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Somatic cells regulate maternal mRNA translation and developmental competence of mouse oocytes

Nature Cell Biology volume 15, pages 1415–1423 (2013)Cite this article

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Abstract

Germ cells divide and differentiate in a unique local microenvironment under the control of somatic cells. Signals released in this niche instruct oocyte reentry into the meiotic cell cycle. Once initiated, the progression through meiosis and the associated programme of maternal messenger RNA translation are thought to be cell autonomous. Here we show that translation of a subset of maternal mRNAs critical for embryo development is under the control of somatic cell inputs. Translation of specific maternal transcripts increases in oocytes cultured in association with somatic cells and is sensitive to EGF-like growth factors that act only on the somatic compartment. In mice deficient in amphiregulin, decreased fecundity and oocyte developmental competence is associated with defective translation of a subset of maternal mRNAs. These somatic cell signals that affect translation require activation of the PI(3)K–AKT–mTOR pathway. Thus, mRNA translation depends on somatic cell cues that are essential to reprogramme the oocyte for embryo development.

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Figure 1: The protein level of the spindle component TPX2 is dependent on the environment in which the oocyte matures.
Figure 2: EGF-like growth factor stimulation of cumulus–oocyte complexes in vitro increases translation in oocytes.
Figure 3: Compromised developmental competence of oocytes from Areg−/− mice.
Figure 4: Altered mRNA translation of a subgroup of maternal mRNAs in Areg−/− oocytes.
Figure 5: PI(3)K–AKT–mTOR signalling is involved in the somatic regulation of oocyte mRNA translation.
Figure 6: Model of the signalling pathways involved in somatic cell control of oocyte translation.

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References

  1. Spradling, A., Fuller, M. T., Braun, R. E. & Yoshida, S. Germline stem cells. Cold Spring Harb. Perspect. Biol. 3, a002642 (2011).

    Article  Google Scholar 

  2. Yoshida, S. Spermatogenic stem cell system in the mouse testis. Cold Spring Harb. Symp. Quant. Biol. 73, 25–32 (2008).

    Article  CAS  Google Scholar 

  3. Matzuk, M. M., Burns, K. H., Viveiros, M. M. & Eppig, J. J. Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science 296, 2178–2180 (2002).

    Article  CAS  Google Scholar 

  4. Gosden, R. & Lee, B. Portrait of an oocyte: our obscure origin. J. Clin. Invest. 120, 973–983 (2010).

    Article  CAS  Google Scholar 

  5. Chian, R. C., Lim, J. H. & Tan, S. L. State of the art in in-vitro oocyte maturation. Curr. Opin. Obstet. Gynecol. 16, 211–219 (2004).

    Article  Google Scholar 

  6. Chang, H. C., Liu, H., Zhang, J., Grifo, J. & Krey, L.C. Developmental incompetency of denuded mouse oocytes undergoing maturation in vitro is ooplasmic in nature and is associated with aberrant Oct-4 expression. Human Reprod. 20, 1958–1968 (2005).

    Article  CAS  Google Scholar 

  7. Luciano, A. M. et al. Developmental capability of denuded bovine oocyte in a co-culture system with intact cumulus-oocyte complexes: role of cumulus cells, cyclic adenosine 3’,5’-monophosphate, and glutathione. Mol. Reproduct. Dev. 71, 389–397 (2005).

    Article  CAS  Google Scholar 

  8. Eppig, J. J. Coordination of nuclear and cytoplasmic oocyte maturation in eutherian mammals. Reprod. Fertil. Dev. 8, 485–489 (1996).

    Article  CAS  Google Scholar 

  9. Park, J. Y. et al. EGF-like growth factors as mediators of LH action in the ovulatory follicle. Science 303, 682–684 (2004).

    Article  CAS  Google Scholar 

  10. Shimada, M., Hernandez-Gonzalez, I., Gonzalez-Robayna, I. & Richards, J. S. Paracrine and autocrine regulation of epidermal growth factor-like factors in cumulus oocyte complexes and granulosa cells: key roles for prostaglandin synthase 2 and progesterone receptor. Mol. Endocrinol. 20, 1352–1365 (2006).

    Article  CAS  Google Scholar 

  11. Hsieh, M. et al. Luteinizing hormone-dependent activation of the epidermalgrowth factor network is essential for ovulation. Mol. Cell Biol. 27, 1914–1924 (2007).

    Article  CAS  Google Scholar 

  12. Panigone, S., Hsieh, M., Fu, M., Persani, L. & Conti, M. Luteinizing hormone signaling in preovulatory follicles involves early activation of the epidermal growth factor receptor pathway. Mol. Endocrinol. 22, 924–936 (2008).

    Article  CAS  Google Scholar 

  13. Hsieh, M., Thao, K. & Conti, M. Genetic dissection of epidermal growth factor receptor signaling during luteinizing hormone-induced oocyte maturation. PLoS ONE 6, e21574 (2011).

    Article  CAS  Google Scholar 

  14. Richter, J. D. CPEB: a life in translation. Trends Biochem. Sci. 32, 279–285 (2007).

    Article  CAS  Google Scholar 

  15. Gruss, O. J. et al. Chromosome-induced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells. Nat. Cell Biol. 4, 871–879 (2002).

    Article  CAS  Google Scholar 

  16. De Luca, M., Lavia, P. & Guarguaglini, G. A functional interplay between Aurora-A, Plk1 and TPX2 at spindle poles: Plk1 controls centrosomal localization of Aurora-A and TPX2 spindle association. Cell Cycle 5, 296–303 (2006).

    Article  CAS  Google Scholar 

  17. Kufer, T. A. et al. Human TPX2 is required for targeting Aurora-A kinase to the spindle. J. Cell Biol. 158, 617–623 (2002).

    Article  CAS  Google Scholar 

  18. Chang, H. et al. The TPX2 gene is a promising diagnostic and therapeutic target for cervical cancer. Oncol. Rep. 27, 1353–1359 (2012).

    CAS  PubMed  Google Scholar 

  19. Ma, Y. et al. Expression of targeting protein for xklp2 associated with both malignant transformation of respiratory epithelium and progression of squamous cell lung cancer. Clin. Cancer Res. 12, 1121–1127 (2006).

    Article  CAS  Google Scholar 

  20. Shigeishi, H. et al. Expression of TPX2 in salivary gland carcinomas. Oncol. Rep. 21, 341–344 (2009).

    CAS  PubMed  Google Scholar 

  21. Brunet, S. et al. Meiotic regulation of TPX2 protein levels governs cell cycle progression in mouse oocytes. PLoS ONE 3, e3338 (2008).

    Article  Google Scholar 

  22. Chen, J. et al. Genome-wide analysis of translation reveals a critical role for deleted in azoospermia-like (Dazl) at the oocyte-to-zygote transition. Genes Dev. 25, 755–766 (2011).

    Article  CAS  Google Scholar 

  23. Brook, M., Smith, J. W. & Gray, N. K. The DAZL and PABP families: RNA-binding proteins with interrelated roles in translational control in oocytes. Reproduction 137, 595–617 (2009).

    Article  CAS  Google Scholar 

  24. Conti, M., Hsieh, M., Park, J. Y. & Su, Y. Q. Role of the epidermal growth factor network in ovarian follicles. Mol. Endocrinol. 20, 715–723 (2006).

    Article  CAS  Google Scholar 

  25. Fan, H. Y. et al. Selective expression of KrasG12D in granulosa cells of the mouse ovary causes defects in follicle development and ovulation. Development 135, 2127–2137 (2008).

    Article  CAS  Google Scholar 

  26. De La Fuente, R., O’Brien, M. J. & Eppig, J. J. Epidermal growth factor enhances preimplantation developmental competence of maturing mouse oocytes. Hum. Reprod. 14, 3060–3068 (1999).

    Article  CAS  Google Scholar 

  27. Prochazka, R., Kalab, P. & Nagyova, E. Epidermal growth factor-receptor tyrosine kinase activity regulates expansion of porcine oocyte-cumulus cell complexes in vitro. Biol. Reprod. 68, 797–803 (2003).

    Article  CAS  Google Scholar 

  28. Richani, D., Ritter, L. J., Thompson, J. G. & Gilchrist, R. B. Mode of oocyte maturation affects EGF-like peptide function and oocyte competence. Mol. Hum. Rreprod. 19, 500–509 (2013).

    Article  CAS  Google Scholar 

  29. Han, S. J. et al. Protein kinase B/Akt phosphorylation of PDE3A and its role in mammalian oocyte maturation. EMBO J. 25, 5716–5725 (2006).

    Article  CAS  Google Scholar 

  30. Kalous, J. et al. PKB/AKT is involved in resumption of meiosis in mouse oocytes. Biol. Cell 98, 111–123 (2006).

    Article  CAS  Google Scholar 

  31. Grzmil, M. & Hemmings, B. A. Translation regulation as a therapeutic target in cancer. Cancer Res. 72, 3891–3900 (2012).

    Article  CAS  Google Scholar 

  32. Jagarlamudi, K. et al. Oocyte-specific deletion of Pten in mice reveals a stage-specific function of PTEN/PI3K signaling in oocytes in controlling follicular activation. PLoS ONE 4, e6186 (2009).

    Article  Google Scholar 

  33. Busser, B., Sancey, L., Brambilla, E., Coll, J. L. & Hurbin, A. Themultiple of amphiregulin in human cancer. Biochim. Biophys. Acta 1816, 119–131 (2011).

    CAS  PubMed  Google Scholar 

  34. Hynes, N. E. & Lane, H. A. ERBB receptors and cancer: the complexity of targeted inhibitors. Nat. Rev. Cancer 5, 341–354 (2005).

    Article  CAS  Google Scholar 

  35. Ma, X. M. & Blenis, J. Molecular mechanisms of mTOR-mediated translational control. Nat. Rev. Mol. Cell Biol. 10, 307–318 (2009).

    Article  Google Scholar 

  36. Inoki, K., Kim, J. & Guan, K. L. AMPK and mTOR in cellular energy homeostasis and drug targets. Annu. Rev. Pharmacol. Toxicol. 52, 381–400 (2012).

    Article  CAS  Google Scholar 

  37. Magnuson, B., Ekim, B. & Fingar, D. C. Regulation and function of ribosomalprotein S6 kinase (S6K) within mTOR signalling networks. Biochem. J. 441, 1–21 (2012).

    Article  CAS  Google Scholar 

  38. Ruggero, D. & Sonenberg, N. The Akt of translational control. Oncogene 24, 7426–7434 (2005).

    Article  CAS  Google Scholar 

  39. Meyuhas, O. Synthesis of the translational apparatus is regulated at the translational level. Eur. J. Biochem. 267, 6321–6330 (2000).

    Article  CAS  Google Scholar 

  40. Hsieh, A. C. et al. The translational landscape of mTOR signalling steers cancer initiation and metastasis. Nature 485, 55–61 (2012).

    Article  CAS  Google Scholar 

  41. Barna, M. et al. Suppression of Myc oncogenic activity by ribosomal protein haploinsufficiency. Nature 456, 971–975 (2008).

    Article  CAS  Google Scholar 

  42. Su, Y. Q. et al. Selective degradation of transcripts during meiotic maturation of mouse oocytes. Dev. Biol. 302, 104–117 (2007).

    Article  CAS  Google Scholar 

  43. Irizarry, R. A. et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249–264 (2003).

    Article  Google Scholar 

  44. Dai, M. et al. Evolving gene/transcriptdefinitions significantly alter the interpretation of GeneChip data. Nucleic Acids Res. 33, e175 (2005).

    Article  Google Scholar 

  45. Huang, D. W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. Nat. Protoc. 4, 44–57 (2009).

    Article  CAS  Google Scholar 

  46. Huang, D. W., Sherman, B. T. & Lempicki, R. A. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists.. Nucleic Acids Res. 37, 1–13 (2009).

    Article  Google Scholar 

  47. P., Machanick & Bailey, T. L. MEME-ChIP: motif analysis of large DNA datasets. Bioinformatics 2712, 1696–1697 (2011.).

    Google Scholar 

  48. Crooks, G. E., Hon, G. & Chandonia, J. M. Brenner SE WebLogo: a sequence logo generator. Genome Res. 14, 1188–1190 (200).

Download references

Acknowledgements

We thank D. Laird, D. Ruggero, T. Nystul and R. Belloch at UCSF for advice during the studies and for critical reading of the manuscript, and A. Susor for assisting with the oocyte confocal microscopy analysis. This work was supported by Eunice Kennedy Shriver National Institute of Child Health and Human Development/NIH cooperative agreement 1U54HD055764-06, as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research, and RO1-GM097165 to M.C.

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Authors and Affiliations

  1. Center for Reproductive Sciences, University of California, San Francisco, California 94143, USA

    Jing Chen, Simona Torcia, Fang Xie, Chih-Jen Lin, Hakan Cakmak, Federica Franciosi, Kathleen Horner, Courtney Onodera, Jun S. Song, Marcelle I. Cedars, Miguel Ramalho-Santos & Marco Conti

  2. Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, California 94143, USA

    Jing Chen, Simona Torcia, Fang Xie, Chih-Jen Lin, Hakan Cakmak, Federica Franciosi, Kathleen Horner, Courtney Onodera, Jun S. Song, Miguel Ramalho-Santos & Marco Conti

  3. Department of Obstetrics and Gynecology and Reproductive Sciences, University of California, San Francisco, California 94143, USA

    Jing Chen, Simona Torcia, Fang Xie, Chih-Jen Lin, Hakan Cakmak, Federica Franciosi, Kathleen Horner, Marcelle I. Cedars, Miguel Ramalho-Santos & Marco Conti

  4. Institute of Human Genetics, University of California, San Francisco, California 94143, USA

    Courtney Onodera & Jun S. Song

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Contributions

J.C. developed the CEO translation assay and carried out the microarray experiments and some of the AKT assays; S.T. carried out the characterization of the Areg null phenotype; F.X. contributed with western blot studies and oocyte isolation for microinjection; C-J.L. helped with immunostaining experiments and the microinjections in cumulus oocyte complexes; H.C. carried out the experiments on protein secretion and contributed to the writing of the manuscript. F.F. carried out microinjections in CEOs; K.H. contributed with the preparation of the translational luciferase reporters; C.O. and J.S.S. carried out the bioinformatic analysis of the microarray data. M.I.C. advised on data analysis and discussed results; M.R-S. provided reagents and constructs and advised in the interpretation of the data; M.C. conceived the project, designed the experiments, analysed the data and wrote the paper.

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Correspondence to Marco Conti.

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Integrated supplementary information

Supplementary Figure 1 AREG-dependent stimulation of reporter translation is absent when meiotic reentry is prevented.

A. Cumulus enclosed oocytes were injected with TPX2 3’UTR Renilla luciferase reporter and cultured overnight in medium that enables maturation to MII or in medium that prevents maturation (2 μM milrinone) with or without AREG. At the end of the incubation oocytes were freed of surrounding cumulus cells and luciferase activity was measured in extracts from oocytes. The data are mean ± SEM of three to four independent experiments. *p<0.05 vs MII, **p<0.01 vs MII+AREG. B. Cumulus enclosed oocytes were injected with TPX2 3’UTR Renilla luciferase reporter (RLuc) and polyadenylated firefly reporter (FLuc) and cultured overnight as described above. Quantitative RT-PCR for RLuc and FLuc was performed from oocyte extracts. Exposure to Milrinone or AREG did not change the stability of the reporter. No significant differences between groups.

Supplementary Figure 2 AREG-dependent stimulation of reporter translation is absent when 3’UTR is truncated.

A. Diagram of the Renilla luciferase constructs injected to oocytes. B. Cumulus enclosed oocytes were injected with TPX2 3’UTR (1-1630) or TPX2 truncated 3’UTR Renilla luciferase reporter and cultured overnight in medium that enables maturation to MII with or without AREG/EGF or in medium that prevents maturation (2 μM milrinone). At the end of the incubation oocytes were freed of surrounding cumulus cells and luciferase activity was measured in extracts from oocytes. The data are mean ± SEM of three independent experiments. *p<0.05 vs control.

Supplementary Figure 3 Enrichment in 3’UTR motifs in transcripts deregulated in the Areg-/- mice.

Unique 3’UTR DNA sequence for all differentially translated transcripts (Supplemental Tables 1 and 2) was downloaded from the mm9 assembly of the UCSC Genome Browser. The sequences were scanned for motifs with MEME-ChIP (ref. 47), searching only the given strand for any number of repetitions and allowing for motifs of length between 6 and 30 bases. Only motifs with evalues less than 0.01 are reported. For each motif discovered by MEME-ChIP, fasta sequences were downloaded and converted to RNA sequence, and the analogous RNA motif sequence logos were generated using WebLogo 3 48. The motif reported in the top panel corresponds to the motif recognized by the Drosophila Hrb87F or Hrb98DE protein. These proteins are homologous to the mammalian A/B-type HnRNP proteins involved in transport and stabilization of mRNAs.

Supplementary Figure 4 Phosphorylation of AKT (Ser 473) in oocytes is dependent on the presence of somatic cells.

A. Representative time course of AKT Ser 473 phosphorylation in oocytes cultured as cumulus enclosed oocytes (CEOs). CEOs were incubated in medium with AREG and 2 μM milrinone to prevent maturation. At each time oocytes were freed of surrounding cumulus and used for Western blot analysis. Total AKT was used as loading control. A representative experiment of the three performed is reported. B. Representative Western blots of phosphorylated AKT in oocytes cultured as CEO or denuded as indicated times with or without AREG. Total AKT was used as loading control.

Supplementary Figure 5 In situ detection of AKT phosphorylation in oocytes stimulated with AREG when in complex with cumulus cells.

A. CEOs were cultured with or without AREG for 150 min; at the end of the incubation, oocytes were freed of cumulus cells and stained for phospho-AKT (green). B. Quantification of the intensity of the phospho-AKT from different experiments (mean ± SEM; n = 3). *p<0.05 vs CEO. The scale bar corresponds to 10 μm.

Supplementary Figure 6 mTOR inhibitors block the reporter translation stimulated by AREG.

A. Representative Western blot of phosphorylation of ribosomal protein S6 (rpS6) in oocytes cultured as CEO in medium containing 50 nM of rapamycin (mTOR inhibitor) or 100nM of INK128 (selective TORC1/2 inhibitor) for 150min. Tubulin was used as loading control. B. Quantification of the intensity of the phospho-rpS6 immunoreactive band from different experiments (mean ± SEM; n = 3) *P<0.05 vs AREG.

Supplementary Figure 7 Uncropped images of the Western blots included in the main text showing the molecular weight markers.

Dashed boxes indicate the portion of the gel included in the figure. The membrane used to generate Fig 1A, 2D and 2E were cut into two sections and developed with different antibodies. The black dotted line indicates the position of the cut. After development, the two sections of the blot were realigned and exposed together.

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Chen, J., Torcia, S., Xie, F. et al. Somatic cells regulate maternal mRNA translation and developmental competence of mouse oocytes. Nat Cell Biol 15, 1415–1423 (2013). https://doi.org/10.1038/ncb2873

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