INTRODUCTION
One of the striking peculiarities of meiotic maturation in mouse
oocytes concerns the first meiotic M phase (MI). In contrast to
all other M phases in vertebrate cells (except the metaphase II
arrest), the first meiotic M phase is long, usually lasting
between 7 and 10 hours. However, the precise timing of MI
seems to be genetically controlled, as shown by significant
variations in the speed of meiotic maturation between different
strains of mice, in particular the strains CBA/Kw and KE
(Polanski, 1986, 1997). CBA/Kw oocytes extrude the first
polar body about 7 hours after germinal vesicle breakdown
(GVBD), whilst the KE oocytes do so approximately 3-4 hours
later, that is 10-11 hours after GVBD.
The major cell cycle kinase, the cyclin/p34
cdc2
kinase
(maturation promoting factor; MPF) is activated just before
GVBD and its activity rises progressively until it reaches a
plateau at MI (Choi et al., 1991; Verlhac et al., 1994). By this
time, the microtubules have formed into a spindle, but exit from
the first meiotic M phase is delayed for a further 3-4 hours.
Mitotic cyclins are the regulatory subunits of the p34
cdc2
kinase that are synthesized throughout the cell cycle and
destroyed at each cell division (Evans et al., 1983). Cyclin
degradation is a proteolytic event involving the ubiquitin
pathway that is regulated throughout the cell cycle, such that
the cyclins remain stable in interphase and are destroyed during
a short interlude just before the metaphase-anaphase transition
(for a review, see Townsley and Ruderman, 1998). It was
reported that the synthesis of cyclin B1 increases gradually
during meiosis I in mouse oocytes (when MPF activity rises)
and the newly synthesized protein becomes complexed
immediately with the p34
cdc2
kinase (Hampl and Eppig, 1995b;
Winston, 1997). Thus, these data suggest that MPF activity is
regulated by a translation-dependent mechanism that
determines the level of cyclin synthesis. Similar increases in
cyclin B synthesis have also been observed during the
transition from G
2
to metaphase I in oocytes from Patella and
Xenopus (Gautier and Maller, 1991; Kobayashi et al., 1991;
van Loon et al., 1991; Galas et al., 1993).
In this work, we have studied the role of cyclin B1
metabolism in the timing of meiotic maturation using oocytes
from the KE and CBA/Kw mouse strains.
4989 Development 125, 4989-4997 (1998)
Printed in Great Britain © The Company of Biologists Limited 1998
DEV4079
To study the mechanisms involved in the progression of
meiotic maturation in the mouse, we used oocytes from two
strains of mice, CBA/Kw and KE, which differ greatly in
the rate at which they undergo meiotic maturation.
CBA/Kw oocytes extrude the first polar body about 7 hours
after breakdown of the germinal vesicle (GVBD), whilst the
oocytes from KE mice take approximately 3-4 hours longer.
In both strains, the kinetics of spindle formation are
comparable. While the kinetics of MAP kinase activity are
very similar in both strains (although slightly faster in
CBA/Kw), the rise of cdc2 kinase activity is very rapid in
CBA/Kw oocytes and slow and diphasic in KE oocytes.
When protein synthesis is inhibited, the activity of the cdc2
kinase starts to rise but arrests shortly after GVBD with a
slightly higher level in CBA/Kw oocytes, which may
correspond to the presence of a larger pool of cyclin B1 in
prophase CBA/Kw oocytes. After GVBD, the rate of cyclin
B1 synthesis is higher in CBA/Kw than in KE oocytes,
whilst the overall level of protein synthesis and the amount
of messenger RNA coding for cyclin B1 are identical in
oocytes from both strains. The injection of cyclin B1
messenger RNA in KE oocytes increased the H1 kinase
activity and sped up first polar body extrusion. Finally,
analysis of the rate of maturation in hybrids obtained after
fusion of nuclear and cytoplasmic fragments of oocytes
from both strains suggests that both the germinal vesicle
and the cytoplasm contain factor(s) influencing the length
of the first meiotic M phase. These results demonstrate that
the rate of cyclin B1 synthesis controls the length of the first
meiotic M phase and that a nuclear factor able to speed up
cyclin B synthesis is present in CBA/Kw oocytes.
Key words: Cyclin, H1 kinase, MAP kinase, Meiosis, Mouse oocyte
SUMMARY
Cyclin synthesis controls the progression of meiotic maturation in mouse
oocytes
Zbigniew Polanski
1,2
, Emilie Ledan
1
, Stéphane Brunet
1
, Sophie Louvet
1
, Marie-Hélène Verlhac
1
,
Jacek Z. Kubiak
1
and Bernard Maro
1,
*
1
Laboratoire de Biologie Cellulaire du Développement, Institut Jacques Monod, CNRS, Université Paris 6 and Université Paris 7,
2 place Jussieu, F-75005 Paris, France
2
Institute of Zoology, Jagiellonian University, Ingardena 6, 30-060 Krakow, Poland
*Author for correspondence (e-mail: maro@ijm.jussieu.fr)
Accepted 7 October; published on WWW 12 November 1998