Advertisement

Journal of Assisted Reproduction and Genetics

, Volume 29, Issue 11, pp 1289–1297 | Cite as

The follicular microenviroment as a predictor of pregnancy: MALDI-TOF MS lipid profile in cumulus cells

  • Daniela Antunes Montani
  • Fernanda Bertuccez Cordeiro
  • Thaís Regiani
  • Amanda Begati Victorino
  • Eduardo Jorge Pilau
  • Fábio Cesar Gozzo
  • Christina Ramires Ferreira
  • Renato Fraietta
  • Edson Guimarães Lo Turco
Technological Innovations

Abstract

Purpose

This research proposed to study the changes in lipid composition in cumulus cells (CCs) from women who achieved pregnancy compared with women who did not, after in vitro fertilization treatment. This approach has the potential to provide novel information on the lipid metabolism of the CCs and as an additional method to predict pregnancy.

Method

Fifty-four samples from couples with tubal and male factor infertility and where the female partner was age 35 or younger were divided in two groups according to their level of hCG 14 days after embryo transfer as follows: (1) 23 samples in pregnant group and (2) 31 samples in non-pregnant group. Lipid extraction was performed by the Bligh-Dyer protocol, and lipid profiles were obtained by MALDI-TOF MS. Mass spectra data were processed with MassLynx, and statistical analysis was performed using MarkerLynx extended statistic. OPLS-DA model was built.

Results

S-plot Analysis revealed three ions as potential markers in the pregnant group, and five ions in the non-pregnant group. These ions were identified in the human metabolome database (HMDB) as phosphatidylcholine in the pregnant group and as phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol species in the non-pregnant group. These lipids might be involved in cell proliferation and differentiation, apoptosis and GAP junction regulation.

Conclusion

We conclude that MALDI-TOF MS can be used as an informative and fast analytical strategy to obtain and study the lipid profile of cumulus cells and can potentially be used as a supporting tool to predict pregnancy based on the metabolic state of the CCs.

Keywords

Cumulus cell Lipid profile MALDI-TOF MS Pregnancy outcome 

Notes

Acknowledgements

Funding for the study was provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) process no 2010/14732-5.

References

  1. 1.
    Adriaenssens T, Wathlet S, Segers I, Verheyen G, De Vos A, Van Der Elst J, et al. Cumulus cell gene expression is associated with oocyte developmental quality and influenced by patient and treatment characteristics. Hum Reprod. 2010;25:1259–70.PubMedCrossRefGoogle Scholar
  2. 2.
    Anderson R, Sciorio R, Kinnell H, Bayne R, Thong K, DeSousa P, et al. Cumulus gene expression as a predictor of human oocyte fertilisation, embryo development and competence to establish a pregnancy. Reproduction. 2009;138:629–37.PubMedCrossRefGoogle Scholar
  3. 3.
    Assidi M, Montag M, Van Der Ven K, Ma S. Biomarkers of human oocyte developmental competence expressed in cumulus cells before ICSI: a preliminary study. J Assist Reprod Genet. 2011;28:173–88.PubMedCrossRefGoogle Scholar
  4. 4.
    Balasubramanian K, Mirnikjoo B, Schroit AJ. Regulated externalization of phosphatidylserine at the cell surface: implications for apoptosis. J Biol Chem. 2007;282:18357–64.PubMedCrossRefGoogle Scholar
  5. 5.
    Blaho VA, Buczynski MW, Brown CR, Dennis EA. Lipidomic analysis of dynamic eicosanoid responses during the induction and resolution of Lyme arthritis. J Biol Chem. 2009;284:21599–612.PubMedCrossRefGoogle Scholar
  6. 6.
    Brison DR, Houghton FD, Falconer D, Roberts SA, Hawkhead J, Humpherson PG, et al. Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod. 2004;19:2319–24.PubMedCrossRefGoogle Scholar
  7. 7.
    Caballero F, Fernandez A, Matias N, et al. Specific contribution of methionine and choline in nutritional nonalcoholic steatohepatitis: impact on mitochondrial S-adenosyl-L-methionine and glutathione. J Biol Chem. 2010;285:18528–36.PubMedCrossRefGoogle Scholar
  8. 8.
    Cui Z, Houweling M, Chen MH, Record M, Chap H, Vance DE, et al. A genetic defect in phosphatidylcholine biosynthesis triggers apoptosis in Chinese hamster ovary cells. J Biol Chem. 1996;271:14668–71.PubMedCrossRefGoogle Scholar
  9. 9.
    Di Paolo G, De Camilli P. Phosphoinositides in cell regulation and membrane dynamics. Nature. 2006;443:651–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Diaz FJ, O'brien MJ, Wigglesworth K, Eppig JJ. The preantral granulosa cell to cumulus cell transition in the mouse ovary: development of competence to undergo expansion. Dev Biol. 2006;299:91–104.PubMedCrossRefGoogle Scholar
  11. 11.
    Elvin JA, Clark AT, Wang P, Wolfman NM, Matzuk MM. Paracrine actions of growth differentiation factor-9 in the mammalian ovary. Mol Endocrinol. 1999;13:1035–48.PubMedCrossRefGoogle Scholar
  12. 12.
    Emoto K, Toyama-Sorimachi N, Karasuyama H, Inoue K, Umeda M. Exposure of phosphatidylethanolamine on the surface of apoptotic cells. Exp Cell Res. 1997;232:430–4.PubMedCrossRefGoogle Scholar
  13. 13.
    Eppig J. Maintenance of meiotic arrest and the induction of oocyte maturation in mouse oocyte-granulosa cell complexes developed in vitro from preantral follicles. Biol Reprod. 1991;45:824–30.PubMedCrossRefGoogle Scholar
  14. 14.
    Eppig J. Oocyte control of ovarian follicular development and function in mammals. Reproduction. 2001;122:829–38.PubMedCrossRefGoogle Scholar
  15. 15.
    Fadok VA, de Cathelineau A, Daleke DL, Henson PM, Dl B. Loss of phospholipid asymmetry and surface exposure of phosphatidylserine is required for phagocytosis of apoptotic cells by macrophages and fibroblasts. J Biol Chem. 2001;276:1071–7.PubMedCrossRefGoogle Scholar
  16. 16.
    Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol. 1992;148:2207–16.PubMedGoogle Scholar
  17. 17.
    Gilchrist RB, Lane M, Thompson JG. Oocyte-secreted factors: regulators of cumulus cell function and oocyte quality. Hum Reprod Update. 2008;14:159–77.PubMedCrossRefGoogle Scholar
  18. 18.
    Giorgetti C, Terriou P, Auquier P, Hans E, Spach JL, Salzmann J, et al. Embryo score to predict implantation after in-vitro fertilization: based on 957 single embryo transfers. Hum Reprod. 1995;10:2427–31.PubMedGoogle Scholar
  19. 19.
    Hamel M, Dufort I, Robert C, Gravel C, Leveille MC, Leader A, et al. Identification of differentially expressed markers in human follicular cells associated with competent oocytes. Hum Reprod. 2008;23:1118–27.PubMedCrossRefGoogle Scholar
  20. 20.
    Hannun YA, Obeid LM. Principles of bioactive lipid signalling: lessons from sphingolipids. Nat Rev Mol Cell Biol. 2008;9:139–50.PubMedCrossRefGoogle Scholar
  21. 21.
    Hillier S. Research challenge: what is the best non-invasive test of oocyte/embryo competence? Mol Hum Reprod. 2008;14:665.PubMedCrossRefGoogle Scholar
  22. 22.
    Kim HY, Akbar M, Lau A, Edsall L. Inhibition of neuronal apoptosis by docosahexaenoic acid (22:6n-3). Role of phosphatidylserine in antiapoptotic effect. J Biol Chem. 2000;275:35215–23.PubMedCrossRefGoogle Scholar
  23. 23.
    Kim JK, Park SY, Na JK, Seong ES, Yu CY. Metabolite profiling based on lipophilic compounds for quality assessment of perilla (Perilla frutescens) cultivars. J Agric Food Chem. 2012;60:2257–63.PubMedCrossRefGoogle Scholar
  24. 24.
    Kruger T, Menkveld R, Stander F, Lombard C, Der Merwe V, Jp VZ, et al. Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil Steril. 1986;46:1118–23.PubMedGoogle Scholar
  25. 25.
    Li Z, Agellon LB, Allen TM, Umeda M, Jewell L, Mason A, et al. The ratio of phosphatidylcholine to phosphatidylethanolamine influences membrane integrity and steatohepatitis. Cell Metab. 2006;3:321–31.PubMedCrossRefGoogle Scholar
  26. 26.
    Liang CG, Huo LJ, Zhong ZS, Chen DY, Schatten H, Sun QY. Cyclic adenosine 3',5'-monophosphate-dependent activation of mitogen-activated protein kinase in cumulus cells is essential for germinal vesicle breakdown of porcine cumulus-enclosed oocytes. Endocrinology. 2005;146:4437–44.PubMedCrossRefGoogle Scholar
  27. 27.
    Lin YH, Hwang JL, Seow KM, Huang LW, Chen HJ, Tzeng CR. Effects of growth factors and granulosa cell co-culture on in-vitro maturation of oocytes. Reprod Biomed Online. 2009;19:165–70.PubMedCrossRefGoogle Scholar
  28. 28.
    Liu Y, Zhou C, Xu Y, Fang C, Zhang M. Pregnancy outcome in preimplantation genetic diagnosis cycle by blastomere biopsy is related to both quality and quantity of embryos on day 3. Fertil Steril. 2009;91(4 Suppl):1355–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Miyoshi T, Otsuka F, Inagaki K, Otani H, Takeda M, Suzuki J, et al. Differential regulation of steroidogenesis by bone morphogenetic proteins in granulosa cells: involvement of extracellularly regulated kinase signaling and oocyte actions in follicle-stimulating hormone-induced estrogen production. Endocrinology. 2007;148:337–45.PubMedCrossRefGoogle Scholar
  30. 30.
    Neshat MS, Raitano AB, Wang HG, Reed JC, Sawyers CL. The survival function of the Bcr-Abl oncogene is mediated by Bad-dependent and -independent pathways: roles for phosphatidylinositol 3-kinase and Raf. Mol Cell Biol. 2000;20:1179–86.32.PubMedCrossRefGoogle Scholar
  31. 31.
    Niebergall L, Vance D. The ratio of phosphatidylcholine to phosphatidylethanolamine does not predict integrity of growing MT58 Chinese hamster ovary cells. Biochim Biophys Acta. 2012;1821:324–34.PubMedCrossRefGoogle Scholar
  32. 32.
    Post JA, Bijvelt JJ, Aj V. Phosphatidylethanolamine and sarcolemmal damage during ischemia or metabolic inhibition of heart myocytes. Am J Physiol. 1995;268:H773–80.PubMedGoogle Scholar
  33. 33.
    Rantalainen M, Cloarec O, Ebbels TM, Lundstedt T, Nicholson JK, Holmes E, et al. Piecewise multivariate modelling of sequential metabolic profiling data. BMC Bioinformatics. 2008;9:105.PubMedCrossRefGoogle Scholar
  34. 34.
    Reich A, Klatsky P, Carson S, Wessel G. The transcriptome of a human polar body accurately reflects its sibling oocyte. J Biol Chem. 2011;286:40743–9.PubMedCrossRefGoogle Scholar
  35. 35.
    Saarikangas J, Zhao H, Lappalainen P. Regulation of the actin cytoskeleton-plasma membrane interplay by phosphoinositides. Physiol Rev. 2010;90:259–89.PubMedCrossRefGoogle Scholar
  36. 36.
    Salomoni P, Wasik MA, Riedel RF, Reiss K, Choi JK, Skorski T, et al. Expression of constitutively active Raf-1 in the mitochondria restores antiapoptotic and leukemogenic potential of a transformation-deficient BCR/ABL mutant. J Exp Med. 1998;187:1995–2007.PubMedCrossRefGoogle Scholar
  37. 37.
    Schiller J, Arnhold J, Benard S, Muller M, Reichl S, Arnold K. Lipid analysis by matrix-assisted laser desorption and ionization mass spectrometry: A methodological approach. Anal Biochem. 1999;267:46–56.PubMedCrossRefGoogle Scholar
  38. 38.
    Sela-Abramovich S, Chorev E, Galiani D, Dekel N. Mitogen-activated protein kinase mediates luteinizing hormone-induced breakdown of communication and oocyte maturation in rat ovarian follicles. Endocrinology. 2005;146:1236–44.PubMedCrossRefGoogle Scholar
  39. 39.
    Shewan A, Eastburn DJ, Mostov K. Phosphoinositides in cell architecture. Cold Spring Harb Perspect Biol. 2011;3:a004796.PubMedCrossRefGoogle Scholar
  40. 40.
    Sirard MA, Dufort I, Coenen K, Tremblay K, Massicotte L, Robert C. The use of genomics and proteomics to understand oocyte and early embryo functions in farm animals. Reprod Suppl. 2003;61:117–29.PubMedGoogle Scholar
  41. 41.
    Tanghe S, Van Soom A, Nauwynck H, Coryn M, De Kruif A. Minireview: Functions of the cumulus oophorus during oocyte maturation, ovulation, and fertilization. Mol Reprod Dev. 2002;61:414–24.PubMedCrossRefGoogle Scholar
  42. 42.
    Vance J. Phosphatidylserine and phosphatidylethanolamine in mammalian cells: two metabolically related aminophospholipids. J Lipid Res. 2008;49:1377–87.PubMedCrossRefGoogle Scholar
  43. 43.
    Verhoven B, Schlegel RA, Williamson P. Mechanisms of phosphatidylserine exposure, a phagocyte recognition signal, on apoptotic T lymphocytes. J Exp Med. 1995;182:1597–601.PubMedCrossRefGoogle Scholar
  44. 44.
    Yu H, Fukami K, Watanabe Y, Ozaki C, Takenawa T. Phosphatidylinositol 4,5-bisphosphate reverses the inhibition of RNA transcription caused by histone H1. Eur J Biochem. 1998;251:281–7.PubMedCrossRefGoogle Scholar
  45. 45.
    Ziebe S, Petersen K, Lindenberg S, Andersen AG, Gabrielsen A, Andersen AN. Embryo morphology or cleavage stage: how to select the best embryos for transfer after in-vitro fertilization. Hum Reprod. 1997;12:1545–9.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Daniela Antunes Montani
    • 1
  • Fernanda Bertuccez Cordeiro
    • 1
  • Thaís Regiani
    • 1
  • Amanda Begati Victorino
    • 1
  • Eduardo Jorge Pilau
    • 2
  • Fábio Cesar Gozzo
    • 2
  • Christina Ramires Ferreira
    • 2
  • Renato Fraietta
    • 1
  • Edson Guimarães Lo Turco
    • 1
  1. 1.Department of Surgery, Division of Urology, Human Reproduction SectionSao Paulo Federal UniversitySao PauloBrazil
  2. 2.Institute of ChemistryUniversity of CampinasCampinasBrazil

Personalised recommendations