Fetus-derived DLK1 is required for maternal metabolic adaptations to pregnancy and is associated with fetal growth restriction


Pregnancy is a state of high metabolic demand. Fasting diverts metabolism to fatty acid oxidation, and the fasted response occurs much more rapidly in pregnant women than in non-pregnant women. The product of the imprinted DLK1 gene (delta-like homolog 1) is an endocrine signaling molecule that reaches a high concentration in the maternal circulation during late pregnancy. By using mouse models with deleted Dlk1, we show that the fetus is the source of maternal circulating DLK1. In the absence of fetally derived DLK1, the maternal fasting response is impaired. Furthermore, we found that maternal circulating DLK1 levels predict embryonic mass in mice and can differentiate healthy small-for-gestational-age (SGA) infants from pathologically small infants in a human cohort. Therefore, measurement of DLK1 concentration in maternal blood may be a valuable method for diagnosing human disorders associated with impaired DLK1 expression and to predict poor intrauterine growth and complications of pregnancy.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: The conceptus is the source of elevated maternal plasma DLK1 levels in late gestation.
Figure 2: Fetus, not placenta, is the source of maternal circulating DLK1.
Figure 3: Maternal genotype and conceptus-derived DLK1 alter maternal metabolism.
Figure 4: Low DLK1 levels in human pregnancy are associated with pathological SGA.


  1. 1

    Butte, N.F. Carbohydrate and lipid metabolism in pregnancy: normal compared with gestational diabetes mellitus. Am. J. Clin. Nutr. 71 (Suppl. 5), 1256S–1261S (2000).

    CAS  Article  Google Scholar 

  2. 2

    Metzger, B.E., Ravnikar, V., Vileisis, R.A. & Freinkel, N. “Accelerated starvation” and the skipped breakfast in late normal pregnancy. Lancet 1, 588–592 (1982).

    CAS  Article  Google Scholar 

  3. 3

    Schmidt, J.V., Matteson, P.G., Jones, B.K., Guan, X.J. & Tilghman, S.M. The Dlk1 and Gtl2 genes are linked and reciprocally imprinted. Genes Dev. 14, 1997–2002 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. 4

    Takada, S. et al. Delta-like and Gtl2 are reciprocally expressed, differentially methylated linked imprinted genes on mouse chromosome 12. Curr. Biol. 10, 1135–1138 (2000).

    CAS  Article  Google Scholar 

  5. 5

    Smas, C.M., Chen, L. & Sul, H.S. Cleavage of membrane-associated pref-1 generates a soluble inhibitor of adipocyte differentiation. Mol. Cell. Biol. 17, 977–988 (1997).

    CAS  Article  Google Scholar 

  6. 6

    Bachmann, E., Krogh, T.N., Højrup, P., Skjødt, K. & Teisner, B. Mouse fetal antigen 1 (mFA1), the circulating gene product of mdlk, pref-1 and SCP-1: isolation, characterization and biology. J. Reprod. Fertil. 107, 279–285 (1996).

    CAS  Article  Google Scholar 

  7. 7

    Floridon, C. et al. Does fetal antigen 1 (FA1) identify cells with regenerative, endocrine and neuroendocrine potentials? A study of FA1 in embryonic, fetal, and placental tissue and in maternal circulation. Differentiation 66, 49–59 (2000).

    CAS  Article  Google Scholar 

  8. 8

    Carlsson, H.E., Persdotter-Hedlund, G., Fries, E., Eriksson, U.J. & Hau, J. Purification, characterization, and biological compartmentalization of rat fetal antigen 1. Biol. Reprod. 63, 30–33 (2000).

    CAS  Article  Google Scholar 

  9. 9

    Smas, C.M. & Sul, H.S. Pref-1, a protein containing EGF-like repeats, inhibits adipocyte differentiation. Cell 73, 725–734 (1993).

    CAS  Article  Google Scholar 

  10. 10

    Charalambous, M. et al. Imprinted gene dosage is critical for the transition to independent life. Cell Metab. 15, 209–221 (2012).

    CAS  Article  Google Scholar 

  11. 11

    Charalambous, M. et al. DLK1/PREF1 regulates nutrient metabolism and protects from steatosis. Proc. Natl. Acad. Sci. USA 111, 16088–16093 (2014).

    CAS  Article  Google Scholar 

  12. 12

    Moraitis, A.A., Wood, A.M., Fleming, M. & Smith, G.C. Birth weight percentile and the risk of term perinatal death. Obstet. Gynecol. 124, 274–283 (2014).

    Article  Google Scholar 

  13. 13

    Raghunandan, R. et al. Dlk1 influences differentiation and function of B lymphocytes. Stem Cells Dev. 17, 495–507 (2008).

    CAS  Article  Google Scholar 

  14. 14

    Chacón, M.R. et al. Human serum levels of fetal antigen 1 (FA1/Dlk1) increase with obesity, are negatively associated with insulin sensitivity and modulate inflammation in vitro. Int. J. Obes. (Lond.) 32, 1122–1129 (2008).

    Article  Google Scholar 

  15. 15

    Rossant, J. & Cross, J.C. Placental development: lessons from mouse mutants. Nat. Rev. Genet. 2, 538–548 (2001).

    CAS  Article  Google Scholar 

  16. 16

    Malik, N.M. et al. Leptin requirement for conception, implantation, and gestation in the mouse. Endocrinology 142, 5198–5202 (2001).

    CAS  Article  Google Scholar 

  17. 17

    Schulz, L.C., Widmaier, E.P., Qiu, J. & Roberts, R.M. Effect of leptin on mouse trophoblast giant cells. Biol. Reprod. 80, 415–424 (2009).

    CAS  Article  Google Scholar 

  18. 18

    Roman, E.A., Ricci, A.G. & Faletti, A.G. Leptin enhances ovulation and attenuates the effects produced by food restriction. Mol. Cell. Endocrinol. 242, 33–41 (2005).

    CAS  Article  Google Scholar 

  19. 19

    da Rocha, S.T. et al. Gene dosage effects of the imprinted delta-like homologue 1 (Dlk1/Pref1) in development: implications for the evolution of imprinting. PLoS Genet. 5, e1000392 (2009).

    Article  Google Scholar 

  20. 20

    Appelbe, O.K., Yevtodiyenko, A., Muniz-Talavera, H. & Schmidt, J.V. Conditional deletions refine the embryonic requirement for Dlk1. Mech. Dev. 130, 143–159 (2013).

    CAS  Article  Google Scholar 

  21. 21

    Tallquist, M.D. & Soriano, P. Epiblast-restricted Cre expression in MORE mice: a tool to distinguish embryonic vs. extra-embryonic gene function. Genesis 26, 113–115 (2000).

    CAS  Article  Google Scholar 

  22. 22

    Muzumdar, M.D., Tasic, B., Miyamichi, K., Li, L. & Luo, L. A global double-fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007).

    CAS  Article  Google Scholar 

  23. 23

    El-Kasti, M.M. et al. The pregnancy-induced increase in baseline circulating growth hormone in rats is not induced by ghrelin. J. Neuroendocrinol. 20, 309–322 (2008).

    CAS  Article  Google Scholar 

  24. 24

    Soares, M.J. The prolactin and growth hormone families: pregnancy-specific hormones/cytokines at the maternal–fetal interface. Reprod. Biol. Endocrinol. 2, 51 (2004).

    Article  Google Scholar 

  25. 25

    Pasupathy, D. et al. Study protocol. A prospective cohort study of unselected primiparous women: the Pregnancy Outcome Prediction Study. BMC Pregnancy Childbirth 8, 51 (2008).

    Article  Google Scholar 

  26. 26

    Sovio, U., White, I.R., Dacey, A., Pasupathy, D. & Smith, G.C. Screening for fetal growth restriction with universal third trimester ultrasonography in nulliparous women in the Pregnancy Outcome Prediction (POP) study: a prospective cohort study. Lancet 386, 2089–2097 (2015).

    Article  Google Scholar 

  27. 27

    Rotham, K.J.G. et al. Modern Epidemiology 3rd edn. (Lippincott-Raven, 2008).

  28. 28

    de Zegher, F. et al. Abundance of circulating preadipocyte factor 1 in early life. Diabetes Care 35, 848–849 (2012).

    Article  Google Scholar 

  29. 29

    Schrey, S. et al. The adipokine preadipocyte factor-1 is downregulated in preeclampsia and expressed in placenta. Cytokine 75, 338–343 (2015).

    CAS  Article  Google Scholar 

  30. 30

    Moore, G.E. et al. The role and interaction of imprinted genes in human fetal growth. Phil. Trans. R. Soc. Lond. B 370, 20140074 (2015).

    CAS  Article  Google Scholar 

  31. 31

    Kappil, M.A. et al. Placental expression profile of imprinted genes impacts birth weight. Epigenetics 10, 842–849 (2015).

    Article  Google Scholar 

  32. 32

    Smith, G.C. Researching new methods of screening for adverse pregnancy outcome: lessons from pre-eclampsia. PLoS Med. 9, e1001274 (2012).

    Article  Google Scholar 

  33. 33

    Ioannides, Y., Lokulo-Sodipe, K., Mackay, D.J., Davies, J.H. & Temple, I.K. Temple syndrome: improving the recognition of an underdiagnosed chromosome 14 imprinting disorder: an analysis of 51 published cases. J. Med. Genet. 51, 495–501 (2014).

    CAS  Article  Google Scholar 

Download references


M.A.M.C. was supported by a PhD studentship from the Cambridge Centre for Trophoblast Research. Research was supported by grants from the MRC (MR/J001597/1 and MR/L002345/1), the Medical College of Saint Bartholomew's Hospital Trust, a Wellcome Trust Investigator Award, EpigeneSys (FP7 Health-257082), EpiHealth (FP7 Health-278414), a Herchel Smith Fellowship (N.T.) and NIH grant RO1 DK89989. The contents are the authors' sole responsibility and do not necessarily represent official NIH views. We thank G. Burton for invaluable support, and M. Constância and I. Sandovici (University of Cambridge) for the Meox2-cre mice. We are extremely grateful to all of the participants in the Pregnancy Outcome Prediction study. This work was supported by the NIHR Cambridge Comprehensive Biomedical Research Centre (Women's Health theme) and project grants from the MRC (G1100221) and Sands (Stillbirth and Neonatal Death Charity). The study was also supported by GE Healthcare (donation of two Voluson i ultrasound systems for this study) and by the NIHR Cambridge Clinical Research Facility, where all research visits took place.

Author information




M.C., M.A.M.C., A.C.F.-S. and G.C.S.S. conceived and designed the experiments. M.C., M.A.M.C., J.A.C., M.H., I.G., N.T., C.L.D. and D.S.C.-J. performed the experiments. M.C., M.A.M.C., A.C.F.-S., C.L.D., F.G. and G.C.S.S. analyzed the data. M.C. and U.S. performed statistical analysis. S.R.B., T.L.P., A.C.F.-S. and G.C.S.S. contributed reagents. M.C., M.A.M.C., A.C.F.-S. and G.C.S.S. wrote the manuscript. M.C., A.C.F.-S. and G.C.S.S. provided supervision.

Corresponding author

Correspondence to Marika Charalambous.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Altered body composition in Null females can be detected during embryogenesis.

(a) Total bodyweight of virgin females at 12 weeks does not differ between genotypes. Null females have larger abdominal WAT deposits (b) and reduced muscle (combined gastrocnemius/soleus) (c) than WT and Mat females at 12 weeks. WT and Null n = 8, Mat n = 7, compared by One-way ANOVA, with Bonferroni's Multiple comparison post-hoc test with WT vs Null and WT vs Mat, *p<0.05. Body weight (d), crown rump (C-R) length (e) and tissue weights (f) in embryos at E18.5. Null embryos have reduced mass and length, and reduced lean (hindlimb and forelimb) mass than embryos expressing Dlk1, WT n > 21, Null n > 13, Mat n > 10, compared by Kruskall-Wallace test with Dunn's Multiple comparison post-hoc test comparing WT vs Null and WT vs Mat, *p<0.05, ***p<0.001. (g) Null mice are born small but catch up in the preweaning period. Serial measurements of pup weight from birth to weaning, Nulls weigh significantly less at birth but not thereafter, n = 4-11 per genotype, each time point compared as above.

Supplementary Figure 2 Summary of experimental crosses

Experimental crosses used to determine which compartment of the conceptus (embryo, placental fetal endothelium or placental trophoblast) is the source of maternal circulating DLK1 in pregnancy.

Supplementary Figure 3 Levels of pituitary Gh mRNA do not differ between the groups.

Expression levels of Gh (a) measured by real-time quantitative PCR in pituitaries from females in the cohort. Gh levels did not differ between the groups. Data was normalised to β-actin levels, then expressed as WT = 1. Groups were compared by One-way ANOVA, with Bonferroni's Multiple comparison post-hoc test with WT vs Null and WT x WT vs Null x WT and Null x Null, ***p<0.001.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3 and Supplementary Tables 1–11. (PDF 1167 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Cleaton, M., Dent, C., Howard, M. et al. Fetus-derived DLK1 is required for maternal metabolic adaptations to pregnancy and is associated with fetal growth restriction. Nat Genet 48, 1473–1480 (2016). https://doi.org/10.1038/ng.3699

Download citation

Further reading