The rate of behavioural decline in the ageing population is remarkably variable among individuals. Despite the considerable interest in studying natural variation in ageing rate to identify factors that control healthy ageing, no such factor has yet been found. Here we report a genetic basis for variation in ageing rates in Caenorhabditis elegans. We find that C. elegans isolates show diverse lifespan and age-related declines in virility, pharyngeal pumping, and locomotion. DNA polymorphisms in a novel peptide-coding gene, named regulatory-gene-for-behavioural-ageing-1 (rgba-1), and the neuropeptide receptor gene npr-28 influence the rate of age-related decline of worm mating behaviour; these two genes might have been subjected to recent selective sweeps. Glia-derived RGBA-1 activates NPR-28 signalling, which acts in serotonergic and dopaminergic neurons to accelerate behavioural deterioration. This signalling involves the SIR-2.1-dependent activation of the mitochondrial unfolded protein response, a pathway that modulates ageing. Thus, natural variation in neuropeptide-mediated glia–neuron signalling modulates the rate of ageing in C. elegans.
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López-Otín, C., Blasco, M. A., Partridge, L., Serrano, M. & Kroemer, G. The hallmarks of aging. Cell 153, 1194–1217 (2013)
Sachidanandam, R. et al. A map of human genome sequence variation containing 1.42 million single nucleotide polymorphisms. Nature 409, 928–933 (2001)
Gems, D. & Partridge, L. Genetics of longevity in model organisms: debates and paradigm shifts. Annu. Rev. Physiol. 75, 621–644 (2013)
Kenyon, C. J. The genetics of ageing. Nature 464, 504–512 (2010)
Herndon, L. A. et al. Stochastic and genetic factors influence tissue-specific decline in ageing C. elegans. Nature 419, 808–814 (2002)
Cohen, E., Bieschke, J., Perciavalle, R. M., Kelly, J. W. & Dillin, A. Opposing activities protect against age-onset proteotoxicity. Science 313, 1604–1610 (2006)
Pinkston, J. M., Garigan, D., Hansen, M. & Kenyon, C. Mutations that increase the life span of C. elegans inhibit tumor growth. Science 313, 971–975 (2006)
Hansen, M. & Kennedy, B. K. Does longer lifespan mean longer healthspan? Trends Cell Biol. 26, 565–568 (2016)
Yin, J. A., Liu, X. J., Yuan, J., Jiang, J. & Cai, S. Q. Longevity manipulations differentially affect serotonin/dopamine level and behavioral deterioration in aging Caenorhabditis elegans. J. Neurosci. 34, 3947–3958 (2014)
Yamazaki, D. et al. The Drosophila DCO mutation suppresses age-related memory impairment without affecting lifespan. Nat. Neurosci. 10, 478–484 (2007)
Erikson, G. A. et al. Whole-genome sequencing of a healthy aging cohort. Cell 165, 1002–1011 (2016)
Bansal, A., Zhu, L. J., Yen, K. & Tissenbaum, H. A. Uncoupling lifespan and healthspan in Caenorhabditis elegans longevity mutants. Proc. Natl Acad. Sci. USA 112, E277–E286 (2015)
Guarente, L. Aging research—where do we stand and where are we going? Cell 159, 15–19 (2014)
Whitehead, A. & Crawford, D. L. Variation within and among species in gene expression: raw material for evolution. Mol. Ecol. 15, 1197–1211 (2006)
Anholt, R. R. H. & Mackay, T. F. C. Quantitative genetic analyses of complex behaviours in Drosophila. Nat. Rev. Genet. 5, 838–849 (2004)
de Bono, M. & Bargmann, C. I. Natural variation in a neuropeptide Y receptor homolog modifies social behavior and food response in C. elegans. Cell 94, 679–689 (1998)
Rockman, M. V., Skrovanek, S. S. & Kruglyak, L. Selection at linked sites shapes heritable phenotypic variation in C. elegans. Science 330, 372–376 (2010)
Greene, J. S. et al. Balancing selection shapes density-dependent foraging behaviour. Nature 539, 254–258 (2016)
Bendesky, A. & Bargmann, C. I. Genetic contributions to behavioural diversity at the gene-environment interface. Nat. Rev. Genet. 12, 809–820 (2011)
Gloria-Soria, A. & Azevedo, R. B. npr-1 regulates foraging and dispersal strategies in Caenorhabditis elegans. Curr. Biol. 18, 1694–1699 (2008)
Loer, C. M. & Kenyon, C. J. Serotonin-deficient mutants and male mating behavior in the nematode Caenorhabditis elegans. J. Neurosci. 13, 5407–5417 (1993)
Canaff, L., Bennett, H. P. & Hendy, G. N. Peptide hormone precursor processing: getting sorted? Mol. Cell. Endocrinol. 156, 1–6 (1999)
Fares, H. & Greenwald, I. Genetic analysis of endocytosis in Caenorhabditis elegans: coelomocyte uptake defective mutants. Genetics 159, 133–145 (2001)
Liu, A. M. et al. Gα 16/z chimeras efficiently link a wide range of G protein-coupled receptors to calcium mobilization. J. Biomol. Screen. 8, 39–49 (2003)
Barr, M.M. & Garcia, L.R. in WormBook (ed. The C. elegans Research Community) https://dx.doi.org/10.1895/wormbook.1.78.1 (Wormbook, 2006)
Chatterjee, I. et al. Dramatic fertility decline in aging C. elegans males is associated with mating execution deficits rather than diminished sperm quality. Exp. Gerontol. 48, 1156–1166 (2013)
Guo, X., Navetta, A., Gualberto, D. G. & Garcia, L. R . Behavioral decay in aging male C. elegans correlates with increased cell excitability. Neurobiol. Aging 33, 1483.e5–1483.e23 (2012)
Lin, S. J., Defossez, P. A. & Guarente, L. Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. Science 289, 2126–2128 (2000)
Tissenbaum, H. A. & Guarente, L. Increased dosage of a sir-2 gene extends lifespan in Caenorhabditis elegans. Nature 410, 227–230 (2001)
Guo, X. & García, L. R. SIR-2.1 integrates metabolic homeostasis with the reproductive neuromuscular excitability in early aging male Caenorhabditis elegans. eLife 3, e01730 (2014)
Mouchiroud, L. et al. The NAD+/sirtuin pathway modulates longevity through activation of mitochondrial UPR and FOXO signaling. Cell 154, 430–441 (2013)
Imai, S. & Guarente, L. NAD+ and sirtuins in aging and disease. Trends Cell Biol. 24, 464–471 (2014)
Wang, Y. & Hekimi, S. Mitochondrial dysfunction and longevity in animals: untangling the knot. Science 350, 1204–1207 (2015)
Lin, Y. F. & Haynes, C. M. Metabolism and the UPR(mt). Mol. Cell 61, 677–682 (2016)
Yoneda, T. et al. Compartment-specific perturbation of protein handling activates genes encoding mitochondrial chaperones. J. Cell Sci. 117, 4055–4066 (2004)
Benedetti, C., Haynes, C. M., Yang, Y., Harding, H. P. & Ron, D. Ubiquitin-like protein 5 positively regulates chaperone gene expression in the mitochondrial unfolded protein response. Genetics 174, 229–239 (2006)
Li, H. A new test for detecting recent positive selection that is free from the confounding impacts of demography. Mol. Biol. Evol. 28, 365–375 (2011)
Andersen, E. C. et al. Chromosome-scale selective sweeps shape Caenorhabditis elegans genomic diversity. Nat. Genet. 44, 285–290 (2012)
Hartigan, J. A. Minimum mutation fits to a given tree. Biometrics 29, 53–65 (1973)
Williams, G. C. Pleiotropy, natural selection, and the evolution of senescence. Evolution 11, 398–411 (1957)
Wong, A., Boutis, P. & Hekimi, S. Mutations in the clk-1 gene of Caenorhabditis elegans affect developmental and behavioral timing. Genetics 139, 1247–1259 (1995)
Gems, D. et al. Two pleiotropic classes of daf-2 mutation affect larval arrest, adult behavior, reproduction and longevity in Caenorhabditis elegans. Genetics 150, 129–155 (1998)
Jenkins, N. L ., McColl, G . & Lithgow, G. J. Fitness cost of extended lifespan in Caenorhabditis elegans. Proc. R. Soc. Lond. B 271, 2523–2526 (2004)
Crawford, D., Libina, N. & Kenyon, C. Caenorhabditis elegans integrates food and reproductive signals in lifespan determination. Aging Cell 6, 715–721 (2007)
Botelho, M. & Cavadas, C. Neuropeptide Y: an anti-aging player? Trends Neurosci. 38, 701–711 (2015)
Aveleira, C. A. et al. Neuropeptide Y stimulates autophagy in hypothalamic neurons. Proc. Natl Acad. Sci. USA 112, E1642–E1651 (2015)
Kallio, J. et al. Altered intracellular processing and release of neuropeptide Y due to leucine 7 to proline 7 polymorphism in the signal peptide of preproneuropeptide Y in humans. FASEB J. 15, 1242–1244 (2001)
Dickinson, D. J., Ward, J. D., Reiner, D. J. & Goldstein, B. Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination. Nat. Methods 10, 1028–1034 (2013)
Bajar, B. T. et al. Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting. Sci. Rep. 6, 20889 (2016)
Hope, I. A. C. elegans: A Practical Approach (Oxford Univ. Press, 1999)
Yoshimura, S., Murray, J. I., Lu, Y., Waterston, R. H. & Shaham, S. mls-2 and vab-3 control glia development, hlh-17/Olig expression and glia-dependent neurite extension in C. elegans. Development 135, 2263–2275 (2008)
Wallace, S. W., Singhvi, A., Liang, Y., Lu, Y. & Shaham, S. PROS-1/Prospero is a major regulator of the glia-specific secretome controlling sensory-neuron shape and function in C. elegans. Cell Reports 15, 550–562 (2016)
Tursun, B., Patel, T., Kratsios, P. & Hobert, O. Direct conversion of C. elegans germ cells into specific neuron types. Science 331, 304–308 (2011)
Egan, C. R. et al. A gut-to-pharynx/tail switch in embryonic expression of the Caenorhabditis elegans ges-1 gene centers on two GATA sequences. Dev. Biol. 170, 397–419 (1995)
Maricq, A. V., Peckol, E., Driscoll, M. & Bargmann, C. I. Mechanosensory signalling in C. elegans mediated by the GLR-1 glutamate receptor. Nature 378, 78–81 (1995)
Sze, J. Y., Victor, M., Loer, C., Shi, Y. & Ruvkun, G. Food and metabolic signalling defects in a Caenorhabditis elegans serotonin-synthesis mutant. Nature 403, 560–564 (2000)
Nass, R., Hall, D. H., Miller, D. M., III & Blakely, R. D. Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. Proc. Natl Acad. Sci. USA 99, 3264–3269 (2002)
Doitsidou, M., Poole, R. J., Sarin, S., Bigelow, H. & Hobert, O. C. elegans mutant identification with a one-step whole-genome-sequencing and SNP mapping strategy. PLoS One 5, e15435 (2010)
Frøkjær-Jensen, C. et al. Single-copy insertion of transgenes in Caenorhabditis elegans. Nat. Genet. 40, 1375–1383 (2008)
Garrison, J. L. et al. Oxytocin/vasopressin-related peptides have an ancient role in reproductive behavior. Science 338, 540–543 (2012)
Lipton, J., Kleemann, G., Ghosh, R., Lints, R. & Emmons, S. W. Mate searching in Caenorhabditis elegans: a genetic model for sex drive in a simple invertebrate. J. Neurosci. 24, 7427–7434 (2004)
Husson, S. J. et al. Approaches to identify endogenous peptides in the soil nematode Caenorhabditis elegans. Methods Mol. Biol. 615, 29–47 (2010)
Bacaj, T. & Shaham, S. Temporal control of cell-specific transgene expression in Caenorhabditis elegans. Genetics 176, 2651–2655 (2007)
Merkwirth, C. et al. Two conserved histone demethylases regulate mitochondrial stress-induced longevity. Cell 165, 1209–1223 (2016)
Calixto, A., Chelur, D., Topalidou, I., Chen, X. & Chalfie, M. Enhanced neuronal RNAi in C. elegans using SID-1. Nat. Methods 7, 554–559 (2010)
Fraser, A. G. et al. Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature 408, 325–330 (2000)
Sneath, P. H. A. & Sokal, R. R . Numerical Taxonomy; The Principles and Practice of Numerical Classification (W. H. Freeman, 1973)
Tamura, K., Nei, M. & Kumar, S. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc. Natl Acad. Sci. USA 101, 11030–11035 (2004)
Saitou, N. & Nei, M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406–425 (1987)
Excoffier, L., Laval, G. & Schneider, S. Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evol. Bioinform. Online 1, 47–50 (2007)
Tajima, F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123, 585–595 (1989)
We thank M.-M. Poo, X. Yu and D. Chen for critical reading of the manuscript; H.-W. Zhu and L.-S. Wang for mass spectrometry analysis; Y. H. Wong and J. Chu for providing the Gα16 and mRuby3 plasmids, respectively; Z. Chen and X. Bai for experimental assistance; and the Caenorhabditis Genetics Center for providing strains. This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB 13000000) and the National Natural Science Foundation of China (31471149 and 81527901).
The authors declare no competing financial interests.
Reviewer Information Nature thanks L. Bianchi, P. McGrath and the other anonymous reviewer(s) for their contribution to the peer review of this work.
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Extended data figures and tables
a, Schematic illustration of generation of SQC0002 strain. CB4856 was crossed with Pbas-1::bas-1::gfp transgenic worms (with N2 genetic background), and about a quarter of aged F2 progeny showed an elevated level of BAS-1::GFP expression. The F2 progeny with high expression levels of BAS-1 were backcrossed with N2 worms eight times; the resulting strain was named SQC0002. b, Left, expression of BAS-1::GFP in SQC0002 and N2 worms at day 9 of adulthood. Scale bar, 10 μm. Representative of n = 5 independent experiments. Right, quantification of BAS-1::GFP fluorescence intensity. GFP fluorescence was normalized to average fluorescence intensity of age-matched N2 worms. c, Mating efficiency of SQC0002, CB4856, and N2 males at a range of ages. d, Whole-genome sequencing of SQC0002 worms identified a 328-kb region in chromosome I that possessed enriched CB4856 alleles. e, Age-dependent changes in mating efficiency in N2, SQC0002, and SQC0002 males with single-copy transgene of empty vector (SQC0002;EV) or rgba-1 (SQC0002;rgba-1). All data shown are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 (b, two-sided t test; c, e, ANOVA with Dunnett’s test). For b, c and e, each data point represents the result of one independent experiment. The numbers of tested worms (b) and mating plates (c, e) are shown beneath the bars.
a, Sequence confirmation of N2;rgba-13V4H, N2;rgba-13G4R, N2;rgba-13V4R, and CB4856;rgba-13G4H worms. The N2;rgba-13V4H, N2;rgba-13G4R, and N2;rgba-13V4R worms were generated by converting the 3G4H rgba-1 allele to 3V4H, 3G4R, and 3V4R, respectively, in N2 worms. The CB4856;rgba-13G4H worms were generated by changing the 3V4R rgba-1 allele to 3G4H in CB4856 worms. Black arrows indicate SNP sites. b, c, Schematic illustrations of molecular details of rgba-1 (b) and npr-28 (c) mutations. Three nucleotides highlighted in purple represent the protospacer-adjacent motif. d, Sequence confirmation of AB3;npr-28N2, AB3;npr-28166L, N2;npr-28AB3, and N2;npr-28166M worms. The AB3;npr-28166L and AB3;npr-28N2 worms were generated by changing the npr-28 allele to the 166L and N2-type npr-28 allele, respectively, in AB3 worms. N2;npr-28166M and N2;npr-28AB3 worms were generated by changing the npr-28 allele to the 166M and the AB3-type npr-28 allele, respectively, in N2 worms. Black arrows indicate SNP sites.
a, Separation of N2 worm neuropeptides by HPLC. Total neuropeptides were isolated from the mixture of N2 males and hermaphrodites. Grey bar indicates fractions selected for further analysis. mAU, milli-absorbance unit. b−f, Tandem mass spectrometry spectrum of RGBA-1-derived peptides. Peptide sequence was confirmed by higher-energy collisional dissociation fragmentation; y-type and b-type ions are shown in the spectrum.
a, Left, representative images show co-localization of mCherry-fused RGBA-1 signal peptides with an endoplasmic reticulum marker protein calnexin, in HEK293T cells. Scale bar, 10 μm; Right, quantitative analysis of cells with normal, mildly defective, and severely impaired endoplasmic reticulum-localization of RGBA-1 signal peptides. b, Cell fractionation and western blot analyses of mCherry-fused RGBA-1 signal peptides. Cell fractionation was performed by density gradient centrifugation. The endoplasmic reticulum fractions were indicated by the presence of calnexin using anti-calnexin, and mCherry-fused RGBA-1 signal peptides were visualized by anti-mCherry. For gel source data, see Supplementary Fig. 1a. c, Schematic of the RGBA-1::mRuby3 secretion assay. d, e, Images showing RGBA-1-fused mRuby3 fluorescence in glia and coelomocytes (labelled by Punc-122::GFP reporter, ccGFP). The expression of Phsp-16.2::mRuby3 was used as negative control (NC). Arrows point to glial cells and dashed circles indicate coelomocytes. Scale bar, 20 μm. f, Quantitative analysis of the ratio of RGBA-1::mRuby3 fluorescence in coelomocytes to that in glial cells. Each data point represents the result of one independent experiment. The total number of tested worms is shown beneath the bar. Data shown are mean ± s.e.m. ***P < 0.001 (ANOVA with Dunnett’s test). For a and b, n = 4 (a) or 3 (b) independent experiments.
Extended Data Figure 5 Cre–LoxP-mediated recombination and Mos1-mediated single-copy insertion of rgba-1 gene.
a, Schematic representation of Cre–LoxP-mediated recombination of rgba-1 gene. b, The cleavage of rgba-1 in various tissues was verified by PCR. c, Mating efficiency of N2 males at day 1 of adulthood with conditional deletion of rgba-1 in glial cells (rgba-1flox/flox;Pptr-10::Cre or rgba-1flox/flox;Pmir-228::Cre), neurons (rgba-1flox/flox;Prab-3::Cre), or intestinal cells (rgba-1flox/flox;Pges-1::Cre). Data shown are mean ± s.e.m. Each data point represents the result of one independent experiment. d, e, Schematic representations of Mos1-mediated single-copy insertion of rgba-1 (d) or empty vector (e). f, PCR validation of Mos1-mediated insertion of rgba-1. For b and f, n = 2 (b) or 3 (f) independent experiments. For gel source data, see Supplementary Fig. 1b.
a, Expression of BAS-1::GFP in N2 worms fed with bacteria that express control or npr-28 double-stranded RNAs. Scale bar, 10 μm. Representative of n = 3 independent experiments. b, Predicted transmembrane topology of NPR-28. The variable residue is highlighted in blue. Alignment of NPR-28 with human somatostatin receptor 5 was used for prediction. c, A phylogenetic tree of NPR-28. d, Expression of NPR-28 in serotonergic, dopaminergic, and motor (or inter-) neurons of N2 males. Serotonergic, dopaminergic, and motor (or inter-) neurons were identified by expression of Ptph-1::mCherry, Pdat-1::mCherry, and Pglr-1::mCherry, respectively. Scale bar, 20 μm. Representative of n = 2 independent experiments. e, Mating efficiency of npr-28-null males selectively expressing N2-type NPR-28 in serotonergic, dopaminergic, and motor (or inter-) neurons at day 1 of adulthood. Two independent transgenic lines per genotype were examined. Data shown are mean ± s.e.m. Each data point represents the result of one independent experiment. f, Alignment of a 30-amino-acid sequence centred at the variable 166th residue from NPR-28 of C. elegans and its Caenorhabditis briggsae, Caenorhadbitis remanei, and Caenorhabditis brenneri homologues. The red asterisk indicates the variation site in wild strains of C. elegans.
a, C. elegans male mating steps. Dashed lines indicate the transport of sperm. b, Number of hermaphrodites contacted by a male before mating initiation. c, d, Turning (c) and vulva location (d) efficiency of males during mating. e, The efficiency of sperm transfer. For b–e, the numbers of tested males are shown beneath the bars. f, g, Age-dependent changes in pharyngeal pumping rate (f) and locomotion speed (g) in N2, rgba-1, npr-28, and rgba-1;npr-28 mutant worms. The numbers of independent experiments are indicated in parentheses. h, Lifespan curves of N2, rgba-1, npr-28, and rgba-1;npr-28 mutant worms. The numbers of tested hermaphrodites are indicated in parentheses. Data shown in b, d, f and g are mean ± s.e.m., and in c, e, and h represent the sum of animals in three (c, e) or four (h) independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 (b, d, f, and g, ANOVA with Dunnett’s test; c, e, Fisher’s exact test; h, two-sided log-rank test).
a, Quantification of UPRmt by measuring the fluorescence intensity of Phsp-6::GFP. n = 3 repeated experiments. Each data point represents the result of one independent experiment. Data shown are mean ± s.e.m. *P < 0.05, ***P < 0.001. (ANOVA with Dunnett’s test). b, Fluorescent images of worms expressing the UPRmt reporter Phsp-6::GFP in the presence of ubl-5 RNAi. Representative of n = 3 repeated experiments.
Extended Data Figure 9 UPGMA trees across 249 natural isolates, and global distribution of rgba-1 and npr-28 alleles.
a, UPGMA trees were generated using DNA polymorphisms within a 20-kb region surrounding rgba-1 (left) or npr-28 (right). The minimum basal branch and its descendants are marked in red, and the size of the minimum basal branch n − ψ = 6 and 5 for rgba-1 and npr-28, respectively. b, Frequency and global distribution of rgba-1 and npr-28 alleles among wild strains. The number of wild strains is indicated inside or near the bar.
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Yin, JA., Gao, G., Liu, XJ. et al. Genetic variation in glia–neuron signalling modulates ageing rate. Nature 551, 198–203 (2017). https://doi.org/10.1038/nature24463
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