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Characterization of a diverse secretome generated by the mouse preimplantation embryo in vitro

By Amanda J Beardsley, Yan Li and Chris O'Neill


This study investigates the suitability of surface-enhanced laser desorption and ionization time-of-flight (SELDI-TOF) and electrospray ionization (ESI) mass spectrometry for analysis of the proteins released by the mouse preimplantation embryo in vitro. SELDI-TOF analysis with CM10 or IMAC30 (but not Q10) protein chips detected a protein peak at m/z ~8570 released by both C57BL6 and hybrid embryos. No other peaks unique to the presence of the embryo were identified with this method. ESI mass spectrometry of tryptic digests of embryo-conditioned media identified a total of 20 proteins released during development from the zygote to blastocyst stage. Four proteins were expressed in at least 7 out of 8 cultures tested, one of these (lactate dehydrogenase B) was in all cultures. A further five proteins were in at least half of the cultures and 11 more proteins were in at least one culture. The expression of two of these proteins is essential for preimplantation embryo development (NLR family, pyrin domain containing 5 and peptidyl arginine deiminase, type VI). A further four proteins detected have roles in redox regulation of cells, and three others are capable of inducing post-translational modifications of proteins. This study shows the feasibility of ESI mass spectrometry for identifying the proteins secreted by the preimplantation embryo in vitro. This analysis identifies a range of targets that now require detailed functional analysis to assess whether their release by the embryo is an important property of early embryo development

Topics: Research
Publisher: BioMed Central
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Provided by: PubMed Central

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  1. (1997). AJ: Bovine oviductal and embryonic insulin-like growth factor binding proteins: possible regulators of "embryotrophic" insulin-like growth factor circuits. Biol Reprod
  2. (2006). Analysis of protein expression (secretome) by human and mouse preimplantation embryos. Fertil Steril
  3. (1990). Buytaert PP: Embryo-derived platelet-activating factor, a marker of embryo quality and viability following ovarian stimulation for invitro fertilization. J in Vitro Fert Emb Transf
  4. (1997). C: The role of albumin in the release of plateletactivating factor by mouse preimplantation embryos in vitro. J Reprod Fertil
  5. (2002). Calciumbinding proteins and calcium-release channels in human maturing oocytes, pronuclear zygotes and early preimplantation embryos. Hum Reprod
  6. (2002). Characterization and biological roles of oviduct-specific, oestrogen-dependent glycoprotein. Reproduction
  7. (2004). CJ: Calreticulin on the mouse egg surface mediates transmembrane signaling linked to cell cycle resumption. Devel Biol
  8. (1994). CM: Identification of the Ped gene at the molecular level: the Q9 MHC class I transgene converts the Ped slow to the Ped fast phenotype. Biol Reprod
  9. (2003). Effect of a null mutation of the oviduct-specific glycoprotein gene on mouse fertilization.
  10. (2003). ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. Dev Biol
  11. (1997). Evidence for the requirement of autocrine growth factors for development of mouse preimplantation embryos in vitro. Biol Reprod
  12. (1998). Hämmerling G: ER-60, a chaperone with thiol-dependent reductase activity involved in MHC class I assembly.
  13. (2004). Impaired p53 Expression, Function, and Nuclear Localization in Calreticulin-deficient Cells. Mol Biol Cell
  14. (2007). JA: Peptidylarginine deiminase (PAD) 6 is essential for oocyte cytoskeletal sheet formation and female fertility. Mol Cell Endocrinol
  15. (1998). JR: Protein Identification at the Low Femtomole Level from Silver-Stained Gels Using a New Fritless Electrospray Interface for Liquid Chromatography-Microspray and Nanospray Mass Spectrometry. Anal Biochem
  16. (2000). Lactate Regulates Pyruvate Uptake and Metabolism in the Preimplantation Mouse Embryo. Biol Reprod
  17. (1996). Librach CL: HLA-G expression during preimplantation human embryo development.
  18. (2004). LM: Developmental Expression and Subcellular Localization of Mouse MATER, an Oocyte-Specific Protein Essential for Early Development. Endocrinology
  19. (2008). Maternally derived FILIAMATER complex localizes asymmetrically in cleavage-stage mouse embryos. Development
  20. (1991). MF: Analysis of litter size and weight in mice differing in Ped gene phenotype and the Q region of the H-2 complex.
  21. (1995). ML: Secretion, Surface Localization, Turnover, and Steady State Expression of Protein Disulfide Isomerase in Rat Hepatocytes.
  22. (2001). Mújica A: Exocytosis of a 60 kDa protein (Calreticulin) from activated hamster oocytes. Mol Reprod Dev
  23. (1986). Non-invasive measurement of nutrient uptake by single cultured pre-implantation mouse embryos. Hum Reprod
  24. (1990). Non-invasive measurement of pyruvate and glucose uptake and lactate production by sigle human preimplantation embryos. Hum Reprod
  25. (1992). O'Neill C: Biosynthesis of platelet-activating factor by the mouse two-embryo.
  26. (1985). Partial characterisation of the embryo-derived platelet activating factor in mice.
  27. (2005). Peptidylarginine deiminase (PAD) is a mouse cortical granule protein that plays a role in preimplantation embryonic development. Reprod Biol Endocrinol
  28. (1993). Preferential survival of mice expressing the Qa-2 antigen. J Reprod Fert
  29. (1997). Preimplantation growth factor physiology. Rev Reprod
  30. (1987). Saunders DM: Use of a bio-assay for embryo-derived platelet activating factor as a means of assessing quality and pregnancy potential of human embryos. Fertil Steril
  31. (1990). SK: Preimplantation embryo development in vitro: Cooperative interactions among embryos and the role of growth factors.
  32. (2005). Soluble human leukocyte antigen G expression in phase I culture media at 46 hours after fertilization predicts pregnancy and implantation from day 3 embryo transfer. Fertil Steril
  33. (2002). Spanos S: Growth factor expression and function in the human and mouse preimplantation embryo.
  34. (1997). Studies of the nature of the binding by albumin of platelet-activating factor released from cells.
  35. (2001). The ins and outs of calreticulin: from the ER lumen to the extracellular space. Trends Cell Biol
  36. (2005). The role of paf in embryo physiology. Hum Reprod Update