Skip to main content
SpringerLink
Log in

Post-fertilization changes in the zona pellucida glycoproteins of rat eggs

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

The zona pellucida (ZP) is the extracellular coat surrounding the mammalian egg. Numerious evidence supports the role of ZP carbohydrate residues as the specific sperm receptors. In this study we used lectins to study different distribution patterns of carbohydrate residues in the rat ZP, and to follow changes at fertilization. ZP were collected from follicular, ovulated, and fertilized eggs, incubated with one of 11 different biotin-labeled lectins, followed by avidin-fluorescein isothiocyanate (FITC) complex, and visualized by epifluorescent microscopy. For electron microscope (EM) histochemistry, eggs were embedded in LR white and ultrathin sections were stained with the complexRicinus communis lectin (RCA-1)-colloidal gold. Some lectins (RCA-I,Glycine max) bound to the entire ZP while others were restricted to the inner or outer zones [Griffonia simplicifolia, Concanovalia ensiformis, Triticum vulgaris (WGA), succinyl-WGA]. Other lectins (Lens culinaris, Ulex europhaeus) were totally excluded. The RCA-1 binding pattern changed following sperm penetration, from homogeneous in ZP of ovulated eggs (57%) to uneven in ZP of fertilized (71%) or activated (68%) eggs. Our results demonstrate an uneven distribution of different sugar residues in the rat ZP, and a post-fertilization change in the distribution of β-galactose, which is specifically recognized by RCA-I, presumably correlated with other changes in the ZP that lead to the block to polyspermy.

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

  • Abdullah M, Kierszenbaum AL (1989) Identification of rat testis galactosyl receptor using antibodies to liver asialoglycoprotein receptor: purification and localization of surfaces of spermatogenic cells and sperm. J Cell Biol 108:367–375.

    Article  PubMed  CAS  Google Scholar 

  • Abdullah M, Widgren EE, O'Rand MG (1991) A mammalian sperm lectin related to rat hepatocyte lectin-2/3. Mol Cell Biochem 103:155–161

    Article  PubMed  CAS  Google Scholar 

  • Ahuja KK, Bolwell GP (1983) Probable asymmetry in the organization of components of the hamster zona pellucida. J Reprod Fertil 69:49–55

    Article  PubMed  CAS  Google Scholar 

  • Araki Y, Orgebin-Crist MC, Tulsiani, DRP (1992) Qualitative characterization of oligosaccharide chains present on the rat zona pellucida glycoconjugates. Biol Repord 46:912–919

    Article  CAS  Google Scholar 

  • Bleil JD, Wassarman PL (1980) Structure and function of the zona pellucida: identification and characterization of the proteins of the mouse oocyte's zona pellucida. Dev Biol 76:185–202

    Article  PubMed  CAS  Google Scholar 

  • Bleil JD, Wassarman PL (1988) Galactose at the nonreducing terminus ofO-linked oligosaccharides of mouse egg zona pellucida glycoprotein ZP3 is essential for the glycoprotein's sperm receptor activity. Proc Natl Acad Sci USA 85:6778–6782

    Article  PubMed  CAS  Google Scholar 

  • Bleil JD, Beall CF, Wassarman PL (1981) Mammlian sperm-egg interaction: fertilization of mouse eggs triggers modification of the major zona pellucida glycoprotein, ZP2. Dev Biol 86:189–197

    Article  PubMed  CAS  Google Scholar 

  • Ducibella T (1991) Mammalian egg cortical granules and the cortical reaction. In: Wassarman PM (ed) Elements of mammalian fertilization, vol I, CRC Press, Boca Raton Boston, pp 205–231

    Google Scholar 

  • Ducibella T, Kurasawa S, Rangarajan S, Kopf GS, Schults RM (1990) Precocious loss of cortical granules during mouse oocyte meiotic maturation and correlation with an egg-induced modification of the zona pellucida. Dev Biol 137:46–55

    Article  PubMed  CAS  Google Scholar 

  • Familiari G, Nottola SA, Macchiarelli G, Micara G, Aragona C, Motta PM (1992) Human zona pellucida during in vitro fertilization: an ultrastructural study using saponin, ruthenium red, and osmium-thiocarboyhdrazide. Mol Reprod Dev 32:51–61

    Article  PubMed  CAS  Google Scholar 

  • Florman HM, Bechtol KB, Wasserman PM (1984) Enzymatic dissection of the functions of the mouse egg's receptor for sperm. Dev Biol 106:243–255

    Article  PubMed  CAS  Google Scholar 

  • Goldstein IJ, Hughes RC, Monsigny M, Osaka T, Sharon N (1980) What should be called a lectin? Nature 285:66

    Article  Google Scholar 

  • Goluboff ET, Mertz JR, Tres LL, Kierszenbaum AL (1995) Galactosyl receptor in human testis and sperm is antigenically related to the minor C-type (Ca2+-dependent) lectin variant of human and rat liver. Mol Reprod Dev 40:460–466.

    Article  PubMed  CAS  Google Scholar 

  • Greve LC, Prody GA, Hedrick JL (1985)N-Acetyl-β-d-glucosea-minidase activity in the cortical granules ofXenopus laevis eggs. Gamete Res 12:305–312

    Article  CAS  Google Scholar 

  • Gwatkin RBL, Williams DT, Hartmann JF, Kniazuk M (1973) The zona reaction of hamster and mouse eggs: production in vitro by a trypsin-like protease from cortical granules. J Reprod Fertil 32:259–265

    Article  PubMed  CAS  Google Scholar 

  • Hedrick JL, Wardrip NJ, Berger T (1987) Rapid communication: differences in the macromolecular composition of the zona pellucida isolated from pig oocytes, eggs, and zygotes. J Exp Zool 241:257–262

    Article  PubMed  CAS  Google Scholar 

  • Hokke CH, Damm JBL, Penninkhof B, Aitken RJ, Kamerling JP, Vliegenthart JFG (1994) Structure of theO-linked carbohydrate chains of porcine zona pellucida glycoproteins. Eur J Biochem 221:491–512

    Article  PubMed  CAS  Google Scholar 

  • Kalab P, Kopf GS, Schultz RM (1991) Modifications of the mouse zona pellucida during oocyte maturation and egg activation: effects of newborn calf serum and fetuin. Biol Reprod 45: 783–787

    Article  PubMed  CAS  Google Scholar 

  • Kan FWK, Roux E, St-Jacques S, Bleau G (1990) Demonstration by lectin-gold cytochemistry of transfer of glycoconjugates of oviductal origin to the zona pellucida of oocytes after ovulation in hamsters. Anat Rec 226:37–47

    Article  PubMed  CAS  Google Scholar 

  • Kaplan-Kraicer R, Bardin CW, Shalgi R (1995) Anordiol and RU486 synergize to produce preimplantation pregnancy loss by increasing embryo transport (rat). Contraception 51:141–146

    Article  PubMed  CAS  Google Scholar 

  • Kaufman MH, Fowler RE, Barratt E, McDougall RD (1989) Ultrastructural and histochemical changes in the murine zona pellucida during the final stages of oocyte maturation prior to ovulation. Gamete Res 24:35–48

    Article  PubMed  CAS  Google Scholar 

  • Keefer CL, Schuetz AW (1982) Spontaneous activation of ovulated rat oocytes during in vitro culture. J Exp Zool 224:371–377

    Article  PubMed  CAS  Google Scholar 

  • Kimura J, Sato K, Tsukise A, Okano M (1991) Appearance of lectin binding affinity to the zona pellucida during hamster oocyte maturation. Cell Mol Biol 37:623–629

    PubMed  CAS  Google Scholar 

  • Kline D, Kline JT (1992) Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg. Dev Biol 149:80–89

    Article  PubMed  CAS  Google Scholar 

  • Lambert CC (1989) Ascidian eggs release glycosidase activity which aids in the block against polyspermy. Development 105:415–420

    PubMed  CAS  Google Scholar 

  • Lindsay LL, Hedrick JL (1989) Proteases released fromXenopus laevis eggs at activation and their role in envelope convertion. Dev Biol 135:202–211

    Article  PubMed  CAS  Google Scholar 

  • Lindsay LL, Larabell CA, Hedrick JL (1992) Brief note. Localization of a chymotrypsin-like protease to the perivitelline space ofXenopus laevis eggs. Dev Biol 154:433–436

    Article  PubMed  CAS  Google Scholar 

  • Lis H, Sharon N (1993) Protein glycosylation, structural and functional aspects. Eur J Biochem 218:1–27

    Article  PubMed  CAS  Google Scholar 

  • Litcher ES, Juntunen K, Seppo A, Penttila L, Niemela R, Renkonen O, Wassarman P (1995) Oligosaccharide constructs with defined structures that inhibit binding of mouse sperm to unfertilized eggs in vitro. Biochemistry 34: 4662–4669

    Article  Google Scholar 

  • Macek MB, Shur BD (1988) Protein-carbohydrate complementarity in mammalian gamete recognition. Gamete Res 20:93–109

    Article  PubMed  CAS  Google Scholar 

  • Miller DJ, Roy LA (1990) Carbohydrates and fertilization in animals. Mol Reprod Dev 26:184–198

    Article  PubMed  CAS  Google Scholar 

  • Miller DJ, Macek MB, Shur BD (1992) Complementarity between sperm surface β-1, 4-galactosyltransferase and egg-coat ZP3 mediates sperm-egg binding. Nature 357:589–592

    Article  PubMed  CAS  Google Scholar 

  • Miller DJ, Gong X, Decker G, Shur BD (1993) Egg cortical granuleN-acetylglucosaminidase is required for the mouse zona block to polyspermy. J Cell Biol 123:1431–1440

    Article  PubMed  CAS  Google Scholar 

  • Moller CC, Wassarman PM (1989) Characterization of a proteinase that cleaves zona pellucida glycoprotein ZP2 following activation of mouse eggs. Dev Biol 132:103–112

    Article  PubMed  CAS  Google Scholar 

  • Moller CC, Bleil JD, Kinloch RA, Wassarman PM (1990) Structural and functional relationship between mouse and hamster zona pellucida glycoproteins. Dev Biol 137:276–286

    Article  PubMed  CAS  Google Scholar 

  • Mori K, Daitoh T, Irahara M, Kamada M, Aono T (1989) Significance ofd-mannose as a sperm receptor site on the zona pellucida in human fertilization. Am J Obstet Gynecol 161:207–211

    PubMed  CAS  Google Scholar 

  • Nagdas SK, Araki Y, Chayco CA, Orgebin-Crist MC, Tulsiani DRP (1994)O-Linked trisaccharide andN-linked poly-N-acetyllactosaminyl glycans are present on mouse ZP2 and ZP3. Biol Reprod 51:262–272

    Article  PubMed  CAS  Google Scholar 

  • Nicolson GL, Yanagimachi R, Yanagimachi Y (1975) Ultrastructural localization of lectin-binding sites on the zone pellucida and plasma membranes of mammalian eggs. J Cell Biol 66:263–274

    Article  PubMed  CAS  Google Scholar 

  • Nikas G, Paraschos T, Psychoyos A, Handyside AH (1994) The zona reaction in human oocytes as seen with scanning electron microscopy. Hum Reprod 9:2135–2138

    PubMed  CAS  Google Scholar 

  • Noguchi S, Nakano M (1993) Structural characterization of theN-linked carbohydrate chains from mouse zona pellucida glycoproteins ZP2 and ZP3. Biochim Biophys Acta 1158:217–226

    PubMed  CAS  Google Scholar 

  • Oikawa T, Yanagimachi R, Nicolson GL (1973) Wheat germ agglutinin blocks mammalian fertilization. Nature 241:256–259

    Article  PubMed  CAS  Google Scholar 

  • Oikawa T, Yanagimachi R, Nicolson GL (1975) Species differences in the lectin-binding sites on the zona pellucida of rodent eggs. J Reprod Fertil 43:137–140

    Article  PubMed  CAS  Google Scholar 

  • Oikawa T, Sendai Y, Kurata S, Yanagimachi R (1988) A glycoprotein of oviductal origin alters biochemical properties of the zona pellucida of hamster egg. Gamete Res 19:113–122

    Article  PubMed  CAS  Google Scholar 

  • Phillips DM, Shalgi R (1980) Surface properties of the zona pellucida. J Exp Zool 213:1–8

    Article  PubMed  CAS  Google Scholar 

  • Roux E, Kan FWK (1991) Changes of glycoconjugate contents of the zona pellucida during oocyte growth and development in the golden hamster: a quantitative cytochemical study. Anat Rec 230:347–360

    Article  PubMed  CAS  Google Scholar 

  • Saling PM (1989) Mammalian sperm interaction with extracellular matrices of the egg. In: Oxford review of reproductive biology, vol 11. Oxford University Press, Oxford, pp 339–388

    Google Scholar 

  • Shabanowitz RB, O'Rand MG (1988) Characterization of the human zona pellucida from fertilized and unfertilized eggs. J Reprod Fertil 82:151–161

    Article  PubMed  CAS  Google Scholar 

  • Shalgi R (1991) Fertilization in the rat: I:: Dumbar BS, O'Rand MG (eds) A comparative overview of mammalian fertilization. Plenum Press, New York, pp 245–255

    Google Scholar 

  • Shalgi R, Matityahu A, Nebel L (1986) The role of carbohydrates in sperm-egg interaction in rats. Biol Reprod 34:446–452

    Article  PubMed  CAS  Google Scholar 

  • Shalgi R, Maymon R, Bar-Shira (Maymon) B, Amihai D, Skutelsky E (1991) Distribution of lectin receptors sites in the zona pellucida of follicular and ovulated rat oocytes. Mol Reprod Dev 29:365–372

    Article  PubMed  CAS  Google Scholar 

  • Shimizu S, Ito M (1986) Glycoproteins of murine zona pellucida from unfertilized eggs and two-cell embryos: comparison of reactivity to lectins. Biochem Int 13:335–341

    PubMed  CAS  Google Scholar 

  • Shimizu S, Ito M, Dean J (1982) Glycoproteins of mouse zona pellucida: analysis of their reactivity to lectins. Biochem Biophys Res Commun 109:449–454

    Article  PubMed  CAS  Google Scholar 

  • Skutelsky E, Goyal V, Alroy J (1987) The use of avidin-gold complex for light microscopic localization of lectin receptors. Histochemistry 86:291–295

    Article  PubMed  CAS  Google Scholar 

  • Skutelsky E, Ranen E, Shalgi R (1994) Variation in the distribution of sugar residues in the zona pellucida as possible species specific determinants of mammalian oocytes. J Reprod Fertil 100:35–41

    Article  PubMed  CAS  Google Scholar 

  • Suprasert A, Hirunagi K, Fujioka T, Yokoyama A (1989) Histochemistry of glycoconjugates in ovarian follicles of the adult house musk shrew,Suncus murinus. Acta Anat 136:269–278

    Article  PubMed  CAS  Google Scholar 

  • Suzuki H, Yang X, Foote RH (1994) Surface alterations of the bovine oocyte and its investments during and after maturation and fertilization in vitro. Mol Reprod Dev 38:421–430

    Article  PubMed  CAS  Google Scholar 

  • Szollosi D (1967) Development of cortical granules and the cortical reaction in rat and hamster eggs. Anat Rec 159:431–446

    Article  PubMed  CAS  Google Scholar 

  • Tawia SA, Lopata A (1992) The fertilization and development of mouse oocytes following cortical granule discharge in the presence of a protease inhibitor. Hum Reprod 7:1004–1009

    PubMed  CAS  Google Scholar 

  • Thall AD, Maly P, Lowe JB (1995) Oocyte Galα1,3Gal epitopes implicated in sperm adhesion to the zona pellucida glycoprotein ZP3 are not required for fertilization in the mouse. J Biol Chem 270:21437–21440

    Article  PubMed  CAS  Google Scholar 

  • Tombes RM, Simerly C, Borisy GG, Schatten G (1992) Meiosis, egg activation, and nuclear envelope breakdown are differentially reliant on Ca2+, whereas germinal vesicle breakdown is Ca2+-independent in the mouse oocyte. J Cell Biol 117:799–811

    Article  PubMed  CAS  Google Scholar 

  • Toyoda Y, Chang MC (1974) Fertilization of rat eggs in vitro by epididymal spermatozoa and the development of eggs following transfer. J Reprod Fertil 36:9–22

    Article  PubMed  CAS  Google Scholar 

  • Tulsiani DRP, Nagdas SK, Cornwall GA, Orgebin-Crist MC (1992) Evidence for the presence of high mannose/hydbrid oligosaccharide chain(s) on the mouse ZP2 and ZP3. Biol Reprod 46:93–100

    Article  PubMed  CAS  Google Scholar 

  • Vincent C, Cheek TR, Johnson MH (1992) Cell cycle progression of parthenogenetically activated mouse oocytes to interphase is dependent on the level of internal calcium. J Cell Sci 103:389–396

    PubMed  CAS  Google Scholar 

  • Wassarman PM (1988) Zona pellucida glycoproteins. Annu Rev Biochem 57:415–442

    Article  PubMed  CAS  Google Scholar 

  • Wassarman PM (1990) Profile of a mammalian sperm receptor. Development 108:1–17

    PubMed  CAS  Google Scholar 

  • Wolf DP, Hamada M (1977) Induction of zonal and egg plasma membrane blocks to sperm penetration in mouse eggs with cortical granule exudate. Biol Reprod 17:350–354

    Article  PubMed  CAS  Google Scholar 

  • Wu TC, Lee MC, Wan YJ, Damjanov I (1984) Lectin binding sites of the mouse ovary, introvarian and ovulated ova. Histochemistry 80:527–533

    Article  PubMed  CAS  Google Scholar 

  • Yanagimachi R (1994) Mammalian fertilization. In: Knobil E, Neil JD (eds) The physiology of reproduction. Raven Press, New York, pp 189–317

    Google Scholar 

  • Yanagimachi R, Nicolson GL (1976) Lectin-binding properties of hamster egg zona pellucida and plasma membrane during maturation and preimplantation development. Exp Cell Res 100:249–257

    Article  PubMed  CAS  Google Scholar 

  • Yurewicz EC, Pack BA, Sacco AG (1991) Isolation, composition, and biological activity of sugar chains of porcine oocyte zona pellucida 55K glycoprotein. Mol Reprod Dev 30:126–134

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Embryology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, 69978, Tel Aviv, Israel

    Tamar Raz & Ruth Shalgi

  2. Department of Pathology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Tel Aviv, Israel

    Ehud Skutelsky

Authors
  1. Tamar Raz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ehud Skutelsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ruth Shalgi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ruth Shalgi.

Additional information

This work is in partial fulfillment of the requirements for the PhD degree of Tamar Raz at the Sackler School of Medicine, Tel Aviv University

Rights and permissions

Reprints and permissions

About this article

Cite this article

Raz, T., Skutelsky, E. & Shalgi, R. Post-fertilization changes in the zona pellucida glycoproteins of rat eggs. Histochem Cell Biol 106, 395–403 (1996). https://doi.org/10.1007/BF02473298

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02473298

Keywords

Search

Navigation