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Oocytes and early embryos selectively express the survival factor BCL2L10

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Abstract

Apoptosis has been reported in oocytes and human preimplantation embryos both in vitro and in vivo. BCL-2 family proteins are likely to play a pivotal role in controlling oocyte and early embryo degeneration. However, no BCL-2-related survival factors have been identified that would specifically function during oocyte maturation, after fertilization and during early embryogenesis. Here, we performed a comprehensive tissue expression pattern analysis of the BCL-2 family at the mRNA level. While expression of various members was detected in human oocytes and during early primate embryogenesis, our data indicate that BCL2L10 is the predominant maternally loaded Bcl-2 family transcript, revealing an evolutionary conserved expression profile at the egg-to-zygote transition. We provide evidence that BCL2L10 is associated with the microtubule binding protein translationally controlled tumor protein and mitochondria, with a stage-specific redistribution along the pericortical regulatory ooplasm. In dying oocytes, BCL2L10 colocalized with proapoptotic BAX and neutralization of BCL2L10 accelerated oocyte death. We propose BCL2L10 as a novel and prime candidate related to oocyte maturation, fertility, and embryo developmental competence.

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References

  1. Jurisicova A, Acton BM (2004) Deadly decisions: the role of genes regulating programmed cell death in human preimplantation embryo development. Reproduction 128:281–291

    Article  PubMed  CAS  Google Scholar 

  2. Levy R, Benchaib M, Cordonier H, Souchier C, Guerin JF (1998) Annexin V labelling and terminal transferase-mediated DNA end labelling (TUNEL) assay in human arrested embryos. Mol Hum Reprod 4:775–783

    Article  PubMed  CAS  Google Scholar 

  3. Jurisicova A, Antenos M, Varmuza S, Tilly JL, Casper RF (2003) Expression of apoptosis-related genes during human preimplantation embryo development: potential roles for the Harakiri gene product and Caspase-3 in blastomere fragmentation. Mol Hum Reprod 9:133–141

    Article  PubMed  CAS  Google Scholar 

  4. Hardy K (1999) Apoptosis in the human embryo. Rev Reprod 4:125–134

    Article  PubMed  CAS  Google Scholar 

  5. Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47–59

    Article  PubMed  CAS  Google Scholar 

  6. Aouacheria A, Brunet F, Gouy M (2005) Phylogenomics of life-or-death switches in multicellular animals: Bcl-2, BH3-Only, and BNip families of apoptotic regulators. Mol Biol Evol 22:2395–2416

    Article  PubMed  CAS  Google Scholar 

  7. Albamonte MS, Willis MA, Albamonte MI, Jensen F, Espinosa MB, Vitullo AD (2008) The developing human ovary: immunohistochemical analysis of germ-cell-specific VASA protein, BCL-2/BAX expression balance and apoptosis. Hum Reprod 23:1895–1901

    Article  PubMed  CAS  Google Scholar 

  8. Exley GE, Tang C, McElhinny AS, Warner CM (1999) Expression of caspase and BCL-2 apoptotic family members in mouse preimplantation embryos. Biol Reprod 61:231–239

    Article  PubMed  CAS  Google Scholar 

  9. Hartley PS, Bayne RA, Robinson LL, Fulton N, Anderson RA (2002) Developmental changes in expression of myeloid cell leukemia-1 in human germ cells during oogenesis and early folliculogenesis. J Clin Endocrinol Metab 87:3417–3427

    Article  PubMed  CAS  Google Scholar 

  10. Jin X, Han CS, Yu FQ, Wei P, Hu ZY, Liu YX (2005) Anti-apoptotic action of stem cell factor on oocytes in primordial follicles and its signal transduction. Mol Reprod Dev 70:82–90

    Article  PubMed  CAS  Google Scholar 

  11. Metcalfe AD, Hunter HR, Bloor DJ, Lieberman BA, Picton HM, Leese HJ, Kimber SJ, Brison DR (2004) Expression of 11 members of the BCL-2 family of apoptosis regulatory molecules during human preimplantation embryo development and fragmentation. Mol Reprod Dev 68:35–50

    Article  PubMed  CAS  Google Scholar 

  12. Tatone C, Carbone MC, Gallo R, Delle Monache S, Di Cola M, Alesse E, Amicarelli F (2006) Age-associated changes in mouse oocytes during postovulatory in vitro culture: possible role for meiotic kinases and survival factor BCL2. Biol Reprod 74:395–402. doi:10.1095/biolreprod.105.046169 biolreprod.105.046169 [pii]

    Article  PubMed  CAS  Google Scholar 

  13. Liu HC, He ZY, Mele CA, Veeck LL, Davis O, Rosenwaks Z (2000) Expression of apoptosis-related genes in human oocytes and embryos. J Assist Reprod Genet 17:521–533

    Article  PubMed  CAS  Google Scholar 

  14. Matikainen T, Perez GI, Jurisicova A, Pru JK, Schlezinger JJ, Ryu HY, Laine J, Sakai T, Korsmeyer SJ, Casper RF, Sherr DH, Tilly JL (2001) Aromatic hydrocarbon receptor-driven Bax gene expression is required for premature ovarian failure caused by biohazardous environmental chemicals. Nat Genet 28:355–360

    Article  PubMed  CAS  Google Scholar 

  15. Perez GI, Jurisicova A, Matikainen T, Moriyama T, Kim MR, Takai Y, Pru JK, Kolesnick RN, Tilly JL (2005) A central role for ceramide in the age-related acceleration of apoptosis in the female germline. FASEB J 19:860–862

    PubMed  CAS  Google Scholar 

  16. Perez GI, Jurisicova A, Wise L, Lipina T, Kanisek M, Bechard A, Takai Y, Hunt P, Roder J, Grynpas M, Tilly JL (2007) Absence of the proapoptotic Bax protein extends fertility and alleviates age-related health complications in female mice. Proc Natl Acad Sci U S A 104:5229–5234

    Article  PubMed  CAS  Google Scholar 

  17. Perez GI, Robles R, Knudson CM, Flaws JA, Korsmeyer SJ, Tilly JL (1999) Prolongation of ovarian lifespan into advanced chronological age by Bax-deficiency. Nat Genet 21:200–203

    Article  PubMed  CAS  Google Scholar 

  18. Rucker EB 3rd, Dierisseau P, Wagner KU, Garrett L, Wynshaw-Boris A, Flaws JA, Hennighausen L (2000) Bcl-x and Bax regulate mouse primordial germ cell survival and apoptosis during embryogenesis. Mol Endocrinol 14:1038–1052

    Article  PubMed  CAS  Google Scholar 

  19. Motoyama N, Wang F, Roth KA, Sawa H, Nakayama K, Nakayama K, Negishi I, Senju S, Zhang Q, Fujii S et al (1995) Massive cell death of immature hematopoietic cells and neurons in Bcl-x-deficient mice. Science 267:1506–1510

    Article  PubMed  CAS  Google Scholar 

  20. Ratts VS, Flaws JA, Kolp R, Sorenson CM, Tilly JL (1995) Ablation of bcl-2 gene expression decreases the numbers of oocytes and primordial follicles established in the post-natal female mouse gonad. Endocrinology 136:3665–3668

    Article  PubMed  CAS  Google Scholar 

  21. Riedlinger G, Okagaki R, Wagner KU, Rucker EB 3rd, Oka T, Miyoshi K, Flaws JA, Hennighausen L (2002) Bcl-x is not required for maintenance of follicles and corpus luteum in the postnatal mouse ovary. Biol Reprod 66:438–444

    Article  PubMed  CAS  Google Scholar 

  22. Print CG, Loveland KL, Gibson L, Meehan T, Stylianou A, Wreford N, de Kretser D, Metcalf D, Kontgen F, Adams JM, Cory S (1998) Apoptosis regulator Bcl-w is essential for spermatogenesis but appears otherwise redundant. Proc Natl Acad Sci U S A 95:12424–12431

    Article  PubMed  CAS  Google Scholar 

  23. Rinkenberger JL, Horning S, Klocke B, Roth K, Korsmeyer SJ (2000) Mcl-1 deficiency results in peri-implantation embryonic lethality. Genes Dev 14:23–27

    PubMed  CAS  Google Scholar 

  24. Kratz E, Eimon PM, Mukhyala K, Stern H, Zha J, Strasser A, Hart R, Ashkenazi A (2006) Functional characterization of the Bcl-2 gene family in the zebrafish. Cell Death Differ 13:1631–1640. doi:10.1038/sj.cdd.4402016 4402016 [pii]

    Article  PubMed  CAS  Google Scholar 

  25. Aouacheria A, Arnaud E, Venet S, Lalle P, Gouy M, Rigal D, Gillet G (2001) Nrh, a human homologue of Nr-13 associates with Bcl-Xs and is an inhibitor of apoptosis. Oncogene 20:5846–5855

    Article  PubMed  CAS  Google Scholar 

  26. Ke N, Godzik A, Reed JC (2001) Bcl-B, a novel Bcl-2 family member that differentially binds and regulates Bax and Bak. J Biol Chem 276:12481–12484

    Article  PubMed  CAS  Google Scholar 

  27. Zhang H, Holzgreve W, De Geyter C (2001) Bcl2-L-10, a novel anti-apoptotic member of the Bcl-2 family, blocks apoptosis in the mitochondria death pathway but not in the death receptor pathway. Hum Mol Genet 10:2329–2339

    Article  PubMed  CAS  Google Scholar 

  28. Zhai D, Jin C, Huang Z, Satterthwait AC, Reed JC (2008) Differential regulation of Bax and Bak by anti-apoptotic Bcl-2 family proteins Bcl-B and Mcl-1. J Biol Chem 283:9580–9586

    Article  PubMed  CAS  Google Scholar 

  29. Arnaud E, Ferri KF, Thibaut J, Haftek-Terreau Z, Aouacheria A, Le Guellec D, Lorca T, Gillet G (2006) The zebrafish bcl-2 homologue Nrz controls development during somitogenesis and gastrulation via apoptosis-dependent and -independent mechanisms. Cell Death Differ 13:1128–1137

    Article  PubMed  CAS  Google Scholar 

  30. Inohara N, Gourley TS, Carrio R, Muniz M, Merino J, Garcia I, Koseki T, Hu Y, Chen S, Nunez G (1998) Diva, a Bcl-2 homologue that binds directly to Apaf-1 and induces BH3-independent cell death. J Biol Chem 273:32479–32486

    Article  PubMed  CAS  Google Scholar 

  31. Song Q, Kuang Y, Dixit VM, Vincenz C (1999) Boo, a novel negative regulator of cell death, interacts with Apaf-1. EMBO J 18:167–178

    Article  PubMed  CAS  Google Scholar 

  32. Burns KH, Owens GE, Ogbonna SC, Nilson JH, Matzuk MM (2003) Expression profiling analyses of gonadotropin responses and tumor development in the absence of inhibins. Endocrinology 144:4492–4507

    Article  PubMed  CAS  Google Scholar 

  33. Hamatani T, Falco G, Carter MG, Akutsu H, Stagg CA, Sharov AA, Dudekula DB, VanBuren V, Ko MS (2004) Age-associated alteration of gene expression patterns in mouse oocytes. Hum Mol Genet 13:2263–2278

    Article  PubMed  CAS  Google Scholar 

  34. Gallardo TD, John GB, Shirley L, Contreras CM, Akbay EA, Haynie JM, Ward SE, Shidler MJ, Castrillon DH (2007) Genomewide discovery and classification of candidate ovarian fertility genes in the mouse. Genetics 177:179–194. doi:10.1534/genetics.107.074823 genetics.107.074823 [pii]

    Article  PubMed  CAS  Google Scholar 

  35. Assou S, Haouzi D, Mahmoud K, Aouacheria A, Guillemin Y, Pantesco V, Reme T, Dechaud H, De Vos J, Hamamah S (2008) A non-invasive test for assessing embryo potential by gene expression profiles of human cumulus cells: a proof of concept study. Mol Hum Reprod 14:711–719. doi:10.1093/molehr/gan067 gan067 [pii]

    Article  PubMed  CAS  Google Scholar 

  36. Zheng P, Patel B, McMenamin M, Reddy SE, Paprocki AM, Schramm RD, Latham KE (2004) The primate embryo gene expression resource: a novel resource to facilitate rapid analysis of gene expression patterns in non-human primate oocytes and preimplantation stage embryos. Biol Reprod 70:1411–1418. doi:biolreprod.103.023788 10.1095/biolreprod.103.023788 [pii]

    Article  PubMed  CAS  Google Scholar 

  37. Lee YS, Latham KE, Vandevoort CA (2008) Effects of in vitro maturation on gene expression in rhesus monkey oocytes. Physiol Genomics 35:145–158. doi:10.1152/physiolgenomics.90281.2008 90281.2008 [pii]

    Article  PubMed  CAS  Google Scholar 

  38. Griffiths-Jones S, Saini HK, van Dongen S, Enright AJ (2008) miRBase: tools for microRNA genomics. Nucleic Acids Res 36:D154–D158. doi:10.1093/nar/gkm952 gkm952 [pii]

    Article  PubMed  CAS  Google Scholar 

  39. Schier AF (2007) The maternal-zygotic transition: death and birth of RNAs. Science 316:406–407. doi:10.1126/science.1140693 316/5823/406 [pii]

    Article  PubMed  CAS  Google Scholar 

  40. Van Blerkom J, Davis P (2006) High-polarized (Delta Psi m(HIGH)) mitochondria are spatially polarized in human oocytes and early embryos in stable subplasmalemmal domains: developmental significance and the concept of vanguard mitochondria. Reprod Biomed Online 13:246–254

    Article  PubMed  Google Scholar 

  41. Bommer UA, Thiele BJ (2004) The translationally controlled tumour protein (TCTP). Int J Biochem Cell Biol 36:379–385

    Article  PubMed  CAS  Google Scholar 

  42. Zhang D, Li F, Weidner D, Mnjoyan ZH, Fujise K (2002) Physical and functional interaction between myeloid cell leukemia 1 protein (MCL1) and fortilin. The potential role of MCL1 as a fortilin chaperone. J Biol Chem 277:37430–37438. doi:10.1074/jbc.M207413200 M207413200 [pii]

    Article  PubMed  CAS  Google Scholar 

  43. Liu H, Peng HW, Cheng YS, Yuan HS, Yang-Yen HF (2005) Stabilization and enhancement of the antiapoptotic activity of mcl-1 by TCTP. Mol Cell Biol 25:3117–3126. doi:10.1128/MCB.25.8.3117-3126.2005 25/8/3117 [pii]

    Article  PubMed  CAS  Google Scholar 

  44. Yang Y, Yang F, Xiong Z, Yan Y, Wang X, Nishino M, Mirkovic D, Nguyen J, Wang H, Yang XF (2005) An N-terminal region of translationally controlled tumor protein is required for its antiapoptotic activity. Oncogene 24:4778–4788. doi:10.1038/sj.onc.1208666 1208666 [pii]

    Article  PubMed  CAS  Google Scholar 

  45. Russell HR, Lee Y, Miller HL, Zhao J, McKinnon PJ (2002) Murine ovarian development is not affected by inactivation of the bcl-2 family member diva. Mol Cell Biol 22:6866–6870

    Article  PubMed  CAS  Google Scholar 

  46. Yoshino O, McMahon HE, Sharma S, Shimasaki S (2006) A unique preovulatory expression pattern plays a key role in the physiological functions of BMP-15 in the mouse. Proc Natl Acad Sci U S A 103:10678–10683

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We wish to thank the ART/PGD Division and Affymetrix™ microarray platform at IRB (CHU Montpellier) and ART Center of Lyon (CECOS) for their assistance. We gratefully acknowledge Dr. Ines Lein and Dr. Kerstin Mätz-Rensing at the German Primate Center (DPZ GmbH), Leibniz Institute for Primate Research (EUPRIM-Net), for the provision of the marmoset ovary slices. We thank Keith Latham, Bela Patel, and Namdori Mtango for their expert assistance in blot hybridizations. The PREGER resource is supported by a grant from the NIH-NCRR (R24 RR-15253). We are grateful to Fabienne Simian and Claire Lionnet (PLATIM, ENS Lyon) and to Marie Teixeira and Denise Aubert (Animal Facility PBES) for their help. Special thanks are extended to Imène Boumela, Séverine Venet, and Stéphane Gasca. YG is a recipient of an MRT fellowship. This work was partially supported by grants from La Ligue Contre le Cancer (Drôme et Rhône) and Ferring and Organon Pharmaceuticals (France).

Conflict of interest statement

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. IBCP, Institut de Biologie et Chimie des Protéines, CNRS, UMR 5086 Université de Lyon 1, IFR 128, CNRS, IBCP, 7 passage du Vercors, Lyon, F-69367, France

    Yannis Guillemin, Philippe Lalle, Germain Gillet & Abdel Aouacheria

  2. Service de Médecine de la Reproduction, Hôpital femme-Mère-Enfant, Bron, France

    Jean-François Guerin

  3. Laboratoire de Biologie de la Reproduction et du Développement, Hôpital Edouard Herriot, Lyon, France

    Jean-François Guerin

  4. INSERM U846, Bron, France

    Jean-François Guerin

  5. INSERM U847, Institut de Recherche en Biothérapie, CHU de Montpellier, Montpellier, France

    Samir Hamamah

  6. Département de Médecine et Biologie de la Reproduction, CHU de Montpellier, Montpellier, France

    Samir Hamamah

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  1. Yannis Guillemin
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  2. Philippe Lalle
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  3. Germain Gillet
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  4. Jean-François Guerin
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  6. Abdel Aouacheria
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Corresponding author

Correspondence to Abdel Aouacheria.

Electronic supplementary material

Supplementary file 1

Microarray data (PDF 3346 kb)

Supplementary file 2–file 4

Supplementary file 2. Expression of BCL2L10 in human oocytes analyzed by immunofluorescence. Detection of BCL2L10 pertaining to different maturational stages of human oocytes. GV (A), MI (B), MII-stage (C) oocytes visualized by immunofluorescence with anti-BCL2L10 antibody (upper panel) and phase contrast (lower panel). Control staining (without primary antibody) was consistently negative (D). Degenerating, unfertilized oocyte (E). For eight BCL-2 family proteins (BCL-2, BCL-X, BFL-1, BCL-W, MCL-1, BID, BIM, BMF, BAD) analyzed in this manner, no appreciable immunofluorescence was detected, except for BCL-X, MCL-1, and BAD (not shown). Supplementary file 3. A Yeast two-hybrid interaction matrix for BCL2L10, BCL2, BAX, and binding partners isolated in the screen. Yeast clones expressing BCL2L10, BCL2, and BAX in bait vector pGILDA were mated by replica plating with clones expressing BCL2L10, BAX, p53, β-actin, γ-actin, HINT1, and TCTP in prey vector pJG4-5. Picture was taken after 48 h incubation at 30°C. Interactions are measured by the development of a blue color following induction of the LacZ reporter gene. BCL2L10 appears to bind to BAX, HINT1, and TCTP as revealed by light blue coloration. BCL2 binds most strongly to BAX and TCTP, as well as to β-actin, γ-actin, and HINT1. BAX binds to itself but not to the other proteins. Expression of the different proteins has been checked by Western blotting (not shown). The BAX–BAX and BAX–BCL2 interactions served as positive controls for the yeast two-hybrid interaction assay. B Histidine pull down assay of the interaction in vitro between GST-BCL2L10 and TCTP-His6. TCTP-His6 was fixed on Ni-NTA beads and GST (lane 2) or GST-BCL2L10 (lane 3) was applied on the complex. After elution, the attached protein complexes were resolved in a 12% polyacrylamide gel. Anti-GST and anti-TCTP antibodies were used in the Western blot. TCTP-His6 bound specifically to GST-BCL2L10 (lane 3), whereas no interaction between TCTP-His6 and GST was detected (lane 2). Lane 1 (control): GST-BCL2L10 did not interact with Ni-NTA resin. C Indirect ELISA binding assay. Increasing concentrations of purified His-tagged TCTP was used to coat wells of a 96-well plate. Serial dilutions of GST-BCL2L10 were incubated with immobilized TCTP-His6. Binding was detected using anti-GST antibody. GST-BCL2L10 demonstrates dose-dependent binding to immobilized TCTP-His6. GST alone did not demonstrate significant binding to immobilized TCTP-His6. Intensity of the linear signal obtained with GST was subtracted from each experimental point. Supplementary file 4. Confocal images of in vitro matured human oocytes showing TCTP/tubulin double-staining. MII-stage human oocytes (C, D) were double-stained with antibodies against TCTP and microtubules. A5–A8, B5–B8 Enlarged images from A1–A4 and B1–B4, respectively. A1–A8, B1–B8 Two different focal planes of the same oocyte. Microtubules and TCTP are abundant in the peripheral region (A4, A8) and around the meiotic apparatus (B4, B8). Scale bar, 10 μm (PPT 3333 kb)

Supplementary file 5

Supplementary material to Figs. 7 and 9 (PPT 486 kb)

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Guillemin, Y., Lalle, P., Gillet, G. et al. Oocytes and early embryos selectively express the survival factor BCL2L10. J Mol Med 87, 923–940 (2009). https://doi.org/10.1007/s00109-009-0495-7

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