We previously demonstrated that an Sp1-dependent reporter gene is preferentially expressed in G2 of the 1-cell mouse embryo following microinjection of the male pronucleus when compared to microinjection of the female pronucleus (P.T. Ram and R.M. Schultz, 1993, Dev. Biol. 156, 552–556). We also noted that expression of the reporter gene is not observed following microinjection of the germinal vesicle of the fully grown oocyte. In the present study, we examined expression of this reporter gene during oocyte growth, as well as the nuclear concentration of two transcription factors, Sp1 and the TATA box-binding protein, TBP, during oocyte growth and the first cell cycle. The extent of reporter gene expression decreases during oocyte growth and this decrease correlates with the decrease in nuclear concentration of Sp1, as determined by confocal immunofluorescent microscopy. In addition, results of immunoblotting experiments also indicate a similar decrease in the total concentration of Sp1 during oocyte growth. The nuclear concentration of TBP also decreases during oocyte growth, as determined by confocal immunofluorescent microscopy. Following fertilization, the pronuclear concentration of these two transcription factors increases in a time-dependent fashion and the concentration of each is greater in the male pronucleus as compared to the female pronucleus. For each pronucleus and for each transcription factor, this increase in nuclear concentration is inhibited by aphidicolin, which inhibits DNA synthesis. Last, the increase in nuclear concentration of these two proteins observed between the 1-cell and 2-cell stages does not require transcription or cytokinesis.
REFERENCES
REFERENCES
Abramczuk
J.
, Sawicki
W.
(1975
) Pronuclear synthesis of DNA in fertilized and parthenogenetically activated mouse eggs.
Exp. Cell Res
92
, 361
–372
Bachvarova
R.
, DeLeon
V.
(1980
) Polyadenylated RNA of mouse ova and loss of maternal RNA in early development.
Dev. Biol
74
, 1
–8
Bachvarova
R.
(1981
) Synthesis, turnover and stability of heterogeneous RNA in growing mouse oocytes.
Dev. Biol
86
, 384
–392
Bachvarova
R.
, Cohen
E. M.
, DeLeon
V.
, Tokunaga
K.
, Sakiyama
S.
, Paynton
B. V.
(1989
) Amounts and modulation of actin mRNAs in mouse oocytes and embryos.
Development
106
, 561
–565
Bachvarova
R.
, DeLeon
V.
, Johnson
A.
, Kaplan
G.
, Paynton
B. V.
(1985
) Changes in total RNA, polyadenylated RNA, and actin mRNA during meiotic maturation of mouse oocytes.
Dev. Biol
108
, 325
–331
Bensaude
O.
, Babinet
C.
, Morange
M.
, Jacob
F.
(1983
) Heat shock proteins, first major products of zygotic gene activity.
Nature
305
, 331
–333
Bevilacqua
A.
, Kinnunen
L. H.
, Mangia
F.
(1992
) Genetic manipulation of mammalian dictyate oocytes: Factors affecting transient expression of microinjected DNA templates.
Mol. Reprod. Dev
33
, 124
–130
Bolton
V. N.
, Oades
P. J.
, Johnson
M. H.
(1984
) The relationship between cleavage, DNA replication, and gene expression in the mouse 2-cell embryos.
J. Embryol. Exp. Morph
79
, 139
–163
Bonnerot
C.
, Vernet
M.
, Grimber
G.
, Briand
P.
, Nicolas
J.-F.
(1991
) Transcriptional selectivity in early mouse embryos: a qualitative study.
Nuc. Acid Res
25
, 7251
–7557
Bornslaeger
E. A.
, Schultz
R. M.
(1985
) Adenylate cyclase activity in zona-free mouse oocytes.
Exp. Cell Res
156
, 277
–281
Briggs
M. R.
, Kadonaga
J. T.
, Bell
S. P.
, Tjian
R.
(1986
) Purification and biochemical characterization of the promoter-specific transcription factor, Sp1.
Science
234
, 47
–52
Brower
P. T.
, Gizang
E.
, Boreen
S. M.
, Schultz
R. M.
(1981
) Biochemical studies of mammalian oogenesis: Synthesis and stability of various classes of RNA during growth of the mouse oocyte in vitro.
Dev. Biol
86
, 373
–383
Burhans
W. C.
, Vassilev
L. T.
, Wu
J.
, Sogo
J. M.
, Nallaseth
F. S.
, DePamphilis
M. L.
(1991
) Emetine allows identification of origins of mammalian DNA replication by imbalanced DNA synthesis, not through conservative nucleosome segregation.
EMBO J
10
, 4351
–4360
Cascio
S. M.
, Wassarman
P. M.
(1982
) Program of early development in the mammal: Post-transcriptional control of a class of proteins synthesized by mouse oocytes and early embryos.
Dev. Biol
89
, 397
–408
Chatot
C. L.
, Ziomek
C. A.
, Bavister
B. D.
, Lewis
J. L.
, Torres
I.
(1989
) An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro.
J. Reprod. Fertil
86
, 679
–688
Clarke
H. J.
(1992
) Nuclear and chromatin composition of mammalian gametes and early embryos.
Biochem. Cell Biol
70
, 856
–866
Debey
P.
, Szöllösi
M. S.
, Szöllösi
D.
, Vautier
D.
, Girousse
A.
, Besombes
D.
(1993
) Competent mouse oocytes isolated from antral follicles exhibit different chromatin organization and follow different maturation dynamics.
Mol. Reprod. Dev
36
, 59
–74
Driever
W.
, Nusslein-Volhard
C.
(1988
) A gradient of bicoid protein in Drosophila embryos.
Cell
54
, 83
–93
Driever
W.
, Nusslein-Volhard
C.
(1989
) The bicoid protein is a positive regulator of hunchback transcription in the early Drosophila embryo.
Nature
337
, 138
–143
Dynan
W. S.
(1986
) Promoters for housekeeping genes.
Trends Gene
2
, 196
–197
Eppig
J. J.
(1977
) Mouse oocyte development in vitro with various culture systems.
Dev. Biol
60
, 371
–388
Felsenfeld
G.
(1992
) Chromatin as an essential part of the transcriptional mechanism.
Nature
355
, 219
–224
Flach
G.
, Johnson
M. H.
, Braude
P. R.
, Taylor
R. A. S.
, Bolton
V. N.
(1982
) The transition from maternal to embryonic control in the 2-cell mouse embryo.
EMBO J
1
, 681
–686
Golbus
M. S.
, Calarco
P. G.
, Epstein
C. J.
(1973
) The effects ofinhibitors of RNA synthesis (-amanitin and actinomycin D) on preimplantation mouse embryogenesis.
J. Exp. Zool
186
, 207
–216
Gottesfeld
J. M.
, Wolf
V. J.
, Dang
T.
, Forbes
D. J.
, Hartl
P.
(1994
) Mitotic repression of RNA polymerase II transcription in vitro mediated by phosphorylation of TFIIB component.
Science
263
, 81
–84
Howlett
S. K.
, Bolton
V. N.
(1985
) Sequence and regulation of morphological and molecular events during the first cell cycle of mouse embryogenesis.
J. Embryol. Exp. Morph
87
, 175
–206
Jackson
S. P.
, Tjian
R.
(1988
) O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation.
Cell
55
, 125
–133
Jackson
S. P.
, MacDonald
J. J.
, Lees-Miller
S.
, Tjian
R.
(1990
) GC box binding induces phosphorylation of Sp1 by a DNA-dependent protein kinase.
Cell
63
, 155
–165
Jahn
C. L.
, Baran
M. M.
, Bachvarova
R.
(1976
) Stability of RNA synthesized by the mouse oocyte during its major growth phase.
J. Exp. Zool
197
, 161
–172
Kadonaga
J. T.
, Jones
K. A.
, Tjian
R.
(1986
) Promoter-specific activation of RNA polymerase II transcription by Sp1.
Trends Biochem. Sci
11
, 20
–23
Kamakaka
R. T.
, Bulger
M.
, Kadonaga
J. T.
(1993
) Potentiation of RNA polymerase II transcription by Gal4-VP16 during but not after DNA replication and chromatin assembly.
Genes Dev
7
, 1779
–1795
Kaplan
G.
, Abreu
S. L.
, Bachvarova
R.
(1982
) rRNA accumulation and protein synthetic patterns in growing mouse oocytes.
J. Exp. Zool
220
, 361
–370
Laemmli
U. K.
(1970
) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
, 680
–685
Latham
K. E.
, Garrels
J. I.
, Chang
C.
, Solter
D.
(1991
) Quantitative analysis of protein synthesis in mouse embryos. I. Extensive reprogramming at the one-and two-cell stages.
Development
112
, 921
–932
Latham
K. E.
, Solter
D.
, Schultz
R. M.
(1992
) Acquisition of a transcriptionally permissive state during the 1-cell stage of mouse embryogenesis.
Dev. Biol
149
, 457
–462
Laybourn
P. J.
, Kadonaga
J. T.
(1991
) Role of nucleosomal cores and histone H1 in regulation of transcription by RNA polymerase II.
Science
254
, 238
–245
Luthardt
F. W.
, Donahue
R. P.
(1973
) Pronuclear DNA synthesis in mouse eggs. An autoradiographic study.
Exp. Cell Res
82
, 143
–151
Manejwala
F. M.
, Logan
C. Y.
, Schultz
R. M.
(1991
) Regulation of hsp70 mRNA levels during oocyte maturation and zygotic gene activation in the mouse.
Dev. Biol
144
, 301
–308
Minie
M. E.
, Kimura
T.
, Felsenfeld
G.
(1992
) The developmental switch in embryonic-globin expression is correlated with erythroid lineage-specific differences in transcription factor levels.
Development
115
, 1149
–1164
Moore
G. P. M.
(1975
) The RNA polymerase activity of the preimplantation mouse embryo.
J. Embryol. Exp. Morph
34
, 291
–298
Moore
G. P. M.
, Lintern-Moore
S.
(1978
) Transcription of the mouse oocyte genome.
Biol. Reprod
17
, 865
–870
Paynton
B. V.
, Rempel
R.
, Bachvarova
R.
(1988
) Changes in state of adenylation and time course of degradation of maternal mRNAs during oocyte maturation and early embryonic development in the mouse.
Dev. Biol
129
, 304
–314
Perreault
S. D.
(1992
) Chromatin remodeling in mammalian zygotes.
Mutat. Res
296
, 43
–55
Poueymirou
W. T.
, Schultz
R. M.
(1987
) Differential effects of activators of cAMP-dependent protein kinase and protein kinase C on cleavage of one-cell mouse embryos and protein synthesis and phosphorylation in one-and two-cell embryos.
Dev. Biol
121
, 489
–498
Poueymirou
W. T.
, Schultz
R. M.
(1989
) Regulation of mouse preimplantation development: Inhibition of synthesis of proteins in the two-cell embryo that require transcription by inhibitors of cAMP-dependent protein kinase.
Dev. Biol
133
, 588
–599
Poueymirou
W. T.
, Schultz
R. M.
(1990
) Regulation of mouse preimplantation development: Inhibitory effect of the calmodulin antagonist W7 on the first cleavage.
Mol. Reprod. Dev
26
, 211
–216
Pugh
B. F.
, Tjian
R.
(1990
) Mechanism of transcriptional activation by Sp1: evidence for coactivators.
Cell
61
, 1187
–1197
Pugh
B. F.
, Tjian
R.
(1991
) Transcription from a TATA-less promoter requires a multisubunit TFIID complex.
Genes Dev
5
, 1935
–1945
Ram
P. T.
, Schultz
R. M.
(1993
) Reporter gene expression in G2 of the 1-cell mouse embryo.
Dev. Biol
156
, 552
–556
Sawicki
J. A.
, Magnuson
T.
, Epstein
C. J.
(1982
) Evidence for the expression of the paternal genome in the two-cell mouse embryo.
Nature
294
, 450
–451
Schultz
R. M.
(1993
) Regulation of zygotic gene activation in the mouse.
BioEssays
15
, 531
–538
Schultz
R. M.
, Wassarman
P. M.
(1977
) Biochemical studies of mammalian oogenesis: Protein synthesis during oocyte growth and meiotic maturation in the mouse.
J. Cell Sci
24
, 167
–194
Schultz
R. M.
, Letourneau
G. A.
, Wassarman
P. M.
(1979
) Program of early development in the mammal: Changes in patterns and absolute rates of tubulin and total protein synthesis during oogenesis and early embryogenesis in the mouse.
Dev. Biol
68
, 341
–359
Schultz
R. M.
, Montgomery
R. R.
, Belanoff
J. R.
(1983
) Regulation of mouse oocyte maturation: Implication of a decrease in oocyte cAMP and protein dephosphorylation in commitment to resume meiosis.
Dev. Biol
97
, 264
–273
Schwartz
D. A.
, Schultz
R. M.
(1992
) Zygotic gene activation in the mouse embryo: Involvement of cyclic adenosine monophosphate-dependent protein kinase and appearance of an AP-1-like activity.
Mol. Reprod. Dev
32
, 209
–216
Sharp
P. A.
(1992
) TATA-binding protein is a classless factor.
Cell
68
, 819
–821
Sorensen
R. A.
, Wassarman
P. M.
(1976
) Relationship between growth and meiotic maturation of the mouse oocyte.
Dev. Biol
50
, 531
–536
Sternlicht
A. L.
, Schultz
R. M.
(1981
) Biochemical studies of mammalian oogenesis:kinetics of accumulation of total and poly (A)-containing RNA during growth of the mouse oocyte.
J. Exp. Zool
215
, 191
–200
Struhl
K.
(1993
) Chromatin and transcription factors: who's on first?.
Current Biol
3
, 220
–222
Svaren
J.
, Hörz
W.
(1993
) Histones, nucleosomes and transcription.
Curr. Opin. Genet. Dev
3
, 219
–225
van Blerkom
J.
(1981
) Structural relationship and post-translational modification of stage-specific proteins synthesized during early preimplantation development in the mouse.
Proc. Natl. Acad. Sci. USA
78
, 7629
–7633
Vernet
M.
, Bonnerot
C.
, Briand
P.
, Nicolas
J.-P.
(1992
) Changes in permissiveness for the expression of microinjected DNA during the first cleavage of mouse embryos.
Mech. Dev
36
, 129
–139
Wassarman
P. M.
, Letourneau
G. E.
(1976
) RNA synthesis in fully-grown mouse oocytes.
Nature
261
, 73
–74
Weis
L.
, Reinberg
D.
(1992
) Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes.
FASEB J
6
, 3300
–3309
Wickramasinghe
D.
, Ebert
K. M.
, Albertini
D. F.
(1991
) Meiotic competence acquisition is associated with the appearance of M-phase characteristics in growing mouse oocytes.
Dev. Biol
143
, 162
–173
Wiekowski
M.
, Miranda
M.
, DePamphilis
M. L.
(1991
) Regulation of gene expression in preimplantation mouse embryos: Effects of the zygotic clock and the first mitosis on promoter and enhancer activities.
Dev. Biol
147
, 403
–414
Wiekowski
M.
, Miranda
M.
, DePamphilis
M. L.
(1993
) Requirements for promoter activity in mouse oocytes and embryos distinguish paternal pronuclei from maternal and zygotic nuclei.
Dev. Biol
159
, 366
–378
Wolffe
A. P.
(1991
) Activating chromatin.
Current Biology
1
, 366
–368
This content is only available via PDF.
© 1994 by Company of Biologists
1994
