Self-sperm induce resistance to the detrimental effects of sexual encounters with males in hermaphroditic nematodes
- Stanford University, United States
- Glenn Laboratories for the Biology of Aging at Stanford University, United States
Figures
Figure 1 with 1 supplement
The length of sexual encounters and age of the sexual partners influences the detrimental effect of males on hermaphrodite lifespan.
(A) Scheme describing the lengths of sexual interactions between C. elegans males and hermaphrodites and the ages of the sexual partners used in this study. Young was defined as the first day of …
Figure 1—figure supplement 1
Effect of long-term exposure to young males and development of the sperm tracking method.
(A) The continuous presence of young males beginning at the onset of adulthood (day 3 of life) shortens hermaphrodite lifespan (p<0.0001). (B-C) Accuracy tests for the presence of fluorescence in …
Figure 2 with 2 supplements
The presence of self-sperm is necessary for the resistance of young hermaphrodites to a brief encounter with males.
(A-B) Young, self-fertile, wild-type hermaphrodites with self-sperm (A, day 3 of life) that received male sperm after a brief interaction with males had a normal lifespan (n.s. vs. no males) whereas …
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Figure 2—source data 1
The DESeq2 output (differential expression) from the RNA-seq analysis.
- https://doi.org/10.7554/eLife.46418.008
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Figure 2—source data 2
The complete list of GO terms whose enrichment was determined using the significantly differentially expressed genes when comparing young hermaphrodites vs. young feminized individuals (selected, enriched GO results are displayed in Figure 2G–I and in Figure 2—figure supplement 2C).
- https://doi.org/10.7554/eLife.46418.009
Figure 2—figure supplement 1
The effect of a brief encounter with males on feminized and sterile individuals.
(A-B) Receiving male sperm (dashed lines) when young (day 3 of life) or middle-aged (day 7 of life) reduced the lifespan of feminized (fog-2[q71]) individuals (p<0.0001 for both young and …
Figure 2—figure supplement 2
RNA-seq of young and middle-aged hermaphrodites and feminized individuals with and without receiving male sperm.
(A) Scheme describing the experimental set up for the RNA-seq experiment. (B) Principal Component Analysis (PCA) of the normalized read counts from the complete transcriptomes of young and …
Figure 3 with 1 supplement
The presence of self-sperm is sufficient for the resistance to mating-induced demise.
(A-D) Hermaphrodites with a masculinized germline (fem-3[q20]) that have self-sperm when young (C, day 3 of life) and middle-aged (D, day 6 of life), were resistant to a brief, 2 hr interaction with …
Figure 3—figure supplement 1
The effect of males on hermaphrodites and masculinized individuals.
(A-B) Middle-aged hermaphrodites but not middle-aged masculinized individuals succumbed to brief mating-induced demise (Figure 3B,D). However, both hermaphrodites and masculinized individuals were …
Figure 4 with 1 supplement
Self-sperm act via repression of a somatic sperm-sensing pathway to mediate resistance to mating-induced demise.
(A) A model for the role of self-sperm in mating-induced demise resistance. The absence of self-sperm (right panel) activates the CEH-18 and VAB-1 sensing pathway in the somatic gonad. (B-C) Young, …
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Figure 4—source data 1
The intersection of the DESeq2 output (differential expression) and the CEH-18 binding sites (Kudron et al., 2018).
- https://doi.org/10.7554/eLife.46418.014
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Figure 4—source data 2
The complete list of GO terms whose enrichment was determined using the significantly differentially expressed genes associated with CEH-18 binding peaks when comparing young hermaphrodites vs. young feminized individuals (selected, enriched GO results are displayed in Figure 4G and Figure 4—figure supplement 1C).
- https://doi.org/10.7554/eLife.46418.015
Figure 4—figure supplement 1
CEH-18 expression and network analysis.
(A) The sperm-sensing gene ceh-18 was not differentially expressed between young and middle-aged hermaphrodites and feminized individuals. Shown here are the VST-normalized read counts for ceh-18. …
Figure 5 with 3 supplements
The importance of self-sperm in protecting young hermaphrodites against males independently evolved in a distantly-related nematode species.
(A) The phylogeny of Caenorhabditis nematodes with the hermaphroditic lineages shown in red. (B) C. brenneri females lived a normal lifespan if they mated with a male during brief, two-hour …
Figure 5—figure supplement 1
The evolution of self-sperm mediated protection from mating-induced demise.
(A) C.brenneri female lifespan was shortened if they interacted with males for a longer period of time. Interacting with males for their entire lifetime significantly shortened lifespan (orange …
Figure 5—figure supplement 2
Conservation of CEH-18 and VAB-1.
(A) Conservation scores for the Caenorhabditis orthologs of CEH-18 and VAB-1. (B) Tissue-specific protein (circles) or RNA (squares) expression levels for Ephrin receptors in C. elegans …
Figure 5—figure supplement 3
RNAi knock-down of the sperm-sensing pathway in C.briggsae.
(A-B) Representative images of the germlines of feminized (mfIs42[Cel-sid-2; Cel-myo-2::DsRed]; she-1[v35]) C. briggsae individuals cultured on control RNAi (empty vector, A) or Cbr-ceh-18 and Cbr-va…
Additional files
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Supplementary file 1
Nematode strains used in this study.
The complete list of all strains used in this study, with their genotype and source listed.
- https://doi.org/10.7554/eLife.46418.020
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Supplementary file 2
Lifespan assay results.
The data for the lifespan assays displayed in the figures as well as lifespan assays whose plots are not shown in the manuscript. Each set of lifespan assays performed together is separated from the other sets of assays by a blank row in the table. ‘Temp.’ describes the temperature at which the worms were grown. Note that 25→20 indicates that the hermaphrodites and feminized individuals were grown during development at the restrictive temperature and then shifted to a lower temperature on day 3 of life as described in the methods. All p-values were determined using Mantel-Cox log-ranking.
- https://doi.org/10.7554/eLife.46418.021
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Supplementary file 3
Mating efficiencies.
The mating efficiencies performed. The individuals whose mating efficiencies were measured are underlined in each set of mating partners. Successful mating was determined by the presence of fluorescent male sperm in the spermatheca or uterus. The presence of fluorescent male sperm is indicative of fertilization (see Figure 1—figure supplement 1B,C) though this was not specifically measured in this assay. See Source data 1 for a complete list of the mating efficiencies for each plate of 20 hermaphrodites (or germline mutants) with 40 males that were used to calculate the median and p-values.
- https://doi.org/10.7554/eLife.46418.022
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Source data 1
The raw data that comprise Supplementary file 3.
- https://doi.org/10.7554/eLife.46418.023
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Transparent reporting form
- https://doi.org/10.7554/eLife.46418.024
