Reprod Dom Anim 43 (Suppl. 2), 224–231 (2008); doi: 10.1111/j.1439-0531.2008.01166.
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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
The mare provides a unique experimental model for studying
follicle development in monovular species. Development of
antral follicles in horses is characterized by the periodic growth
of follicular waves which often involve the selection of a single
dominant follicle. If properly stimulated, the dominant follicle
will complete development and eventually ovulate a fertile
oocyte. Regulation of follicular wave emergence and follicle
selection involves an interplay between circulating gonadotro-
pins and follicular factors that ensures that individual follicles
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
life in the mare and is influenced by factors such as stage of
oestrous cycle, season, pregnancy, age, breed and individual so
that different types of follicular waves (minor or major,
ovulatory or anovulatory) and different levels of activity
within waves may develop under different physiological
conditions. Changes in gonadotropin levels and ⁄ or in the
sensitivity of follicles to circulating gonadotropins seem
to account largely for these physiological variations in follicle
development.
Introduction
Although scientific interest in equine follicles existed by
the 1920s (Seaborne 1925), detailed studies on equine
follicle dynamics did not begin until 50 years later under
the pioneering lead of OJ Ginther at the University of
Wisconsin (reviewed in Ginther 1979). During the early
1980s, ultrasonography became available for equine
theriogenology (Palmer and Driancourt 1980). Trans-
rectal ultrasonography combined with systemic hor-
mone measurements were subsequently used extensively
to understand equine follicle dynamics. The more recent
introduction (mid-1990s) of the technique of ultrasound-
guided transvaginal ovarian puncture provided scientists
unprecedented access to the live equine ovary and
allowed the targeting of individual follicles for collection
of follicle samples, injection of test substances or
experimental manipulation of follicles (Gastal et al.
1997; Gerard and Monget 1998; Donadeu and Ginther
2001; Martoriati et al. 2003). The experimental use of
these procedures has led to extraordinary advances in
the knowledge of equine ovarian physiology during the
last 15 years (reviewed in Beg and Ginther 2006;
Donadeu and Watson 2007). Two important outcomes
of this knowledge have been an improvement of repro-
ductive efficiency in mares (Squires 2006) and a greater
understanding of follicular physiology in monovular
species in general. In regard to the latter, consideration
of the mare as a useful model for studying ovarian
processes during human health and disease has recently
increased (Ginther et al. 2004c; Carnevale 2008).
 2008 The Authors. Journal compilation  2008 Blackwell Verlag
This review summarizes current knowledge on equine
follicle development. Information on pre-antral and
early antral follicles is particularly scarce in the horse;
therefore, relatively more focus has been placed on the
late stages of antral follicle development up to the pre-
ovulatory stage. Details on physiological and practical
aspects of the ovulatory process and subsequent forma-
tion of the corpus luteum can be found elsewhere
(Ginther 1992; Boerboom and Sirois 2001; Squires 2006).
Preantral Stages of Follicle Development
During early fetal life, primordial germ cells migrate to
the primitive gonadal ridge where they proliferate and
subsequently enter meiotic division to become arrested
in prophase I as primary oocytes (Ginther 1992).
Meiotic arrest continues until atresia or until the
primary oocyte is stimulated by an LH surge during
post-natal life. Based on histological studies of the
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
phase between days 73 and 150 of pregnancy with a
peak at approximately day 100. Development of oocytes
into primordial follicles is not apparent until day 150
(Deanesly 1975) and the number of primordial follicles
continues to increase during subsequent fetal life so that
several thousand of these follicles are contained within
the ovaries at birth (Ginther 1992). Proliferation of the
somatic cell layer surrounding the primary oocyte leads
to the sequential development of primordial follicles
into primary, secondary and, finally, antral follicles,
a process that begins in the fetus and continues
throughout post-natal life. It is not known how follicles
are selected to grow from the pool of primordial, resting
follicles in the mare. Adult equine ovaries were reported
to contain approximately 36 000 primordial follicles and
100 growing follicles at any one time, with large
variability in actual follicle numbers between individual
mares (Driancourt et al. 1982). Comparatively higher
follicle numbers (threefold) were found in the adult
bovine ovary. Proliferative activity of small pre-antral
follicles in pony mares was higher at the beginning of the
ovulatory season than during anoestrus or at the end of
the ovulatory season, whereas follicular atresia was
unaffected by season (Driancourt et al. 1983).
Antral Follicle Development and Follicular
Waves
The equine follicle develops an antrum when it reaches
approximately 0.3 mm in diameter (Driancourt et al.
Follicles and Mares
1982). Similar to pre-antral stages, little is known on the
development of antral follicles before they reach
approximately 2 mm, the smallest diameter that can
usually be detected by transrectal ultrasonography. It
has been reported that the growth of an equine follicle
from 0.1 to 1 mm takes approximately two oestrous
cycles (Driancourt 1979) and that atresia during
this early phase of antral development is rare (Ginther
1992).
As in other farm species and humans, the develop-
ment of antral follicles in the horse is characterized by
the periodic growth of cohorts of follicles or follicular
waves (Sirois et al. 1989; Bergfelt and Ginther 1993).
Characterization of follicular waves has involved the
ultrasonic day-to-day identification of individual folli-
cles (Bergfelt and Ginther 1993; Gastal et al. 1997) and
the use of a statistical method that avoids the need to
maintain the identities of individual follicles during
serial ultrasound examinations (Ginther and Bergfelt
1992; Donadeu and Ginther 2002b). Follicular waves
in the horse can be identified in relation to follicles
2 mm in diameter and larger; however, it is not known
whether earlier antral and pre-antral stages are also
characterized by wave-like patterns of growth,
a question that has also not been clarified in other
species (Mizunuma et al. 1999; McGee and Hsueh
2000). Follicular waves and their regulation with an
emphasis on normal oestrous cycles will be described in
this and the next section. Follicular wave patterns
characteristic of other reproductive states will be
described in a separate section.
Characteristics of a follicular wave. Follicular wave
emergence has been normally defined for experimental
purposes as occurring when the largest follicle reaches 6
or 13 mm, depending on the study (Ginther et al.
2003a). Identification of wave emergence often requires
day-to-day ultrasonic evaluation of the ovary usually
after aspiration of all follicles from previous waves.
A follicular wave initially involves the simultaneous
growth of a variable number of follicles at a common
rate of between 2 and 3 mm ⁄ day. A recent study
involving ablation of all follicles during the middle of
an oestrous cycle in pony mares reported a mean of
approximately 12 follicles emerging during the common
growth phase of the ablation-induced wave (Gastal
et al. 2004). Approximately two follicles emerged each
day during the first 4 days after wave emergence with a
progressive decrease in the numbers of follicles emerging
thereafter. The exact number of follicles emerging within
waves is affected by factors such as season (Donadeu
and Ginther 2003).
The phase of common follicle growth is followed by
the selection of a single follicle (occasionally two
follicles) which is manifested as a deviation in diameter
between the two largest follicles of the wave beginning
when the largest follicle reaches approximately 22 mm
(Ginther et al. 2003a). Deviation begins a mean of
7 days before ovulation and is characterized by the
continuous growth (at an unchanged rate) of the largest
(selected) follicle as a dominant follicle and the
simultaneous cease in growth and subsequent regression
of smaller, subordinate follicles. Once it grows to
 2008 The Authors. Journal compilation  2008 Blackwell Verlag
225
approximately 35–45 mm, the dominant follicle nor-
mally either ovulates or ceases to grow and begins to
regress, depending on whether an ovulatory LH surge
occurs. The establishment of dominance is mediated by
a differential increase in trophic support to the largest
follicle of a wave by mechanisms that will be explained
in more detail in the next section. This leads to profound
changes in follicular cell function that are necessary for
the eventual full maturation of the follicle to an
ovulatory-competent state and that are reflected in
dramatic changes in global gene expression, changes
that have begun to be characterized in other species,
most notably cattle (Sisco et al. 2003; Fayad et al. 2004;
Mihm et al. 2008).
Dominance does not seem to be a pre-determined
trait among the follicles growing in a wave because
follicle ablation studies have demonstrated that all
follicles have similar capacity to become dominant,
a capacity that in subordinate follicles is lost within
approximately 48 h after the beginning of deviation
(Gastal et al. 2004). The same studies showed that in
approximately 61% of waves the first follicle to
emerge at 6 mm maintains its size advantage over
smaller follicles through the common growth phase
and becomes dominant. The likelihood of the largest
follicle becoming dominant increases as it approaches
the expected diameter at the beginning of deviation
(Gastal et al. 2004). In a few instances, yet, the largest
follicle ceases or slows down its growth during the
common growth phase and is replaced by the second
largest follicle (or sometimes even a smaller follicle)
which then becomes the dominant follicle.
Types of follicular waves. Follicular waves have been
classified as major waves (referred to in the literature
and throughout this review simply as follicular waves)
or minor waves, depending on whether they involve the
development of a readily identifiable dominant follicle
or they produce only smaller follicles, respectively
(Ginther 1993; Donadeu and Ginther 2002b). The
number of follicular waves during an oestrous cycle
varies between species. In the horse, as in humans, only
one or two major follicular waves develop during each
cycle (Ginther et al. 2004c). A major wave (named
primary wave) always emerges during the middle of the
equine oestrous cycle and produces the ovulatory
follicle. Approximately 25% of interovulatory intervals
involve an additional major wave (secondary wave) that
develops during the first half of the cycle (Sirois et al.
1989; Bergfelt and Ginther 1993). The incidence of
secondary waves is significantly higher in some breeds
such as Thoroughbreds, and some of these waves may
produce ovulations. The ability to ovulate in the
presence of high progesterone levels during dioestrus
seems to be unique to the horse (Ginther 1992). Minor
waves (largest follicle usually <30 mm) have been
identified at different stages of the oestrous cycle and
their incidence has been reported to be low, 25% or less
of oestrous cycles (Bergfelt and Ginther 1993; Ginther
1993). Yet, a study considering follicles as small as
2 mm concluded that relatively high levels of underlying
activity may involve the smallest follicles of a wave
(Ginther and Bergfelt 1993).
226
Regulation of Follicular Wave Development
Emergence of a follicular wave and subsequent selection
of a dominant follicle are under finely-tuned regulation
by systemic mechanisms involving changes in gonado-
tropin levels and local mechanisms that involve changes
in follicular factor levels.
Systemic regulation of follicular wave develop-
ment. Unlike pre-antral follicles, antral follicles cannot
develop without adequate gonadotropin stimulation and
this has been experimentally shown in the mare
(Pedersen et al. 2002; Imboden et al. 2006).
Follicular waves in the mare, as in other species, are
temporally preceded by a stimulatory surge in circulat-
ing FSH (Ginther et al. 2003a). The acute dependence of
follicular waves on FSH has been shown in mares by the
inability of follicles to grow beyond a diameter of 15 mm
after suppression of circulating FSH by systemic injec-
tion of follicular fluid (Bergfelt and Ginther 1985). Close
functional relationships between FSH surges and follic-
ular waves involve not only positive effects of FSH on
follicles but also negative effects of follicles on FSH. The
wave-associated FSH surge reaches a peak when the
largest follicle is approximately 13 mm in diameter
(Gastal et al. 1997; Donadeu and Ginther 2001). The
following decline in FSH results from an increase in
circulating inhibin, presumably inhibin-A (Watson and
Al-zi’abi 2002), contributed mainly by the three largest
follicles of the wave as they grow above 13 mm
(Donadeu and Ginther 2001). The declining FSH
concentrations continue to support growth of the
follicles of the wave until the largest (future dominant)
follicle reaches the expected diameter at the beginning of
deviation (approximately 22 mm). At that time, circu-
lating FSH levels become too low to maintain the
growth of the follicles of the wave. The low FSH, yet,
does not restrict the growth of the future dominant
follicle which by that time has acquired the ability to
more efficiently use circulating gonadotropins for
growth (Ginther et al. 2003a). The differential responses
of the largest and smaller follicles of a wave to
gonadotropins result in the initiation of diameter
deviation. Continuous suppression of FSH throughout
deviation ensures the morphological and functional
demise of subordinate follicles and is attributable to
production of high levels of inhibin and oestradiol by
the dominant follicle (Gastal et al. 1999; Donadeu and
Ginther 2001). The critical role of low FSH levels in the
deviation mechanism in mares is illustrated by the
disruption of the deviation mechanism after adminis-
tration of FSH (Squires 2006) or immunization against
inhibin (McCue et al. 1992) leading to the development
of multiple ovulatory follicles.
While FSH is particularly important for follicular
growth before deviation, LH becomes more critical
during deviation. This has been demonstrated by studies
showing that experimental suppression of LH in cycling
mares leads to the regression of the dominant follicle
early during its development (Bergfelt et al. 2001), and is
consistent with an increase in circulating LH before the
beginning of deviation (Bergfelt et al. 2001). The study
of LH–follicle interrelationships during the anovulatory
 2008 The Authors. Journal compilation  2008 Blackwell Verlag
FX Donadeu and HG Pedersen
season has revealed a direct temporal relationship
between an increase in circulating LH, but not FSH,
and the development of major waves, further indicating
that an increase in LH plays a major role in stimulating
the development of dominant follicles (reviewed in
Donadeu and Watson 2007). An additional conclusion
was that circulating LH concentrations above those
required for growth of the dominant follicle are required
for the development of ovulatory competence, i.e. for
the dominant follicle to become fully responsive to an
LH surge (Donadeu and Watson 2007).
Studies in cattle have shown that LH receptor
expression differentially increases in granulosa and theca
cells of the early dominant follicle (Bao and Garverick
1998; Beg and Ginther 2006). These findings are
consistent with those in other species (Richards 2001).
Although not critically examined in relation to the
beginning of deviation, higher LH receptor levels have
been reported in dominant-size follicles than in smaller
follicles in the horse (Fay and Douglas 1987; Goudet
et al. 1999). Taken together, the findings on LH levels
and LH receptor expression during a follicular wave in
mares are consistent with the conclusion that deviation
involves a critical increase in the dependence of the
dominant follicle on LH.
Limited data exist on the involvement of substances
other than gonadotropins in the endocrine regulation of
antral follicles in mares. Based on observed direct effects
on follicle growth or on the presence of specific receptors
in equine ovaries, positive roles on follicle growth have
been suggested for circulating levels of substances
including growth hormone (Cochran et al. 1999), dopa-
mine (King et al. 2005) and prolactin (Thompson et al.
1997).
Although follicular insulin-like growth factor-1
(IGF-1) in mares largely derives from the systemic
circulation, its bioactivity is regulated mostly through
local mechanisms (Watson et al. 2004) and the role of
IGF-1 is therefore considered in the next section.
Local regulation of follicular wave development. Local
regulation of follicle development in monovular species
has been studied extensively in relation to follicle
selection (reviewed in Fortune et al. 2004; Beg and
Ginther 2006; Knight and Glister 2006). A variety of
protein and steroid factors including members of the
IGF family, oestradiol, inhibins and activins, follistatin
and vascular endothelial growth factor (VEGF) are
involved. In general, these factors act, often in a
paracrine manner, to either enhance or diminish the
trophic effects of gonadotropins on follicular cells
through a variety of mechanisms. Differential changes
in the levels of specific factors between follicles thus
ensure the continuous development of the dominant
follicle and the regression of subordinate follicles during
deviation.
A differential increase in the levels of oestradiol, IGF-1,
activin-A and inhibin-A in the future dominant follicle
was associated with the beginning of deviation in mares,
whereas differentially elevated levels of progesterone
and inhibin-B occurred later during the development of
the dominant follicle (Donadeu and Ginther 2002a).
Follicles and Mares
Studies involving intrafollicular factor injection in mares
have provided insight into the complex interrelation-
ships between these factors during follicle deviation
(Ginther et al. 2005). It has been concluded that
although all these factors are likely involved in the
development of the dominant follicle after the beginning
of deviation, only IGF-1 is involved in the initiation of
deviation by, among other actions, regulating the levels
of other growth factors in the dominant follicle (Beg and
Ginther 2006). This is consistent with the particularly
important role of the IGF-1 system in follicle selection
in other species (Mazerbourg et al. 2003). Results from
a series of in vivo experiments have convincingly
confirmed the critical role of the IGF system in follicle
selection in mares. Injection of IGF-1 into the second
largest or a smaller follicle at the beginning of deviation
changed its fate from subordinate to co-dominant
resulting in the development of multiple ovulatory
follicles (Ginther et al. 2004a; b; Gastal et al. 2007).
Conversely, injection of IGF binding protein (IGFBP)-3
into the largest follicle at the beginning of deviation
resulted in the follicle regressing and being replaced by
the second largest follicle which became dominant
(Ginther et al. 2004a). At least four types of IGFBPs
(IGFBP-2, 3, 4 and 5), which negatively regulate IGF
activity, have been identified in equine follicles and the
concentrations of three of these (IGFBP-2, 4 and 5) are
correlated negatively with those of IGF-1 during follicle
development (Gerard and Monget 1998; Bridges et al.
2002). Consistent with the stimulatory role of IGF-1 in
the development of ovulatory follicles, reduced levels of
bioactive IGF-1 are thought to be involved in the
developmental deficiencies of dominant follicles during
the spring transition that prevent them from acquiring
ovulatory competence (Acosta et al. 2004; Watson et al.
2004).
An additional factor that has begun to be explored in
relation to follicle selection in horses is VEGF. This
angiogenic factor has been shown to be necessary for
follicle development in other species (Fraser and Wulff
2001). VEGF levels differentially increase in the dom-
inant follicle in horses (Ginther et al. 2004b), and this
increase is thought to be mediated, at least partly, by
IGF-1. VEGF is likely involved in the reported increase
in vascularization of the future dominant follicle before
the beginning of deviation (Acosta et al. 2004) which
presumably increases the availability of circulating
gonadotropins to the follicle. The reduced levels of
follicular VEGF and low vascularity of the wall of
dominant follicles during the spring transition relative to
the ovulatory season (Watson and Al-zi’abi 2002)
underscore the critical role of VEGF in the development
of the ovulatory follicle in horses.
Effects of Different Physiological Conditions on
Follicle Development in Mares
Effects of season. The effects of season on follicular
activity in mares have been studied in considerable
detail. Important variations in follicular activity occur
not only between the ovulatory and anovulatory seasons
but also between different periods within each season.
 2008 The Authors. Journal compilation  2008 Blackwell Verlag
227
Studies in pony mares using follicle ablation to facilitate
the identification of individual follicular waves revealed
that, as during the ovulatory season, follicular waves
periodically occur during the anovulatory season despite
the reduced levels of follicle development (Donadeu and
Ginther 2002b, 2003; Ginther et al. 2003b). Only minor
waves (largest follicle <21 mm) occurred during the
middle of the anovulatory season or deep anoestrus,
whereas major waves were often detected during the first
2 months (fall transition) and the last 2 months (spring
transition) of the anovulatory season, in agreement with
previous studies (Ginther 1990; Watson et al. 2002). The
onset of the spring transitional period involved an
abrupt increase in follicular activity from the low
activity typical of deep anoestrus which was reflected
in a distinct increase in the diameter of follicles and,
most notably, a fourfold increase in the numbers of
follicles >12 mm within waves (means of 3.2 and 11.5
follicles in waves developing during deep anoestrous and
the early spring transition, respectively). Although
dominant follicles during transition may not grow to
the diameters typical of ovulatory follicles, in the same
study transitional waves produced more follicles than
waves developing during the ovulatory season (means of
11.5 and 6.0 follicles >12 mm, respectively) attesting to
the high levels of follicular activity even in the absence
of ovulation in transitional mares (Donadeu and Gin-
ther 2003).
Based on consistent temporal relationships between
follicles and circulating hormones, it has been concluded
that the differences in follicle development between
different periods of the anovulatory season as well as the
deficient development of dominant follicles during the
spring transition relative to the ovulatoy season are not
attributable to deficient circulating FSH levels but
rather to changes in LH and, possibly, differences in
follicular sensitivity to gonadotropins (reviewed in
Donadeu and Watson 2007).
Season-related effects on follicular activity have also
been reported between the two halves of the ovulatory
season, with higher levels of activity during the first half
of the season due to higher incidence of both secondary
waves and minor waves associated with higher gonado-
tropin levels (Ginther 1992, 1993).
Effects of pregnancy and parturition. Considerable
research work is needed to better characterize follicular
dynamics and associated regulatory mechanisms during
and following pregnancy in the mare. Based on com-
bined data from ultrasound and rectal palpation studies,
follicular dynamics during the first half of pregnancy
are similar to those occurring during the first half of
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,
more commonly, development of sporadic major waves
or only minor waves (Ginther and Bergfelt 1992),
followed by a pronounced decrease in follicular activity
in all mares between days 50 and 140 of pregnancy so
that the diameter of the largest follicle does not exceed
15 mm by day 140 (Squires et al. 1974). A decrease in
follicular activity has also been reported after the first
one-third of pregnancy in cattle (Ginther et al. 1996).
228
Based on reported hormone–follicle associations, defi-
cient FSH levels do not seem to be responsible for the
reduction in follicular growth in some mares between
days 11 and 40 of pregnancy (Ginther and Bergfelt
1992), an effect that may instead be accounted for by
reduced levels of LH (due to persistent progesterone
negative feedback), similar to the effects of the seasonal
reduction in circulating LH on follicle growth during the
fall transitional period (Ginther et al. 2003b).
The mechanisms responsible for the dramatic reduc-
tion in follicular growth during mid and late pregnancy
in mares have not been clarified but likely involves a
temporally associated decrease in circulating FSH
(reviewed in Ginther 1992). This is different from the
anovulatory season during which changes in FSH levels
do not seem to play a major role in the decrease in
follicle growth during deep anoestrus (Donadeu and
Watson 2007). Reduced follicle numbers during mid-
pregnancy are also likely attributable to ovulation or
luteinization of follicles into accessory corpora lutea
under the influence of chorionic gonadotropin (Ginther
1992). Further complexity into the regulation of follicle
growth during pregnancy is provided by the observation
that the effects of season on hormones and follicles
continue to occur during pregnancy (Ginther 1992).
The natural pressure to produce a foal each year in a
species with an 11-month-long pregnancy is reflected in
an early post-partum ovulation in the mare (foal heat,
typically within 2 weeks of parturition). A steady
increase in the diameter of the largest follicle and
in the numbers of follicles after parturition resulting
in ovulation 14 days later was recently shown in
Arabian mares (Gunduz et al. 2008). The post-partum
increase in follicular growth is induced by an increase in
gonadotropin secretion at the time of parturition (Hines
et al. 1987; Ginther et al. 1994). Studies have shown that
the follicular response to parturition may vary in
individual horses or different types of horses, and may
not readily occur in primiparous mares (Nagy et al.
1998), in the presence of a nursing foal (Ginther 1992) or
during the winter, when the negative effects of season
may prevail over the positive effects of parturition
(Ginther et al. 1994).
Follicle development before puberty. Follicle develop-
ment in spring-born pre-puberal pony fillies has recently
been studied in detail (Nogueira and Ginther 2004),
adding to limited information on follicle profiles from
earlier studies (reviewed in Ginther 1992). Follicular
activity during 2–10 months of life was characterized by
a progressive increase in mean follicle diameter (from
approximately 6 to 10 mm) and mean follicle numbers
(from 3 to 17 follicles) between 2 and 5 months of age,
a short plateau in activity coinciding with the winter
months and a re-initiation of follicle growth after
7 months of age leading to the onset of the first
ovulatory season in spring (Nogueira and Ginther
2004). Changes in follicle activity during the first year
of life were positively correlated with changes in
circulating gonadotropins, consistent with a regulatory
role of season on gonadotropin and follicular activity
beginning early during life in mares. Follicular growth
before puberty was characterized by the development of
 2008 The Authors. Journal compilation  2008 Blackwell Verlag
FX Donadeu and HG Pedersen
(minor) follicular waves. Remarkably, these waves were
not temporally associated with statistically significant
circulating FSH surges, an observation that warrants
further investigation.
Effects of aging on follicular activity. As highlighted in a
recent review, many age-related changes in follicular
activity in the horse resemble those occurring in humans
(Carnevale 2008). Follicular activity during oestrous
cycles begins to decrease in mares 20 years of age or
older eventually leading to a cease in ovarian activity
(Carnevale et al. 1993, 1994). Interovulatory intervals
first become longer in these mares due to longer
follicular phases associated with a primary follicular
wave that emerges later and contains less follicles. In
addition, the ovulatory LH surge is less pronounced in
the older mares, and there is a higher incidence of
ultrasonically atypical ovulations characterized by a
central hypoechogenic area at the ovulatory site. The
reduction in follicular activity and frequency of ovula-
tion in mares ‡20 years old is associated with an overall
elevation in concentrations of FSH and LH during the
follicular phase, a phenomenon that also occurs during
the peri-menopausal period in women. This is eventually
associated with persistent ovarian inactivity with folli-
cles <5 mm and no ovulations in the presence of
continuously elevated concentrations of FSH and LH.
Reportedly, approximately 40% of horses 24 years or
older have no ovarian activity (Vanderwall 1990)
whereas 50% of ponies 20 years or older have irregular
or absent ovulations (Carnevale et al. 1994). It has been
suggested that an age-related elevation in FSH concen-
trations may hasten the eventual depletion of the
primordial follicle pool in older mares, similar to what
occurs in women (Carnevale 2008). Age-related differ-
ences in follicular activity have also been reported in
mares during the period of transition into the ovulatory
season during which progressively lower follicle diam-
eters and numbers of follicles were found in mares 3–7,
17–19 and ‡20 years old (Carnevale et al. 1997).
Conclusions
The possibility of experimentally targeting equine folli-
cles at very precise stages of development in vivo using
a relatively noninvasive ultrasound-guided technique
makes the horse a very attractive model to study in
detail the dynamics of antral follicles. In addition, the
seasonal nature of ovarian activity in mares offers a
unique model to study the factors naturally regulating
follicle development in a monovular species.
As in other monovular species, the development of
antral follicles in horses is characterized by the periodic
emergence of follicular waves and the selection of
usually a single dominant follicle that under the appro-
priate hormonal milieu will ovulate a fertile oocyte.
Follicular wave emergence and follicle selection are
driven by systemic changes in levels of FSH (wave
emergence and follicle selection) and LH (follicle selec-
tion), and by local changes in follicular factors that,
through a variety of mechanisms, are responsible for
fine-tuning the response of follicular cells to gonadotro-
pin stimuli to ensure the continuous growth of the
Follicles and Mares
dominant follicles and the simultaneous regression of
subordinate follicles during follicle deviation.
Follicular waves occur throughout post-natal life in
the horse until follicles irreversibly cease to grow at
approximately ‡ 20 years of age. The levels of follicular
activity are affected by factors such as stage of the
oestrous cycle, season, pregnancy, age, breed and
individual. This results in variations in follicular wave
patterns that most notably include the production or
absence of a dominant follicle (major or minor waves)
and the development of ovulatory-competent or ovula-
tory-incompetent dominant follicles. Available informa-
tion indicates that these physiological variations in
follicular wave patterns are driven by changes in
circulating levels of FSH and ⁄ or LH and by local
changes in responsiveness of follicles to gonadotropins.
Yet, considerable work is needed to better understand
the mechanisms by which follicle development is regu-
lated during different physiological conditions outside
the oestrous cycle in mares.
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