Role of sperm surface proteins with epididymal origin on fertilization

Sadeghi, Mohammad Reza; Akhondi, Mohammad Mehdi; Zahraei, Mahin

Role of sperm surface proteins with epididymal origin on fertilization

Vol. 1, Issue 3, / July-September 2000
(pages 43-54)

Mohammad Reza Sadeghi: - Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Mohammad Mehdi Akhondi: - Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Mahin Zahraei: - Medical Biochmistry Department, Faculty of Medicine, Tehran Medical Sciences University, Tehran, Iran

Received: 7/1/2000 Accepted: 7/1/2000 - Publisher : Avicenna Research Institute

Other Format

Abstract

During transit of spermatozoa through the epididymis, where develop its capacity to fertilize oocytes, several new antigenic determinants appear on surface of spermatozoa. Many of these are proteins and glycoproteins with epididymal origin. The aim of this study was evaluation the role of epididymal secretory proteins on fertilizing ability of spermatozoa. In This study, epithelial cells from proximal portion of rat epididymis were cultured in RPMI medium supplemented with growth factors and androgens. To identify epididymal secretory proteins, pulse labeling with [35S]-(Met, Cys) was used. Labeled conditioned medium recovered and subjected to SDS-PAGE and flurography. Antiserum against 8 secretory proteins were produced in Rabbits. The immunogens were lyophilized narrow strip of polyacrylamide gel from preparative electrophoresis with ultrapure proteins. Incubation of intact rat spermatozoa with these antisera was revealed secretory proteins on surface of spermatozoa. The effects of these antisera on in vitro fertilization were evaluated. The results showed that epithelial cells secreted about 20-30 proteins in culture medium. High titer antisera for 8 secretory proteins were produced. Three antisera were reacted with surface of rat spermatozoa (20, 24 & 72 kD proteins), and only 20 kD protein antiserum decreased fertilization rate. (28% in treatment group against 89% for control group). In Conclusion, the epididymal maturation process of spermatozoa was associated with the secretion of a number proteins by epididymal epithelial cells, many of these proteins bound to plasma membrane of spermatozoa. Only secreted 20 kD protein has a critical role in fertilizing ability of spermatozoa.

Keywords: Secretory protein, Epididymis, Rat, IVF, Sperm maturation

To cite this article:

Sadeghi MR, Akhondi MM, Zahraei M. Role of sperm surface proteins with epididymal origin on fertilization. J Reprod Infertil. 2000;1(3):43-54.

References

Brooks DE. Secretion of proteins and glycoproteins by the rat epididymis: regional differences, androgen-dependence and effects of protease inhibitors, procaine and tunicamycin. Biol Reprod. 1981, 25:1099-1117.
Hegde UC. Epididymal sperm maturation proteins. J Bioch Biophy. 1996, 33:103-110.
Iusem ND, Pineiro L, Blaquier J, et al. Identification of a major secretory glycoprotein from rat epididymis: Interaction with spermatozoa. Biol Reprod. 1989, 40:307-316.
Cooper TG. Epididymis and sperm function. J Androl. 1996, 1:57-59.
Frank B, Bruno B, deLmirande E, et al. Human Sperm-Zona Pellucida Interaction Is Inhibited by an Antiserum against a Hamster Sperm Protein. Biol Reprod. 1994, 51:577-587.
Perez-Martinez S, Conesa D, Cuasnicu P. Potential contraceptive use of epididymal proteins: evidence for the participation of specific antibodies against rat epididymal protein DE in male and female fertility inhibition. J Reprod Immunol. 1995, 29:31-45.
Amero S.A, James T. C, Elgin SCR. Production of antibodies using proteins in gel bands. In: Walker JM(ed)The protein protocols handbook. Totowa, New Jersey. Hum Press.1996, 717-720.
Miyoshi K, Kono T, Niwa K. Stage-dependent development of rat 1-cell embryos in a chemically defined medium after fertilization in vivo and in vitro. Biol Reprod. 1997, 56:180-185.
Wassarman PM, Litscher ES. Sperm-egg recognition mechanisms in mammals. Curr Top Dev Biol. 1995, 30:1-19.
Myles D, 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, 56:320-327.
Bedford J. The status and the state of the human epididymis. Hum Reprod. 1994, 9: 2187-2199.
Weaver F, Dino J, Germain B, et al. G. Biochemical characterization and epididymal localization of the maturation-dependent ram sperm surface antigen ESA152. Mol Reprod Dev. 1993, 35:293-301.
Boue F, Duquenne C, Lassalle B, et al. FLB1, a human protein of epididymal origin that is involved in the sperm-oocyte recognition process. Biol Reprod. 1995, 52:267-278.
Boue F, Blais J, Sullivan R. Surface localization of P34H an epididymal protein, during maturation, capacitation,and acrosome reaction of human spermatozoa. Biol Reprod. 1996, 54:1009-1017.
Focarelli R, Giuffrida A, Capparelli S, et al. Specific localization in the equatorial region of gp20, a 20 kDa sialylglycoprotein of the capacitated human spermatozoon acquired during epididymal transit which is necessary to penetrate zona-free hamster eggs. Mol Hum Reprod. 1998, 4: 119-125.
Bailey G. Raising of polyclonal antisera. In: Walker, J(ed)The protein protocols handbook. Totowa, New Jersey Hum Press. 1996, 695-698.
Walker JM. SDS polyacrylamide gel electrophoresis of proteins. In: Walker, JM(ed). The protein protocols handbook.Totowa, New Jersey Hum Press. 1996, 55-62.
Harding C, Williams P, Wagner E, et al. Mice with genetic gamma-glutamyl transpeptidase deficiency exhibit glutathionuria, severe growth failure, reduced life spans and infertility. J Biol Chem. 1997, 272:12560-12567.
Baba T, Azuma S, Kashiwabara S, et al. Sperm from mice carrying a targeted mutation of the acrosin gene can penetrate the oocyte zona pellucida and effect fertilization. J Biol Chem. 1994, 269:31845-31849.
Lu Q, Shur B. Sperm from B1, 4-galactosyl transferase-null mice are refractory to ZP3-induced acrosome reactions and penetrate the zona pellucida poorly. Dev. 1997, 124: 4121-4131.
Xu W.Wang L, Miao S, et al. Identification of a rabbit epididymal protein gene. Arch Androl. 1996, 37:135-141.
Xu W, Miao S, Zhang M, et al. Expression of the BE-20 epididymal protein gene:in situ hybridization. Arch Androl. 1997, 38:1-6.
Holland M, Nixon B. The specificity of epididymal secretory proteins. J Reprod Fertil.1998, 53:197-210.
Nixon B, Hardy C, Clarke H, et al. REP38:a rabbit epididymal secretory protein present on spermatozoa. The Epididymis Cellular Molecular Aspects Boden. 1998, 20.
Kirchhoff C. Gene expression in the epididymis. Int Rev Cytol. 1999, 188:133-202
Sdeghi MR, Akhondi MA, Zahraie M. Identification of proteins shared between epididymal secretions and spermatozoa epithelium. J Res Med Sci. 2001;25(1):43-54.
Moore HDM. Contribution of epididymal factors to sperm maturation and storage. J Androl. 1998, 30: 233-239.
Moore HDM. The influence of the epididymis on human and animal sperm maturation and storage. Hum Reprod. 1996, 11:103-110.
Orgebin-Crist MC. The epididymis across 24 centuries. J Reprod Fertil. 1998, 53:285-292.
Dacheux J, Druart X, Fouchecourt S, et al. Role of epididymal secretory proteins in sperm maturation with particular reference to the boar. J Reprod Fertil. 1998, 53:99-107
Druart X, Gatti JL, Dacheux F, et al. Analysis by two-dimensional gel electrophoresis of ram epididymal secreted proteins. Cell Mol Biol. 1994, 40:91-93.
Jones R. Plasma membrane structure and remodelling during sperm maturation in the epididymis. J Reprod Fertil. 1998, 53:73-84.
Fornes M, Burgos MH. Epididymal glycoprotein involved in rat sperm association. Mol Reprod Dev. 1994, 38:43-47.
Cooper TG. Interactions between epididymal secretions and spermatozoa. J Reprod Fertil. 1998, 53:119-136.
Beagley K, Wu ZL. Pomering M, et al. Jones RC., Immune responses in the epididymis:implications for immunocontraception. J Reprod Fertil. 1998, 53:235-245.