Skip to main content
SpringerLink
Log in

Effect of monoclonal antibody to bull sperm sp18 on murine fertilization and embryos development

  • Notes
  • Published:
Chinese Science Bulletin

Abstract

Western blot analysis revealed that one IgG1 monoclonal antibody (mAb) to sp18 family membrane proteins (Mr. 14, 16 and 18 ku) of bovine sperm reacted faintly with protein bands of 14, 18, 22, 30 and 60 ku (reducing) in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence (IIF) showed that the sp18 antigens were present in the posterior head of murine sperm. In murinein vitro fertilization (IVF) and embryo development trails, a total of 426 oocytes from C57,BL/6 and F1 hybrid strain (CD1 × C57BL/6 cross) of 12 female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μ/mL of sp18 mAb in the modified TYH IVF medium for 15–20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 rnAb groups was 77.1 %, which was not significantly (P > 0.05) different from the nonspecific mouse IgG (79.2 %) and non-lgG (80.3 %) control groups. Fertilized oocytes had been continuously cultured in modifed CZB medium. 100%, 100% and 97.9% of 1cell embryos developed to 2-cell stage in sp18 mAb, nonspecific mouse IgG and non-lgG group 30 h after the start of fertilization, respectively. In the nonspecific mouse IgG and nonlgG groups, 64.1% and 64.3% of embryos developed to the 4-cell stage, respectively, but all developing eggs in sp18 mAb groups arrested developmentin vitro at 2-cell stage. After zonae of 2-cell blocked embryos were enzymatically removed with 0.5% pronase, detection of sp18 antigens by IIF indicated that the fluorescence scattered on two embryonic cells. For embryos fertilizedin vivo and co-cultured with 182 μg/mL sp18 mAb, the numbers of 1-cell embryos reaching the 2-cell and 4-cell stage were 95.2% and 70.5%, which were not significantly (P> 0.05) different from the control group (92.9% and 77.9%). These results indicate that the sp18 antigens on posterior head of mouse sperm were incorporated into the egg plasma membrane during fertilization, and played an active role in development of murine preimplantation embryo.

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

  1. Lin, Y., Mahan, K., Lathop, W. F.et al., A hyaluronidase activity of the sperm plasma membrane protein PH-20 enables sperm to penetrate the cumulus cell layer surrounding the egg,J. Cell Biol., 1994, 125(5): 1157.

    Article  Google Scholar 

  2. Snell, W. J., White, J. M., The molecules of mammalian fertilization,Cell., 1996, 85(5): 629.

    Article  Google Scholar 

  3. Myles, D. G., Primakoff, P., Why did the sperm cross the cumulus? To get to the oocyte, functions of the sperm surface proteins PH-20 and fertilin in arriving at, and fusing with, the egg,Biol. Reprod., 1997, 56(2): 320.

    Article  Google Scholar 

  4. Gou, K. M., Lin, A. X., An, X. R,et al., Characteristics of sp18 family membrane proteins on posterior head of bull sperm,Acta Veterinaria et Zootechnica Sinica (in Chinese), 1999, in press.

  5. Gou, K. M., Shang, L. J., An, X. R.,et al., Extraction of sp18 family membrane proteins on posterior head of bull sperm and the effect of anti-spl8 IgG on sperm motility and murinein vitro fertilization,Anim. Biotech., 1998, 9(2): 149.

    Article  Google Scholar 

  6. Gou, K. M., Li, Y., An, X. R.et al., Production of anti-bull sperm monoclonal antibody: sp18, inProceedings of International Conference on Animal Biotechnology, Beijing: International Academic Publishers, 1997, 325.

    Google Scholar 

  7. Chatot, C. L., Ziomek, C. A., Bavister, B. D.et al., An improved medium supports development of random-bred 1-cell mouse embryosin vitro, J. Reprod Fertil, 1989, 86: 679.

    Google Scholar 

  8. Haraguchi, S., Naito, K., Azuma, S.et al., Effects of phosphate onin vitro 2-cell block of AKR/N mouse embryos based on changes in cdc2 kinase activity and phosphorylation states,Biol. Reprod., 1996, 55: 598.

    Article  Google Scholar 

  9. Leyton, L., Saling, P., Evidence that aggregation of mouse sperm receptors by ZP3 triggers the acrosome reaction,J. Cell Biol., 1989, 108(6): 2163.

    Article  Google Scholar 

  10. Yanagimachi, R., Sperm-egg fusion, inCurret Topics in Membranes and Transport (eds. Duzgunes, N., Bronner, F.) 32nd ed., San Diego: Ca. Academic Press, 1988, 3.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Biotechnology, Academy of Military Medical Sciences, 100071, Beijing, China

    Kemian Gou, Jixian Deng & Peitang Huang

  2. State Key Laboratory of Agrobiotechnology, Chinese Agricultural University, 100094, Beijing, China

    Lijuan Shang, Xiaorong An & Yongfu Chen

Authors
  1. Kemian Gou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lijuan Shang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaorong An
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jixian Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yongfu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Peitang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Gou, K., Shang, L., An, X. et al. Effect of monoclonal antibody to bull sperm sp18 on murine fertilization and embryos development. Chin.Sci.Bull. 44, 236–240 (1999). https://doi.org/10.1007/BF02896282

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02896282

Keywords

Search

Navigation