Reprod Dom Anim 43 (Suppl. 2), 224–231 (2008); doi: 10.1111/j.1439-0531.2008.01166.

x
ISSN 0936-6768

Follicle Development in Mares

FX Donadeu1 and HG Pedersen2
1
Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, UK; 2Veterinary Reproduction and Obstetrics,
Department of Large Animal Sciences, Faculty of Life Sciences, University of Copenhagen, Copenhagen, Denmark

Contents This review summarizes current knowledge on equine

The mare provides a unique experimental model for studying follicle development. Information on pre-antral and
follicle development in monovular species. Development of early antral follicles is particularly scarce in the horse;
antral follicles in horses is characterized by the periodic growth therefore, relatively more focus has been placed on the
of follicular waves which often involve the selection of a single late stages of antral follicle development up to the pre-
dominant follicle. If properly stimulated, the dominant follicle ovulatory stage. Details on physiological and practical
will complete development and eventually ovulate a fertile aspects of the ovulatory process and subsequent forma-
oocyte. Regulation of follicular wave emergence and follicle
tion of the corpus luteum can be found elsewhere
selection involves an interplay between circulating gonadotro-
pins and follicular factors that ensures that individual follicles (Ginther 1992; Boerboom and Sirois 2001; Squires 2006).
are properly stimulated to grow (or to regress) at any given
stage of follicular wave development. Periodic development of
follicular waves continuously occurs during most of post-natal Preantral Stages of Follicle Development
life in the mare and is influenced by factors such as stage of During early fetal life, primordial germ cells migrate to
oestrous cycle, season, pregnancy, age, breed and individual so the primitive gonadal ridge where they proliferate and
that different types of follicular waves (minor or major, subsequently enter meiotic division to become arrested
ovulatory or anovulatory) and different levels of activity in prophase I as primary oocytes (Ginther 1992).
within waves may develop under different physiological Meiotic arrest continues until atresia or until the
conditions. Changes in gonadotropin levels and ⁄ or in the
primary oocyte is stimulated by an LH surge during
sensitivity of follicles to circulating gonadotropins seem
to account largely for these physiological variations in follicle post-natal life. Based on histological studies of the
development. developing fetal horse ovary, meiotic divisions begin at
about day 70 of pregnancy (Deanesly 1975). Widespread
atresia characterizes the first oocytes to enter the meiotic
Introduction phase between days 73 and 150 of pregnancy with a
Although scientific interest in equine follicles existed by peak at approximately day 100. Development of oocytes
the 1920s (Seaborne 1925), detailed studies on equine into primordial follicles is not apparent until day 150
follicle dynamics did not begin until 50 years later under (Deanesly 1975) and the number of primordial follicles
the pioneering lead of OJ Ginther at the University of continues to increase during subsequent fetal life so that
Wisconsin (reviewed in Ginther 1979). During the early several thousand of these follicles are contained within
1980s, ultrasonography became available for equine the ovaries at birth (Ginther 1992). Proliferation of the
theriogenology (Palmer and Driancourt 1980). Trans- somatic cell layer surrounding the primary oocyte leads
rectal ultrasonography combined with systemic hor- to the sequential development of primordial follicles
mone measurements were subsequently used extensively into primary, secondary and, finally, antral follicles,
to understand equine follicle dynamics. The more recent a process that begins in the fetus and continues
introduction (mid-1990s) of the technique of ultrasound- throughout post-natal life. It is not known how follicles
guided transvaginal ovarian puncture provided scientists are selected to grow from the pool of primordial, resting
unprecedented access to the live equine ovary and follicles in the mare. Adult equine ovaries were reported
allowed the targeting of individual follicles for collection to contain approximately 36 000 primordial follicles and
of follicle samples, injection of test substances or 100 growing follicles at any one time, with large
experimental manipulation of follicles (Gastal et al. variability in actual follicle numbers between individual
1997; Gerard and Monget 1998; Donadeu and Ginther mares (Driancourt et al. 1982). Comparatively higher
2001; Martoriati et al. 2003). The experimental use of follicle numbers (threefold) were found in the adult
these procedures has led to extraordinary advances in bovine ovary. Proliferative activity of small pre-antral
the knowledge of equine ovarian physiology during the follicles in pony mares was higher at the beginning of the
last 15 years (reviewed in Beg and Ginther 2006; ovulatory season than during anoestrus or at the end of
Donadeu and Watson 2007). Two important outcomes the ovulatory season, whereas follicular atresia was
of this knowledge have been an improvement of repro- unaffected by season (Driancourt et al. 1983).
ductive efficiency in mares (Squires 2006) and a greater
understanding of follicular physiology in monovular
Antral Follicle Development and Follicular
species in general. In regard to the latter, consideration
of the mare as a useful model for studying ovarian
Waves
processes during human health and disease has recently The equine follicle develops an antrum when it reaches
increased (Ginther et al. 2004c; Carnevale 2008). approximately 0.3 mm in diameter (Driancourt et al.

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

Follicles and Mares 225

1982). Similar to pre-antral stages, little is known on the approximately 35–45 mm, the dominant follicle nor-
development of antral follicles before they reach mally either ovulates or ceases to grow and begins to
approximately 2 mm, the smallest diameter that can regress, depending on whether an ovulatory LH surge
usually be detected by transrectal ultrasonography. It occurs. The establishment of dominance is mediated by
has been reported that the growth of an equine follicle a differential increase in trophic support to the largest
from 0.1 to 1 mm takes approximately two oestrous follicle of a wave by mechanisms that will be explained
cycles (Driancourt 1979) and that atresia during in more detail in the next section. This leads to profound
this early phase of antral development is rare (Ginther changes in follicular cell function that are necessary for
1992). the eventual full maturation of the follicle to an
As in other farm species and humans, the develop- ovulatory-competent state and that are reflected in
ment of antral follicles in the horse is characterized by dramatic changes in global gene expression, changes
the periodic growth of cohorts of follicles or follicular that have begun to be characterized in other species,
waves (Sirois et al. 1989; Bergfelt and Ginther 1993). most notably cattle (Sisco et al. 2003; Fayad et al. 2004;
Characterization of follicular waves has involved the Mihm et al. 2008).
ultrasonic day-to-day identification of individual folli- Dominance does not seem to be a pre-determined
cles (Bergfelt and Ginther 1993; Gastal et al. 1997) and trait among the follicles growing in a wave because
the use of a statistical method that avoids the need to follicle ablation studies have demonstrated that all
maintain the identities of individual follicles during follicles have similar capacity to become dominant,
serial ultrasound examinations (Ginther and Bergfelt a capacity that in subordinate follicles is lost within
1992; Donadeu and Ginther 2002b). Follicular waves approximately 48 h after the beginning of deviation
in the horse can be identified in relation to follicles (Gastal et al. 2004). The same studies showed that in
2 mm in diameter and larger; however, it is not known approximately 61% of waves the first follicle to
whether earlier antral and pre-antral stages are also emerge at 6 mm maintains its size advantage over
characterized by wave-like patterns of growth, smaller follicles through the common growth phase
a question that has also not been clarified in other and becomes dominant. The likelihood of the largest
species (Mizunuma et al. 1999; McGee and Hsueh follicle becoming dominant increases as it approaches
2000). Follicular waves and their regulation with an the expected diameter at the beginning of deviation
emphasis on normal oestrous cycles will be described in (Gastal et al. 2004). In a few instances, yet, the largest
this and the next section. Follicular wave patterns follicle ceases or slows down its growth during the
characteristic of other reproductive states will be common growth phase and is replaced by the second
described in a separate section. largest follicle (or sometimes even a smaller follicle)
which then becomes the dominant follicle.
Characteristics of a follicular wave. Follicular wave
emergence has been normally defined for experimental Types of follicular waves. Follicular waves have been
purposes as occurring when the largest follicle reaches 6 classified as major waves (referred to in the literature
or 13 mm, depending on the study (Ginther et al. and throughout this review simply as follicular waves)
2003a). Identification of wave emergence often requires or minor waves, depending on whether they involve the
day-to-day ultrasonic evaluation of the ovary usually development of a readily identifiable dominant follicle
after aspiration of all follicles from previous waves. or they produce only smaller follicles, respectively
A follicular wave initially involves the simultaneous (Ginther 1993; Donadeu and Ginther 2002b). The
growth of a variable number of follicles at a common number of follicular waves during an oestrous cycle
rate of between 2 and 3 mm ⁄ day. A recent study varies between species. In the horse, as in humans, only
involving ablation of all follicles during the middle of one or two major follicular waves develop during each
an oestrous cycle in pony mares reported a mean of cycle (Ginther et al. 2004c). A major wave (named
approximately 12 follicles emerging during the common primary wave) always emerges during the middle of the
growth phase of the ablation-induced wave (Gastal equine oestrous cycle and produces the ovulatory
et al. 2004). Approximately two follicles emerged each follicle. Approximately 25% of interovulatory intervals
day during the first 4 days after wave emergence with a involve an additional major wave (secondary wave) that
progressive decrease in the numbers of follicles emerging develops during the first half of the cycle (Sirois et al.
thereafter. The exact number of follicles emerging within 1989; Bergfelt and Ginther 1993). The incidence of
waves is affected by factors such as season (Donadeu secondary waves is significantly higher in some breeds
and Ginther 2003). such as Thoroughbreds, and some of these waves may
The phase of common follicle growth is followed by produce ovulations. The ability to ovulate in the
the selection of a single follicle (occasionally two presence of high progesterone levels during dioestrus
follicles) which is manifested as a deviation in diameter seems to be unique to the horse (Ginther 1992). Minor
between the two largest follicles of the wave beginning waves (largest follicle usually <30 mm) have been
when the largest follicle reaches approximately 22 mm identified at different stages of the oestrous cycle and
(Ginther et al. 2003a). Deviation begins a mean of their incidence has been reported to be low, 25% or less
7 days before ovulation and is characterized by the of oestrous cycles (Bergfelt and Ginther 1993; Ginther
continuous growth (at an unchanged rate) of the largest 1993). Yet, a study considering follicles as small as
(selected) follicle as a dominant follicle and the 2 mm concluded that relatively high levels of underlying
simultaneous cease in growth and subsequent regression activity may involve the smallest follicles of a wave
of smaller, subordinate follicles. Once it grows to (Ginther and Bergfelt 1993).

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

226 FX Donadeu and HG Pedersen

Regulation of Follicular Wave Development season has revealed a direct temporal relationship
Emergence of a follicular wave and subsequent selection between an increase in circulating LH, but not FSH,
of a dominant follicle are under finely-tuned regulation and the development of major waves, further indicating
by systemic mechanisms involving changes in gonado- that an increase in LH plays a major role in stimulating
tropin levels and local mechanisms that involve changes the development of dominant follicles (reviewed in
in follicular factor levels. Donadeu and Watson 2007). An additional conclusion
was that circulating LH concentrations above those
Systemic regulation of follicular wave develop- required for growth of the dominant follicle are required
ment. Unlike pre-antral follicles, antral follicles cannot for the development of ovulatory competence, i.e. for
develop without adequate gonadotropin stimulation and the dominant follicle to become fully responsive to an
this has been experimentally shown in the mare LH surge (Donadeu and Watson 2007).
(Pedersen et al. 2002; Imboden et al. 2006). Studies in cattle have shown that LH receptor
Follicular waves in the mare, as in other species, are expression differentially increases in granulosa and theca
temporally preceded by a stimulatory surge in circulat- cells of the early dominant follicle (Bao and Garverick
ing FSH (Ginther et al. 2003a). The acute dependence of 1998; Beg and Ginther 2006). These findings are
follicular waves on FSH has been shown in mares by the consistent with those in other species (Richards 2001).
inability of follicles to grow beyond a diameter of 15 mm Although not critically examined in relation to the
after suppression of circulating FSH by systemic injec- beginning of deviation, higher LH receptor levels have
tion of follicular fluid (Bergfelt and Ginther 1985). Close been reported in dominant-size follicles than in smaller
functional relationships between FSH surges and follic- follicles in the horse (Fay and Douglas 1987; Goudet
ular waves involve not only positive effects of FSH on et al. 1999). Taken together, the findings on LH levels
follicles but also negative effects of follicles on FSH. The and LH receptor expression during a follicular wave in
wave-associated FSH surge reaches a peak when the mares are consistent with the conclusion that deviation
largest follicle is approximately 13 mm in diameter involves a critical increase in the dependence of the
(Gastal et al. 1997; Donadeu and Ginther 2001). The dominant follicle on LH.
following decline in FSH results from an increase in Limited data exist on the involvement of substances
circulating inhibin, presumably inhibin-A (Watson and other than gonadotropins in the endocrine regulation of
Al-zi’abi 2002), contributed mainly by the three largest antral follicles in mares. Based on observed direct effects
follicles of the wave as they grow above 13 mm on follicle growth or on the presence of specific receptors
(Donadeu and Ginther 2001). The declining FSH in equine ovaries, positive roles on follicle growth have
concentrations continue to support growth of the been suggested for circulating levels of substances
follicles of the wave until the largest (future dominant) including growth hormone (Cochran et al. 1999), dopa-
follicle reaches the expected diameter at the beginning of mine (King et al. 2005) and prolactin (Thompson et al.
deviation (approximately 22 mm). At that time, circu- 1997).
lating FSH levels become too low to maintain the Although follicular insulin-like growth factor-1
growth of the follicles of the wave. The low FSH, yet, (IGF-1) in mares largely derives from the systemic
does not restrict the growth of the future dominant circulation, its bioactivity is regulated mostly through
follicle which by that time has acquired the ability to local mechanisms (Watson et al. 2004) and the role of
more efficiently use circulating gonadotropins for IGF-1 is therefore considered in the next section.
growth (Ginther et al. 2003a). The differential responses
of the largest and smaller follicles of a wave to Local regulation of follicular wave development. Local
gonadotropins result in the initiation of diameter regulation of follicle development in monovular species
deviation. Continuous suppression of FSH throughout has been studied extensively in relation to follicle
deviation ensures the morphological and functional selection (reviewed in Fortune et al. 2004; Beg and
demise of subordinate follicles and is attributable to Ginther 2006; Knight and Glister 2006). A variety of
production of high levels of inhibin and oestradiol by protein and steroid factors including members of the
the dominant follicle (Gastal et al. 1999; Donadeu and IGF family, oestradiol, inhibins and activins, follistatin
Ginther 2001). The critical role of low FSH levels in the and vascular endothelial growth factor (VEGF) are
deviation mechanism in mares is illustrated by the involved. In general, these factors act, often in a
disruption of the deviation mechanism after adminis- paracrine manner, to either enhance or diminish the
tration of FSH (Squires 2006) or immunization against trophic effects of gonadotropins on follicular cells
inhibin (McCue et al. 1992) leading to the development through a variety of mechanisms. Differential changes
of multiple ovulatory follicles. in the levels of specific factors between follicles thus
While FSH is particularly important for follicular ensure the continuous development of the dominant
growth before deviation, LH becomes more critical follicle and the regression of subordinate follicles during
during deviation. This has been demonstrated by studies deviation.
showing that experimental suppression of LH in cycling A differential increase in the levels of oestradiol, IGF-1,
mares leads to the regression of the dominant follicle activin-A and inhibin-A in the future dominant follicle
early during its development (Bergfelt et al. 2001), and is was associated with the beginning of deviation in mares,
consistent with an increase in circulating LH before the whereas differentially elevated levels of progesterone
beginning of deviation (Bergfelt et al. 2001). The study and inhibin-B occurred later during the development of
of LH–follicle interrelationships during the anovulatory the dominant follicle (Donadeu and Ginther 2002a).

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

Follicles and Mares 227

Studies involving intrafollicular factor injection in mares Studies in pony mares using follicle ablation to facilitate
have provided insight into the complex interrelation- the identification of individual follicular waves revealed
ships between these factors during follicle deviation that, as during the ovulatory season, follicular waves
(Ginther et al. 2005). It has been concluded that periodically occur during the anovulatory season despite
although all these factors are likely involved in the the reduced levels of follicle development (Donadeu and
development of the dominant follicle after the beginning Ginther 2002b, 2003; Ginther et al. 2003b). Only minor
of deviation, only IGF-1 is involved in the initiation of waves (largest follicle <21 mm) occurred during the
deviation by, among other actions, regulating the levels middle of the anovulatory season or deep anoestrus,
of other growth factors in the dominant follicle (Beg and whereas major waves were often detected during the first
Ginther 2006). This is consistent with the particularly 2 months (fall transition) and the last 2 months (spring
important role of the IGF-1 system in follicle selection transition) of the anovulatory season, in agreement with
in other species (Mazerbourg et al. 2003). Results from previous studies (Ginther 1990; Watson et al. 2002). The
a series of in vivo experiments have convincingly onset of the spring transitional period involved an
confirmed the critical role of the IGF system in follicle abrupt increase in follicular activity from the low
selection in mares. Injection of IGF-1 into the second activity typical of deep anoestrus which was reflected
largest or a smaller follicle at the beginning of deviation in a distinct increase in the diameter of follicles and,
changed its fate from subordinate to co-dominant most notably, a fourfold increase in the numbers of
resulting in the development of multiple ovulatory follicles >12 mm within waves (means of 3.2 and 11.5
follicles (Ginther et al. 2004a; b; Gastal et al. 2007). follicles in waves developing during deep anoestrous and
Conversely, injection of IGF binding protein (IGFBP)-3 the early spring transition, respectively). Although
into the largest follicle at the beginning of deviation dominant follicles during transition may not grow to
resulted in the follicle regressing and being replaced by the diameters typical of ovulatory follicles, in the same
the second largest follicle which became dominant study transitional waves produced more follicles than
(Ginther et al. 2004a). At least four types of IGFBPs waves developing during the ovulatory season (means of
(IGFBP-2, 3, 4 and 5), which negatively regulate IGF 11.5 and 6.0 follicles >12 mm, respectively) attesting to
activity, have been identified in equine follicles and the the high levels of follicular activity even in the absence
concentrations of three of these (IGFBP-2, 4 and 5) are of ovulation in transitional mares (Donadeu and Gin-
correlated negatively with those of IGF-1 during follicle ther 2003).
development (Gerard and Monget 1998; Bridges et al. Based on consistent temporal relationships between
2002). Consistent with the stimulatory role of IGF-1 in follicles and circulating hormones, it has been concluded
the development of ovulatory follicles, reduced levels of that the differences in follicle development between
bioactive IGF-1 are thought to be involved in the different periods of the anovulatory season as well as the
developmental deficiencies of dominant follicles during deficient development of dominant follicles during the
the spring transition that prevent them from acquiring spring transition relative to the ovulatoy season are not
ovulatory competence (Acosta et al. 2004; Watson et al. attributable to deficient circulating FSH levels but
2004). rather to changes in LH and, possibly, differences in
An additional factor that has begun to be explored in follicular sensitivity to gonadotropins (reviewed in
relation to follicle selection in horses is VEGF. This Donadeu and Watson 2007).
angiogenic factor has been shown to be necessary for Season-related effects on follicular activity have also
follicle development in other species (Fraser and Wulff been reported between the two halves of the ovulatory
2001). VEGF levels differentially increase in the dom- season, with higher levels of activity during the first half
inant follicle in horses (Ginther et al. 2004b), and this of the season due to higher incidence of both secondary
increase is thought to be mediated, at least partly, by waves and minor waves associated with higher gonado-
IGF-1. VEGF is likely involved in the reported increase tropin levels (Ginther 1992, 1993).
in vascularization of the future dominant follicle before
the beginning of deviation (Acosta et al. 2004) which Effects of pregnancy and parturition. Considerable
presumably increases the availability of circulating research work is needed to better characterize follicular
gonadotropins to the follicle. The reduced levels of dynamics and associated regulatory mechanisms during
follicular VEGF and low vascularity of the wall of and following pregnancy in the mare. Based on com-
dominant follicles during the spring transition relative to bined data from ultrasound and rectal palpation studies,
the ovulatory season (Watson and Al-zi’abi 2002) follicular dynamics during the first half of pregnancy
underscore the critical role of VEGF in the development are similar to those occurring during the first half of
of the ovulatory follicle in horses. the anovulatory season, with an initial period of variable
activity (between days 11 and 40 of pregnancy) charac-
terized by the periodic development of major waves or,
Effects of Different Physiological Conditions on more commonly, development of sporadic major waves
Follicle Development in Mares or only minor waves (Ginther and Bergfelt 1992),
followed by a pronounced decrease in follicular activity
Effects of season. The effects of season on follicular in all mares between days 50 and 140 of pregnancy so
activity in mares have been studied in considerable that the diameter of the largest follicle does not exceed
detail. Important variations in follicular activity occur 15 mm by day 140 (Squires et al. 1974). A decrease in
not only between the ovulatory and anovulatory seasons follicular activity has also been reported after the first
but also between different periods within each season. one-third of pregnancy in cattle (Ginther et al. 1996).

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

228 FX Donadeu and HG Pedersen

Based on reported hormone–follicle associations, defi- (minor) follicular waves. Remarkably, these waves were
cient FSH levels do not seem to be responsible for the not temporally associated with statistically significant
reduction in follicular growth in some mares between circulating FSH surges, an observation that warrants
days 11 and 40 of pregnancy (Ginther and Bergfelt further investigation.
1992), an effect that may instead be accounted for by
reduced levels of LH (due to persistent progesterone Effects of aging on follicular activity. As highlighted in a
negative feedback), similar to the effects of the seasonal recent review, many age-related changes in follicular
reduction in circulating LH on follicle growth during the activity in the horse resemble those occurring in humans
fall transitional period (Ginther et al. 2003b). (Carnevale 2008). Follicular activity during oestrous
The mechanisms responsible for the dramatic reduc- cycles begins to decrease in mares 20 years of age or
tion in follicular growth during mid and late pregnancy older eventually leading to a cease in ovarian activity
in mares have not been clarified but likely involves a (Carnevale et al. 1993, 1994). Interovulatory intervals
temporally associated decrease in circulating FSH first become longer in these mares due to longer
(reviewed in Ginther 1992). This is different from the follicular phases associated with a primary follicular
anovulatory season during which changes in FSH levels wave that emerges later and contains less follicles. In
do not seem to play a major role in the decrease in addition, the ovulatory LH surge is less pronounced in
follicle growth during deep anoestrus (Donadeu and the older mares, and there is a higher incidence of
Watson 2007). Reduced follicle numbers during mid- ultrasonically atypical ovulations characterized by a
pregnancy are also likely attributable to ovulation or central hypoechogenic area at the ovulatory site. The
luteinization of follicles into accessory corpora lutea reduction in follicular activity and frequency of ovula-
under the influence of chorionic gonadotropin (Ginther tion in mares ‡20 years old is associated with an overall
1992). Further complexity into the regulation of follicle elevation in concentrations of FSH and LH during the
growth during pregnancy is provided by the observation follicular phase, a phenomenon that also occurs during
that the effects of season on hormones and follicles the peri-menopausal period in women. This is eventually
continue to occur during pregnancy (Ginther 1992). associated with persistent ovarian inactivity with folli-
The natural pressure to produce a foal each year in a cles <5 mm and no ovulations in the presence of
species with an 11-month-long pregnancy is reflected in continuously elevated concentrations of FSH and LH.
an early post-partum ovulation in the mare (foal heat, Reportedly, approximately 40% of horses 24 years or
typically within 2 weeks of parturition). A steady older have no ovarian activity (Vanderwall 1990)
increase in the diameter of the largest follicle and whereas 50% of ponies 20 years or older have irregular
in the numbers of follicles after parturition resulting or absent ovulations (Carnevale et al. 1994). It has been
in ovulation 14 days later was recently shown in suggested that an age-related elevation in FSH concen-
Arabian mares (Gunduz et al. 2008). The post-partum trations may hasten the eventual depletion of the
increase in follicular growth is induced by an increase in primordial follicle pool in older mares, similar to what
gonadotropin secretion at the time of parturition (Hines occurs in women (Carnevale 2008). Age-related differ-
et al. 1987; Ginther et al. 1994). Studies have shown that ences in follicular activity have also been reported in
the follicular response to parturition may vary in mares during the period of transition into the ovulatory
individual horses or different types of horses, and may season during which progressively lower follicle diam-
not readily occur in primiparous mares (Nagy et al. eters and numbers of follicles were found in mares 3–7,
1998), in the presence of a nursing foal (Ginther 1992) or 17–19 and ‡20 years old (Carnevale et al. 1997).
during the winter, when the negative effects of season
may prevail over the positive effects of parturition
(Ginther et al. 1994). Conclusions
The possibility of experimentally targeting equine folli-
Follicle development before puberty. Follicle develop- cles at very precise stages of development in vivo using
ment in spring-born pre-puberal pony fillies has recently a relatively noninvasive ultrasound-guided technique
been studied in detail (Nogueira and Ginther 2004), makes the horse a very attractive model to study in
adding to limited information on follicle profiles from detail the dynamics of antral follicles. In addition, the
earlier studies (reviewed in Ginther 1992). Follicular seasonal nature of ovarian activity in mares offers a
activity during 2–10 months of life was characterized by unique model to study the factors naturally regulating
a progressive increase in mean follicle diameter (from follicle development in a monovular species.
approximately 6 to 10 mm) and mean follicle numbers As in other monovular species, the development of
(from 3 to 17 follicles) between 2 and 5 months of age, antral follicles in horses is characterized by the periodic
a short plateau in activity coinciding with the winter emergence of follicular waves and the selection of
months and a re-initiation of follicle growth after usually a single dominant follicle that under the appro-
7 months of age leading to the onset of the first priate hormonal milieu will ovulate a fertile oocyte.
ovulatory season in spring (Nogueira and Ginther Follicular wave emergence and follicle selection are
2004). Changes in follicle activity during the first year driven by systemic changes in levels of FSH (wave
of life were positively correlated with changes in emergence and follicle selection) and LH (follicle selec-
circulating gonadotropins, consistent with a regulatory tion), and by local changes in follicular factors that,
role of season on gonadotropin and follicular activity through a variety of mechanisms, are responsible for
beginning early during life in mares. Follicular growth fine-tuning the response of follicular cells to gonadotro-
before puberty was characterized by the development of pin stimuli to ensure the continuous growth of the

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

Follicles and Mares 229

dominant follicles and the simultaneous regression of Cochran RA, Leonardi-Cattolica AA, Sullivan MR, Kincaid
subordinate follicles during follicle deviation. LA, Leise BS, Thompson DL Jr, Godke RA, 1999: The
Follicular waves occur throughout post-natal life in effects of equine somatotropin (eST) on follicular develop-
the horse until follicles irreversibly cease to grow at ment and circulating plasma hormone profiles in cyclic
mares treated during different stages of the estrous cycle.
approximately ‡ 20 years of age. The levels of follicular
Domest Anim Endocrinol 16, 57–67.
activity are affected by factors such as stage of the Deanesly R, 1975: Germ cell development and the meiotic
oestrous cycle, season, pregnancy, age, breed and prophase in the fetal horse ovary. J Reprod Fertil Suppl 23,
individual. This results in variations in follicular wave 547–552.
patterns that most notably include the production or Donadeu FX, Ginther OJ, 2001: Effect of number and
absence of a dominant follicle (major or minor waves) diameter of follicles on plasma concentrations of inhibin
and the development of ovulatory-competent or ovula- and FSH in mares. Reproduction 121, 897–903.
tory-incompetent dominant follicles. Available informa- Donadeu FX, Ginther OJ, 2002a: Changes in Concentrations
tion indicates that these physiological variations in of follicular fluid factors during follicle selection in mares.
follicular wave patterns are driven by changes in Biol Reprod 66, 1111–1118.
Donadeu FX, Ginther OJ, 2002b: Follicular waves and
circulating levels of FSH and ⁄ or LH and by local
circulating concentrations of gonadotrophins, inhibin and
changes in responsiveness of follicles to gonadotropins. oestradiol during the anovulatory season in mares. Repro-
Yet, considerable work is needed to better understand duction 124, 875–885.
the mechanisms by which follicle development is regu- Donadeu FX, Ginther OJ, 2003: Interactions of follicular
lated during different physiological conditions outside factors and season in the regulation of circulating concen-
the oestrous cycle in mares. trations of gonadotrophins in mares. Reproduction 125,
743–750.
Donadeu FX, Watson ED, 2007: Seasonal changes in ovarian
References activity: lessons learnt from the horse. Anim Reprod Sci 100,
Acosta TJ, Beg MA, Ginther OJ, 2004: Aberrant blood flow 225–242.
area and plasma gonadotropin concentrations during the Driancourt MA, 1979: Follicular kinetics in the mare ovary.
development of dominant-sized transitional anovulatory Ann Biol Anim Biochim Biophys 19, 1443–1453.
follicles in mares. Biol Reprod 71, 637–642. Driancourt MA, Paris A, Roux C, Mariana JC, Palmer E,
Bao B, Garverick HA, 1998: Expression of steroidogenic 1982: Ovarian follicular populations in pony and saddle-
enzyme and gonadotropin receptor genes in bovine follicles type mares. Reprod Nutr Dev 22, 1035–1047.
during ovarian follicular waves: a review. J Anim Sci 76, Driancourt MA, Prunier A, Palmer E, Mariana J, 1983:
1903–1921. Seasonal effects on ovarian follicular development in pony
Beg MA, Ginther OJ, 2006: Follicle selection in cattle and mares. Reprod Nutr Dev 23, 207–215.
horses: role of intrafollicular factors. Reproduction 132, Fay JE, Douglas RH, 1987: Changes in thecal and granulosa
365–377. cell LH and FSH receptor content associated with follicular
Bergfelt DR, Ginther OJ, 1985: Delayed follicular develop- fluid and peripheral plasma gonadotrophin and steroid
ment and ovulation following inhibition of FSH with equine hormone concentrations in preovulatory follicles of mares. J
follicular fluid in the mare. Theriogenology 24, 99–108. Reprod Fertil Suppl 35, 169–181.
Bergfelt DR, Ginther OJ, 1993: Relationships between FSH Fayad T, Levesque V, Sirois J, Silversides DW, Lussier JG, 2004:
surges and follicular waves during the estrous cycle in mares. Gene expression profiling of differentially expressed genes in
Theriogenology 39, 781–796. granulosa cells of bovine dominant follicles using suppression
Bergfelt DR, Gastal EL, Ginther OJ, 2001: Response of subtractive hybridization. Biol Reprod 70, 523–533.
estradiol and inhibin to experimentally reduced luteinizing Fortune JE, Rivera GM, Yang MY, 2004: Follicular develop-
hormone during follicle deviation in mares. Biol Reprod 65, ment: the role of the follicular microenvironment in selection
426–432. of the dominant follicle. Anim Reprod Sci 82–83, 109–126.
Boerboom D, Sirois J, 2001: Equine P450 cholesterol side-chain Fraser HM, Wulff C, 2001: Angiogenesis in the primate ovary.
cleavage and 3 beta-hydroxysteroid dehydrogenase ⁄ delta(5)- Reprod Fertil Dev 13, 557–566.
delta(4) isomerase: molecular cloning and regulation of their Gastal EL, Gastal MO, Bergfelt DR, Ginther OJ, 1997: Role
messenger ribonucleic acids in equine follicles during the of diameter differences among follicles in selection of a
ovulatory process. Biol Reprod 64, 206–215. future dominant follicle in mares. Biol Reprod 57, 1320–
Bridges TS, Davidson TR, Chamberlain CS, Geisert RD, 1327.
Spicer LJ, 2002: Changes in follicular fluid steroids, insulin- Gastal EL, Donadeu FX, Gastal MO, Ginther OJ, 1999:
like growth factors (IGF) and IGF-binding protein concen- Echotextural changes in the follicular wall during follicle
tration, and proteolytic activity during equine follicular deviation in mares. Theriogenology 52, 803–814.
development. J Anim Sci 80, 179–190. Gastal EL, Gastal MO, Beg MA, Ginther OJ, 2004: Interre-
Carnevale EM, 2008: The mare model for follicular matura- lationships among follicles during the common-growth
tion and reproductive aging in the woman. Theriogenology phase of a follicular wave and capacity of individual follicles
69, 23–30. for dominance in mares. Reproduction 128, 417–422.
Carnevale EM, Bergfelt DR, Ginther OJ, 1993: Aging effects Gastal EL, Gastal MO, Donadeu FX, Acosta TJ, Beg MA,
on follicular activity and concentrations of FSH, LH, and Ginther OJ, 2007: Temporal relationships among LH,
progesterone in mares. Anim Reprod Sci 31, 287–299. estradiol, and follicle vascularization preceding the first
Carnevale EM, Bergfelt DR, Ginther OJ, 1994: Follicular compared with later ovulations during the year in mares.
activity and concentrations of FSH and LH associated with Anim Reprod Sci 102, 314–321.
senescence in mares. Anim Reprod Sci 35, 231–246. Gerard N, Monget P, 1998: Intrafollicular insulin-like growth
Carnevale EM, Hermenet MJ, Ginther OJ, 1997: Age and factor-binding protein levels in equine ovarian follicles
pasture effects on vernal transition in mares. Theriogenology during preovulatory maturation and regression. Biol
47, 1009–1018. Reprod 58, 1508–1514.

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

230 FX Donadeu and HG Pedersen

Ginther OJ, 1979: Reproductive Biology of the Mare: Basic Knight PG, Glister C, 2006: TGF-{beta} superfamily members
and Applied Aspects, 1st edn. Equiservices Cross Plains, and ovarian follicle development. Reproduction 132, 191–
WI, USA. 206.
Ginther OJ, 1990: Folliculogenesis during the transitional Martoriati A, Duchamp G, Gerard N, 2003: In vivo effect of
period and early ovulatory season in mares. J Reprod Fertil epidermal growth factor, interleukin-1{beta}, and interleu-
90, 311–320. kin-1RA on equine preovulatory follicles. Biol Reprod 68,
Ginther OJ, 1992: Reproductive Biology of the Mare: Basic 1748–1754.
and Applied Aspects, 2nd edn. Equiservices Publishing Mazerbourg S, Bondy CA, Zhou J, Monget P, 2003: The
Cross Plains, WI, USA. insulin-like growth factor system: a key determinant role in
Ginther OJ, 1993: Major and minor follicular waves during the the growth and selection of ovarian follicles? A comparative
equine estrous cycle. J Eq Vet Sci 13, 18–25. species study. Reprod Domest Anim 38, 247–258.
Ginther OJ, Bergfelt DR, 1992: Associations between FSH McCue PM, Carney NJ, Hughes JP, Rivier J, Vale W, Lasley
concentrations and major and minor follicular waves in BL, 1992: Ovulation and embryo recovery rates following
pregnant mares. Theriogenology 38, 807–821. immunization of mares against an inhibin alpha-subunit
Ginther OJ, Bergfelt DR, 1993: Growth of small follicles and fragment. Theriogenology 38, 823–831.
concentrations of FSH during the equine oestrous cycle. J McGee EA, Hsueh AJW, 2000: Initial and cyclic recruitment
Reprod Fert 99, 105–111. of ovarian follicles. Endocrine Rev 21, 200–214.
Ginther OJ, Baucus KL, Bergfelt DR, 1994: Follicular and Mihm M, Baker PJ, Fleming LM, Monteiro AM,
FSH responses to parturition during the anovulatory season O’Shaughnessy PJ, 2008: Differentiation of the bovine
in mares. Theriogenology 41, 613–627. dominant follicle from the cohort upregulates mRNA
Ginther OJ, Kot K, Kulick LJ, Martin S, Wiltbank MC, 1996: expression for new tissue development genes. Reproduction
Relationships between FSH and ovarian follicular waves 135, 253–265.
during the last six months of pregnancy in cattle. J Reprod Mizunuma H, Liu X, Andoh K, Abe Y, Kobayashi J, Yamada
Fert 108, 271–279. K, Yokota H, Ibuki Y, Hasegawa Y, 1999: Activin from
Ginther OJ, Beg MA, Donadeu FX, Bergfelt DR, 2003a: secondary follicles causes small preantral follicles to
Mechanism of follicle deviation in monovular farm species. remain dormant at the resting stage. Endocrinology 140,
Anim Reprod Sci 78, 239–257. 37–42.
Ginther OJ, Woods BG, Meira C, Beg MA, Bergfelt DR, Nagy P, Huszenicza G, Juhasz J, Kulcsar M, Solti L, Reiczigel J,
2003b: Hormonal mechanism of follicle deviation as indi- Abavary K, 1998: Factors influencing ovarian activity and
cated by major versus minor follicular waves during the sexual behavior of postpartum mares under farm conditions.
transition into the anovulatory season in mares. Reproduc- Theriogenology 50, 1109–1119.
tion 126, 653–660. Nogueira GP, Ginther OJ, 2004: Dynamics of follicle popu-
Ginther OJ, Bergfelt DR, Beg MA, Meira C, Kot K, 2004a: lations and gonadotropin concentrations in fillies age two to
In vivo effects of an intrafollicular injection of insulin-like ten months. Equine Vet J 32, 482–488.
growth factor 1 on the mechanism of follicle deviation in Palmer E, Driancourt MA, 1980: Use of ultrasonic echography
heifers and mares. Biol Reprod 70, 99–105. in equine gynecology. Theriogenology 13, 203–216.
Ginther OJ, Gastal EL, Gastal MO, Checura CM, Beg MA, Pedersen HG, Berrocal B, Thomson SRM, Watson ED, 2002:
2004b: Dose–response study of intrafollicular injection of Gonadotropic control of follicular growth in the mare.
insulin-like growth factor-I on follicular fluid factors Theriogenology 58, 465–468.
and follicle dominance in mares. Biol Reprod 70, 1063– Richards JS, 2001: Perspective: the ovarian follicle –
1069. a perspective in 2001. Endocrinology 142, 2184–2193.
Ginther OJ, Gastal EL, Gastal MO, Bergfelt DR, Baerwald Seaborne E, 1925: The oestrous cycle of the mare and some
AR, Pierson RA, 2004c: Comparative study of the dynamics associated phenomena. Anat Rec 30, 277–286.
of follicular waves in mares and women. Biol Reprod 71, Sirois J, Ball BA, Fortune JE, 1989: Patterns of growth and
1195–1201. regression of ovarian follicles during the oestrous cycle and
Ginther OJ, Gastal EL, Gastal MO, Beg MA, 2005: In vivo after hemiovariectomy in mares. Equine Vet J 8(Suppl.), 43–
effects of pregnancy-associated plasma protein-A, activin-A 48.
and vascular endothelial growth factor on other follicular- Sisco B, Hagemann LJ, Shelling AN, Pfeffer PL, 2003:
fluid factors during follicle deviation in mares. Reproduc- Isolation of genes differentially expressed in dominant and
tion 129, 489–496. subordinate bovine follicles. Endocrinology 144, 3904–3913.
Goudet G, Belin F, Bezard J, Gerard N, 1999: Intrafollicular Squires EL, 2006: Superovulation in mares. Veter Clin N Am:
content of luteinizing hormone receptor, {alpha}-inhibin, Eq Pract 22, 819–830.
and aromatase in relation to follicular growth, estrous cycle Squires EL, Garcia MC, Ginther OJ, 1974: Effects of
stage, and oocyte competence for in vitro maturation in the pregnancy and hysterectomy on the ovaries of pony. J
mare. Biol Reprod 60, 1120–1127. Anim Sci 38, 823–830.
Gunduz MC, Kasikci G, Ekiz B, 2008: Follicular and steroid Thompson DL Jr, Hoffman R, DePew CL, 1997: Prolactin
hormone changes in Arabian mares in the postpartum administration to seasonally anestrous mares: reproductive,
period. Anim Reprod Sci (in press). metabolic, and hair-shedding responses. J Anim Sci 75,
Hines KK, Fitzgerald BP, Loy RG, 1987: Effect of pulsatile 1092–1099.
gonadotrophin release on mean serum LH and FSH Vanderwall DKW, GL, 1990: Age-related subfertility in the
in peri-parturient mares. J Reprod Fertil Suppl 35, 635– mare. In: Proc 35th Annual Conv American Assoc Equine
640. Practitioners, 85–89.
Imboden I, Janett F, Burger D, Crowe MA, Hassig M, Thun R, Watson ED, Al-zi’abi MO, 2002: Characterization of mor-
2006: Influence of immunization against GnRH on repro- phology and angiogenesis in follicles of mares during spring
ductive cyclicity and estrous behavior in the mare. Theriog- transition and the breeding season. Reproduction 124, 227–
enology 66, 1866–1875. 234.
King SS, Campbell AG, Dille EA, Roser JF, Murphy LL, Watson ED, Heald M, Tsigos A, Leask R, Steele M, Groome
Jones KL, 2005: Dopamine receptors in equine ovarian NP, Riley SC, 2002: Plasma FSH, inhibin A and inhibin
tissues. Domest Anim Endocrinol 28, 405–415. isoforms containing pro- and -alphaC during winter anoes-

 2008 The Authors. Journal compilation  2008 Blackwell Verlag

Follicles and Mares 231

trus, spring transition and the breeding season in mares. Author’s address (for correspondence): FX Donadeu, Roslin Institute,
Reproduction 123, 535–542. Royal (Dick) School of Veterinary Studies, University of Edinburgh,
Watson ED, Bae SE, Thomassen R, Thomson SR, Woad K, Edinburgh, UK. E-mail: [email protected]
Armstrong DG, 2004: Insulin-like growth factors-I and -II
and insulin-like growth factor-binding protein-2 in domi- Conflict of interest: All authors declare no conflict of interests.
nant equine follicles during spring transition and the
ovulatory season. Reproduction 128, 321–329.

 2008 The Authors. Journal compilation  2008 Blackwell Verlag