BIOLOGY OF REPRODUCTION 59, 476–482 (1998)
M i n i r e v i ew
Hormonal Control of the Cell Cycle in Ovarian Cells: Proliferation
Versus Differentiation
Rebecca L. Robker and JoAnne S. Richards1
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030
INTRODUCTION
The growth of ovarian follicles, ovulation, and the for-
mation of the corpus luteum are complex processes that
involve dramatic changes in granulosa cell function. The
changes are sequential and are dictated by specific, tightly
regulated responses to gonadotropins, steroids, and growth
factors [1–3]. One of the most dramatic changes in granu-
losa cell function is the rapid switch from the highly pro-
liferative stage characterizing granulosa cells of preovula-
tory follicles to the nonproliferative, terminally differenti-
ated phase of luteal cells. Some of the cell cycle regulatory
mechanisms mediating this switch, as well as their control
by hormones, are the topics of this minireview.
In primordial follicles, the oocyte is surrounded by a
single layer of nondividing granulosa cells arrested in Go
phase of the cell cycle. Primordial follicles leave this qui-
escent state and initiate a phase of slow growth in which
the granulosa cells have entered the cell cycle but prolif-
eration is exceedingly slow [1]. However, as these slowly
dividing granulosa cells acquire enhanced responsiveness
to FSH and LH and begin producing estradiol [4, 5], ex-
posure to these hormones triggers a rapid burst of prolif-
eration that results in the formation of large preovulatory
follicles [6]. This rapid phase of growth is characterized by
a marked increase in the labeling of granulosa cells by tri-
tiated thymidine [1, 6], as well as by 5-bromodeoxyuridine
(BrdU) [7]. Granulosa cells of these preovulatory follicles
not only are highly proliferative but are also differentiating
and acquire LH receptors [8]. The LH surge then triggers
dramatic changes in both follicular structure and function.
LH terminates follicular growth by causing granulosa cells
of preovulatory follicles to exit the cell cycle [1, 6] and
rapidly initiates a program of terminal differentiation (lu-
teinization) in which the cells cease to divide [9, 10]. As
shown herein, the exit of preovulatory granulosa cells from
the cell cycle occurs within about 4 h after the LH surge
(Fig. 1), and this related to dramatic changes in specific
molecules regulating cell cycle progression (see Figs. 4 and
5). In addition, follicular rupture (ovulation) occurs and
granulosa cells luteinize to form mature corpora lutea. In-
terestingly, granulosa cells are completely reprogrammed to
luteinize by 7 h after exposure to the LH surge [4, 10].
Accepted March 26, 1998.
Received January 22, 1998.
1
Correspondence: JoAnne S. Richards, Department of Cell Biology,
Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. FAX:
(713) 790–1275; e-mail: joanner@bcm.tmc.edu
476
Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023
This pattern of granulosa cell proliferation and differ-
entiation that characterizes the natural growth of follicles
can be mimicked in hypophysectomized rats by a specific
hormonal regimen [4, 5, 11]. Injections of estradiol (1.5 mg
for 3 days) followed by FSH (1.0 mg for 2 days) stimulate
granulosa cell proliferation and follicle growth to the preo-
vulatory stage. A subsequent ovulatory dose of hCG (10
IU) triggers ovulation and luteinization. In the hypophy-
sectomized rat [4, 11], as well as in mutant mice lacking
either gonadotropins (hypogonadal [12, 13]) or FSH [14],
follicular development is arrested at the preantral stage
(Fig. 2). Mice lacking estrogen receptor a (ERa) also ex-
hibit impaired follicular growth and infertility [15], but the
follicles develop to the antral stage presumably due to the
presence of estrogen receptor b (ERb) in these cells [16].
These models illustrate that although the early, slow stages
of granulosa cell proliferation and preantral follicle growth
occur in the absence of gonadotropins and estradiol, these
hormones (and their receptors) are required for normal
growth—especially the final rapid stages of development
that form preovulatory follicles and permit these cells to
luteinize.
Cell cycle progression and proliferation are controlled
by a balance of positive and negative regulators converging
on cell cycle kinase cascades (reviewed in [17–23]; see
schematics, Figs. 3 and 7). Interestingly, specific roles for
cell cycle regulatory molecules in the control of granulosa
cell proliferation and differentiation during follicular de-
velopment have been elucidated by the altered ovarian phe-
notypes described in mice null for cyclin D2 [24] and
p27Kip1 [25–27]. Cyclin D2 [28, 29] acts as a positive reg-
ulator of cell cycle progression by its ability to bind cyclin-
dependent kinases (cdks) 4 or 6 and thereby activate a cas-
cade of events that permits progression through G1 phase
of the cell cycle ([30]; Fig. 3). Cyclin E also acts as a
positive regulator of cell cycle progression [31, 32]. By
binding and activating cdk 2, it regulates the G1 to S phase
transition. In contrast, p27Kip1 blocks cell cycle progression
by inactivating these same cdk cascades, and cells remain
in G1 phase [33]. In mice null for cyclin D2, granulosa cell
proliferation is impaired, the ovarian follicles remain small,
and ovulation fails to occur [24]. In mice null for p27Kip1,
follicular growth is not compromised but granulosa cells do
not luteinize properly in response to LH [25–27]. There-
fore, in order to better understand the control of cellular
proliferation in the ovary, we analyzed the expression of
these cell cycle regulatory molecules and their regulation
 REGULATION OF THE CELL CYCLE IN GRANULOSA CELLS 477

Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023

FIG. 1. Changes in granulosa cell proliferation in the ovary detected by BrdU labeling. Preovulatory follicles in mice 48 h after treatment with 4 IU
eCG have many proliferating granulosa cells, seen as cells with black nuclei. In response to the LH surge (5 IU hCG), these cells exit the cell cycle,
as seen by the absence of labeled granulosa cells in large preovulatory follicles. Methods as described previously [7]. 3200 (reproduced at 88%).

by hormones during follicular development, specifically
when granulosa cell proliferation is rapid and during lu-
teinization when cell division has terminated. The results
presented highlight the ability of the LH surge to acutely
regulate cyclin D2 and p27Kip1 in an inverse manner. Meth-
ods for the analysis of cyclin E protein done in the studies
presented herein were the same as previously described
[34].
REGULATION OF CYCLIN D2, CYCLIN E, AND P27Kip1
To characterize the hormonal regulation of cyclin D2,
cyclin E, and p27 in the ovary, we used the hypophysec-
tomized rat, in which the effects of estradiol, FSH, and LH
on granulosa cell proliferation [4–6] and differentiation [4,
5, 9, 11] have been well characterized. In situ hybridization
of ovarian sections from hormone-treated hypophysecto-
mized (H) rats showed that cyclin D2 and p27 exhibit dis-
tinctly regulated patterns of expression in granulosa cells
(Fig. 4). The results were verified by Western analysis of
protein obtained from granulosa cells or luteal cells of these
same animals. Specifically, cyclin D2 mRNA and protein
are expressed at high levels in granulosa cells of preovu-
latory follicles of H rats treated with estradiol and FSH
(HEF). When HEF rats are injected with hCG, cyclin D2
is down-regulated within 4 h and remains low throughout
luteinization. As shown by in situ localization, the down-
regulation of cyclin D2 mRNA occurs specifically in gran-
ulosa cells of preovulatory follicles that are destined to ovu-
late and undergo terminal differentiation or luteinization.
Cyclin D2 mRNA continues to be expressed in smaller
growing follicles that lack LH receptors [3, 4, 8].
The p27 cdk inhibitor is expressed in the preovulatory
granulosa cells of HEF rats, and like cyclin D2, is reduced
to low levels after 4 h of hCG treatment (Fig. 4). However,
after this time its expression pattern differs dramatically
from that of cyclin D2. By 24 h after hCG, p27 mRNA and

FIG. 2. Schematic of ovarian follicular

development and models that disrupt this
process. Granulosa cell proliferation leads
to follicular growth and the formation of
large preovulatory follicles. LH triggers
ovulation and the formation of corpora lu-
tea. Hypophysectomized rats, as well as
mice null for GnRH (hypogonadal), FSHb
subunit, and cyclin D2, exhibit impaired
follicular growth with follicles arrested at
the preantral stage of growth. Mice null
for ERa, but not ERb, have antral follicles
that do not ovulate or luteinize. Mice
lacking p27Kip1 exhibit abnormal corpus lu-
teum formation.
478 ROBKER AND RICHARDS

FIG. 3. Cyclin D2/cyclin E and p27Kip1 exert opposing actions on cdk

activity and cell cycle progression. Increased synthesis and binding of

Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023

cyclin D2 activate cdk4/6 activity. Subsequently, cyclin E binds and ac- FIG. 5. Regulation of cyclin E in granulosa cells. Western analysis of
tivates cdk2, enabling progression through G1 phase of the cell cycle. cyclin E expression in granulosa/luteal cells isolated from ovaries of hor-
Conversely, binding of p27KIP1 blocks cdk activity, and the cell cycle is mone-treated H rats.
arrested in G1.

the G1 checkpoint. Western analysis (Fig. 5) of whole cell

protein are expressed at very high levels and, as shown by
the in situ hybridization, are localized to the luteinizing
granulosa cells of the corpus luteum.
On the basis of the observations that both FSH and es-
tradiol induce cyclin D2 mRNA and protein expression in
granulosa cells [34], we sought to determine whether these
hormones also affect the expression of cyclin E, a down-
stream mediator critical for cell cycle progression through
extracts showed that levels of cyclin E were low in gran-
ulosa cells isolated from ovaries of H rats. Treatment in
vivo with FSH increased cyclin E levels within 2 h, after
which cyclin E remained elevated at this level. Interesting-
ly, treatment with estradiol caused a greater increase in cy-
clin E within 2 h than did FSH. Cyclin E levels then con-
tinued to increase progressively in the estradiol-treated rats
for 48 h. Additional experiments showed that cyclin E ex-

FIG. 4. Expression of cyclin D2 and p27Kip1 in the ovary. In situ hybridiza-

tion of rat ovarian sections shows that cy-
clin D2 mRNA is expressed at high levels
in the granulosa cells of preovulatory folli-
cles and subsequently down-regulated by
hCG treatment. Expression of p27 mRNA
is initially low following an hCG surge,
but then is elevated to high levels by 24 h.
Western analysis performed on protein
from granulosa cells and luteal cells isolat-
ed from ovaries of these same animals
shows similar results. Data are modified
from results presented in [34] with permis-
sion from The Endocrine Society. 3100
(reproduced at 92%).
 REGULATION OF THE CELL CYCLE IN GRANULOSA CELLS
pression is also regulated as cells luteinize in response to
an ovulatory stimulus of hCG. As in the previous experi-
ment, granulosa cells from untreated H rats contained low
levels of cyclin E that were increased in response to estra-
diol (HE) or estradiol followed by FSH (HEF). When HEF
rats were injected with an ovulatory dose of hCG, cyclin
E levels remained high at 4 h and 24 h; however, by 48 h
after hCG, cyclin E was low in the luteinized granulosa
cells.
Collectively, these data suggest that one putative mech-
anism by which the LH surge terminates granulosa cell pro-
liferation involves the rapid inhibition of cyclin D2 tran-
scription. As shown in Figure 1, granulosa cell exit from
the cell cycle occurs within 4 h of the LH surge, coinciding
with the drastic down-regulation of cyclin D2, but prior to
the down-regulation of cyclin E and the induction of p27.
Regulation of cyclin D2 is highly probable as the primary
regulatory event controlling granulosa cell proliferation,
since 1) cyclin D2, but not cyclin D1 or cyclin D3 (which
have redundant functions [30]), is expressed in granulosa
cells [34] and 2) the absence of cyclin D2, but not the
absence of cyclin D1 [35], markedly impairs granulosa cell
proliferation [24]. Additionally, cyclin E, a downstream
mediator of cell cycle progression, continues to be ex-
pressed in luteinizing granulosa cells long after the rapid
disappearance of cyclin D2. Therefore, the down-regulation
of cyclin D2 in response to LH would presumably prevent
the first step in cell cycle progression, thereby initiating
granulosa cell exit from the cell cycle before reaching the
cyclin E-regulated checkpoint. The temporal expression
pattern for p27 suggests that a second mechanism by which
LH terminates granulosa cell proliferation is by increasing
the level of this cdk inhibitor. In addition, the increase in
p27 may control some aspect of granulosa cell differenti-
ation or maintenance of luteal cell differentiation [25–27].
In summary, these data indicate that the LH surge termi-
nates granulosa cell proliferation and initiates differentia-
tion by inverting the balance of these positive and negative
regulators of cell cycle progression (Fig. 6). LH coordi-
nately down-regulates expression of cyclin D2, followed by
cyclin E, as it increases the levels of p27. The ovarian phe-
notypes of the mice lacking cyclin D2 or p27 support such
a model. The loss of cyclin D2 results in the absence of
FSH- and estradiol-stimulated granulosa cell proliferation
[24], which normally leads to large, preovulatory follicles.
The loss of p27 results in impaired differentiation as seen
by the inability of granulosa cells to luteinize normally and
produce sufficient progesterone to support pregnancy [25–
27].
MECHANISMS OF CELL CYCLE CONTROL
Regulation of the cell cycle within any cell type is com-
plex, involves the balance of many regulatory molecules,
and can be altered by numerous external signals acting at
multiple steps in the cycle. In the ovary, estradiol, FSH,
and LH are essential signals for the growth of preovulatory
follicles and their subsequent terminal differentiation as
corpora lutea. Each hormone acts via specific receptors and
intracellular signaling pathways. Additionally, FSH and LH
act by controlling distinct levels of cAMP [36] and the
activation of the A-kinase pathway [37]. The pivotal roles
of cyclin D2 and p27 in ovarian follicular growth and dif-
ferentiation are indicated by their selective expression and
regulation in the ovary and their critical and opposing ef-
fects on cdk activity that controls entry and progression
through G1 of the cell cycle. However, as briefly summa-
479
Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023
FIG. 6. The LH surge inverts the balance of positive versus negative cell
cycle regulators and triggers granulosa cell exit from the cell cycle con-
current with luteinization.
rized below (see reviews [17–23]), other regulatory mole-
cules control progression through additional checkpoints of
the cell cycle, and some of these are likely to be critical in
the ovary as well (Fig. 7).
Early in G1 phase, the presence of D-type cyclins acti-
vates cdk4 or cdk6, while the accumulation of cyclin E later
in G1 phase activates cdk2. Progression through S phase is
regulated by cyclin A complexes followed by the initiation
of M by cyclin B-cdc2 complexes. The binding of cyclin
D and cdk4/6 results in the formation of a complex that is
then phosphorylated by cdk-activating kinase (CAK). The
active cyclin-cdk complex in turn phosphorylates cellular
substrates that regulate DNA synthesis. The best-known ex-
ample is the retinoblastoma protein, Rb, which in a hypo-
phosphorylated state acts as a suppressor of cell division
by binding to DP/E2F transcription factors, thereby pre-
venting the transcription of genes necessary for replication
and cell division. The repression of transcription by Rb
appears to be mediated by its ability to recruit histone de-
acetylase [38, 39]. However, hyperphosphorylation of Rb
by cyclin D-cdk4/6 and cyclin E-cdk2 relieves its suppres-
sive abilities, and the cell cycle is able to progress past the
restriction point (R), from G1 to S phase, at which time the
cell is irreversibly committed to divide.
The Cip/Kip family of cdk inhibitors, which includes
p27, acts to block cell cycle progression by binding and
inhibiting the activity of cdks. The Cip/Kips (p21Cip1,
p27Kip1, p57Kip2) have relatively broad specificity and are
able to bind not only cdk4/6, but also cdk2 and cdc2, en-
abling them to inhibit the activity of several kinase cascades
and thereby block cell cycle progression at multiple points.
In contrast, the Ink4 family of cdk inhibitors bind only cdk4
and cdk6, making them specific inhibitors of G1 phase.
Even in this basic scheme it is clear that regulation of
the cell cycle is complex and occurs at many levels. Not
only are many different types of molecules involved, but
many are families of molecules that are expressed in over-
lapping as well as tissue-specific patterns. In the ovary, cy-
clin D2 is highly expressed specifically in granulosa cells,
as is cdk4 [40], while cyclin D1 and cyclin D3 are barely
detectable [34] and mice lacking cyclin D1 exhibit normal
ovarian function [35]. In contrast, cyclin D1 and cyclin D3
are expressed in an overlapping pattern, with both ex-
pressed at the highest levels in theca cells [34]. Both p27
and a related family member, p21Cip1, are highly expressed
in the corpus luteum with only slightly different patterns of
induction [34]. However, they appear to play unique roles
480 ROBKER AND RICHARDS
FIG. 7. Schematic of known cell cycle
events and factors regulated in ovarian
cells by gonadotropins and estradiol. Solid
arrows depict events shown herein to be
regulated by FSH, LH, and estrogen during
follicular growth and luteinization. Dashed
arrows show events likely to be controlled
by FSH/cAMP and estrogen on the basis of
studies described in the text. Other arrows
indicate the sequence of regulatory steps
in the cell cycle as summarized from nu-
merous reviews cited in the text. T-shaped
lines indicate inhibitory actions.
in this tissue, since mice lacking p27 exhibit an altered
ovarian phenotype [25–27] while mice lacking p21 do not
[41]. Rb is also a member of a family of molecules (in-
cluding p107 and p130), and there are five mammalian iso-
forms of E2F. Mice lacking p107 [42], p130 [43], or E2F-
1 [44, 45] are fertile; thus there may exist overlapping ex-
pression and functional redundancy within members of
these families in granulosa cells. Precedence for this comes
from the observation that phosphatase cdc25, which acti-
vates the cyclin B-cdc2 complex, is also expressed as three
isoforms that are differentially expressed within the ovary
[46].
Another layer of complexity is added to this scenario
when one considers the fact that hormonal control of ex-
pression and activity of cell cycle regulators not only is
achieved by different hormones, but also depends on the
dose of hormone. Our studies have shown that cAMP and
protein kinase A play a role in accelerating granulosa cell
proliferation as well as in terminating follicle growth and
initiating terminal differentiation. Specifically, the low lev-
els of cAMP generated in response to FSH induce high
levels of cyclin D2 [34] and also increase cyclin E (Fig. 5).
In contrast, the LH surge and high levels of cAMP rapidly
turn off expression of cyclin D2 followed by cyclin E. Con-
versely, the LH surge markedly increases levels of p27 as
well as p21 [34]. High levels of cAMP have been known
to terminate cell division in other cell types [47], and in
glial cells this is associated with the induction of p27 ex-
pression [48]. Additionally, the cAMP/protein kinase A
cascade controls the exit from mitosis by regulating the
degradation of cyclin B [49]. Thus, not only can the pres-
ence or level of hormone, i.e., FSH versus LH, alter the
amount of a regulatory molecule; it can also exert effects
at multiple points within the cell cycle machinery, at G1-S
and M-G1 (Fig. 7).
Estrogens are known to be potent mitogens and are often
associated with cancer. Indeed, most studies that have been
designed to determine the mechanism by which estrogens,
principally estradiol, regulate proliferation have been per-
Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023
formed in breast cancer cell lines, in which the cell cycle
machinery and the signals impinging on the cell cycle may
be abnormal. In these cell lines, estradiol is able to increase
the activity of cdk4 and cdk2 [50, 51] by inducing expres-
sion of cyclin D1 [50] as well as by decreasing the levels
of cdk inhibitors [51]. Similar mechanisms have also been
observed in the uterus, where estradiol induces cyclins D1,
D3, E, and A [52]. We have shown that estradiol induces
the expression of cyclin D2 and cyclin E in granulosa cells,
concurrent with a reduction in levels of p27 [34]. Simul-
taneous down-regulation of a cdk inhibitor and induction
of cyclins may account for the greater mitogenicity of es-
tradiol compared to FSH in these cells [5, 6]. Studies have
also shown that cyclin D1 can directly bind the estrogen
receptor and enhance transcription of specific genes [53].
It would be interesting to determine whether such a mech-
anism occurs in granulosa cells, which selectively express
the beta subtype of the estrogen receptor (ERb; [16]) and
in which estradiol induces cyclin D2 [34].
Lastly, there are many additional hormones that act in
the ovary to affect cellular proliferation and differentiation.
For instance, activin, which is produced at high levels in
preovulatory granulosa cells, has been shown to stimulate
granulosa cell DNA synthesis [54]. Therefore, it is possible
that activin along with estradiol and FSH regulates cyclin
D2. Paradoxically, activin has been shown to down-regulate
cyclin D2 in plasmacytic cells [55], and targeted deletion
of the activin bB subunit in mice did not result in an overt
ovarian phenotype [56]. Conversely, mice null for the het-
erodimeric molecule inhibin (a/bA or a/bB) develop ovar-
ian tumors at the time of puberty [57] that are dependent
on gonadotropin support [58]. These observations indicate
that the unopposed actions of activin in the presence of
cAMP or steroid are tumorigenic. Insulin-like growth fac-
tor-1 (IGF-1) has also been implicated in granulosa cell
proliferation [59]. However, large antral follicles are present
in the ovaries of mice null for IGF-1, indicating that IGF-
1 may be more important for differentiation than for pro-
liferation [60].
 REGULATION OF THE CELL CYCLE IN GRANULOSA CELLS
In summary, the relationship of cell proliferation to dif-
ferentiation is fundamental to all biological processes. Pro-
liferation precedes differentiation; differentiation often pre-
cludes further cell division (exceptions are metastasis—tis-
sue repair); nonproliferating and nondifferentiated cells are
usually excluded by apoptosis (programmed cell death, a
topic not covered herein). The ovarian phenotypes of the
cyclin D2 and p27 null mice provide intriguing insights into
the relationship between proliferation and differentiation of
follicular granulosa cells. In the absence of p27, differen-
tiation characteristic of luteinization appears impaired.
However, despite the key role of p27 in checking cell cycle
progression and its presence in granulosa cells and luteal
cells, the absence of p27 does not lead to rampant uncon-
trolled proliferation of these cells. Thus, other inhibitors of
cell division appear to be more critical during follicular
growth. Conversely, in the absence of cyclin D2, the mitotic
activity of granulosa cells is markedly impaired, and grow-
ing follicles remain small with few (usually only one or
two) layers of granulosa cells. Despite the reduced number
of cells, these cells respond to LH in a normal pattern of
differentiation. For instance, progesterone receptor and
prostaglandin synthase-2, two regulators of ovulation [9,
61, 62], are induced by the LH surge in a pattern similar
to that of normal ovulating follicles [34]. Yet the ‘‘pre-
ovulatory’’ follicles of the cyclin D2 mice do not ovulate.
This raises the intriguing question whether or not the num-
ber of granulosa cells is critical for stimulating some event
associated with ovulation. Are there other situations in
which cell number dictates some physiological process?
These and many other questions will be answered as we
learn more about the control of the cell cycle and cell dif-
ferentiation in the ovary and other tissues.
REFERENCES
1. Hirshfield AN. Development of follicles in the mammalian ovary. Int
Rev Cytol 1991; 124:43–101.
2. Pederson T. Follicular kinetics in the ovary of the cyclic mouse. Acta
Endocrinol 1970; 64:304–323.
3. Richards JS. Hormonal control of follicular growth and maturation in
mammals. In: Jones RE (ed.) The Vertebrate Ovary: Comparative Bi-
ology and Evolution. New York: Plenum Press; 1978: 331–360.
4. Richards JS. Maturation of ovarian follicles: actions and interactions
of pituitary and ovarian hormones on follicular cell differentiation.
Physiol Rev 1980; 60:51–89.
5. Richards JS. Estradiol receptor content in rat granulosa cells during
follicular development. Endocrinology 1975; 97:1174–1184.
6. Rao MC, Midgley AR Jr, Richards JS. Hormonal regulation of ovarian
cellular proliferation. Cell 1978; 14:71–78.
7. Gaytan F, Morales C, Bellido C, Aguilar E, Sanchez-Criado JE. Pro-
liferative activity in the different ovarian compartments in cycling rats
estimated by the 5-bromodeoxyuridine technique. Biol Reprod 1996;
54:1356–1365.
8. Uilenbroek JThJ, Richards JS. Ovarian follicular development during
the rat estrous cycle: gonadotropin receptors and follicular respon-
siveness. Biol Reprod 1979; 20:1159–1165.
9. Richards JS. Hormonal control of gene expression in the ovary. En-
docr Rev 1994; 15:725–751.
10. Richards JS, Hedin L, Caston L. Differentiation of rat ovarian cells:
evidence for functional luteinization. Endocrinology 1986; 118:1660–
1668.
11. Richards JS, Midgley AR Jr. Protein hormone action: a key to under-
standing ovarian follicular and luteal cell development. Biol Reprod
1976; 14:82–94.
12. Mason AJ, Hayflick JS, Zoeller RT, Young WS, Phillips HS, Nikolics
K, Seeburg PH. A deletion truncating the gonadotropin-releasing hor-
mone gene is responsible for hypogonadism in the hpg mouse. Science
1986; 234:1366–1371.
13. Mason AJ, Pitts SL, Nikolics K, Szonyi E, Wilcox JN, Seeburg PH,
Stewart TA. The hypogonadal mouse: reproductive functions restored
by gene therapy. Science 1986; 234:1372–1378.
481
14. Kumar TR, Wang Y, Lu N, Matzuk MM. Follicle stimulating hormone
is required for ovarian follicle maturation but not male fertility. Nat
Genet 1997; 15:201–204.
15. Lubahn DB, Moyer JS, Golding TS, Couse JF, Korach KS, Smithies
O. Alteration of reproductive function but not prenatal sexual devel-
opment after insertional disruption of the mouse estrogen receptor
gene. Proc Natl Acad Sci USA 1993; 90:11162–11166.
16. Byers M, Kuiper GGJM, Gustafsson J-A, Park-Sarge O-K. Estrogen
receptor-b mRNA expression in rat ovary: down-regulation by go-
nadotropins. Mol Endocrinol 1997; 11:172–182.
17. Sherr CJ. Cancer cell cycles. Science 1996; 274:1672–1677.
18. Sherr CJ. G1 progression: cycling on cue. Cell 1994; 79:551–555.
19. Sherr CJ, Roberts JM. Inhibitors of mammalian G1 cyclin-dependent
kinases. Genes Dev 1995; 9:1149–1163.
20. Hunter T, Pines J. Cyclins and cancer II: cyclin D and cdk inhibitors
come of age. Cell 1994; 79:573–582.
Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023
21. Elledge SJ. Cell cycle checkpoints: preventing an identity crisis. Sci-
ence 1996; 274:1664–1671.
22. Weinberg RA. The retinoblastoma protein and cell cycle control. Cell
1995; 81:323–330.
23. Weinberg RA. E2F and cell proliferation: a world turned upside down.
Cell 1996; 85:457–459.
24. Sicinski P, Donaher JL, Geng Y, Parker SB, Gardner H, Park MY,
Robker RL, Richards JS, McGinnis LK, Biggers JD, Eppig JJ, Bron-
son RT, Elledge SJ, Weinberg RA. Cyclin D2 is an FSH-responsive
gene involved in gonadal cell proliferation and oncogenesis. Nature
1996; 384:470–474.
25. Nakayama K, Ishida N, Shirane M, Inomata A, Inoue T, Shishido N,
Horii I, Loh DY, Nakayama K. Mice lacking p27Kip1 display increased
body size, multiple organ hyperplasia, retinal dysplasia, and pituitary
tumors. Cell 1996; 85:707–720.
26. Kiyokawa H, Kineman RD, Manova-Todorova KO, Soares VC, Hoff-
man ES, Ono M, Khanam D, Hayday AC, Frohman LA, Koff A.
Enhanced growth of mice lacking the cyclin-dependent kinase inhib-
itor function of p27Kip1. Cell 1996; 85:721–732.
27. Fero ML, Rivkin M, Tasch M, Porter P, Carow CE, Firpo E, Polyak
K, Tsai LH, Broudy V, Perlmutter RM, Kaushansky K, Roberts JM.
A syndrome of multiorgan hyperplasia with features of gigantism,
tumorigenesis, and female sterility in p27Kip1-deficient mice. Cell
1996; 85:733–744.
28. Inaba T, Matsushime H, Valentine M, Roussel MF, Sherr CJ, Look
AT. Genomic organization, chromosomal localization, and indepen-
dent expression of human cyclin D genes. Genomics 1992; 13:565–
574.
29. Xiong Y, Menninger J, Beach D, Ward DC. Molecular cloning and
chromosomal mapping of CCND genes encoding human D-type cy-
clins. Genomics 1992; 13:575–584.
30. Xiong Y, Zhang H, Beach D. D type cyclins associate with multiple
protein kinases and the DNA replication and repair factor PCNA. Cell
1992; 71:505–514.
31. Lew DJ, Vjekoslav D, Reed SI. Isolation of three novel human cyclins
by rescue of G1 cyclin (Cln) function in yeast. Cell 1991; 66:1197–
1206.
32. Koff A, Cross F, Fisher A, Schumacher J, Leguellec K, Philippe M,
Roberts JM. Human cyclin E, a new cyclin that interacts with two
members of the CDC2 gene family. Cell 1991; 66:1217–1228.
33. Polyak K, Lee M, Erdjument-Bromage H, Koff A, Roberts JM,
Tempst P, Massague J. Cloning of p27Kip1, a cyclin-dependent kinase
inhibitor and a potential mediator of extracellular antimitotic signals.
Cell 1994; 78:59–66.
34. Robker RL, Richards JS. Hormone-induced proliferation and differ-
entiation of granulosa cells: a coordinated balance of the cell cycle
regulators cyclin D2 and p27KIP1. Mol Endocrinol 1998; 12: 924–940.
35. Sicinski P, Donaher JL, Parker SB, Li T, Fazell A, Gardner H, Haslam
SZ, Bronson RT, Elledge SJ, Weinberg RA. Cyclin D1 provides a link
between development and oncogenesis in the retina and breast. Cell
1995; 82:621–630.
36. Jonassen JA, Richards JS. Granulosa cell desensitization: effects of
gonadotropins on antral and preantral follicles. Endocrinology 1980;
106:1786–1794.
37. Richards JS, Kirchick HJ. Changes in the content and phosphorylation
of cytosol proteins in luteinizing ovarian follicles and corpora lutea.
Biol Reprod 1984; 30:737–751.
38. Brehm A, Miska EA, McCance DJ, Reid JL, Bannister AJ, Kouzarides
T. Retinoblastoma protein recruits histone deacetylase to repress tran-
scription. Nature 1998; 391:597–601.
39. Magnaghi-Jaulin L, Groisman R, Naguibneva I, Robin P, Lorain S,
482 ROBKER AND RICHARDS
LeVillain JP, Troalen F, Trouche D, Harel-Bellan A. Retinoblastoma
protein represses transcription by recruiting a histone deacetylase. Na-
ture 1998; 391:601–605.
40. Rhee K, Wolgemuth DJ. Cdk family genes are expressed not only in
dividing but also in terminally differentiated mouse germ cells, sug-
gesting their possible function during both cell division and differ-
entiation. Dev Dynam 1995; 204:406–420.
41. Deng C, Zhang P, Harper JW, Elledge SJ, Leder P. Mice lacking
p21CIP1/WAF1 undergo normal development, but are defective in G1
checkpoint control. Cell 1995; 82:675–684.
42. Lee M-Y, Williams BO, Mulligan G, Mukai S, Bronson RT, Dyson
N, Harlow E, Jacks T. Targeted disruption of p107: functional overlap
between p107 and Rb. Genes Dev 1996; 10:1621–1632.
43. Cobrinik D, Lee M-Y, Hannon G, Mulligan G, Bronson RT, Dyson
N, Harlow E, Beach D, Weinberg RA, Jacks T. Shared role of the
pRB-related p130 and p107 proteins in limb development. Genes Dev
1996; 10:1633–1644.
44. Yamasaki L, Jacks T, Bronson R, Goillot E, Harlow E, Dyson NJ.
Tumor induction and tissue atrophy in mice lacking E2F-1. Cell 1996;
85:537–548.
45. Field SJ, Tsai F-Y, Kuo F, Zubianga AM, Kaelin WG Jr, Livingston
DM, Orkin SH, Greenberg ME. E2F-1 functions in mice to promote
apoptosis and suppress proliferation. Cell 1996; 85:549–561.
46. Wu S, Wolgemuth DJ. The distinct and developmentally regulated
patterns of expression of members of the mouse cdc25 gene family
suggests differential functions during gametogenesis. Dev Biol 1995;
170:195–206.
47. Kato J, Matsuoka M, Polyak K, Massague J, Sherr CJ. Cyclic AMP-
induced G1 phase arrest mediated by an inhibitor (p27Kip1) of cyclin-
dependent kinase 4 activation. Cell 1994; 79:487–496.
48. Tikoo R, Casaccia-Bonnefil P, Chao MV, Koff A. Changes in cyclin-
dependent kinase 2 and p27Kip1 accompany glial cell differentiation
of central glial-4 cells. J Biol Chem 1997; 272:442–447.
49. Grieco D, Porcellini A, Avvedimento EV, Gottesman ME. Require-
ment for cAMP-PKA pathway activation by M phase-promoting fac-
tor in the transition from mitosis to interphase. Science 1996; 271:
1718–1723.
50. Prall OWJ, Sarcevic B, Musgrove EA, Watts CKW, Sutherland RL.
Estrogen-induced activation of cdk4 and cdk2 during G1-S phase pro-
gression is accompanied by increased cyclin D1 expression and de-
creased cyclin-dependent kinase inhibitor association with cyclin E-
cdk2. J Biol Chem 1997; 272:10882–10894.
51. Foster JS, Wimalasena J. Estrogen regulates activity of cyclin-depen-
dent kinases and retinoblastoma protein phosphorylation in breast can-
cer cells. Mol Endocrinol 1996; 10:488–498.
52. Altucci L, Addeo R, Cicatiello L, Germano D, Pacilio C, Battista T,
Cancemi M, Petrizzi VB, Bresciani F, Weisz A. Estrogen induces early
and timed activation of cyclin-dependent kinases 4, 5, and 6 and in-
creases cyclin messenger ribonucleic acid expression in rat uterus.
Endocrinology 1997; 138:978–984.
53. Zwijsen RML, Wientjens E, Klompmaker R, van der Sman J, Ber-
nards R, Michalides RJAM. CDK-independent activation of estrogen
receptor by cyclin D1. Cell 1997; 88:405–415.
54. Miro F, Hillier SG. Modulation of granulosa cell deoxyribonucleic
acid synthesis and differentiation by activin. Endocrinology 1996;
137:464–468.
55. Yamato K, Koseki T, Ohguchi M, Kizaki M, Ikeda Y, Nishihara T.
Downloaded from https://academic.oup.com/biolreprod/article/59/3/476/2740757 by guest on 10 February 2023
Activin A induction of cell-cycle arrest involves modulation of cyclin
D2 and p21CIP1/WAF1 in plasmacytic cells. Mol Endocrinol 1997; 11:
1044–1052.
56. Vassali A, Matzuk MM, Gardner HAR, Lee KF, Jaenisch R. Activin/
inhibin bB subunit disruption leads to defects in eyelid development
and female reproduction. Genes Dev 1994; 8:414–427.
57. Matzuk MM, Finegold MJ, Su J-GJ, Hsueh AJW, Bradley A. a-In-
hibin is a tumour-suppressor gene with gonadal specificity in mice.
Nature 1992; 360:313–319.
58. Kumar TR, Wang Y, Matzuk MM. Gonadotropins are essential mod-
ifier factors for gonadal tumor development in inhibin-deficient mice.
Endocrinology 1996; 137:4210–4216.
59. Zhou J, Refuerzo J, Bondy C. Granulosa cell DNA synthesis is strictly
correlated with the presence of insulin-like growth factor I and ab-
sence of c-fos/c-jun expression. Mol Endocrinol 1995; 9:924–931.
60. Baker J, Hardy MP, Zhou J, Bondy C, Lupu F, Bellve AR, Efstratiadis
A. Effects of an Igf1 gene null mutation on mouse reproduction. Mol
Endocrinol 1996; 10:903–918.
61. Sirois J, Richards JS. Purification and characterization of a novel,
distinct isoform of prostaglandin endoperoxide synthase induced by
human chorionic gonadotropin in granulosa cells of rat preovulatory
follicles. J Biol Chem 1992; 267:6382–6388.
62. Park O-K, Mayo KE. Transient expression of progesterone receptor
messenger RNA in ovarian granulosa cells after the preovulatory lu-
teinizing hormone surge. Mol Endocrinol 1991; 5:967–978.