Advertisement

Cytology and Genetics

, Volume 52, Issue 1, pp 80–85 | Cite as

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

  • Sandeep Kumar
  • Sunny Dholpuria
  • Gaurav Kumar Chaubey
  • Rakesh Kumar
  • Tirtha Kumar DattaEmail author
Article

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.

Keywords

chromatin nuclear lamin buffalo oocyte GVBD inhibition 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 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.PubMedGoogle Scholar
  2. 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.CrossRefPubMedGoogle Scholar
  3. 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.CrossRefPubMedGoogle Scholar
  4. 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.CrossRefPubMedGoogle Scholar
  5. 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.CrossRefPubMedGoogle Scholar
  6. 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.CrossRefPubMedGoogle Scholar
  7. 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.CrossRefPubMedGoogle Scholar
  8. 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.CrossRefGoogle Scholar
  9. 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.CrossRefPubMedGoogle Scholar
  10. 10.
    Gerace, L. and Burke, B., Functional organization of the nuclear envelope, Annu. Rev. Cell Biol., 1988, vol. 4, pp. 335–374.CrossRefPubMedGoogle Scholar
  11. 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.CrossRefPubMedGoogle Scholar
  12. 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.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 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.Google Scholar
  14. 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.CrossRefPubMedGoogle Scholar
  15. 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.CrossRefPubMedGoogle Scholar
  16. 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.CrossRefPubMedGoogle Scholar
  17. 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. 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.CrossRefGoogle Scholar
  19. 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.CrossRefPubMedGoogle Scholar
  20. 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.CrossRefPubMedGoogle Scholar
  21. 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.CrossRefPubMedGoogle Scholar
  22. 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.CrossRefGoogle Scholar

Copyright information

© Allerton Press, Inc. 2018

Authors and Affiliations

  • Sandeep Kumar
    • 1
  • Sunny Dholpuria
    • 1
  • Gaurav Kumar Chaubey
    • 1
  • Rakesh Kumar
    • 1
  • Tirtha Kumar Datta
    • 1
    Email author
  1. 1.Animal Biotechnology CentreNational Dairy Research InstituteKarnalIndia

Personalised recommendations