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Follicular wave dynamics after estradiol-17β treatment

of heifers with or without a progestogen implant

Article  in  Theriogenology · June 1994

DOI: 10.1016/0093-691X(94)90821-Y

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FOLLICULAR WAVE DYNAMICS AFTER ESTRADIOL-17P TREATMENT OF HEIFERS
WITH OR WITHOUT A PROGESTOGEN IMPLANT

G.A. Bo,l G.P. Adams2 R.A. Pierson3 HE. Tribulo,4 M. Caccia4 and R.J. Mapletoftl

Departments of ‘Herd Medicine and Theriogenology, 2Veterinary Anatomy, and

30bstetrics and Gynecology, University of Saskatchewan, Saskatoon, Sask, Canada, S7N OWO
4Facultad de Ciencias Agropecuarias, Universidad Catolica de Cordoba, Cordoba, Argentina

Received for publication: Octiober 20, 1993

Accepted: March 9, 2994

ABSTRACT

Two experiments were designed to evaluate the effects of estradiol-17a (E-17P) on follicular
wave dynamics and gonadotropin in cattle. The first experiment was designed to evaluate the
effect of 5 mg E-17P administered on Day 1 (ovulation=Day 0) in heifers with or without a
progestogen (SMB) ear implant. The dominant follicle in heifers treated with E-17@+SMB ceased
to grow 1 d after E-17p treatment and subsequently regressed resulting in early emergence of the
next follicular wave. Conversely, E-17P treatment of non-implanted heifers resulted in transient
or incomplete suppression of the dominant follicle, and delayed emergence of the next follicular
wave (P&.05). A post-treatment surge in plasma LH concentration was detected in S of 6 heifers
treated with E-17@ alone versus 1 of 6 treated with E-17P+SMB (PcO.05). In all but 1 heifer,
the LH surge was accompanied by a concurrent FSH surge (18 to 36 h after E-17@. The second
experiment was designed to determine an effective dosage regimen of E-17B for suppression of
follicular growth in SMB-implanted heifers and to test the hypothesis that estradiol-induced
follicle suppression will result in a synchronous emergence of the subsequent follicular wave. On
Day 0, 48 heifers were implanted with SMB and allocated to 1 of 7 treatment groups: control
heifers, and those that received 10 mg or 5 mg E-17/3 i.m. on Day 1 or Day 3; or 2.5 mg E-17P
b.i.d. on Days 1 to 3 or Days 3 to 5. The growth of the dominant follicle of the first follicular
wave was suppressed in all the E-17fi treated groups. Emergence of the second follicular wave
was later (P<O.OS) in control heifers than heifers treated with a single injection of E-17B on Day
1, but was not different from heifers treated on Day 3. Furthermore, the day of wave emergence
was less variable (P<O.OSf in heifers treated with 5 mg of E-l@ than in control heifers. It was
concluded that E-17P and progestogen trea~ent may be used to suppress follicular growth and
cause the synchronous emergence of a new follicular wave in cattle.

Key words: bovine, estradiol, dominant follicle, follicular dynamics, ultrasonography

Acknowledgments
We thank Sanofi Inc., Overland Park, KS, USA, for Syncro-Mate-B and Coopers Agropharm
Inc, Ajax, ON, Canada for Estrumate. Research was supported by the Catholic University
of Cordoba, Argentina, the University of Saskatchewan and the Natural Sciences and
Engineering Research Council of Canada. Special thanks to R. Rabella, A. Gomez, A.
Alonso, G. Brogliatti, L. Nasser, J.W. Mapletoft and W.B. Kerr for technical assistance.

Copyright 0 1994 Buttetworth-Heinemann

 1556 Theriogenology

INTRODUCTION

The objective of superstimulation treatments in cattle is to obtain the maximum number of

viable embryos by stimulating antral follicles with exogenous gonadotropin. However, the
variable and unpredictable superovulatory response of donor animals is one of the most limiting
factors to successful embryo transfer (4). While many studies have focused on gonadotropin and
dosage regimens (4,8,29), individual animal variability with regard to the stage of follicular
development may be a more important factor affecting superovulatory response (16,18,30,3 1,32).
In addition, variation in the interval to estrus and reduced pregnancy rates following fixed-time
inseminations have been impediments to the use of estrus synchronization programs in large scale
operations (36). It has been recently shown that in prostaglandin-treated cattle, variability in
return to estrus is determined by the viability of the dominant follicle at the time of treatment
(22,38). Heifers with viable dominant follicles returned to estrus 48 to 60 h after prostaglandin
treatment, whereas those in which the dominant follicle was in its static or regression phases
exhibited estrus 5 to 7 d later; a reflection of the time required for a follicle from the new wave
to develop to a preovulatory state. The phase of follicular development may be assessed by serial
examinations using transrectal ultrasonography, but multiple examinations are time consuming
and labor and time intensive. An alternative involves artificial synchronization of follicular wave
emergence so that treatments may be initiated when a population of responsive growing follicles
is available.

Ovarian follicular development in cattle occurs in waves, usually involving either 2 or 3

follicular waves within an estrous cycle (14,15,26,39,42). During each wave one follicle is
selected to become dominant while subordinate follicles in the cohort undergo atresia (14,15).
Through the production of steroidal and non-steroidal substances (12,20,27,33), the dominant
follicle of a wave is responsible for suppression of its subordinates and prevention of the
emergence of a new wave (2,23,25). A dominant follicle of a wave produces SOO-to lOOO-fold
more estradiol than the smaller follicles and persists to produce large amounts of estrogen while
all others regress (20). Estrogens have been shown to induce follicle atresia (19) and the effect
of estrogens on gonadotropin and preovulatory follicles have been reported in several studies
(9,10,24,34,35,41). However, the effect of estrogens on follicular wave dynamics have not been
extensively investigated. Evidence regarding the effect of estrogens on follicular dynamics was
provided by a preliminary study in which estradiol valerate was given to cows with a progestogen
ear implant (SMB;a 7). In a subsequent study, estradiol valerate treatment during the early
growing phase (Day l), suppressed the growth of the dominant follicle of the first wave and
hastened the emergence of the second follicular wave in non-implanted heifers (6). However,
estradiol valerate treatment during the mid- (Day 3) or late-(Day 6) growing phase resulted in
delayed emergence of the next wave. This was attributed to incomplete suppression (Day 3
treatment) or no suppression (Day 6 treatment) of the dominant follicle and the prolonged effect
of the valerate form of estradiol.

Mechanisms by which estrogens affect ovarian follicles may include FSH suppression
(9,24), LH release (41) or a direct effect of estradiol within the ovary (19). Exogenous estrogens

a Syncro-Mate-B, Sanofi Inc., Overland Park, KS, USA.

 Theriogenology 1557

have been shown to induce LH release 16 to 20 h after administration (41) and GnRH-induced
LH release has been reported to induce follicle luteinization and atresia in the cow (28).
However, results of recent studies have not supported the notion that LH release induced follicle
atresia (6).

Two experiments were designed to evaluate the effect of a short acting estrogen, estradiol-
17p (E-17P), on circulating concentrations of gonadotropin and follicular wave dynamics in
cattle. The purpose of Experiment 1 was to evaluate the effect of E-17@ administered alone or
in combination with a progestogen ear implant. Three hypotheses were tested: 1) estradiol-17p
more effectively suppresses dominant follicle growth when combined with progestogen treatment,
2) progestogen blocks estradiol-induced LH release, and 3) follicle suppression is not a result of
estradiol-induced LH release. Experiment 2 was designed to determine an effective dosage
regimen of E-170 for suppression of follicular growth in SMB implanted heifers and to test the
hypothesis that estradiol-induced follicle suppression will result in a synchronous emergence of
the subsequent follicular wave.

MATERIALS AND METHODS

Experiment 1

Twelve cross-bred beef heifers, 18 to 22 mo of age and weighing 450 to 500 kg were used.
All heifers bearing a CL were injected im with 500 ug of cloprostenolb At the time of ovulation
(Day 0), heifers were randomly allocated to a group in which an SMB ear implant (containing
6 mg of norgestomet) was inserted and removed on Day 9 or to a non SMB-treated control group
(n=6 per group). On Day 1, all heifers received a single im injection of 5 mg E-17PC dissolved
in 2 ml of sesame oil.

Experiment 2

Forty-six beef heifers, 20 to 24 mo of age and weighing 300 to 450 kg were used. As in
Experiment 1, heifers bearing a CL received 500 ug cloprostenol im. On Day 0 (ovulation) all
heifers were given SMB ear implants and were randomly allocated to 1 of 7 treatment groups:
control heifers (n=7; no E-17P) and those that received im injections of 10 mg E-17P on Day
1 (n=6) or Day 3 (n=7), 5 mg E-17P on Day 1 (n=7) or Day 3 (n=6), or 2.5 mg E-17P b.i.d. on
Days 1 to 3 (n=6) or Days 3 to 5 (n=7). Implants were removed on Day 6. The twice daily
injections were intended to imitate the prolonged effect of the estradiol valerate injection used
in previously reported experiments (6,7).

Ovarian Ultrasonography

Ovarian ultrasonographic examinations were performed as previously described (1) using

a real time, B-mode scanner equipped with a 5 Mhz linear-array transducer.d Examinations were

b Estrumate, Coopers Agropharm, Ajax, ON, Canada.

’ Sigma Chemical Company, St. Louis, MO, USA.
d Aloka SSDSOO, Overseas Monitor Corporation Ltd., Richmond, BC, Canada.
 1558 Theriogenology

done daily by a single operator without knowledge of the treatment groups, initially to detect
ovulation, and thereafter until the fourth day after the emergence of the second follicular wave
(Experiment 1) or the subsequent ovulation (Experiment 2). During each examination a sketch
of the ovaries was made recording the location and diameter of the CL and of individually
identified follicles 24 mm (26). The dominant follicle of a wave was defined as the one that
reached the largest diameter and subordinate follicles were defined as those that appeared to
originate from the same pool of follicles (15). The day of emergence of a follicular wave was
defined as the day that the dominant follicle was retrospectively identified at a diameter of 4 to
5 mm (26,lS). The growing phase of the dominant follicle was defined as the period from the
day that the follicle was first identified until the day that it appeared to cease a progressive
increase in diameter (day of cessation of growth of the dominant follicle; 1.5). The onset of
regression of the dominant follicle was defined as the first day that the follicle appeared to begin
a progressive decrease in diameter (15).

Hormone Assays

In Experiment 1, blood was collected by jugular venipuncture into heparinized tubes. Within
30 min of collection, plasma was harvested and stored at -20°C until RIA was performed.
Samples were taken every 12 h on Day 0, every 6 h from Day 1 to Day 4 and every 12 h until
the fourth day after the emergence of the next follicular wave.

Plasma concentrations of estradiol were measured in all samples from Day 0 until Day 4
using a validated RIA (21). Standards were prepared in charcoal-stripped bovine serum and the
standard curve ranged from 2 to 200 pg/ml. When values were higher than 100 pg, 100 pl of the
samples were diluted in 400 yl of charcoal-stripped serum and re-analyzed. The minimum
detectable limit of the assay was 1 pg/ml. The intra-assay CV was 10% (n=8) and the inter-assay
CV was 13% (n=24).

Plasma concentrations of FSH were measured in all samples taken using a validated RIA
(11,2 1). The range of the standard curve was 0.13 to 16 ng/ml. The minimum detectable limit
of the assay was 0.2 @ml. The first antibody was NIAMDD-anti-oFSH-I, and FSH
concentrations were expressed in terms of USDA-bFSH-II. The intra-assay CV was 8.5% (n=8)
and the inter-assay CV was 12% (n=16). On an individual-animal basis, a post-treatment FSH
surge was defined as an increase in concentration of >2 SD above the mean FSH concentration
for that animal (averaged over all days). The day of maximum FSH concentration prior to each
follicular wave was defined as the day of the highest concentration within 5 d of emergence of
a wave (2,6). The day of initial increase in FSH concentration prior to each follicular wave was
defined as the first day that 2 sequential increases occurred leading to the maximum
concentration (2,6).

Plasma concentrations of LH were measured in all samples taken on Days 0 to 4, using a

validated RIA (11,21). The range of the standard curve was 0.13 to 16 ng/ml; the minimum
detectable limit was 0.1 ng/ml. Concentrations were expressed in terms of NIAMDD-bLH4. The
intra-assay CV was 7.1% (n=8) and the inter-assay CV was 11.3% (n=16). On an individual-
animal basis, a post-treatment LH surge was defined as an increase in concentration of >2 SD
above the mean LH concentration for that animal (averaged over all days).
 Theriogenology 1559

Statistical Analyses

In Experiment 1, single point-measurements for follicular characteristics, indicated in Table

1, were compared using Student’s t-tests (37,43). Bartlett’s test of homogeneity of variance was
used to evaluate the variability in the day of emergence of the second follicular wave (43).
Profiles of the diameter of the dominant and largest subordinate follicles in the first wave, plasma
concentrations of LH, FSH and estradiol were examined by split-plot ANOVA to determine the
effects of time and treatment, and the time-by-treatment interaction (13,37). For analysis and
illustration of the follicular profiles, data for each wave were normalized to begin on the mean
day of wave emergence for the group represented (2,14). Proportions were compared by Chi-
square test.

In Experiment 2, single point measurements were compared among groups by ANOVA. If

main effects or their interactions were significant, means were compared by the method of
protected least significant difference (37). As in Experiment 1, Bartlett’s test was used to
compare variability in the day of wave emergence between control heifers and E-17P treated
heifers. Profiles of the diameter of the dominant and largest subordinate follicles were analyzed
as in Experiment 1.

RESULTS

Experiment I

Mean plasma estradiol, LH and FSH concentrations are depicted in Figure 1. In both
groups, estradiol concentrations increased dramatically 6 h after E-17P administration and
subsequently decreased to basal levels 42 h after treatment. A surge (> 2 SD above the mean)
in plasma LH concentration was detected in 5 of 6 heifers in which SMB was not implanted (18
to 24 h after E-17P treatment), and in 1 of 6 heifers implanted with SMB (36 h after E-17P
treatment; Chi-square PcO.05). A time-by-treatment interaction (PcO.05) was detected in the post-
treatment FSH profiles. A surge (>2 SD) in plasma FSH concentration was detected in 5 of 6
heifers in which SMB was not implanted. In 4 of these heifers the FSH surge was concurrent
with the LH surge (18 to 24 h after E-17P). The remaining heifer had a post-treatment FSH surge
42 h after E-17P). In the E-17P+SMB treated group, 3 heifers had a FSH surge 30, 36 and 72
h after E-170 treatment, respectively, with 1 having a concurrent LH surge (36 h after E-17P).

The effects of E-17P and SMB on follicular characteristics and wave emergence are
depicted in Table 1. The mean day of cessation of growth and onset of regression of the
dominant follicle was earlier (PcO.05) in heifers treated with E-17P+SMB than those treated with
E-17P alone. Furthermore, mean maximum diameter of the dominant follicle was smaller
(P<O.O5) in heifers treated with E-17P+SMB. Mean day of emergence of the second follicular
wave was earlier (PcO.01) and the day of emergence was less variable (PcO.01) in the heifers
treated with E-17P+SMB than those treated with E-17P alone (Table 1). There was a time-by-
treatment interaction (P<O.Ol) in the mean diameter profiles of the dominant follicle in the first
wave, which was attributed to a smaller follicle diameter after Day 4 in heifers treated with
E- 17fi+SMB.
 1560 Theriogenology

7001

---0--- E-17fi (nz6)

- E-170+SMB (n=6)

Time P~0.0 1
Treatment Pd.73
Time x Treatment P~0.52

3.0.

2.5.

-i 2.0-
Time P~0.01
ae ,j I1 Treatment P~0.04
1.5. 0:
;; , Time x Treatment P~0.01

3 l.O-

0.5

0.0’

1.50-I

1.25-

2
-$ l.oo-
x
2 oe’5.
Time P~0.01
0.50. Treatment P~0.20
Time x Treatment P<O.O4
0.257
-1 0 1 2 3 4
Days after ovulation

Figure 1. Mean (GEM) plasma estradiol, LH and FSH concentrations in heifers treated with
5 mg estradiol-17P (E-17P) on Day 1 with or without a progestogen (Sh4B) ear
implant (Experiment 1; Day O=ovulation).

* Means differed (P<O.O5).

 Theriogenology

Analysis of FSH concentrations normalized to the emergence of the second follicular wave
revealed a significant time effect (P<O.Ol) for each group (Figure 2). The mean day of initial
increase and the day of maximum FSH concentration prior to the emergence of the second wave
was earlier (P&05) in the heifers treated with E-17P+SMB than those treated with E-17P alone
(Table 1).

Table 1. Comparison (meanGEM) of follicular characteristics and wave emergence in heifers

treated with 5 mg estradiol-17s (E-17P) on Day 1, with or without a progestogen ear
implant (SMEl; Day O=ovulation)

End Point E- 17p+SMB E-17@

(n=6) (n=6)

Wave emergence (Day)

First wave -0.5iO.2 -0.2kO.2
Second wave 5.2j~O.2~ 9.8k1.1b
Range (second wave) 5-6 5-10

Dominant follicle
Maximum diameter (mm) 7.2*1.0a 11.0*1.2b
Cessation of growth (Day) 1.5i0.4a 5.0-t1.0b
Onset of regression (Day) 4.2ti.5a 9.8+1.gb

First Subordinate follicle

Maximum diameter (mm) 6.7 so.9 6.8 *0.3
Maximum diameter (Day) 1.2 kO.3 1.8 rt0.3

FSH orior to the second wave

Initial increase (Day) 3.3 k0.4a 8.6 *1.2b
Maximum FSH (Day) 4.1 +0.4a 9.5 +1.2b

ab Means within rows with different superscripts are different (PcO.05).

Experiment 2

Follicular characteristics and wave emergence of SMl3 implanted heifers treated with
different doses of E-17P are depicted in Table 2. Mean maximum diameter of the dominant
follicle was smaller (PcO.05) in heifers treated with E-17B than in control heifers, except for
those treated with 10 mg on Day 3 which was not different from controls. Mean day of cessation
of growth and onset of regression of the dominant follicle was earlier (PcO.05) in all heifers
treated with E-17P than in control heifers.
 1562 Theriogenology

14 E-178 (n=6)
-1.6

1 T

lo- -1.2

Ii - 1.0
_
B8
s - 0.8
ii 6
3 - 0.6

z -
._
24‘ -0.4 _

5Ai, i
r1.6 x
2
-1.4

- 1.2

- 1.0
8-
- 0.8
6-
- 0.6

4- Dominant follicle - 0.4

----+--- FSH

2
0
, 2
(
4
,
6
,
8
,
10
,
12
,
14
10.2

Number of days from ovulation

Figure 2. Relationship between profiles of FSH (ng/ml) and the diameter of the dominant follicle
of the first and second follicular waves in heifers treated with 5 mg estradiol-17P (E-
17p) on Day 1, with (E-17P+SMB) or without (E-17@ a progestogen ear implant
(Day O=ovulation). Follicle data were normalized to the mean day of wave emergence
in each treatment group. The day of FSH values correspond to the normalized
follicular data (Experiment 1).
 Theriogenology 1563

Mean day of emergence of the second follicular wave was earlier (PcO.05) in heifers treated
with a single injection of E-17P on Day 1 than control heifers. Mean day of emergence of the
second follicular wave in heifers treated with a single injection of E-17P on Day 3 and heifers
treated with twice daily injections on Days 1 to 3 were not different from that of control heifers.
The emergence of the second follicular wave was delayed in the group treated with 2.5 mg E-17P
on Days 3 to 5 compared to all the other treatment groups. The day of emergence of the second
follicular wave was less variable (P<O.Ol) in heifers treated with 5 mg of E-17P on Day 1 or Day
3 than in control heifers, and were not different from the remaining treatment groups.

The mean length of the interovulatory interval was not different among groups. However,
more (PcO.05) heifers treated with 2.5 mg E-17P on Days 3 to 5 (4 of 7, 57%) or 5 mg E-17P
on Day 3 (3 of 6, 50%) had short interovulatory intervals (< 17 d) compared to the controls
(0%). The incidence of heifers with short interovulatory intervals was intermediate in the
remaining treatment groups (range 14 to 33%). The proportion of heifers with three-wave
interovulatory intervals was not different between the control heifers and heifers treated with a
single injection of E-17P or 2.5 mg b.i.d on Days 1 to 3. Conversely, only 1 heifer treated with
2.5 mg E-17P on Days 3 to 5 had a three-wave interovulatory period of 23 d.

Diameter profiles of the dominant follicle in the first wave are shown in Figure 3. A time-
by-treatment interaction (PcO.01) was attributed primarily to a larger diameter (PcO.05) after Day
4 in control heifers than in those of the Day 1 or Days 1 to 3 treatment groups, and in those of
the Day 3 treatment groups after Day 5. The mean diameter profiles in heifers treated on Day
3 or Days 3 to 5 were larger that those of heifers treated on Day 1 or Days 1 to 3 (P<O.OS).
Mean diameter profiles were not different between Day 1 groups and between Day 3 groups,
respectively.

DISCUSSION

Results were consistent with those of previous studies which suggest that estradiol
suppresses the growing phase of the dominant follicle (6,7). The hypothesis that E-17P more
effectively suppresses the growth of the dominant follicle when administered with a progestogen
implant than when administered alone was supported. When E-17P (with SMB) was given on
Day 1 (Experiments 1 and 2), the dominant follicle of the first wave regressed and the emergence
of the second wave was hastened; a finding similar to that observed previously with estradiol
valerate (without SMB; 6). However, unlike estradiol valerate treatment, E-17b given on Day 3
(in the presence of SMB; Experiment 2) effectively suppressed the dominant follicle of the first
follicular wave and did not delay the emergence of the second follicular wave. The dominant
follicle stopped growing within 24 h of E-17P+SMB treatment in 10 of 13 (77%) heifers treated
on Day 1 and 5 of 6 (83%) heifers treated on Day 3. These results can be interpreted to suggest
that E-17p plus progestogen given in combination is effective in inducing follicle regression.

The hypothesis that E-17P treatment in SMB implanted heifers reduced variability in the
emergence of the second follicular wave was also supported. The day of emergence of the second
follicular wave was less variable in heifers treated with a single S-mg dose of E-17P than in
control heifers (Table 1). From a total of 13 SMB-implanted heifers treated with the 5 mg dose
of E-17P on Day 1 (Experiments 1 and 2), the emergence of the second wave occurred on Day
Table 2. Effect (mean*SEM) of exogenous estradiol-17P (E-17P) on follicle characteristics and wave emergence in heifers given a
progestogen (SMB) ear implant on Day 0 (day of ovulation; Experiment 2) ;;t
z

10 mg E-17P 5 mg E-17P 2.5 mg E-17P b.i.d.

Control Day 1 Day 3 Day 1 Day 3 Day l-3 Day 3-5
End point (n=7) (n=6) (n=7) (n=7) (n=6) (ll=6) (n=7)

InterovulatorV interval 2O.lrto.4 20.7+1 .O l&7+-1.0 17.Okl.3 19.2zk1.9 19.9kl.3 17.121.3

Prouortion of heifers with 3 waves 5Ra 5i6a .s/7a 4/7ab 4/6ab 5f6ab Ii+

Wave emergence (Day’)

First wave o.ozk0.2 o.o+o.o -0.1ko.l 0.020.0 0.3kO.3 o.ok0.2 0.3ztO.3

Second wave 7.7zkOsb 5.5k0.3a 6.9e0.3b 5.6ti.2a 6.8&.2b 7.6~0.2~ 8.8~0.2~

(Range) (6-10) (4-6) (6-8) (5-6) (6-7) (7-8) (8-10)

Third wave 14.4io.6 1 I .6&0.7 12.4kO.9 11.31to.7 14.0ztOto.6 15.2ti.3 16.0

Dominant follicle of first wave

Maximum diameter (mm) 11.2~.4c 9.8+0.4bC

Cessation of growth (Day) 5.6%0.Sd 3.040.1bc

Onset of regression (Day) 9.7s1.2= 5.0kO.5ab

E.
0

abc Means or proporti ons within a row with no common superscripts are different (P<O.OS). 9
 Theriogenology 1565

121 a)

lo-

8-
z
g
Q 6-
“u
z
Control (n=7)
z
2 4 -C- 10 mg E-170 Day 1 (n=6)
*...0”. 5 mg E-17RDay 1 (n=7)
.I
z -‘-.F” 2.5 mg E-170 b.i.d.Days l-3 (II=@
~2~“~‘*Q’1”““”
‘El 12
b)
3
3 10‘
;

8-

6-

4- - 10 mg E-17RDay 3 (n=7)
..
..“‘o”“” 5 mg E-170 Day 3 (n=6)
-‘-‘m-” 2.5 mg E-170 b.i.d.Days 3-5 (n=7)
2, ” 1 . 0. 1”. 0 ‘I”
-1 1 3 5 7 9 11 13 15
Number of days from ovulation

Figure 3. Diameter profiles (mean6EM) of the dominant follicle of the first wave in control
heifers and heifers treated with estradiol-17P (E-17P; Experiment 2). Graph a)
represents the control heifers and heifers treated with 5 mg or 10 mg E-17P on Day
1 or 2.5 mg b.i.d. on Days 1 to 3. Graph b) represents the control heifers and heifers
treated with 5 mg or 10 mg E-17P on Day 3 or 2.5 mg b.i.d. on Days 3 to 5. A time-
by-treatment interaction (P<O.Ol) was attributed primarily to a larger diameter (P<O.O5)
after Day 4 in control heifers than in those of the Day 1 or Days 1 to 3 treatment
groups, and in those of the Day 3 treatment groups after Day 5.
 1566 Theriogenology

5 (8 heifers) or Day 6 (5 heifers). Although both estradiol valerate and E-17P (with SMB) given
on Day 1 hastened the emergence of the second wave, the day of emergence in cows treated with
estradiol valerate in the previous study ranged from Day 5 to Day 9 (6). A single 5-mg injection
of E-17P given on Day 3 resulted in emergence of the second wave on Day 7 in 5 of 6 heifers
and on Day 6 in the remaining heifer (Experiment 2), whereas the emergence of the second wave
ranged from Day 9 to Day 14 in heifers treated with estradiol valerate on Day 3 (6). These
results supported the conclusion that delayed wave emergence in heifers treated with estradiol
valerate on Day 3 was a result of incomplete suppression of the dominant follicle and prolonged
suppressive effect of the valerate form of estradiol (6). Twice daily treatments of 2.5 mg E-17P
given on Days 3 to 5, which was intended to induce prolonged elevation in plasma estradiol
similar to estradiol valerate (approximately 5 d), also delayed wave emergence. Single estradiol
valerate treatment resulted in elevated plasma estradiol concentration for a period of 5 to 7 d (6);
whereas single E-17p treatment resulted in elevated plasma estradiol concentrations for only 42
h (1.75 d) in the present study (Figure 1).

The hypotheses that progestogen blocks the estradiol-induced LH release and that estradiol-
induced LH release is not associated with follicle suppression were supported. Despite a post-
treatment surge in LH, the dominant follicle was not completely suppressed in non-SMB
implanted heifers. Furthermore, the dominant follicle regressed in heifers that did not exhibit an
LH surge after E-17P injection. Blockage of the estradiol-induced LH surge by exogenous
progestogen, observed in the present study, is consistent with previous studies in which estrogen
treatments did not induce a preovulatory-like surge of LH during the luteal phase of the estrous
cycle (17,41) or in progestogen-treated cows (9). Concurrent post-treatment surges of LH and
FSH in 4 heifers treated with E-l7fl alone were likely mediated through the hypothalamus by
causing an increased release of GnRH (24).

The effect of E- 17p on post-treatment FSH concentrations differed between SMB-implanted

and non-implanted heifers. In SMB implanted heifers, in which the dominant follicle was
suppressed by E-17P treatment on Day 1, plasma FSH levels decreased by 6 h after E-17P
treatment and gradually increased over a period of approximately 24 to 42 h (Figure 1). In non-
SMB implanted heifers, plasma FSH levels also decreased for 6 h after E-17P treatment but
increased dramatically 12 h later. In association with the changes in FSH concentrations, the
growth of the dominant follicle in heifers treated with E-17P alone (no progestogen) was
apparently suppressed for 1 or 2 d but resumed thereafter (Figure 3). From these observations,
it appears that the suppression induced by E-17P alone was not prolonged enough to cause the
demise of the dominant follicle. This is consistent with the concept that follicle regression is not
an all-or-none process, but rather a result of a prolonged suppressive effect (4). Other
mechanisms, such as the negative effect of estradiol and progesterone on the pulsatile secretion
of LH must also be considered (33). Exogenous progesterone given during the growing phase
suppressed the dominant follicle (1) and high levels of progesterone/progestogen have been
associated with decreased LH pulse frequency and follicle regression (1,40,44). Collectively,
these observations provide the rationale for the hypothesis that the suppressive effect of estradiol
and progestogen in combination is due to suppression of both FSH and LH secretion and that
gonadotropin suppression must occur for at least 24 h, to elicit complete follicle regression.
 Theriogenology 1567

Profiles of plasma FSH concentrations and follicular wave emergence were temporally
related (Figure 2). In SMB-implanted heifers, early suppression of the dominant follicle allowed
an early increase in circulating FSH concentrations, which in turn elicited an early emergence
of the second follicular wave. Conversely, incomplete suppression of the dominant follicle
following E-17P treatment in heifers without SMB implants resulted in a delayed pre-wave FSH
surge and delayed emergence of the second follicular wave. These results are consistent with
those in which termination of the dominance phase by cauterization of the dominant follicle (25)
or treatment with a proteinaceous fraction of follicular fluid (23) was followed by a premature
FSH surge and early emergence of the next follicular wave. The association between increases
in FSH concentration and wave emergence observed in the present study, is consistent with
previous studies in which a surge in plasma FSH was detected prior to the emergence of each
wave in heifers with 2-wave or 3-wave cycles (2).

In summary, exogenous estradiol suppressed growth of the dominant follicle, but the effect
was more consistent when combined with a progestogen ear implant. A single 5 mg dose of E-
17p was as effective as higher or repeated dosage regimens in inducing follicular suppression and
resulted in consistent emergence of a new follicular wave 4 to 5 d later. The mechanism by
which exogenous estradiol induces follicle suppression remains to be elucidated; however results
of this study provide rationale for the hypothesis that the effect is mediated through suppression
of gonadotropin. Estradiol-induced LH release was not associated with follicle suppression, but
a direct effect of estradiol on ovarian follicles cannot be ruled out. Results from this study
suggest that treatment with E-17P and progestogen, in combination, may be useful to manipulate
ovarian follicular development in cattle. Results provide impetus for a study to evaluate the
efficacy of E-17P plus progestogen treatment for synchronizing wave emergence in randomly
selected cycling cattle.

REFERENCES
1. Adams GP, Matteri RL, Ginther OJ. Effect of progesterone on growth of ovarian follicles, emergence of
follicular waves and circulating FSH in heifers. J Reprod Fertil 1992; 95:627-640.
2. Adams GP, Matteri RL, Kastelic JP, Ko JCH, Ginther OJ. Association between surges of follicle
stimulating hormone and the emergence of follicular waves in heifers. J Reprod Fertil 1992; 94: 177.188.
3. Adams GP, Kot K, Smith CA, Ginther OJ. Selection of a dominant follicle and suppression of follicular
growth in heifers. Anim Reprod Sci 1993; 30:259-271.
4. Adams GP, Kot K, Smith CA, Ginther OJ. Effect of the dominant follicle on regression of its subordinates.
Can J Anim Sci 1993; 73:267-275.
5 Armstrong DT. Recent advances in superovulation of cattle. Theriogenology 1993; 39:7-24.
6: Bo GA, Nasser LF, Adams GP, Pierson RA. Mapletoft RJ. Effect of estradiol valerate on ovarian follicles
emergence of follicular waves and circulating gonadotropin in heifers. Theriogenology 1993; 40:225-239.
I. Bo GA, Pierson RA, Mapletoft RJ. The effect of estradiol valerate on follicular dynamics and
superovulatory response in cows with Syncro-Mate-B implants. Theriogenology 1991; 36:169-183.
8. Boland M, Gouldin D, Roche JF. Alternative gonadotropin for superovulation in cattle. Theriogenology
1991; 35:5-18.
9. Bolt DJ, Scott V, Kiracofe GH. Plasma LH and FSH after estradiol, norgestomet and Gn-RH treatment
in ovariectomized beef heifers. Anim Reprod Sci 1990; 23:263-271.
 1566 Theriogenology

10. Engelhart H, Walton JS, Miller RB, King GJ. Estradiol induced blockade of ovulation in the cow: effect
of luteinizing hormone release and follicular fluid steroids. Biol Reprod 1989; 40:1287-1297.
11. Evans ACO, Currie WD, Rawlings NC. Effects of naloxone on circulating gonadotropin concentrations
in prepubertal heifers. J Reprod Fertil 1992; 96: 847855.
12. Findlay JK, Robertson DM, Clarke IJ, Klein R, Doughton BW, Xiao S, Russel DL, Shukovski L.
Hormonal regulation of reproduction: General concepts. Anim Reprod Sci 1992; 28:3 19-328.
13. Gill JL, Hafs HD. Analysis of repeated measurements in animals. J Anim Sci 1971; 33:331-336.
14. Ginther OJ, Kastelic JP, Knopf L. Temporal associations among ovarian events in cattle during estrous
cycles with two and three follicular waves. J Reprod Fertil 1989; 87:223-230.
15. Ginther OJ, Kastelic JP, Knopf L. Composition and characteristics of follicular waves during the bovine
estrous cycle. Anim Reprod Sci 1989; 20: 187-200.
16. Guilbault LA, Grass0 F, Lussier JG, Matton P, Rouillier P. Decreased superovulatory responses in heifers
superovulated in presence of a dominant follicle. J Reprod Fertil 1991; 91:81-89.
17. Hobson WC, Hansel W. Plasma LH levels after ovariectomy, corpus luteum removal and estradiol
administration in cattle. Endocrinology 1972; 91: 185190.
18. Huhtinen M, Raino V, Aalto J, Bredbacka P. Increased ovarian responses in the absence of the dominant
follicle in superovulated cows. Theriogenology 1992; 37:457-463.
19 Hutz RJ, Diershke DJ, Wolf RC. Induction of atresia of the dominant follicle in rhesus monkeys by the
local application of estradiol-17b. Am J Primatol 1988; 15:69-77.
20 Ireland JJ. Control of follicular growth and development. J Reprod Fertil Suppl 1987; 34:39-54.
21 Joseph IBJK, Currie WD, Rawlings NC. Effects of time after ovariectomy, season and &radio1 on
luteinizing hormone and follicle stimulating hormone secretion in ovariectomized ewes, J Reprod Fertil
1992; 94:511-523.
22 Kastelic JP, Knopf L, Ginther OJ. Effect of day of prostaglandin F2 alpha treatment on selection and
development of the ovulatory follicle in heifers. Anim Reprod Sci 1990; 23: 169-180.
23 Kastelic JP, Ko JCH and Ginther OJ. Suppression of dominant and subordinate ovarian follicles by a
proteinaceous fraction of follicular fluid in heifers. Theriogenology 1990; 34:499-509.
24 Kesner JS, Convey EM. Interaction of estradiol and luteinizing hormone releasing hormone on follicle
stimulating hormone release in cattle. J Anim Sci 1982; 54:817-821.
25 Ko JCH, Kastelic JP, Del Campo MR and Ginther O.J. Effects of a dominant follicle on ovarian follicular
dynamics during the estrous cycle in heifers. 1991; J Reprod Fertil 91:51 l-519.
26 Knopf L, Kastelic JP, Schallenberger E, Ginther OJ. Ovarian follicular dynamics in heifers: test of two-
wave hypothesis by ultrasonically monitoring individual follicles. Dom Anim Endocr 1989; 6: 11 l-l 19.
21. Loob DK, Dorrington J. Intraovarian regulation of follicular development, Anim Reprod Sci 1992; 28:343-
354.
28 MacMillan KL, Thatcher WW. Effects of an agonist of gonadotropin releasing hormone on ovarian
follicles in cattle. Biol Reprod 1991; 45:883-889.
29 Mapletoft RJ, Bo GA, Murphy BD. The effect of biological activity of gonadotropin on superovulation
in the cow. Proc IX Congreso Brasileiro de Reproducao Animal 1991; 1:74-92.
30 Monniaux D, Chupin D, Saumande J. Superovulatory responses of cattle. Theriogenology 1983; 19:55-81.
31 Nasser LF, Adams GP, Bo GA, Mapletoft RJ. Ovarian superstimulatory response relative to follicular
wave emergence in heifers. Theriogenology 1993; 40:713-724.
32 Pierson RA, Ginther OJ. Follicular populations during the estrous cycle in heifers III. Time of selection
of the ovulatory follicle. Anim Reprod Sci 1988; 16:81-95.
33 Price CA, Webb R. Steroid control of gonadotropin secretion and ovarian function in heifers.
Endocrinology 1988; 122:2222-223 1.
 Theriogenolog y 1569

34. Rajamahendran R, Walton JS. Effect of treatment with estradiol valerate on endocrine changes and ovarian
follicle populations in dairy cows. Theriogenology 1990; 33:441-452.
35. Refsal KR, Jarrim-Maldonado JH, Nachreiner RF. Endocrine profiles in cows with ovarian cysts
experimentally induced by treatment with exogenous estradiol or adrenocorticotrophic hormone.
Theriogenology 1987; 28:87 1-889.
36. Roche RJ, Boland MP. Turnover of dominant follicles in cattle of different reproductive states,
Theriogenology 1991; 35:81-90.
37. SAS User’s Guide: Statistics, SAS Institute Inc., Cary, NC, 1989.
38. Savio JD, Boland MP, Hynes N, Mattiacei MR. Roche JF. Will the first dominant follicle of the estrous
cycle of heifers ovulate following luteolysis on Day 7? Theriogenology 1990; 33:677-687.
39. Savio JD, Keenan L, Boland MP, Roche JF. Pattern of growth of dominant follicles during the oestrous
cycle in heifers. J Reprod Fertil 1988; 83:663-671.
40. Savio JD, Thatcher WW, Badinga L, De la Sota RL, Wolfenson D. Regulation of dominant follicle
turnover during the oestrus cycle in cows. J Reprod Fertil 1993; 97:197-203.
41. Short RJS, Randel RD, Staigmiller RB, Bellows RA. Factors affecting estrogen-induced LH release in the
cow. Biol Reprod 1979; 21:683-689.
42. Sirois J, Fortune JE. Ovarian follicular dynamics during the estrous cycle in heifers monitored by real-time
ultrasonography. Biol Reprod 1988; 39:308-317.
43. Steel RGD, Torrie JH. Principles procedures of Statistics. MC Graw-Hill Inc., New York, 1980; 471-472.
44. Stock AE, Fortune JE. Ovarian follicular dominance in cattle: Relationship between prolonged growth of
the ovulatory follicle and endocrine parameters. Endocrinology 1993; 132: 1108-l 114.

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