Lecture 2: REPRODUCTIVE ENDOCRINOLOGY
Dr. Jen Lesiguez | February 2021
OUTLINE:
I. HYPOTHALAMUS AND GnRH
II. ANTERIOR PITUITARY AND GONADOTROPINS
III. OVARIES
IV. OVARIAN-HYPOTHALAMIC-PTUITARY FEEDBACK
LOOPS
V. THE MENSTRUAL CYCLE
VI. THE MENSTRUAL CYCLE AND THE
ENDOMETRIUM
VII. MENSTRUAL CYCLE AND CERVICAL GLANDS
VIII. REFERENCES
IX. APPENDIX
HYPOTHALAMUS AND GnRH
• Reproductive process starts in the brain
• Initial hormone GnRH stimulates pituitary gland to
secrete gonadotropins>ovaries to secrete estrogen
• GnRH synthesizing neurons are not found in the brain
• Derive from progenitor cells of the embryonic olfactory
placode
• Migrate in early fetal life to anterior hypothalamus
• Failure result to hypogonadotropic hypogonadism
accompanied by anosmia » (this is called Kallmann
Syndrome)
• A decapeptide (10 amino acids)
ß Several forms of GnRH decapeptide have been
identified, the principal of which is GnRH-2, which
differs from GnRH by 3 amino acids. It is found in
several areas of the body, where it may subserve
functions unrelated to those of GnRH.
ß Functional connections between GnRH neurons
and the hypophyseal portal system that will transport
GnRH to the anterior pituitary gland are established
by about 16 weeks of fetal life.
ß The majority of GnRH neurons controlling the HPO
axis are located within the anterior hypothalamus and
primarily within the medial basal hypothalamus and
primarily within the medial basal hypothalamus, with
the greatest number in the primate within the arcuate
nucleus.
ß A substantial number of GnRH axons terminate
within the external zone of the median eminence
(infundibulum) where GnRH is released. This area is
the site of an important capillary plexus, with
fenestrated epithelium similar to that of peripheral
capillaries, which allows passage of large molecules.
(These capillaries differ from brain capillaries, which
are not fenestrated. Thus the median eminence is
viewed as an area outside BBB). This pathway is the
most relevant one in regard to the control of the
pituitary-ovarian axis.
Transport to Anterior Pituitary
• GnRH, after it is released, collects into several
hypophyseal portal vessels > descends along
pituitary stalk > terminate in another capillary plexus
within anterior lobe of pituitary
• Short half-life (2-4 mins.)
Berino | Gobenciong
• GnRH is not measurable in periph blood
Physiology
• Intermittent pulsatile release of GnRH by a ‘pulsatile
generator’
• Hourly intervals
• Inherent pace-making activity of the GnRH neuron
itself
• Key role of kisspeptin (KISS1) – directly innervate &
stimulate GnRH neurons
- Foremost modulatory influence on frequency &
amplitude of GnRH pulses is exerted by the
ovarian steroid hormones through their feedback
loop actions –
Estrogen = amplitude
Progesterone= frequency
ß In humans, mutations or targeted deletions of
KISS1 or of its receptor cause hypogonadotropic
hypogonadism. Patients however do not have
anosmia unlike in Kallmann syndrome
• Inhibitory inputs = GABA, Dopamine, endogenous B-
endorphins and CRH neurons – (either totally or
conditionally)
• Nutritional deprivation & abnormal eating habits
interfere with normal reproductive process e.g.,
anorexia nervosa & obesity
ANTERIOR PITUITARY GLAND
AND THE GONADOTROPINS
• aka adenohypophysis
• Ant. Pit. originates at about 3rd week of life
• Only vascularization is through the hypothalamic-
hypophyseal portal system
• Gonadotropes produce gonadotropins
• Stimulation is by GnRH>2 gonadotropins released
into the general circulation which regulate function in
the ovaries & testes
ß The anterior pituitary derives from the Rathke
pouch, a depression in the roof of the developing
mouth in front of the buccopharyngeal membrane
Physiology
• GnRH released by GnRH neurons in the arcuate
nucleus > GnRH receptors on the gonadotropes in the
anterior pituitary > stimulate both synthesis & release
of both gonadotropins subunit gene transcription:
Low GnRH pulse frequency = favors FSH synthesis
High GnRH pulse frequency= favors LH synthesis
• GnRH activation requires the release of containing
intramolecular bonds, which maintain the receptor in
the inactive configuration
ß In contrast to the response to the normal pulsatile
mode of GnRH release, sustained exposure of the
GnRH to constant GnHR concentrations drastically
reduces the response of the gonadotrope to
subsequent stimulation with GnRH. This phenomenon
is referred to as homologous desensitization or
downregulation of the receptor, which denotes a
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 Lecture 2: REPRODUCTIVE ENDOCRINOLOGY
Dr. Jen Lesiguez | February 2021
reduction in the ability of GnRH to elicit gonadotropin
release after prior continuous exposure to GnRH.
• Once activated cellular production of specific
membrane-associated lipid-like glycerols acting as
second messengers are stimulated
• Activate several cellular proteins: protein kinase c
(PKC) & mitogen-activated protein kinase (ERK)
• Transcription of gonadotropin subunits > gonadotropin
synthesis
• GnRH binding to receptor > calcium release >
exocytosis> release of LH & FSH
• GnRH agonist – initial stimulation of gonadotropin
release followed by desensitization blocking the
releasing effect on the gonadotropins
- “medical castration” state by shutting down the
pituitary-gonadal axis
• GnRH Antagonist – competes with GnRH for receptor
sites thereby never activating a stimulatory signal
- Rapidly decreasing LH & FSH release
THE GONADOTROPINS (LH & FSH)
• hCG is also a gonadotropin
• LH & FSH are glycoproteins of high molecular weight
containing 2 monomeric units (subunits)
• Both have similar L-subunit. Shared with hCG
& TSH
»LH & FSH are both produced by the gonadotropes.
They are glycoproteins that has high molecular weight
and it contains 2 subunits. Both of them, LH & FSH, will
have the same alpha-subunit and this alpha-subunit is
being shared with hCG and TSH. This alpha-subunit of
the LH, FSH, hCG, and TSH will be similar. This is why
when we do pregnancy testing, we test for the beta-
subunit especially in serum studies.
• B-subunits have different structures (amino acids &
carbohydrates) & encoded by separate genes
• B-subunit that confers specific biologic activity of each
hormone
• FSH acts primarily on the granulosa cells of the
ovarian follicles to stimulate follicular growth
• LH acts primarily on the theca cells of the ovarian
follicles and on the luteal cells to stimulate ovarian
steroid hormone production
OVARIES
• OOGENESIS begins in fetal life when primordial
germ cells/oogonia migrate to the genital ridge
• The number of oogonia increase to 600,000 at 2nd
month of fetal life >7 million by the 7th month
• With meiotic division – primary oocytes
• By apoptosis & atresia > 2-4 million at birth > 90%
depleted at puberty
• By 37 yrs old – 25,000; by 50-1000 oocytes remain
• Only about 400 follicles complete maturation process
“Don’t waste time. If you think you are called to become a
parent, think hard about it.” – Dr JLL
OVARIAN STEROIDS
Berino | Gobenciong
• One primary function of the ovary is secretion of
ovarian steroids after LH & FSH bind to their
respective receptors
1. Estradiol (primary estrogen)- maturing follicle
2. Progesterone – corpus luteum
3. Androstenedione – ovarian stroma
4. Estrone
5. Pregnenolone
6. 17-hydroxyprogesterone
7. Testosterone
8. DHEA
• Androgens are converted to estrone or estradiol by
the enzyme aromatase
• The aromatase enzyme is found in many tissues
besides the gonads – endometrium, brain, placenta,
bone, skin & others
• Also synthesized in adipose tissues – in
postmenopausal women becomes the major site of
estrogen biosynthesis
• SHBG – transport proteins (syn by liver) for steroid
hormones > to non-steroid specific albumin. Others
are ‘free’
• Albumin – high capacity but binds with low affinity
• Steroids can readily dissociate from its binding and
enter target cells
• SHBG binds dihydrotestosterone, testosterone, &
estradiol
PROSTAGLANDINS
• Play an important role in ovarian physiology
• Help control early follicular growth by increasing blood
supply to certain follicles and inducing FSH receptors
in granulosa cells of preovulatory follicles
• May assist in the process of follicular rupture by
facilitating proteolytic enzyme activity in the follicular
walls
• May help regulate myometrial contractility and may
also play a role in regulating the process of
menstruation
OVARIAN-HYPOTHALAMIC-PITUITARY FEEDBACK
LOOPS
• FSH and LH act on the ovaries to induce morphologic
changes and ovarian steroid secretion
• Morphologic processes: folliculogenesis and formation
of corpus luteum
• It is important for the brain and pituitary gland to
modulate their secretion in response to the minute-to-
minute activity status of the ovary
• Estradiol and progesterone play a major role in these
feedback communications
NEGATIVE STEROID FEEDBACK LOOP
• Inhibitory
• Small increase in the levels of the hormone induce a
decrease in gonadotropins
• As circulating estradiol levels increase during the
follicular phase, gonadotropin concentrations
decrease
• In postmenopausal women, the lack in estradiol
secretion causes sustained increases in LH and FSH
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release because of the lack of active negative
feedback loop
OVARIAN PEPTIDE FEEDBACK LOOP
• Inhibin
- Secreted by the ovaries
- Preferential inhibition of FSH over LH through
negative feedback loop
- At menopause/POF decreased secretion of
inhibin > increase in FSH
• Activin
- Stimulates FSH release
THE MENSTRUAL CYCLE
• Ovulatory cycle
• Coordination of hormonal secretion and morphologic
changes
• Cyclic process occurring at about monthly intervals
• 2 phases: follicular phase and luteal phase
• Separated by ovulatory period
• Mean duration 28 +/- 7 days (21-35 days)
• Length of follicular phase – variable
• Life span of corpus luteum – 14 days
FOLLICULAR PHASE
Subdivided into 3 periods:
1. Successive recruitment of a cohort antral follicles
• FSH provides critical signal
• FSH signal is the major survival factor that rescues
the follicles from their programmed death 9atresia)
• 3-7 secondary preantral follicles
2. Selection of a dominant follicle
• Only 1 is selected to complete growth to maturity
• Not well understood
• Dominant follicle has a well-vascularized theca layer
allowing a better access of the gonadotropins to their
target receptors
• Results in greater estradiol secretion > increases the
density of gonadotropin receptors and promote cell
multiplication
• Elevated estradiol levels activate negative feedback
loop > decrease in circulating FSH
• Selection is completed by day 5
3. Growth of the dominant follicle
• GnRH pulse frequency is at maximum > production of
estradiol > continue to stimulate growth of the
dominant follicle
• Stimulation by FSH of its receptors activates the
enzyme aromatase
• Stimulation of LH receptor synthesis within stromal
cells of theca interna
• LH – promotes steroid biosynthesis and production of
androgens
• Androgens biotransformed to estradiol by aromatase
> increased intraovarian estradiol levels and
increased estradiol secretion to peripheral circulation
Berino | Gobenciong
• At preovulatory stage, the number of granulosa cells
has increased from about 50 at primordial stage to
5x107
• Formation of ANTRUM (cavity) into which follicular
fluid accumulates
• At maturation, the dominant follicle mean diameter =
18-25 mm
• Within the dominant follicle, oocyte develops and
surrounded by zona pellucida – a
mucopolysaccharide coat containing specific protein
sites that late will allow only one spermatozoa to
penetrate the fertilized ovum
• At the end of follicular phase, antral follicle contain
oocytes that are fully grown but not yet able t undergo
full activation
• Activation will wait for the ovulatory LH surge
OVULATORY GONADOTROPIN SURGE AND OVULATION
• When threshold estradiol level is reached > positive
feedback loop > hypothalamus and anterior pituitary
signaled that follicle is ready for ovulation >
Gonadotropin surge > LH surge (ovulatory surge)
• LH levels increase 10-fold over 2-3 days, FSH levels
increase 4-fold
• This surge is an ABSOLUTE requirement for the final
maturation of the oocyte and the initiation of follicular
rupture
• As oocyte enters metaphase II, the 1st polar appears
• At ovulation, meiosis is arrested again (2nd meiotic
arrest)
• The 2nd meiotic division will only be completed during
fertilization
• OVULATION = 32 hours after initial rise of LH surge
*16 hours after peak
• Molecular events and changes in the matured follicle
• Acute inflammatory-like reaction and release of
enzymes which lead to degradation of follicular layers
and wall > follicular rupture
LUTEAL PHASE
• Corpus luteum formation results from 2 events
initiated at ovulation
1. Granulosa and theca cells hypertrophy, take up
increasing amounts of lipids, acquire organelles
associated with steroidogenesis
• Activation of new steroidogenic enzymes
• Hallmark of corpus luteum: secretion of progesterone
2. The basal lamina which separates the granulosa
and theca cell layers, is disrupted, and capillaries
from the theca interna invade the granulosa layer
• Each steroidogenic cell within the corpus luteum is in
close proximity to blood vessels
• Normal function of the corpus luteum depends
primarily on LH stimulation throughout the luteal
phase
• Progesterone dominance results in significant
activation of the progesterone negative feedback loop
on the GnRH pulse generator which also decreases
GnRH pulse frequency from 1/90 mins to 1/3 hrs
• Progesterone dominance also affects hypothalamic
thermoregulatory center > small increase in BBT
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• Corpus luteum reaches maturity 8-9 days after
ovulation after which it starts to degenerate
• Only rapidly rising hCG can rescue corpus luteum
following conception
THE MENSTRUAL CYCLE AND THE ENDOMETRIUM
• Primary goal is to ensure an appropriate environment
for the implantation of the developing conceptus
• Stratum basale and stratum functionale
• Upper layer serves as the site of blastocyst
implantation and provide metabolic environment for it
• Lower layer maintains the integrity of the mucosa
• Changes in hormones affect mainly the stratum
Functionale
ENDOMETRIUM IN THE PROLIFERATIVE/FOLLICULAR
PHASE
• Immediately after menstruation, endometrium is 1-2
mm thick, mainly stratum basale and few glands
• Increased estradiol levels > increase estradiol
receptors and proliferation of stratum functionale
• Towards late follicular phase, the glands become
more voluminous and tortuous
• At time of onset of LH surge and before ovulation >
subnuclear vacuoles – first indication of progesterone
effect
• Endometrial thickness by ultrasound – 4 mm in early
follicular phase to 12 mm at ovulation
• PMN and monocytes infiltrate glands and stroma >
autolysis of stratum functionale > desquamation
• Degradation lead to loss of integrity of blood vessels,
destruction of endometrial interstitial matrix > bleeding
• Usually last 3-5 days (2-7 days)
• Average blood loss 35 ml (10-80 ml)
• Shedding of endometrial lining (tissue mixed with
blood)
MENSTRUAL CYCLE AND CERVICAL GLANDS
• Changes in production and property of mucus –
important for fertility
• Increased production of mucus facilitate transport
and storage of spermatozoa during midcycle
• Clear water-like appearance, acellular, “fern” when
dried and seen in microscope “stringy” –
“spinnbarkeit” (cervical mucus that can stretch on a
slide at least 6 cm)
• Signifies “fertile period”
• Progesterone during luteal phase thickens cervical
mucus > less conductive for sperm transport
END OF TRANSCRIPTION
REFERENCES
• Comprehensive Gynecology
• Dr. JLL’s Lecture
APPENDIX

ENDOMETRIUM IN THE SECRETORY (LUTEAL) PHASE

• Well-developed subnuclear glycogen-rich vacuoles

• As progesterone levels increase during the 1st part of
the luteal phase, the glycogen-containing vacuoles
ascend progressively toward the gland lumen
• Contents of the glands are released into the
endometrial lumen = coincides with the arrival of the
free-floating blastocyst which reaches the uterine
cavity about 3-5 days after fertilization (implantation
occurs about 1 week after fertilization)
• After 1st week of luteal phase, stromal changes
become more important and relevant
• Stroma becomes more edematous due to increased
capillary permeability
• Endothelial proliferation > coiling of capillaries and
vessels
• Predecidual stromal cells are precursors of
gestational decidual cells – in nonpregnant
endometrium, engaged in phagocytosis and digestion
of extracellular collagen matrix > breakdown of
Legend:
endometrium (menses)
• Supportive role to the endometrial mucosa, control the
Important Audio Book UERM Previous
invasive nature of the normal trophoblast (absence
may lead to placenta accrete) Record Trans Year
• Decidualization succeeds predecidualization should Trans
pregnancy occur
 » ß ₪
MENSTRUATION
• If implantation does not occur and hCG is not
produced to maintain corpus luteum, endometrial
glands collapse and fragment

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