditis elegans hermaphrodites from mating-induced death
- Princeton University, United States
- Stanford University, United States
Figures
Figure 1 with 1 supplement
Brief mating-induced death is dependent on self-sperm and acts through male seminal fluid.
(A) Schematic and representative survival curves illustrating survival after a brief (2 hr) mating; C. elegans hermaphrodite lifespan is dependent on the presence of self-sperm. (B) Wild-type N2 …
Figure 1—figure supplement 1
The presence of self-sperm protects the hermaphrodites from brief mating-induced death.
(A) Both self-spermless fog-2(q71) and fem-1(hc17) hermaphrodites are susceptible to 2 hr mating on day 3 of adulthood. fog-2 unmated: 16.3 ± 0.7 days; mated: 13.4 ± 0.5 days, p=0.0010. fem-1 …
Figure 2
IIS/FOXO in brief mating-induced death.
(A) daf-16(mu86) mutants lived shorter after 2hr mating on Day 3 of adulthood. Unmated: 12.2±0.5 days; mated: 9.5±0.5 days, p=0.0003. (B) daf-2(e1370) mutants are resistant to 2 hr mating in the …
Figure 3 with 2 supplements
INS-37 is required for self-sperm protection from brief mating-induced death.
(A) Heatmap of all the insulin genes from microarray-based transcriptome comparison between N2 and fog-2 L4 hermaphrodites. (Four biological replicates) (B) qRT-PCR reveals over 25 fold …
Figure 3—figure supplement 1
Transcriptomic comparison between self-spermless, wild type, and excess self-sperm mutants reveals HLH-30 binding motif.
L4 hermaphrodites of each genotype were collected for RNA extraction and microarray-based transcriptome analysis. One-class SAM was used to determine genes with significant changes in expression …
Figure 3—figure supplement 2
Ins-7 is highly up-regulated in L4 hermaphrodites with self-sperm.
(A) qRT-PCR of ins-37. Ins-37 has an over 25-fold increase in N2 L4 hermaphrodites compared to fog-2 L4 hermaphrodites (same figure shown in Figure 3B). Mating has very little impact on ins-37 …
Figure 4 with 2 supplements
HLH-30 protects against brief mating-induced death.
(A) hlh-30(tm1978) mutants live shorter after brief mating on day 3, unlike wild type (N2) hermaphrodites, which are resistant to 2 hr mating on day 3 of adulthood (Figure 4—figure supplement 1L). hl…
Figure 4—figure supplement 1
Quantitation of HLH-30::GFP in worms at different ages and after mating for various lengths of time.
(A–K) HLH-30::GFP becomes more diffuse with age, even if the hermaphrodites have not been mated; however, mating accelerates this process. All worms were mated on Day 3 of adulthood. (B, C; E–K) 2 …
Figure 4—figure supplement 2
Mating also decreases nuclear HLH-30::GFP in fog-2 self-spermless hermaphrodites.
Left: representative images of HLH-30::GFP in unmated and mated fog-2 hermaphrodites (Day 4). Quantitation of HLH-30::GFP in fog-2. (p=0.0328) Each worm was assigned a category based on HLH-30::GFP …
Figure 5 with 2 supplements
TOR signaling components promote Seminal Fluid-induced killing/brief mating-induced death.
(A) Most positive regulators of TOR signaling (e.g., Rictor/rict-1) were modestly downregulated in conditions in which worms have self-sperm and are protected from brief mating-induced death (left …
Figure 5—figure supplement 1
TOR signaling mediates self-sperm protection from brief mating.
(A–B) Knockdown of let-363 has no impact on lifespan of N2 upon brief mating. (A) N2 on vector control RNAi unmated: 14.9 ± 0.3 days; mated: 14.9 ± 0.4 days, p=0.7888. (B) N2 on TOR/let-363 RNAi …
Figure 5—figure supplement 2
Insulin and mTOR signaling pathways influence HLH-30 and DAF-16 localization in mated hermaphrodites.
(A–B) Knocking down pqm-1, insulins, and Tor/let-363 does not affect HLH-30::GFP localization in unmated worms; (C) By contrast, knocking down pqm-1, ins-7, ins-8, or let-363 largely prevent …
Figure 6 with 1 supplement
PQM-1 is required for SF and brief mating-induced killing.
(A–B) pqm-1(ok485) mutant hermaphrodites are resistant to short term (2 hr) mating-induced death (A), as well as long term (24 hr) mating (B) 2 hr pqm-1 unmated: 10.5 ± 0.3 days; mated: 11.1 ± 0.4 …
Figure 6—figure supplement 1
PQM-1 is required for SF killing.
(A–B) pqm-1 mutants are resistant to both long-term (24 hr; B) and brief (2 hr) mating (C). (C) Knocking down hlh-30 in pqm-1 mutants does not make them susceptible to brief mating-induced death. hlh…
Figure 7
Insulins mediate seminal fluid killing.
(A) Heatmap of insulin gene expression comparisons between fog-2 mated worms (short-lived after mating) and pqm-1 mated worms (no lifespan decrease after mating). (B) Both mean and maximum Pins-8::gf…
Figure 8
Model of self-sperm mediated protection from seminal fluid killing Prior to mating, the presence of self-sperm maintains high ins-37 expression.
INS-37 antagonizes DAF-2 activity and promotes DAF-16 and HLH-30 maintenance in the nucleus. Self-sperm also inhibits TOR signaling, which regulates HLH-30 nuclear localization. Seminal fluid …
Additional files
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Supplementary file 1
Lifespan results summary.
Summary of all the lifespan results in this study.
- https://doi.org/10.7554/eLife.46413.019
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Supplementary file 2
N2 vs fog2(q71) L4 unmated hermaphrodites microarray analysis.
List of significantly up- and down-regulated genes in N2 vs fog2(q71) L4 unmated hermaphrodites transcriptome comparison identified by Significant Analysis of Microarrays (SAM), and functional enrichment analysis of these genes by g:Profiler.
- https://doi.org/10.7554/eLife.46413.020
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Supplementary file 3
fem-3(q20) vs N2 L4 unmated hermaphrodites microarray analysis.
List of significantly up- and down-regulated genes in fem-3(q20) vs N2 L4 unmated hermaphrodites transcriptome comparison identified by Significant Analysis of Microarrays (SAM), and functional enrichment analysis of these genes by g:Profiler.
- https://doi.org/10.7554/eLife.46413.021
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Supplementary file 4
fog-2(q71) vs pqm-1(ok485) mated hermaphrodites (Day 3) microarray analysis.
List of significantly up- and down-regulated genes in fog-2(q71) vs pqm-1(ok485) mated hermaphrodites (Day 3) transcriptome comparison identified by Significant Analysis of Microarrays (SAM), and functional enrichment analysis of these genes by g:Profiler.
- https://doi.org/10.7554/eLife.46413.022
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Transparent reporting form
- https://doi.org/10.7554/eLife.46413.023
