Skip to main content
SpringerLink
Log in

Expression of ganglioside GT1b in mouse embryos at different developmental stages after cryopreservation

  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

Gangliosides are a family of sialic acid-containing glycosphingolipids that are abundant in neurons and have a variety of functions in developing and mature tissues. We examined the expression of ganglioside GT1b in the embryonic preimplantation stage after freezing and thawing processes to determine the regulatory roles of ganglioside GT1b in early embryonic development. ICR mouse embryos at the two-cell stage obtained by flushing the oviducts were frozen by two cryopreservation procedures, slow freezing using a programmable freezer or vitrification by direct plunging into liquid nitrogen. Slow freezing was conducted with equilibration in 1.5 M 1,2-propanediol or 5% equilibration glycerol. Vitrification was applied with a 10–15 min equilibration in 7.5% ethylene glycol (EG), 7.5% dimethylsulfoxide (DMSO), and 30 sec in a solution of 15% EG, 15% DMSO and 0.5 M sucrose. Immediately after thawing, the survival rate of the embryos was assessed by their morphology and ability to develop to blastocysts in culture. The survival rate of vitrified and thawed embryos (92%) was significantly higher than that of slow frozen and thawed embryos (76%) (P<0.05). A tendency of higher blastocyst rate was found in the vitrified and thawed embryos compared to that of the slow frozen and thawed embryos. Confocal immunofluorescence staining confirmed that surviving embryos expressed ganglioside GT1b, with the strongest expression at the compacted eight-cell or later stage embryos. Ganglioside GT1b was not observed in the TUNEL-positive, apoptotic embryos, suggesting that cryopreservation had induced DNA breaks in them. These results suggest that ganglioside GT1b may play an important role in embryo survival or development.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aono, N., Naganuma, T., Abe, Y., Hara, K., Sasada, H., Sato, E., and Yoshida, H., Successful production of blastocysts following ultrarapid vitrification with step-wise equilibriation of germinal vesicle-stage mouse oocytes. J. Reprod. Dev., 49, 501–506 (2003).

    Article  PubMed  Google Scholar 

  • Bouvier, J. D. and Seyfired, T. N., Ganglioside composition of normal and mutant mouse embryos. J. Neurochem., 52, 460–466 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Bremer, E. G., Schlessinger, J., and Hakomori, S., Ganglioside-mediated modulation of cell growth. Specific effects of GM3 on tyrosine phosphorylation of the epidermal growth factor receptor. J. Biol. Chem., 261, 2434–2440 (1986).

    PubMed  CAS  Google Scholar 

  • Choo, Y. K., Distribution of ganglioside GM3 in the rat ovary after gonadotropin stimulation. Mol. Cells., 9, 365–375 (1999).

    PubMed  CAS  Google Scholar 

  • Cooper, A., O’Neil, L., Bernard, A., Fuller, B., and Shaw, R., The effect of protein source and temperature on the damage to mouse oocyte cytoskeleton after exposure to a vitrification solution. Cryo. Lett., 17, 149–156 (1996).

    Google Scholar 

  • Draper, J. S., Pigott, C., Thomson, J. A., and Andrews, P. W., Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J. Anat., 200, 249–258 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Dreyfus, H., Urban, P. F., and Edel-Harth, S., Developmental patterns of gangliosides and phospholipids in chick retina and brain. J. Neurochem., 25, 245–250 (1975).

    Article  PubMed  CAS  Google Scholar 

  • Duchemin, A. M., Ren, Q., Mo, L., Neff, N. H., and Hadjiconstantinou, M., GM1 ganglioside induces phosphorylation and activation of Trk and Erk in brain. J. Neurochem., 81, 696–707 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Fugger, E. F., Bustillo, M., Katz, L. P., Dorfmann, A. D., Bender, S. D., and Schulman, J. D., Embryonic development and pregnancy from fresh and cryopreserved sibling pronucleate human zygotes.. Fertile. Steril., 50, 273–278 (1998).

    Google Scholar 

  • Fujino, Y., Ozaki, K., Yamamasu, S., Ito, F., Matsuoka, I., Hayashi, E., Nakamura, H., Ogita, S., Sato, E., and Inoue, M., DNA fragmentation of oocytes in aged mice. Hum. Reprod., 11, 1480–1483 (1996).

    PubMed  CAS  Google Scholar 

  • Glenister, P. H., Whittingham, D. G., and Wood, M. J., Genome cryopreservation: a valuable contribute to mammalian genetic research. Genet. Res., 56, 564–570 (1990).

    Article  Google Scholar 

  • Hakomori, S., Glycosphingolipids in cellular interaction, differentiation, and oncogenesis. Annu. Rev. Biochem., 50, 733–764 (1981).

    Article  PubMed  CAS  Google Scholar 

  • Hakomori, S., Bifunctional role of glycosphingolipids Modulators for transmembrane signaling and mediators for cellular interactions. J. Biol. Chem., 265, 18713–18716 (1990).

    PubMed  CAS  Google Scholar 

  • Hakomori, S. and Igarashi, Y., Functional role of glycosphingolipids in cell recognition and signaling. J. Biochem. (Tokyo), 118, 1091–1103 (1995).

    CAS  Google Scholar 

  • Hanada, K., Nishijima, M., Kiso, M., Hasegawa, A., Fujita, S., Ogawa, T., and Akamatsu, Y., Sphingolipids are essential for the growth of Chinese hamster ovary cells. Restoration of the growth of a mutant defective in sphingoid base biosynthesis by exogenous sphingolipids. J. Biol. Chem., 267, 23527–23533 (1992).

    PubMed  CAS  Google Scholar 

  • Hidari, K. I., Functional glycoconjugates involved in cellular interaction. Yakugaku Zasshi, 123, 315–322 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Higashi, H. and Chen, N. H., Ganglioside/protein kinase signals triggering cytoskeletal actin reorganization. Glycoconj. J., 20, 49–58 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Irwin, L. N., Micheal, D. B., and Irwin, C. C., Ganglioside pattern of fetal rat and mouse brain. J. Neurochem., 34, 1527–1520 (1980).

    Article  PubMed  CAS  Google Scholar 

  • Ishida, G. M., Saito, H., Ohta, N., Takashi, T., Ito, M. M., Saito, T., Nakahara, K., and Hiroi, M., The optimal equilibration time for mouse embryos frozen by vitrification with trehalose. Hum. Rerpod., 12, 1259–1262 (1997).

    Article  CAS  Google Scholar 

  • Janich, P. and Corbeil, D., GM1 and GM3 gangliosides highlight distinct lipid microdomains within the apical domain of epithelial cells. FEBS. Lett., 581, 1783–1787 (2007).

    Article  PubMed  CAS  Google Scholar 

  • Jurisicova, A., Varmuza, S., and Casper, R. F., Programmed cell death and human embryo fragmentation. Mol. Hum. Reprod., 2, 93–98 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Kasai, M., Komi, J. H., Takakamo, A., Tsudera, H., Sakurai, T., and Machida, T., A simple method for mouse embryo cryopreservation in al low toxicity vitrification solution, without appreciable loss of viability. J. Reprod. Fertil., 89, 91–97 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Kim, S. M., Kwak, D. H., Kim, S. M., Jung, J. U., Lee, D. H., Lee, S., Jung, K. Y., Do, S. I., and Choo, Y. K., Differential expression of gangliosides in the ovary and uterus of streptozotocin-induced and db/db diabetic mice. Arch. Pharm. Res., 29, 666–676 (2006).

    Article  PubMed  CAS  Google Scholar 

  • Kwak, D. H., Yu, K., Kim, S. M., Lee, D. H., Kim, S. M., Jung, J. U., Seo, J. W., Kim, N., Lee, S., Jung, K. Y., You, H. K., Kim, H. A., and Choo, Y. K., Dynamic changes of gangliosides expression during the differentiation of embryonic and mesenchymal stem cells into neural cells. Exp. Mol. Med., 38, 668–676 (2006).

    PubMed  CAS  Google Scholar 

  • Lassalle, B., Testart, J., and Remard, J. P., Human embryo features that influence the success of cryopreservation with the use of 1,2 propanediol. Fertil. Sterril., 44, 645–651 (1985).

    CAS  Google Scholar 

  • Lee, D. H., Koo, D. B., Ko, K., Ko, K., Kim, S. M., Jung, J. U., Ryu, J. S., Jin, J. W., Yang, H. J., Do, S. I., Jung, K. Y., and Choo, Y. K., Effects of daunorubicin on ganglioside expression and neuronal differentiation of mouse embryonic stem cells. Biochem. Biophys. Res. Commun., 362, 313–318 (2007).

    Article  PubMed  CAS  Google Scholar 

  • Leibo, S. P., Mazur, P., and Jackowski, S. J., Factors affecting survival of mouse embryos during freezing and thawing. Exp. Cell. Res., 89, 79–88 (1974).

    Article  PubMed  CAS  Google Scholar 

  • Li, R., Manela, J., Kong, Y., and Ladisch, S., Cellular gangliosides promote growth factor-induced proliferation of fibroblasts. J. Biol. Chem., 275, 34213–34223 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Liu, L., Trimarchi, J. R., and Keefe, D. L., Involvement of mitochondria in oxidative stress-induced cell death in mouse zygotes. Biol. Reprod., 62, 1745–1753 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Lloyd, K. O. and Furukawa, K., Biosynthesis and functions of gangliosides: recent advances. Glycoconj. J., 15, 627–636 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Luyet, B. J., The vitrification of organic colloids and protoplasm. Biodynamica., 11–14 (1937).

  • Maccioni, H. J. F., Panzetta, P., Arrieta, D., and Caputto, R., Ganglioside glycosyltransferase activities in the cerebral hemispheres from developing rat embryos. Int. J. Dev. Neurosci., 2, 13–19 (1984).

    Article  CAS  Google Scholar 

  • Miljan, E. A. and Bremer, E. G., Regulation of growth factor receptors by gangliosides. Sci. STKE., 160, RE15 (2002).

    Google Scholar 

  • Miyamoto, H. and Ishibashi, T., Survival of frozen-thawed mouse and rat embryos in the presence of ethylene glycol. J. Reprod. Fertil., 50, 373–375 (1977).

    Article  PubMed  CAS  Google Scholar 

  • Okada, M., Itoh, Mi. M., Haraguchi, M., Okajima, T., Inoue, M., Oishi, H., Matsuda, Y., Iwamoto, T., Kawano, T., Fukumoto, S., Miyazaki, H., Furukawa, K., Aizawa, S., and Furukawa, K., b-series Ganglioside deficiency exhibits no definite changes in the neurogenesis and the sensitivity to Fas-mediated apoptosis but impairs regeneration of the lesioned hypoglossal nerve. J. Biol. Chem., 277, 1633–1636 (2002).

    Article  PubMed  CAS  Google Scholar 

  • O’Neil, L., Paynter, S. J., Fuller, B. J., and Shaw, R. W., Murine oocytes cytoskeleton changes, fertilization and embryonic development following exposure to a vitrification solution. Cryo. Lett., 18, 17–26 (1997).

    CAS  Google Scholar 

  • Panzett, P. H. J., Maccioni, H. J., and Caputto, R., Synthesis of retinal gangliosides during chick embryonic development. J. Neurochem., 35, 100–108 (1980).

    Article  Google Scholar 

  • Rall, W. F. and Fahy, G. M., Ice-free cryopreservation of mouse embryos at-196 degrees C by vitrification. Nature, 313, 573–575 (1985).

    Article  PubMed  CAS  Google Scholar 

  • Rall, W. F., Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology, 24, 387–402 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Rosner, H., Ganglioside changes in the chicken optic lobes as biochemical indicators of brain development and maturation. Brain Res., 236, 49–51 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Selick, C. E., Hofmann, G. E., and Albano, C., Embryo quality and pregnancy potential of fresh compared with frozen embryos freezing detrimental to quality embryos. Hum. Reprod., 10, 392–395 (1995).

    PubMed  CAS  Google Scholar 

  • Seyfried, T. N., Yu, R. K., Satio, M., and Albert, M. D., Ganglioside abnormalities associated with failed neural differentiation in a T-locus mutant mouse embryo. Dev. Biol., 123, 286–291 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Sonna, L. A., Fujita, J., Gaffin, S. L., and Lilly, C. M., Molecular biology of thermoregulation invited review: Effects of heat and cold stress on mammalian gene expression. J. Appl. Physiol., 92, 1725–1742 (2002).

    PubMed  CAS  Google Scholar 

  • Svennerholm, L., Gangliosides and synaptic transmission. Adv. Exp. Med. Biol., 125, 533–544 (1980).

    PubMed  CAS  Google Scholar 

  • Toledo, M. S., Suzuki, E., Handa, K., and Hakomori, S., Effect of ganglioside and tetraspanins in microdomains on interaction of integrins with fibroblast growth factor receptor. J. Biol. Chem., 280, 16227–16234 (2005).

    Article  PubMed  CAS  Google Scholar 

  • Uechi, H., Tsutsmi, O., Morita, Y., and Taketani, Y., Cryopreservation of mouse embryos affects later embryonic development possibly through reduced expression of the glucose transporter GLUT1. Mol. Reprod. Dev., 48, 496–500 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Van Wagtendonk-De Leeuw, A. M., Den Daas, J. H. G., Kruip, A., and Rall, W. F., Comparison of the efficiently of conventional slow freezing and rapid cryopreservation methods for bovine embryos. Cryobiology, 35, 93–105 (1995).

    Google Scholar 

  • Vancetto, M. D., Curi, L. C., and Pereira, C. A., Neutralization of the effect of Crotalus durissus terrificus venom by gangliosides. Braz. J. Med. Biol. Res., 28, 553–556 (1995).

    PubMed  CAS  Google Scholar 

  • Whittingham, D. G., Leibo, S. P., and Mazur, P., Survival of mouse embryos frozen to 196 degrees and 169 degrees C. Science, 178, 411–414 (1972).

    Article  PubMed  CAS  Google Scholar 

  • Wiegandt, H., The chemical constitution of gangliosides of the vertebrate nervous system. Behav. Brain Res., 66, 85–97 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto, H. A. and Mohanan, P. V., Ganglioside GT1B and melatonin inhibit brain mitochondrial DNA damage and seizures induced by kainic acid in mice. Brain Res., 964, 100–106 (2003a).

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto, H. A. and Mohanan, P. V., In vivo and in vitro effects of melatonin or ganglioside GT1B on L-cysteine-induced brain mitochondrial DNA damage in mice. Toxicol. Sci., 73, 416–422 (2003b).

    Article  PubMed  CAS  Google Scholar 

  • Yu, R. K., Macala, L. J., Taki, T., Weinfield, H. M., and Yu, F. S., Development changes in ganglioside composition and synthesis in embryonic rat brain. J. Neurochem., 50, 1825–1829 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Zurita, A. R., Crespo, P. M., Koritschoner, N. P., and Daniotti, J. L., Membrane distribution of epidermal growth factor receptors in cells expressing different gangliosides. Eur. J. Biochem., 271, 2428–2437 (2004).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Obstetric and gynecology clinic, Mirae-Heemang Hospital, Seoul 135-888, Korea

    Hyuck-Chan Kwon

  2. College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University, Cheongju, 361-763, Korea

    Sang-Yoon Nam & Dong-Hoon Kwak

  3. Department of Biotechnology, Catholic University, Bucheon, 420-749, Korea

    Yong-II Park

  4. Center for Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 305-806, Korea

    Deog-Bon Koo

  5. Department of Biological Science, College of Natural Sciences, Wonkwang University, 344-2 Shinyong-dong, Iksan, Jeonbuk, 570-749, Korea

    Bo-Hyun Kim, Ji-Ung Jung, Kisung Ko, Won-Sin Kim, Sun-Mi Kim, Jae-Sung Ryu, Jung-Woo Jin, Hyo-Jung Yang, Ji-Su Kim & Young-Kug Choo

Authors
  1. Bo-Hyun Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ji-Ung Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kisung Ko
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Won-Sin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sun-Mi Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jae-Sung Ryu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jung-Woo Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hyo-Jung Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ji-Su Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hyuck-Chan Kwon
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Sang-Yoon Nam
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Dong-Hoon Kwak
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Yong-II Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Deog-Bon Koo
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Young-Kug Choo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Young-Kug Choo.

Additional information

These authors contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, BH., Jung, JU., Ko, K. et al. Expression of ganglioside GT1b in mouse embryos at different developmental stages after cryopreservation. Arch. Pharm. Res. 31, 88–95 (2008). https://doi.org/10.1007/s12272-008-1125-6

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12272-008-1125-6

Key words

Search

Navigation