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Effects of a Unilateral or Bilateral Ovariectomy on a

Female Rats Reproductive Endocrinology

Alexandra Digenakis
Biology 473
TA: Aaron Montani
06 April 2014

Introduction:
A relationship between the brain and the gonads exists in which the hypothalamus,
pituitary gland and gonads interact in the regulation of sperm production (spermatogenesis) in
males and oocyte release (ovulation) in females.1 The experiment being addressed focuses
specifically on the anatomy and physiology of a female rat. In females the hypothalamus
produces gonadotropin releasing hormone (GnRH) for a brief time.1 The release of GnRH
stimulates the anterior pituitary gland to secrete the luteinizing hormone (LH) and the follicle
stimulating hormone (FSH).1 Both of these secreted hormones are important in the regulation of
female and male gonads. Specifically in females, the FSH and LH hormones are responsible for
beginning ovulation and stimulating estrogen production in the ovary through the development
of the ovarian follicle.1 After ovulation occurs the remaining part of the follicle remains inside
the ovary to go through luteinization in order to form the corpus luteum.1 The corpus luteum
functions in maintaining estrogen and progesterone levels as well as maintaining these hormones
until the placenta (as long as fertilization occurred) is capable to take over the corpus luteums
role.1 Estrogen and progesterone hormones function in the regulation of FSH, LH and GnRH
production; also, estrogen and progesterone function in the preparation and maintenance of the
uterus so a fertilized egg can safely implant.1 Throughout the ovarian cycle a negative feedback
effect occurs when rising levels of estrogen and inhibin act on the hypothalamus and anterior
pituitary which inhibits further GnRH, FSH or LH release.1
This experiment is important in understanding how an ovarectomy and loss of the ovarian
hormones affects the hypothalamic-pituitary-ovarian axis organs. The investigation into the
effect of a unilateral or bilateral ovarectomy is important to physicians who have patients that
had an ovary removed for health reasons. Its also important to investigate how an ovarectomy
affects hormone release and to determine if fertility is impacted.
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It is hypothesized that after a unilateral ovarectomy the remaining ovary of the rat will try
to compensate for the loss of the ovary and the ovarian hormones; therefore, this will cause the
remaining ovary to increase in weight. Also, its hypothesized that there will not be a significant
difference in the weight of the uterine horn after a unilateral ovarectomy or if neither of the
ovaries are removed. This is because the uterine horns function will not be hindered after the
removal of one ovary. However, it is hypothesized that the uterine horn after a bilateral
ovarectomy will weigh less compared to the weight of a uterine horn after a unilateral
ovarectomy or a sham surgery. The reasoning for the hypothesis is that when both ovaries are
removed the uterine horn is no longer functional in the development of the fetus and therefore
will begin to atrophy from lack of use.
Its also hypothesized that the pituitary gland will be the heaviest after the sham surgery
because no changes in the physiology are made. Its predicted that the pituitary gland after the
unilateral ovarectomy will weigh less compared to the control pituitary gland because the
removal of an ovary decreases the amount of estrogen released, in turn decreasing the amount of
GnRH, FSH and LH secreted; this results in a decrease in the weight of the pituitary gland. The
pituitary gland after the bilateral ovarectomy will weigh the least amount compared to the control
and unilateral groups because the estrogen release is inhibited from the gonad. Therefore this
causes the GnRH, FSH and LH to be inhibited, further decreasing the weight of the pituitary
gland.
Methods and Materials:
The protocol from this experiment was followed from the Physiology Lab Manual.1 The
possible surgical treatments included: a sham surgery with neither gonad removed, a unilateral
ovarectomy with one gonad removed or a bilateral ovarectomy with the removal of both gonads.1

Sterile procedure was used during preparation of the Sprague-Dawley female rats and during the
surgical procedures.1 The rats were placed under anesthesia during the duration of the surgical
preparation and throughout the surgery. The rats remained alive for two weeks and then were
placed under anesthesia and euthanized through cervical dislocation. After the rats were
euthanized the remaining ovary from a unilateral ovarectomy or both ovaries from a sham
surgery were autopsied, weighed and the average weight for one ovary was calculated. The
uterine horn and pituitary gland from all the rats were autopsied and weighed. Graphs comparing
the average weight of the three organs autopsied and the different procedures were created in
excel. Data analysis in order to test for significance was completed in Minitab using a 2-sample,
2-tailed t-test with a 95% confidence level and a 0.05 alpha value.
Results:
Table 1: T-test results of mean ovarian weights comparing a sham and unilateral surgery
Control vs. unilateral
calculated t-value:
2.21
critical t-value:
2.037
df:
32
p-value:
0.034
significance:
significant
Table 1 displays calculated t-value, critical t-value, degrees of freedom, p-value and significance
from a t-test of mean ovarian weights when comparing a control (sham) and a unilateral
ovariectomy.

Table 2: T-test results of mean uterine horn weights comparing different procedures
Control vs.
Control vs.
Unilateral vs.
unilateral
bilateral
bilateral
calculated t-value:
0.22
7.32
9.34
critical t-value:
2.028
2.042
2.093
df:
36
30
19
p-value:
0.83
0
0
significance:
not significant
significant
significant
Table 2 displays the calculated t-value, critical t-value, degrees of freedom, p-value and
significance from a t-test of mean uterine horn weights from a control and a unilateral
ovariectomy; a control and bilateral ovariectomy; and a unilateral and bilateral ovariectomy.
Table 3: T-test results of mean pituitary gland weights comparing different procedures
Control vs.
Control vs.
Unilateral vs.
unilateral
bilateral
bilateral
calculated t-value:
0.88
1.08
1.38
critical t-value:
2.086
2.069
2.11
df:
20
23
17
p-value:
0.391
0.292
0.184
significance:
not significant
not significant
not significant
Table 3 displays the calculated t-value, critical t-value, degrees of freedom, p-value and
significance from a t-test of mean pituitary gland weights from a control and a unilateral
ovariectomy; a control and bilateral ovariectomy; and a unilateral and bilateral ovariectomy.

Average gonad weight vs. type of procedure

Weight of gonad (g)

0.16
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
Control

Unilateral
Type of procedure

Figure 1 displays a graph of average gonad weight after a control (sham) surgery and after a
unilateral ovariectomy. The graph also includes standard error bars.

Average uterine horn weight vs. type of procedure


0.7
0.6
0.5
0.4
Weight of uterine horn (g)

0.3
0.2
0.1
0
Control

Unilateral

Bilateral

Type of procedure

Figure 2 displays a graph of average uterine horn weight after a control (sham) surgery, a
unilateral ovariectomy or a bilateral ovariectomy. The graph also includes standard error bars.

Average pituitary weight vs. type of procedure


0.03
0.02
0.02
Weight of pituitary (g)

0.01
0.01
0
Control

Unilateral

Bilateral

Type of procedure

Figure 3 displays the graph of average pituitary weight after a control (sham) surgery, a
unilateral ovariectomy or a bilateral ovariectomy. The graph also includes standard error bars.
The results in figure 1 show that the average ovarian weight for the rats was slightly
heavier after a unilateral surgery compared to a sham surgery. Table 1 shows that the results of
the ovarian weights for the rats in the control and unilateral treatments were significantly
different. Figure 2 displays that the average weight of the uterine horn was about the same for a
control and unilateral surgery; the average weight of the uterine horn was much lighter in
comparison to the control and unilateral surgeries. Table 2 shows that there was no significant
difference in the mean uterine horn weight for the control and unilateral treatments. The results
in table 2 display that there was a significant difference between the mean uterine horn weights
between the control and bilateral treatments; also there was a significant difference between the
unilateral and bilateral treatments. The average pituitary weight as displayed in figure 3
displayed that the pituitary gland weight is heaviest after the unilateral surgery; the weight of the
pituitary glands were very similar after the control and bilateral surgeries. Table 3 displays that
there is no significant difference in the mean pituitary gland weight for the control and unilateral
treatments; the control and bilateral treatments; and the unilateral and bilateral treatments.
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Discussion:
The purpose of this experiment was to investigate the affect that removing one or both
ovaries would have on a remaining ovary, the uterine horn and the pituitary gland. The results
support some of the hypotheses made prior to the experiment. It was hypothesized that the
remaining gonad after a unilateral ovarectomy would weigh more than an ovary from a rat with
neither ovary removed. Table 1 shows that there was a significant difference with a p-value less
than 0.05 between mean ovarian weights comparing a sham and unilateral surgery. The data
strongly supports this hypothesis with a p-value of 0.034; also figure 1 show that the ovary after
the unilateral surgery weighed more compared to the ovary of the sham surgery.
Another hypothesis was made that said that there would not be a significant difference in
the weight of the uterine horn after a unilateral ovarectomy or when neither ovary was removed.
The data strongly supports this hypothesis because the p-value was 0.83 indicating that there was
not a significant difference in the weight of the uterine horn after these two procedures. In
comparison, another hypothesis was made stating that the uterine horn following a bilateral
ovarectomy would weigh less compared to after both the unilateral ovarectomy and the sham
surgery. This hypothesis was strongly supported by the results found in table 2 which indicated
that there was a significant difference between bilateral and unilateral as well as bilateral and
control with p-values of 0. Figure 2 visually shows that there was not a difference between the
uterine horn weight within the control and unilateral groups; however, the bilateral group had a
significantly smaller weight in comparison.
It was originally hypothesized that the pituitary gland would be heaviest after the sham
surgery in comparison to after a unilateral or bilateral ovarectomy. However, the results did not

support this hypothesis and strongly indicated that there was not a significant difference for
either comparison. Figure 3 shows that the average pituitary weight was slightly heavier after the
unilateral ovarectomy compared to the control group; this indicates that the removal of one ovary
has some effect on the pituitary weight, although the results did not turn out to be significant.
In addition, it was also hypothesized that the pituitary gland after the bilateral
ovarectomy would weigh the least compared to the unilateral ovarectomy and sham surgery
results. The results strongly refuted this hypothesis with non-significant p-values of 0.184 and
0.292. It was hypothesized that if both ovaries were removed, estrogen would not be released
from the gonads; therefore, preventing the pituitary gland from producing LH and FSH. The
logic was that this would in turn cause the pituitary gland weight to decrease. A possible reason
that there was not a significant difference with any of the pituitary gland data was because the
time after the surgery was only two weeks. Its possible that waiting a longer period of time after
the surgery in order to autopsy the rats would result in more significant outcomes.
Obvious sources of error occurred in this experiment; for instance, when the organs were
weighed they still contained some fat. This couldve caused an inflated weight because it was
difficult to remove all of the fat before weighing it. Another error occurred because the amount
of subjects in the control, unilateral and bilateral groups were very uneven. For example, the
bilateral group had a lot fewer subjects tested compared to the control group. This likely caused
inaccuracies in the data.
In order to minimize error, an equal number of rats per test group should be used. It
should also be taken into account if a rat dies from a specific test group then data from one more
rat in each of the two test groups should be randomly removed in order to ensure there is an

equal number of subjects per test group. Also, error can be reduced by taking more care to
remove excess fat from the organs before weighing them.
It can be concluded that removing one ovary does not have a significant effect on the
function of the other ovary or the uterine horn. This information would be important for doctors
who consult patients with a unilateral ovarectomy to understand and inform their patients. The
results suggested that removing both ovaries however would have a significant impact on the
uterine horn. This could be important for patients who are considering being a surrogate whove
previously had a bilateral ovarectomy. Its likely that having both ovaries removed would hinder
their ability to carry a fetus in the uterus because as seen in this experiment a bilateral
ovarectomy causes a significant decrease in the weight of the uterine horn
References:
1

Waters, J.; Tomicek, N. Physiology: Laboratory Manual,1st ed; Hayden-McNeil: Michigan,


2014; pg. 76-101.

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