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Dysregulation of bisphosphoglycerate mutase during in vitro maturation of oocytes

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Oxygen is vital for oocyte maturation; however, oxygen regulation within ovarian follicles is not fully understood. Hemoglobin is abundant within the in vivo matured oocyte, indicating potential function as an oxygen regulator. However, hemoglobin is significantly reduced following in vitro maturation (IVM). The molecule 2,3-bisphosphoglycerate (2,3-BPG) is essential in red blood cells, facilitating release of oxygen from hemoglobin. Towards understanding the role of 2,3-BPG in the oocyte, we characterized gene expression and protein abundance of bisphosphoglycerate mutase (Bpgm), which synthesizes 2,3-BPG, and whether this is altered under low oxygen or hemoglobin addition during IVM.

Methods

Hemoglobin and Bpgm expression within in vivo matured human cumulus cells and mouse cumulus-oocyte complexes (COCs) were evaluated to determine physiological levels of Bpgm. During IVM, Bpgm gene expression and protein abundance were analyzed in the presence or absence of low oxygen (2% and 5% oxygen) or exogenous hemoglobin.

Results

The expression of Bpgm was significantly lower than hemoglobin when mouse COCs were matured in vivo. Following IVM at 20% oxygen, Bpgm gene expression and protein abundance were significantly higher compared to in vivo. At 2% oxygen, Bpgm was significantly higher compared to 20% oxygen, while exogenous hemoglobin resulted in significantly lower Bpgm in the COC.

Conclusion

Hemoglobin and 2,3-BPG may play a role within the maturing COC. This study shows that IVM increases Bpgm within COCs compared to in vivo. Decreasing oxygen concentration and the addition of hemoglobin altered Bpgm, albeit not to levels observed in vivo.

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Data availability

All data generated or analyzed during this study are included in this published article and are available from the corresponding author on reasonable request.

Code availability

N/A

References

  1. Thompson JG, Brown HM, Kind KL, Russell DL. The ovarian antral follicle: living on the edge of hypoxia or not? Biol Reprod. 2015;92(6):1–6. https://doi.org/10.1095/biolreprod.115.128660.

    Article  CAS  Google Scholar 

  2. Fischer B, Kunzel W, Kleinstein J, Gips H. Oxygen tension in follicular fluid falls with follicle maturation. Eur J Obstet Gynecol Reprod Biol. 1992;43(1):39–43.

    Article  CAS  Google Scholar 

  3. Redding GP, Bronlund JE, Hart AL. Theoretical investigation into the dissolved oxygen levels in follicular fluid of the developing human follicle using mathematical modelling. Reprod Fertil Dev. 2008;20(3):408–17. https://doi.org/10.1071/rd07190.

    Article  CAS  PubMed  Google Scholar 

  4. Hashimoto S, Minami N, Takakura R, Yamada M, Imai H, Kashima N. Low oxygen tension during in vitro maturation is beneficial for supporting the subsequent development of bovine cumulus-oocyte complexes. Mol Reprod Dev. 2000;57(4):353–60.

    Article  CAS  Google Scholar 

  5. Kim J, Bagchi IC, Bagchi MK. Signaling by hypoxia-inducible factors is critical for ovulation in mice. Endocrinology. 2009;150(7):3392–400. https://doi.org/10.1210/en.2008-0948.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. van den Driesche S, Myers M, Gay E, Thong KJ, Duncan WC. HCG up-regulates hypoxia inducible factor-1 alpha in luteinized granulosa cells: implications for the hormonal regulation of vascular endothelial growth factor A in the human corpus luteum. Mol Hum Reprod. 2008;14(8):455–64. https://doi.org/10.1093/molehr/gan040.

    Article  CAS  PubMed  Google Scholar 

  7. Kind KL, Tam KK, Banwell KM, Gauld AD, Russell DL, Macpherson AM, et al. Oxygen-regulated gene expression in murine cumulus cells. Reprod Fertil Dev. 2015;27(2):407–18. https://doi.org/10.1071/RD13249.

    Article  CAS  PubMed  Google Scholar 

  8. Lim M, Thompson JG, Dunning KR. HYPOXIA AND REPRODUCTIVE HEALTH: Hypoxia and ovarian function: follicle development, ovulation, oocyte maturation. Reproduction. 2021;161(1):F33–40. https://doi.org/10.1530/rep-20-0509.

    Article  CAS  PubMed  Google Scholar 

  9. Kind KL, Banwell KM, Gebhardt KM, Macpherson A, Gauld A, Russell DL, et al. Microarray analysis of mRNA from cumulus cells following in vivo or in vitro maturation of mouse cumulus-oocyte complexes. Reprod Fertil Dev. 2013;25(2):426–38. https://doi.org/10.1071/RD11305.

    Article  CAS  PubMed  Google Scholar 

  10. Brown HM, Anastasi MR, Frank LA, Kind KL, Richani D, Robker RL, et al. Hemoglobin: a gas transport molecule that is hormonally regulated in the ovarian follicle in mice and humans. Biol Reprod. 2015;92(1):26. https://doi.org/10.1095/biolreprod.114.124594.

    Article  CAS  PubMed  Google Scholar 

  11. Benesch R, Benesch RE. The effect of organic phosphates from the human erythrocyte on the allosteric properties of hemoglobin. Biochem Biophys Res Commun. 1967;26(2):162–7.

    Article  CAS  Google Scholar 

  12. Pritlove DC, Gu M, Boyd CAR, Randeva HS, Vatish M. Novel placental expression of 2,3-bisphosphoglycerate mutase. Placenta. 2006;27(8):924–7. https://doi.org/10.1016/j.placenta.2005.08.010.

    Article  CAS  PubMed  Google Scholar 

  13. Lim JM, Hansel W. Improved development of in vitro-derived bovine embryos by use of a nitric oxide scavenger in a cumulus-granulosa cell coculture system. Mol Reprod Dev. 1998;50(1):45–53. https://doi.org/10.1002/(SICI)1098-2795(199805)50:1<45::AID-MRD6>3.0.CO;2-X.

    Article  CAS  PubMed  Google Scholar 

  14. Banwell KM, Lane M, Russell DL, Kind KL, Thompson JG. Oxygen concentration during mouse oocyte in vitro maturation affects embryo and fetal development. Hum Reprod. 2007;22(10):2768–75. https://doi.org/10.1093/humrep/dem203.

    Article  CAS  PubMed  Google Scholar 

  15. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001;25(4):402–8. https://doi.org/10.1006/meth.2001.1262.

    Article  CAS  PubMed  Google Scholar 

  16. Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper – Excel-based tool using pair-wise correlations. Biotechnol Lett. 2004;26(6):509–15. https://doi.org/10.1023/B:BILE.0000019559.84305.47.

    Article  CAS  PubMed  Google Scholar 

  17. Dunning KR, Watson LN, Zhang VJ, Brown HM, Kaczmarek AK, Robker RL, et al. Activation of mouse cumulus-oocyte complex maturation in vitro through EGF-like activity of versican. Biol Reprod. 2015;92(5):116. https://doi.org/10.1095/biolreprod.114.127274.

    Article  CAS  PubMed  Google Scholar 

  18. Sutton-McDowall ML, Purdey M, Brown HM, Abell AD, Mottershead DG, Cetica PD, et al. Redox and anti-oxidant state within cattle oocytes following in vitro maturation with bone morphogenetic protein 15 and follicle stimulating hormone. Mol Reprod Dev. 2015;82(4):281–94. https://doi.org/10.1002/mrd.22470.

    Article  CAS  PubMed  Google Scholar 

  19. Saha D, Patgaonkar M, Shroff A, Ayyar K, Bashir T, Reddy K. Hemoglobin expression in nonerythroid cells: novel or ubiquitous? Int J Inf Secur. 2014;2014:1–8.

    Google Scholar 

  20. Gorr TA, Wichmann D, Pilarsky C, Theurillat JP, Fabrizius A, Laufs T, et al. Old proteins - new locations: myoglobin, haemoglobin, neuroglobin and cytoglobin in solid tumours and cancer cells. Acta Physiol (Oxford). 2011;202(3):563–81. https://doi.org/10.1111/j.1748-1716.2010.02205.x.

    Article  CAS  Google Scholar 

  21. Emara M, Turner AR, Allalunis-Turner J. Hypoxia differentially upregulates the expression of embryonic, fetal and adult hemoglobin in human glioblastoma cells. Int J Oncol. 2014;44(3):950–8.

    Article  CAS  Google Scholar 

  22. Lim M, Brown HM, Kind KL, Thompson JG, Dunning KR. Hemoglobin: potential roles in the oocyte and early embryo†. Biol Reprod. 2019;101(2):262–70. https://doi.org/10.1093/biolre/ioz078.

    Article  PubMed  Google Scholar 

  23. Lim M, Brown HM, Kind KL, Breen J, Anastasi MR, Ritter LJ, et al. Haemoglobin expression in in vivo murine preimplantation embryos suggests a role in oxygen-regulated gene expression. Reprod Fertil Dev. 2018;31:724–34. https://doi.org/10.1071/RD17321.

    Article  CAS  Google Scholar 

  24. Brown HM, Dunning KR, Sutton-McDowall M, Gilchrist RB, Thompson JG, Russell DL. Failure to launch: aberrant cumulus gene expression during oocyte in vitro maturation. Reproduction. 2017;153(3):R109–R20. https://doi.org/10.1530/rep-16-0426.

    Article  CAS  PubMed  Google Scholar 

  25. Lee YS, Latham KE, Vandevoort CA. Effects of in vitro maturation on gene expression in rhesus monkey oocytes. Physiol Genomics. 2008;35(2):145–58. https://doi.org/10.1152/physiolgenomics.90281.2008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

ML is supported by an Australian Postgraduate Award; JGT is supported by a National Health & Medical Research Council Research Fellowship (ID 1077694), and KRD is supported by a Mid-Career Fellowship from the Hospital Research Foundation (C-MCF-58-2019). This study was funded by the Australian Research Council Centre of Excellence for Nanoscale Biophotonics (CE140100003).

Author information

Authors and Affiliations

  1. Robinson Research Institute, The University of Adelaide, Adelaide, South Australia, Australia

    Megan Lim, Ryan D. Rose, Jeremy G. Thompson & Kylie R. Dunning

  2. Australian Research Council Centre of Excellence for Nanoscale BioPhotonics, The University of Adelaide, Adelaide, South Australia, Australia

    Megan Lim, Jeremy G. Thompson & Kylie R. Dunning

  3. Institute for Photonics and Advanced Sensing, The University of Adelaide, Adelaide, South Australia, Australia

    Megan Lim, Jeremy G. Thompson & Kylie R. Dunning

  4. The Victorian Heart Institute, Monash University, Melbourne, Victoria, Australia

    Hannah M. Brown

  5. Fertility SA, St. Andrews Hospital, South Terrace, Adelaide, South Australia, Australia

    Ryan D. Rose

Authors
  1. Megan Lim
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  2. Hannah M. Brown
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  3. Ryan D. Rose
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  4. Jeremy G. Thompson
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  5. Kylie R. Dunning
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Contributions

ML performed the experiments, analyzed data, wrote and edited the manuscript. HMB, RDR, and JGT provided critical feedback on the final manuscript. JGT conceived the original idea. KRD supervised the project, conceived and planned the experiments, and aided in interpreting results and in editing of the manuscript.

Corresponding author

Correspondence to Kylie R. Dunning.

Ethics declarations

Ethics approval

Ethics approval for the study was obtained from the Human Research Ethics Committee, University of Adelaide, Adelaide, Australia (approval number H-2018-205) and the University of Adelaide Animal Ethics Committee (M-2016-147).

Consent to participate

Consent for cumulus cells to be stored and used for research purposes (St Andrews Hospital, Human Research Ethics Committee Project No. 93).

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The authors consent for publication of this article.

Conflict of interest

The authors declare no competing interests.

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Lim, M., Brown, H.M., Rose, R.D. et al. Dysregulation of bisphosphoglycerate mutase during in vitro maturation of oocytes. J Assist Reprod Genet 38, 1363–1372 (2021). https://doi.org/10.1007/s10815-021-02230-0

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  • DOI: https://doi.org/10.1007/s10815-021-02230-0

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