Skip to main content
Log in

Assessment of nuclear membrane dynamics using anti-lamin staining offers a clear cut evidence of germinal vesicle breakdown in buffalo oocytes

  • Published:
Cytology and Genetics Aims and scope Submit manuscript

Abstract

Importance of germinal vesicle breakdown (GVBD) in the development chronology of oocyte development can be gauged from the fact that it has been the target of several studies attempting to improve oocyte competence. Manipulation of this event rests on its precise and explicit documentation. Pertinently, we decided to compare three different methods for their efficacy and precision in assessing GVBD. Immature buffalo oocytes were subjected to GVBD inhibition using Roscovitine (Ros) and Cilostamide (Cil) under different concentrations of each as well as their combination, along with control. After 24 h of inhibition, chromatin assessment was done using aceto-orcein staining, hoechst staining and anti-lamin staining. Hoechst staining and anti-lamin staining proved to be easier and less time consuming when compared to aceto-orcein, which was cumbersome and lengthy. Aceto-orcein and hoechst staining were equally ambiguous, while antilamin staining was most precise, accurate and clear in characterizing an oocyte as germinal vesicle (GV) or GVBD. We conclude by stating that anti-lamin staining is an easy and specific technique for assessing GVBD in buffalo oocytes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Terasaki, M., Redistribution of cytoplasmic components during germinal vesicle breakdown in starfish oocytes, J. Cell Sci., 1994, vol. 107, no. 7, pp. 1797–1805.

    PubMed  CAS  Google Scholar 

  2. Metwally, M., Cutting, R., Tipton, A., Skull, J., Ledger, W.L., and Li, T.C., Effect of increased body mass index on oocyte and embryo quality in IVF patients, Reprod. Biomed. Online, 2007, vol. 15, no. 5, pp. 532–538.

    Article  PubMed  CAS  Google Scholar 

  3. Leibfried, L. and First, N.L., Characterization of bovine follicular oocytes and their ability to mature in vitro, J. Anim. Sci., 1979, vol. 48, no. 1, pp. 76–86.

    Article  PubMed  CAS  Google Scholar 

  4. Lonergan, P., Dinnyes, A., Fair, T., Yang, X., and Boland, M., Bovine oocyte and embryo development following meiotic inhibition with butyrolactone I, Mol. Reprod. Dev., 2000, vol. 57, no. 2, pp. 204–209.

    Article  PubMed  CAS  Google Scholar 

  5. Hashimoto, S., Minami, N., Takakura, R., and Imai, H., Bovine immature oocytes acquire developmental competence during meiotic arrest in vitro, Biol. Reprod., 2002, vol. 66, no. 6, pp. 1696–1701.

    Article  PubMed  CAS  Google Scholar 

  6. Prentice-Biensch, J.R., Singh, J., Alfoteisy, B., and Anzar, M., A simple and high-throughput method to assess maturation status of bovine oocytes: Comparison of anti-lamin A/C-DAPI with an aceto-orcein staining technique, Theriogenology, 2012, vol. 78, no. 7, pp. 1633–1638.

    Article  PubMed  CAS  Google Scholar 

  7. Bézard, J., Bøgh, I.B., Duchamp, G., Hyttel, P., and Greve, T., Comparative evaluation of nuclear morphology of equine oocytes aspirated in vivo and stained with Hoechst and orcein, Cells Tissues Organs, 2002, vol. 170, no. 4, pp. 228–236.

    Article  PubMed  Google Scholar 

  8. Bézard, J., Mekarska, A., Goudet, G., Duchamp, G., and Palmer, E., Timing of in vivo maturation of equine preovulatory oocytes and competence for in vitro maturation of immature oocytes collected simultaneously, Equine Vet. J., 1997, vol. 29, no. S25, pp. 33–37.

    Article  Google Scholar 

  9. Goudet, G., Leclercq, L., Bezard, J., Duchamp, G., Guillaume, D., and Palmer, E., Chorionic gonadotropin secretion is associated with an inhibition of follicular growth and an improvement in oocyte competence for in vitro maturation in the mare, Biol. Reprod., 1998, vol. 58, no. 3, pp. 760–768.

    Article  PubMed  CAS  Google Scholar 

  10. Gerace, L. and Burke, B., Functional organization of the nuclear envelope, Annu. Rev. Cell Biol., 1988, vol. 4, pp. 335–374.

    Article  PubMed  CAS  Google Scholar 

  11. Hall, V.J., Cooney, M.A., Shanahan, P., Tecirlioglu, R.T., Ruddock, N.T., and French, A.J., Nuclear lamin antigen and messenger RNA expression in bovine in vitro produced and nuclear transfer embryos, Mol. Reprod. Dev., 2005, vol. 72, no. 4, pp. 471–482.

    Article  PubMed  CAS  Google Scholar 

  12. Lénárt, P., Rabut, G., Daigle, N., Hand, A.R., Terasaki, M., and Ellenberg, J., Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes, J. Cell Biol., 2003, vol. 160, no. 7, pp. 1055–1068.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Ogushi, S., Fulka, J., Jr., and Miyano, T., Germinal vesicle materials are requisite for male pronucleus formation but not for change in the activities of CDK1 and MAP kinase during maturation and fertilization of pig oocytes, Dev. Biol., 2005, vol. 286, no. 1, pp. 287–298.

    CAS  Google Scholar 

  14. Arnault, E., Doussau, M., Pesty, A., Lefevre, B., and Courtot, A.M., Lamin A/C, caspase-6, and chromatin configuration during meiosis resumption in the mouse oocyte, Reprod. Sci., 2010, vol. 17, no. 2, pp. 102–115.

    Article  PubMed  CAS  Google Scholar 

  15. Kalous, J., Solc, P., Baran, V., Kubelka, M., Schultz, R.M., and Motlik, J., PKB/AKT is involved in resumption of meiosis in mouse oocytes, Biol. Cell., 2006, vol. 98, no. 2, pp. 111–123.

    Article  PubMed  CAS  Google Scholar 

  16. Keefer, C.L., Stice, S.L., and Matihews, D.L., Bovine inner cell mass cells as donor nuclei in the production of nuclear transfer embryos and calves, Biol. Reprod., 1994, vol. 50, no. 4, pp. 935–939.

    Article  PubMed  CAS  Google Scholar 

  17. Datta, T.K. and Goswami, S.L., Feasibility of harvesting oocytes from buffalo (Bubalus bubalis) ovaries by different methods, Buffalo J., 1998, vol. 14, pp. 277–284.

    Google Scholar 

  18. Nagai, T., Ebihara, M., Onishi, A., and Kubo, M., Germinal vesicle stages in pig follicular oocytes collected by different methods, J. Reprod. Dev., 1997, vol. 43, no. 4, pp. 339–343.

    Article  Google Scholar 

  19. Motlik, I., Koefoed-Johnson, H.H., and Fulka, J., Breakdown of the germinal vesicle in bovine oocytes cultivated in vitro, J. Exp. Zool., 1978, vol. 205, no. 2, pp. 377–384.

    Article  PubMed  CAS  Google Scholar 

  20. Sun, X.S., Liu, Y., Yue, K.Z., Ma, S.F., and Tan, J.H., Changes in germinal vesicle (GV) chromatin configurations during growth and maturation of porcine oocytes, Mol. Reprod. Dev., 2004, vol. 69, no. 2, pp. 228–234.

    Article  PubMed  CAS  Google Scholar 

  21. Hinrichs, K., Choi, Y.H., Love, L.B., Varner, D.D., Love, C.C., and Walckenaer, B.E., Chromatin configuration within the germinal vesicle of horse oocytes: changes post mortem and relationship to meiotic and developmental competence, Biol. Reprod., 2005, vol. 72, no. 5, pp. 1142–1150.

    Article  PubMed  CAS  Google Scholar 

  22. Viuff, D., Madison, V., Hyttel, P., Avery, B., and Greve, T., Fluorescent intravital staining of bovine oocytes and zygotes, Theriogenology, 1991, vol. 35, no. 1, pp. 291–291.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tirtha Kumar Datta.

Additional information

The article is published in the original.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kumar, S., Dholpuria, S., Chaubey, G.K. et al. Assessment of nuclear membrane dynamics using anti-lamin staining offers a clear cut evidence of germinal vesicle breakdown in buffalo oocytes. Cytol. Genet. 52, 80–85 (2018). https://doi.org/10.3103/S0095452718010061

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0095452718010061

Keywords

Navigation