ABSTRACT
Mouse sp56 is considered as one of the candidates for mouse zona pellucida 3 (mZP3) receptor. Up to date, its homologue has only been cloned from guinea pig, namely AM67. Based on the cDNA sequence of mouse sp56, we designed a pair of primer to amplify its homologue from rat testis cDNA. Using RT-PCR, two fragments of 743 bp and 938 bp were amplified. The PCR products show very high homology to mouse sp56. However, the 743 bp product completely lacks one of the seven Sushi domains of mouse sp56. Using the 743 bp product as the probe to detect the expression profile of rat sp56 in rat tissues, Northern blot shows that a ∼ 2.0 kb mRNA expresses specifically in testis. Employed the RACE method, two full cDNA sequences of rat sp56 were obtained. A Mr ∼ 42 KD band was detected in denatured and non-reducing protein sample of rat testis and sperm with anti-mouse sp56 monoclonal antibody by Western blot method. Rat sp56 was localized on rat sperm head by the indirect immunofluorescence method. Rat sp56 immunoreactivity was detected from the early pachytene spermatocytes and throughout the spermatogenesis. Its cloning will further our understanding of the mechanism of the sperm-egg recognition and binding.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Wassarman PM, Jovine L, Litscher ES . A profile of fertilization in mammals. Nat Cell Biol 2001; 3(2):E59–64.
Cohen N, Wassarman PM . Association of egg zona pellucida glycoprotein mZP3 with sperm protein sp56 during fertilization in mice. Int J Dev Biol 2001; 45(3 Spec No)(3–Spec–No):569–76.
Miller DJ, Macek MB, Shur BD . Complementarity between sperm surface beta-1, 4-galactosyltransferase and egg-coat ZP3 mediates sperm-egg binding. Nature 1992; 357(6379):589–93.
Kalab P, Visconti P, Leclerc P, Kopf GS . p95, the major phosphotyrosine-containing protein in mouse spermatozoa, is a hexokinase with unique properties. J Biol Chem. 1994; 269(5):3810–7.
Bleil JD, Wassarman PM . Identification of a ZP3-binding protein on acrosome-intact mouse sperm by photoaffinity crosslinking. Proc Natl Acad Sci USA 1990; 87(14):5563–7.
Cheng A, Le T, Palacios M, et al. Sperm-egg recognition in the mouse: characterization of sp56, a sperm protein having specific affinity for ZP3. J Cell Biol 1994; 125(4):867–78.
Bookbinder LH, Cheng A, Bleil JD . Tissue- and species-specific expression of sp56, a mouse sperm fertilization protein. Science 1995; 269(5220):86–9.
Kim KS, Cha MC, Gerton GL . Mouse sperm protein sp56 is a component of the acrosomal matrix. Biol Reprod 2001; 64(1):36–43.
Foster JA, Friday BB, Maulit MT, et al. AM67, a secretory component of the guinea pig sperm acrosomal matrix, is related to mouse sperm protein sp56 and the complement component 4-binding proteins. J Biol Chem 1997; 272(19):12714–22.
Schmell ED, Gulyas BJ . Mammalian sperm-egg recognition and binding in vitro. I. Specificity of sperm interactions with live and fixed eggs in homologous and heterologous inseminations of hamster, mouse, and guinea pig oocytes. Biol Reprod 1980; 23(5):1075–85.
Janatova J, Reid KB, Willis AC . Disulfide bonds are localized within the short consensus repeat units of complement regulatory proteins: C4b-binding protein. Biochemistry 1989; 28(11):4754–61.
Hu JH, Yan YC . Identification of gamma1 subunit of GABA(A) receptor in rat testis. Cell Res 2002; 12(1):33–7.
Hu JH, He XB, Yan YC . Identification of gamma-aminobutyric acid transporter (GAT1) on the rat sperm. Cell Res 2000; 10(1):51–8.
Acknowledgements
This project was supported by the grant from National Key Basic Research Project, “973”, No. G199905592.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
HE, X., YAN, Y., LI, Y. et al. Cloning of rat sp56, the homologue of mouse sperm ZP3 receptor–sp56. Cell Res 13, 121–129 (2003). https://doi.org/10.1038/sj.cr.7290156
Issue date:
DOI: https://doi.org/10.1038/sj.cr.7290156
Keywords
This article is cited by
-
Acrosome markers of human sperm
Anatomical Science International (2016)
