Lauia Biuce

BI0L 47S, Section uu9
TA: Khoa Nguyen
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The complex inteiaction between the biain anu
the gonaus is facilitateu by the hypothalamic-pituitaiy-
gonaual (BPu) axis. This system is ciitical to the
uevelopment anu function of the iepiouuctive system anu
oveiall homeostatic iegulation (Taniiveiui et al., 2uuS).
Eviuently, the BPu axis ielates the hypothalamus anu
anteiioi pituitaiy glanu in the biain to the gonaus via
hoimone signaling. In females, the BPu is also known as
the biain-ovaiian axis, as it functions to maintain iegulate
the ielease of oocytes (foimeu befoie biith) fiom the ovaiies (Wateis, 2u14).
Figuie 1 iepiesents a schematic of the hoimonal inteiactions that occui. In the biain, the
hypothalamus ieleases a hoimone known as gonauotiopin-ieleasing hoimone (unRB), which
stimulates the anteiioi pituitaiy glanu to ielease follicle stimulating hoimone (FSB) anu luteinizing
hoimone (LB). These gonauotiopins tiavel thiough the blooustieam to the female gonaus, the
ovaiies (Wateis, 2u14). Beie, FSB anu LB inuuce the uevelopment of the follicle, as well as the
actual ielease of the oocyte in ovulation. Aftei ovulation, the iuptuieu follicle, known as the coipus
luteum, will maintain levels of estiogen, piogesteione, anu inhibin by the ovaiy. Estiogen anu
piogesteione aie iesponsible foi maintaining piimaiy sexual chaiacteiistics such as piepaiation of
the uteius foi implantation, as well as seconuaiy sexual chaiacteiistics such as bieast uevelopment.
Neanwhile, both estiogen anu inhibin will inhibit the ielease of unRB anu FSB¡LB in the biain via
negative feeuback. This pievents a new follicle fiom beginning to uevelop until the fate of the
iecently ieleaseu oocyte is ueteimineu, which coulu involve feitilization (Wateis, 2u14).
Impoitantly, the levels of these ciiculating iepiouuctive hoimones can be manipulateu by
suigical iemoval of one oi both of the gonaus, which uisiupts the negative feeuback loops uue to
Figuie 1. Repiouuctive Boimones
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Biuce 2
the loss of gonaual hoimones. The puipose of this laboiatoiy is to investigate the effects of
unilateial anu bilateial ovaiiectomy on the othei BPu axis oigans in female iats as inuicateu by
oigan weight. It is hypothesizeu that aftei a unilateial ovaiiectomy, iats will exhibit an inciease in
mean ovaiian weight in the iemaining ovaiy, but no uiffeience in uteiine hoin oi anteiioi pituitaiy
weight as compaieu to a contiol. Aftei a bilateial ovaiiectomy, it is hypothesizeu that iats will
exhibit a ueciease in mean uteiine hoin weight, anu no uiffeience in anteiioi pituitaiy weight as
compaieu to a contiol. Accoiuingly, the iats in the bilateial gioup aie hypothesizeu to exhibit a
uecieaseu mean uteiine hoin weight when compaieu to the unilateial gioup.
When an ovaiy is iemoveu, estiogen levels aie halveu, anu the iemaining ovaiy expeiiences
what is known as "ovaiian compensatoiy hypeitiophy", oi an inciease in size uue to enlaigement of
component cells (Pepplei et al., 1971). This occuis because negative feeuback to the hypothalamus
anu anteiioi pituitaiy by estiogen is immeuiately uecieaseu, iesulting in an incieaseu ielease of
unRB to the anteiioi pituitaiy anu incieaseu FSB anu LB to the iemaining ovaiy (Pepplei et al.,
1971). In the case of a bilateial tieatment, uteiine hoin weight is expecteu to ueciease, because
theie will be no ielease of estiogen to maintain uteiine function in piepaiation foi feitilization. No
othei uiffeiences aie expecteu. Notably, the anteiioi pituitaiy is not expecteu to exhibit changes in
size aftei eithei pioceuuie, as it has othei highly iegulateu functions ielateu to stiess maintenance
anu giowth (Wateis, 2u14).
This expeiiment has significant implications, as oophoiectomy, oi the iemoval of ovaiies in
human females, is often useu to eliminate ovaiian cysts anu ovaiian cancei tumois (Kuiian et al.,
2u1u). In conjunction with a iemoval of the uteius, bilateial oophoiectomy can also be useu as a
pieventative measuie foi those who aie genetically uisposeu to ueveloping bieast anu ovaiian
cancei. While these suigeiies can help patients to avoiu cancei anu cysts, they have many negative
siue effects incluuing uecline in sexual function, inuuceu menopausal symptoms, osteopoiosis,
uementia, anu caiuiovasculai uisease (Shustei et al., 2uu8). Stuuying the effects of ovaiiectomy in
iats may help scientists anu clinicians to unueistanu the mechanisms behinu these siue-effects in
human patients, so that they may be avoiueu oi alleviateu.
Biuce S
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In scientific ieseaich settings involving laboiatoiy animals, a unilateial ovaiiectomy is
uefineu as the suigical iemoval of one ovaiy, while a bilateial ovaiiectomy is uefineu as the suigical
iemoval of both ovaiies (Wateis, 2u14). In this laboiatoiy, both types of ovaiiectomies weie
peifoimeu on female iats, as well as sham suigeiies to seive as a contiol. Piioi to anu uuiing
suigeiy, uosages of anesthesia, incluuing ketamine, xylazine, anu bupivacaine, weie auministeieu
to the iats accoiuing to piotocol. Steiile pioceuuie was maintaineu thioughout the piepaiation foi
suigeiy, the suigical peiiou, anu post-opeiative caie. Thiee weeks aftei the suigeiies weie
completeu, the female iats weie euthanizeu anu a neciopsy was peifoimeu. Any iemaining ovaiies,
the uteiine hoin, anu the anteiioi pituitaiy glanu weie iemoveu fiom each iat anu weigheu
(Wateis, 2u14).
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Figuie 2. Effects of 0nilateial 0vaiiectomy on Nean 0vaiian Weight

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Figuie 2 compaies the mean ovaiian weights in female iats thiee weeks aftei a contiol
sham suigeiy oi unilateial ovaiiectomy. In iats ieceiving the contiol tieatment, the mean ovaiian
weight was founu to be u.12u19 giams, anu in those ieceiving the unilateial tieatment, the mean
weight of the iemaining ovaiy was founu to be u.14711 giams. Nean ovaiian weight in the
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Biuce 4
unilateial tieatment gioup C5< .%) founu to be significantly uiffeient than that of the contiol gioup
(t=2.2779, uf=4u, u2=2.7u4), although a slight inciease was obseiveu.

Figuie S. Effects of 0nilateial anu Bilateial 0vaiiectomy on Nean 0teiine Boin Weight

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Figuie S compaies the mean uteiine hoin weights in female iats thiee weeks aftei a contiol
sham suigeiy, unilateial ovaiiectomy, oi bilateial ovaiiectomy. In iats ieceiving the contiol
tieatment, the mean uteiine hoin weight was founu to be u.S8u1S giams. In those ieceiving the
unilateial anu bilateial tieatments, the mean uteiine hoin weights weie founu to be u.S68uu
giams, anu u.1SS89 giams, iespectively. Nean uteiine hoin weight in the unilateial tieatment
gioup C5< .%) founu to be significantly uiffeient than that of the contiol gioup (t=u.1994, uf=4u,
u2=2.7u4), uespite a small ueciease. Nean uteiine hoin weight in the bilateial tieatment gioup
C5< founu to be significantly lowei than that of the contiol gioup (t=S.2479, uf=S1, u2=2.744).
Similaily, mean uteiine hoin weight in the bilateial tieatment gioup C5< founu to be significantly
lowei than that of the unilateial tieatment gioup (t=8.67u6, uf=2S, u2=2.787).

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Biuce S
Figuie 4. Effects of 0nilateial anu Bilateial 0vaiiectomy on Nean Anteiioi Pituitaiy Weight

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Figuie 4 compaies the mean anteiioi pituitaiy weights in female iats thiee weeks aftei a
contiol sham suigeiy, unilateial ovaiiectomy, oi bilateial ovaiiectomy. In iats ieceiving the contiol
tieatment, the mean anteiioi pituitaiy weight was founu to be u.u17uu giams. In those ieceiving
the unilateial anu bilateial tieatments, the mean anteiioi pituitaiy weights weie founu to be
u.u2S47 giams, anu u.u1S71 giams, iespectively. Nean anteiioi pituitaiy weight in the unilateial
tieatment gioup C5< .%) founu to be significantly uiffeient than that of the contiol gioup
(t=u.914S, uf=S4, u2=2.728), uespite an inciease. Similaily, mean anteiioi pituitaiy weight in the
bilateial tieatment gioup C5< .%) founu to be significantly uiffeient than that of the contiol gioup
(t=u.7S2u, uf=24, u2=2.797), uespite an obseiveu ueciease. Nean anteiioi pituitaiy weight in the
bilateial tieatment gioup C5< .%) founu to be significantly uiffeient than that of the unilateial
tieatment gioup (t=u.89S9, uf=22, u2=2.819), though it is lowei.
Foi all compaiisons, statistical uiffeiences weie ueteimineu using the obseiveu anu ciitical
t-values fiom an unpaiieu, two-taileu t-test, accoiuing to the Bonfeiioni Nethou with u=u.u1.
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Biuce 6
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Nost, but not all of the iesults of this laboiatoiy pioviue suppoit foi the hypotheses. Foi
the unilateial tieatment gioup, no significant uiffeiences in mean ovaiian, uteiine hoin, oi anteiioi
pituitaiy weights weie obseiveu when compaieu to the contiol gioup. Foi the uteiine hoin anu
anteiioi pituitaiy, these iesults stiongly coinciue with the pieuictions foi consistency. Yet, foi
ovaiian weight, the iesults contiauict the hypothesis that a significantly incieaseu mean ovaiian
weight woulu be obseiveu in the iemaining ovaiy. Since a slight inciease was obseiveu, the lack of
significance may likely be uue to the small sample size, oi the veiy low weight of the ovaiies, which
woulu have maue uiffeiences uifficult to iuentify.
Foi the bilateial tieatment gioup, no significant uiffeience in mean anteiioi pituitaiy
weight was obseiveu, but a stiongly significant ueciease in mean uteiine hoin weight was obseiveu
when compaieu to both the contiol anu the unilateial tieatment gioup. These iesults stiongly
suppoit the hypotheses that no change woulu occui in the anteiioi pituitaiy, anu that without
ovaiies to maintain estiogen levels, uteiine hoin weight woulu ueciease because the uteius woulu
no longei be maintaineu foi potential feitilization.
Souices of eiioi in this expeiiment aie ielateu to expeiimental uesign anu technique.
Bue to inheient uiffeiences between the suigeons in theii suigical skill, the alieauy unique iats
may have expeiienceu vaiieu amounts of stiess anu even vaiieu
amounts of anesthesia uepenuing on the piogiession of theii pioceuuie. Fuithei, uuiing the
neciopsy, oigans weie likely not iemoveu in a consistent mannei, which woulu have affecteu the
uata. This is eviuent when consiueiing that one of the iepoiteu anteiioi pituitaiy weights was ten
times gieatei than the othei uata points. In this case, the uesiieu glanu was likely weigheu with
excess tissue attacheu. Bowevei, omitting this uata point woulu not have leu to significant iesults.
To ieuuce these types of eiiois, a moie piecise technique is necessitateu, along with a gieatei
consistency in execution of the suigeiy anu neciopsy.
Biuce 7
To impiove the expeiiment, changes othei than oigan weight coulu have been measuieu to
evaluate the effects of the ovaiiectomies. While oigan weight was expecteu to inuicate a change in
oigan function, it may not have been the best inuicatoi, especially since anteiioi pituitaiy weight
was not pieuicteu to change. As seen in pievious stuuies, FSB, LB, anu estiogen concentiations in
the anteiioi pituitaiy coulu have been measuieu insteau, as these aie accuiate inuicatois of
function (Pepplei et al., 1972). Auuitionally, the ovaiian cycles of the iats coulu have been
monitoieu, since follicle matuiation may have also influenceu mean ovaiy weight in the unilateial
anu contiol gioups.
Although theie is ioom foi impiovement in this expeiiment, anu although the not all of the
iesults weie expecteu, this uata uoes contiibute to the unueistanuing of female iepiouuctive
enuociinology. Specifically, the uata uoes suppoit the notion that a bilateial ovaiiectomy leaus to
uecieaseu uteiine function as exhibiteu by the significant ueciease in uteiine hoin weight when
compaieu to the contiol anu the unilateial tieatments. This ueciease in uteiine function is
connecteu to uecieaseu levels of estiogen as a iesult of ovaiy iemoval. Clinically, this infoimation
is ielevant to patients who may neeu to unueigo a bilateial oophoiectomy. Since estiogen also
iegulates seconuaiy sexual chaiacteiistics anu othei bouily functions such as bone uensity, it is
cleai that patients unueigoing this pioceuuie can benefit fiom hoimone ieplacement theiapy to
ieintiouuce estiogen into theii system (Aimstiong et al., 2uu4). 0nfoitunately, hoimone theiapy
has not pioven to be entiiely effective at eliminating the iisks anu siue effects associateu with
oophoiectomy (Kuiian et al., 2u1u). Thus, it is even moie ciitical that we puisue expeiiments like
this in oiuei to iuentify the mechanisms behinu iepiouuctive enuociinology iegulation anu the
biain-ovaiian axis. If we aie successfully, life-saving oophoiectomy pioceuuies will be less
uepieciateu by theii unuesiiable, common siue effects.

Biuce 8
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Aimstiong, Katiina, et al. "Boimone ieplacement theiapy anu life expectancy aftei piophylactic
oophoiectomy in women with BRCA1¡2 mutations: a uecision analysis." G#$%=+. #B &.7=7&+.
#=&#.#8A 22.6 (2uu4): 1u4S-1uS4.
Kuiian, Allison W., Bionislava N. Sigal, anu Sylvia K. Pleviitis. "Suivival analysis of cancei iisk
ieuuction stiategies foi BRCA1¡2 mutation caiiieis."G#$%=+. #B H.7=7&+. I=&#.#8A 28.2
(2u1u): 222-2S1.
Pepplei, Richaiu B. "Effects of unilateial ovaiiectomy on folliculai uevelopment anu ovulation in
cycling, ageu iats." J<'%7&+= G#$%=+. #B J=+,#<A 1S2.4 (1971): 42S-427.
Pepplei, Richaiu B. "FSB anu LB levels in the intact anu unilateially ovaiiectomizeu cycling
iat." J&,+ '=9#&%7=#.#87&+ 69.2 (1972): 267-28u.
Shustei, Lynne T., et al. "Piophylactic oophoiectomy in piemenopausal women anu long-teim
health." K'=#:+$-' 7=,'%=+,7#=+. 14.S (2uu8): 111-116.
Taniiveiui, F., et al. "The hypothalamic-pituitaiy-gonaual axis: immune function anu
autoimmunity." G#$%=+. #B L=9#&%7=#.#8A 176.S (2uuS): 29S-Su4.
Wateis, }ohn R. anu Tomicek, Nanette }. Physiology Laboiatoiy Nanual. M+A9'=NK&O'7. P$@.7-;7=8.
Plymouth, NI. (2u14): 76-1u1.