The mammalian zona pellucida is an extracellular matrix that surrounds growing oocytes, ovulated eggs and early embryos. The mouse zona is composed of three sulfated glycoproteins: ZP1, ZP2 and ZP3. Each is critically involved in fertilization, the postfertilization block to polyspermy and protection of the preimplantation embryo. We have previously isolated cDNAs encoding mouse ZP2 and ZP3 and now report the isolation of a full-length cDNA encoding ZP1. Mouse ZP1 is composed of a 623 amino acid polypeptide chain with a signal peptide and a carboxyl terminal transmembrane domain, typical of all zona proteins. Sequence comparison demonstrate that mouse ZP1 is an orthologue of a rabbit zona protein, R55. The expression of R55 has been reported previously in both oocytes and granulosa cells. However, by northern analysis and in situ hybridization with 33P-labelled antisense probes to each of the three mouse zona mRNAs, we have determined that the expression of each mouse zona gene is restricted to the oocyte. ZP2 transcripts, but not ZP1 or ZP3, are detected in resting (15 microns diameter) oocytes, and all three zona transcripts coordinately accumulate as oocytes begin to grow. Together they represent approximately 1.5% of the total poly(A)+ RNA in 50–60 microns oocytes. In the latter stages of oogenesis, their abundance declines and each zona transcript is present in ovulated eggs at less than 5% of its maximal level. No zona transcripts were detected above background signal in granulosa cells. We conclude that, in mice, the three zona pellucida genes are expressed in a coordinate, oocyte-specific manner during the growth phase of oogenesis. Our data support the hypothesis that the transcription of the zona genes is controlled, in part, by shared regulatory element(s).

Reference

Arcellana-Panlilio
M. Y.
,
Schultz
G. A.
(
1994
)
Temporal and spatial expression of major histocompatibility complex class I H-2K in the early mouse embryo.
Biol. Reprod
51
,
169
183
Beebe
S. J.
,
Leyton
L.
,
Burks
D.
,
Fuerst
T.
,
Dean
J.
,
Saling
P. M.
(
1992
)
Recombinant ZP3 inhibits sperm binding and induces the acrosome reaction.
Dev. Biol
151
,
48
54
Bleil
J. D.
,
Greve
J. M.
,
Wassarman
P. M.
(
1988
)
Identification of a secondary sperm receptor in the mouse egg zona pellucida: role in maintenance of binding of acrosome-reacted sperm to eggs.
Dev. Biol
128
,
376
385
Bleil
J. D.
,
Wassarman
P. M.
(
1980
)
Mammalian sperm-egginteraction: Identification of a glycoprotein in mouse egg zonae pellucidae possessing receptor activity for sperm.
Cell
20
,
873
882
Bleil
J. D.
,
Wassarman
P. M.
(
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
Bleil
J. D.
,
Wassarman
P. M.
(
1980
)
Synthesis of zona pellucida proteins by denuded and follicle-enclosed mouse oocytes during culture in vitro.
Proc. Natl. Acad. Sci. USA
77
,
1029
1033
Bousquet
D.
,
Leveille
M. C.
,
Roberts
K. D.
,
Chapdelaine
A.
,
Bleau
G.
(
1981
)
The cellular origin of the zona pellucida antigen in the human and hamster.
J. Exp. Zool
215
,
215
218
Burnette
W. N.
(
1981
)
‘Western blotting:: electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
Anal. Biochem
112
,
195
203
Chamberlin
M. E.
,
Dean
J.
(
1990
)
Human homolog of the mouse sperm receptor.
Proc. Natl. Acad. Sci. USA
87
,
6014
6018
Cheng
A.
,
Le
T.
,
Palacios
M.
,
Bookbinder
L. H.
,
Wassarman
P. M.
,
Suzuki
F.
,
Bleil
J. D.
(
1994
)
Sperm-egg recognition in the mouse: characterization of sp56, a sperm protein having specific affinity for ZP3.
J. Cell Biol
125
,
867
878
Chiquoine
A. D.
(
1954
)
The identification, origin and migration of the primordial germ cells in the mouse embryo.
Anat. Rec
118
,
135
146
Derman
E.
,
Krauter
K.
,
Walling
L.
,
Weinberger
C.
,
Ray
M.
,
Darnell
J. E.
Jr
(
1981
)
Transcriptional control in the production of liver-specific mRNAs.
Cell
23
,
731
739
Dunbar
B. S.
,
Liu
C.
,
Sammons
D. W.
(
1981
)
Identification of the three major proteins of porcine and rabbit zonae pellucidae by high resolution two-dimensional gel electrophoresis: comparison with serum, follicular, fluid, and ovarian cell proteins.
Biol. Reprod
24
,
1111
1124
Dunbar
B. S.
,
Avery
S.
,
Lee
V.
,
Prasad
S.
,
Schwahn
D.
,
Schwoebel
E.
,
Skinner
S.
,
Bilins
B.
(
1995
)
The mammalian zona pellucida: its biochemistry, immunochemistry, mulecular biology, and developmental expression.
Reprod. Fert. Develop
6
,
331
347
East
I. J.
,
Dean
J.
(
1984
)
Monoclonal antibodies as probes of the distribution of ZP-2, the major sulfated glycoprotein of the murine zona pellucida.
J. Cell Biol
98
,
795
800
Flechon
J. E.
,
Pavlok
A.
,
Kopecny
V.
(
1984
)
Dynamics of zona pellucida formation by mouse oocyte. An autoradiographic study. Biol.
Cell
51
,
403
406
Florman
H. M.
,
Wassarman
P. M.
(
1985
)
O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity.
Cell
41
,
313
324
Greve
J. M.
,
Wassarman
P. M.
(
1985
)
Mouse egg extracellular coat is a matrix of interconnected filaments possessing a structural repeat.
J. Mol. Biol
181
,
253
264
Haddad
A.
,
Nagai
E. M. T.
(
1977
)
Radioautographic study of glycoprotein biosynthesis and renewal in the ovarian follicles of mice and the origin of the zona pellucida.
Cell Tissue Res
177
,
347
369
Hadek
R.
(
1965
)
The structure of the mammalian egg.
Internat. Rev. Cyt
18
,
29
68
Haines
D. S.
,
Gillespie
D. H.
(
1992
)
RNA abundance measured by a lysate RNase protection assay.
BioTechniques
12
,
736
741
Hedrick
J. L.
,
Wardrip
N. J.
(
1987
)
On the macromolecular composition of the zona pellucida from porcine oocytes.
Dev. Biol
121
,
478
488
Hosaka
M.
,
Nagahama
M.
,
Kim
W. S.
,
Watanabe
T.
,
Hatsuzawa
K.
,
Ikemizu
J.
,
Murakami
K.
,
Nakayama
K.
(
1991
)
Arg-X-Lys/Arg-Arg motif as a signal for precursor cleavage catalyzed by furin within the constitutive secretory pathway.
J. Biol. Chem
266
,
12127
12130
Huarte
J.
,
Belin
D.
,
Vassalli
A.
,
Strickland
S.
,
Vassalli
J. D.
(
1987
)
Meiotic maturation of mouse oocytes triggers the translation and polyadenylation of dormant tissue-type plasminogen activator mRNA.
Genes Dev
1
,
1201
1211
Kang
Y.-H.
(
1974
)
Development of the zona pellucida in the rat oocyte.
Am. J. Anat
139
,
535
566
Kinloch
R. A.
,
Mortillo
S.
,
Stewart
C. L.
,
Wassarman
P. M.
(
1991
)
Embryonal carcinoma cells transfected with ZP3 genes differentially glycosylate similar polypeptides and secrete active mouse sperm receptor.
J. Cell Biol
115
,
655
664
Kinloch
R. A.
,
Sakai
Y.
,
Wassarman
P. M.
(
1995
)
Mapping the mouse ZP3 combining site for sperm by exon swapping and site-directed mutagenesis.
Proc. Natl. Acad. Sci. USA
92
,
263
267
Kozak
M.
(
1991
)
Structural features in eukaryotic mRNAs that modulate the initiation of translation.
J. Biol. Chem
266
,
19867
19870
Kyte
J.
,
Doolittle
R. F.
(
1982
)
A simple method for displaying the hydropathic character of a protein.
J. Mol. Biol
157
,
105
132
Lee
V. H.
,
Dunbar
B. S.
(
1993
)
Developmental expression of the rabbit 55-kDa zona pellucida protein and messenger RNA in ovarian follicles.
Dev. Biol
155
,
371
382
Leveille
M. C.
,
Roberts
K. D.
,
Chevalier
S.
,
Chapdelaine
A.
,
Bleau
G.
(
1987
)
Formation of the hamster zona pellucida in relation to ovarian differentiation and follicular growth.
J. Reprod. Fertil
79
,
173
183
Leyton
L.
,
LeGuen
P.
,
Bunch
D.
,
Saling
P. M.
(
1992
)
Regulation of mouse gamete interaction by a sperm tyrosine kinase.
Proc. Natl. Acad. Sci. USA
89
,
11692
11695
Liang
L.-F.
,
Chamow
S. M.
,
Dean
J.
(
1990
)
Oocyte-specific expression of mouse Zp-2: Developmental regulation of the zona pellucida genes.
Mol. Cell. Biol
10
,
1507
1515
Liang
L.-F.
,
Dean
J.
(
1993
)
Conservation of the mammalian secondary sperm receptor genes results in promoter function of the human homologue in heterologous mouse oocytes.
Dev. Biol
156
,
399
408
Lyons
C. E.
,
Payette
K. L.
,
Price
J. L.
,
Huang
R. C. C.
(
1993
)
Expression and structural analysis of a teleost homolog of a mammalian zona pellucida gene.
J. Biol. Chem
268
,
21351
21358
Manova
K.
,
Nocka
K.
,
Besmer
P.
,
Bachvarova
R. F.
(
1990
)
Gonadal expression of c-kit encoded at the W locus of the mouse.
Development
110
,
1057
1069
Matsudaira
P.
(
1987
)
Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.
J Biol. Chem
262
,
10035
10038
McLaughlin
S. K.
,
Margolskee
R. F.
(
1993
)
32P is preferable to 35S for labeling probes used in in situ hybridization.
BioTechniques
15
,
506
511
Millar
S. E.
,
Chamow
S. M.
,
Baur
A. W.
,
Oliver
C.
,
Robey
F.
,
Dean
J.
(
1989
)
Vaccination with a synthetic zona pellucida peptide produces long-term contraception in female mice.
Science
246
,
935
938
Millar
S. E.
,
Lader
E.
,
Liang
L.-F.
,
Dean
J.
(
1991
)
Oocyte-specific factors bind a conserved upstream sequence required for mouse zona pellucida promoter activity.
Mol. Cell. Biol
12
,
6197
6204
Millar
S. E.
,
Lader
E. S.
,
Dean
J.
(
1993
)
ZAP-1 DNA binding activity is first detected at the onset of zona pellucida gene expression in embryonic mouse oocytes.
Dev. Biol
158
,
410
413
Moos
M.
Jr.
,
Nguyen
N. Y.
,
Liu
T. Y.
(
1988
)
Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support.
J. Biol. Chem
263
,
6005
6008
Needleman
S. B.
,
Wunsch
C. D.
(
1970
)
A general method applicable to the search for similarities in the amino acid sequence of two proteins.
J. Mol. Biol
48
,
443
453
Phillips
D. M.
,
Shalgi
R.
(
1980
)
Surface architecture of the mouse and hamster zona pellucida and oocyte.
J. Ultrastruct. Res
72
,
1
12
Philpott
C. C.
,
Ringuette
M. J.
,
Dean
J.
(
1987
)
Oocyte-specific expression and developmental regulation of ZP3, the sperm receptor of the mouse zona pellucida.
Dev. Biol
121
,
568
575
Ringuette
M. J.
,
Sobieski
D. A.
,
Chamow
S. M.
,
Dean
J.
(
1986
)
Oocyte-specific gene expression: molecular characterization of a cDNA coding for ZP-3, the sperm receptor of the mouse zona pellucida.
Proc. Natl. Acad. Sci. USA
83
,
4341
4345
Ringuette
M. J.
,
Chamberlin
M. E.
,
Baur
A. W.
,
Sobieski
D. A.
,
Dean
J.
(
1988
)
Molecular analysis of cDNA coding for ZP3, a sperm binding protein of the mouse zona pellucida.
Dev. Biol
127
,
287
295
Sacco
A. G.
,
Yurewicz
E. C.
,
Subramanian
M. G.
,
Matzat
P. D.
(
1989
)
Porcine zona pellucida: association of sperm receptor activity with the alpha-glycoprotein component of the M r= 55,000 family.
Biol. Reprod
41
,
523
532
Saling
P. M.
(
1991
)
How the egg regulates sperm function during gamete interaction: facts and fantasies.
Biol. Reprod
44
,
246
251
Schwoebel
E.
,
Prasad
S.
,
Timmons
T. M.
,
Cook
R.
,
Kimura
H.
,
Niu
E.-M.
,
Cheung
P.
,
Skinner
S.
,
Avery
S. E.
,
Wilkins
B.
,
Dunbar
B. S.
(
1991
)
Isolation and characterization of a full-length cDNA encoding the 55 kDa rabbit zona pellucida protein.
J. Biol. Chem
266
,
7214
7219
Seto
D.
(
1990
)
An improved method for sequencing double stranded plasmid DNA from minipreps using DMSO and modified template preparation.
Nucleic Acids. Res
18
,
5905
5906
Shabanowitz
R. B.
,
O'Rand
M. G.
(
1988
)
Characterization of the human zona pellucida from fertilized and unfertilized eggs.
J. Reprod. Fertil
82
,
151
161
Shimizu
S.
,
Tsuji
M.
,
Dean
J.
(
1983
)
In vitro biosynthesis of three sulfated glycoproteins of murine zonae pellucidae by oocytes grown in follicle culture.
J. Biol. Chem
258
,
5858
5863
Stone
K. L.
,
Williams
K. R.
(
1986
)
High-performance liquid chromatographic peptide mapping and amino acid analysis in the sub-nanomole range.
J Chromatogr
359
,
203
212
Tempst
P.
,
Riviere
L.
(
1989
)
Examination of automated polypeptide sequencing using standard phenyl isothiocyanate reagent and subpicomole high-performance liquid chromatographic analysis.
Anal. Biochem
183
,
290
300
Von Heijne
G.
(
1985
)
Signal sequences: The limits of variation.
J. Mol. Biol
184
,
99
105
Von Heijne
G.
(
1986
)
A new method for predicting signal sequence cleavage sites.
Nucleic Acids Res
14
,
4683
4690
Wolgemuth
D. J.
,
Celenza
J.
,
Bundman
D. S.
,
Dunbar
B. S.
(
1984
)
Formation of the rabbit zona pellucida and its relationship to ovarian follicular development.
Dev. Biol
106
,
1
14
Youakim
A.
,
Hathaway
H. J.
,
Miller
D. J.
,
Gong
X.
,
Shur
B. D.
(
1994
)
Overexpressing sperm surface beta 1,4-galactosyltransferase in transgenic mice affects multiple aspects of sperm-egg interactions.
J. Cell Biol
126
,
1573
1583
Yurewicz
E. C.
,
Pack
B. A.
,
Sacco
A. G.
(
1991
)
Isolation, composition, and biological activity of sugar chains of porcine oocyte zona pellucida 55K glycoproteins.
Mol. Reprod. Dev
30
,
126
134
Yurewicz
E. C.
,
Hibler
D.
,
Fontenot
G. K.
,
Sacco
A. G.
,
Harris
J.
(
1993
)
Nucleotide sequence of cDNA encoding ZP3 alpha, a sperm-binding glycoprotein from zona pellucida of pig oocyte.
Biochim. Biophys. Acta
1174
,
211
214
Zamboni
L.
,
Upadhyay
S.
(
1983
)
Germ cell differentiation in mouse adrenal gland.
J. Exp. Zool
228
,
173
194
This content is only available via PDF.