PENGANTAR
FISIOLOGI REPRODUKSI
Kuliah 1
Rahmatina B. Herman
Bagian Fisiologi
Reproduction
Reproduction is process
to maintain
continuation of species
by which
-
new individuals
of a species are produced
-
genetic material
is passed from generation
to generation
Cell division in a multicellular organism is
necessary for growth and it involves passing of
genetic material from parent cells to daughter
cells
The Reproductive System
does not contribute to homeostasis
is not essential for survival of an individual
But still plays a i porta t i a perso ’s life,
e.g. the manner:
- in which people relate as sexual beings
contributes in significant ways to
psychosocial behavior
- how people
view themselves
The Reprodu tive Syste …..
Reproductive function also has a profound
effect on society:
- universal organization of societies into
family units provide a stable environment
that is
conducive for perpetuating
our
species
- on other hand,
population explosion
and its
resultant drain on dwindling resources
The Reprodu tive Syste …..
Reproductive capability depends on
intricate
relationship among hypothalamus, anterior
pituitary, reproductive organs, and target cells
of sex hormones
The Reprodu tive Syste …..
Sexual behavior and attitudes are deeply
influenced by emotional factors and
socio-cultural mores of the society in which the
individual lives
However, Reproductive Physiology will
The Reprodu tive Syste …..
The organ of male and female may be grouped
by function
Testes and ovaries (gonads), function in
production of gametes: sperm and ova
Gonads also secrete hormones
The ducts of reproductive systems transport,
receive, and store gametes
The Reproductive System
…..
In females, the breasts are also considered
accessory reproductive organs
The externally visible portions of reproductive
system are known as external genitalia
The production of gametes and fluid, and their
discharge into ducts classify the gonads as
exocrine glands
Secondary Sexual Characteristic
Secondary sexual characteristic are many external
characteristics
not directly involved in
reproduction
That
distinguish male and female
Development and maintenance governed by
testosterone in males and estrogen in females
Progesterone has no influence on secondary
sexual characteristic
Secondary Sexual Characteristic
..…
In some species, secondary sexual
characteristic are great importance in courting
a d ati g ehavior e.g. to attra t fe ale’s
attention)
Overview of Functions and Organs
of Male Reproductive System
The essential reproductive functions of
male
are:
1. Production
of sperm (spermatogenesis) by
testes (in skin-covered sac: scrotum)
2. Delivery
of sperm to female
–
semen by
- male reproductive tract: epididymis, vas
deferens, ejaculatory duct
- urethra (in penis)
3. Male
accessory
sex glands: providing bulk of
semen: seminal vesicle, prostate,
Overview of Functions and Organs
of Female Reproductive System
Female
’s role i reprodu tio is ore o pli ated:
1. Production of ova (oogenesis) by ovaries
2. Reception of sperm: vagina-cervix
3. Reception of sperm and ovum to a common site for union (fertilization or conception): Fallopian tube
4. Maintenance of the developing fetus until it can survive in outside world (gestation or pregnancy), including formation of placenta (organ exchange between mother and fetus): uterus
5. Giving birth to the baby (parturition)
Overview of Functions and Organs
of Fe ale Reprodu tive Syste …..
Product of fertilization:
embryo
During first 2 months of intrauterine
development when tissue differentiation is
taking place
Developing living being is recognizable as
human:
fetus
-
no further tissue differentiation
Overview of Functions and Organs
of Fe ale Reprodu tive Syste …..
Female
reproductive tract consists of:
Ovaries
Oviduct s (Fallopian tubes)
- pick up ova on ovulation and serve as fertilization site
Uterus, thick-walled hollow: responsible for
- maintaining fetus during development , and - expelling it at the end of pregnancy
Cervical canal
- small opening of cervix
- pathway for sperm to uterus then to oviduct - passageway for delivery of baby from uterus
Cervix
Overview of Functions and Organs
of Fe ale Reprodu tive Syste ……
Vagina
- expandable tube
- connects uterus to external environment
Vaginal opening
- located in perineal region
- between urethral opening and anal opening
Hymen
- thin mucus membrane partially covering vaginal opening
• Labia minora and labia majora
- skin folds surrounding vaginal and urethral openings
• Clitoris
Sex Determination and Differentiation
Reproductive cells each contain a half set of
chromosomes
Gametogenesis is accomplished by
meiosis
The sex of and individual is determined by
combination of sex chromosomes
Parents with diploid (46 chr) somatic cells
Mother
Father
Meiotic division of germ cells
Meiotic division of germ cells
Haploid Ovum Haploid Sperm
Fertilization
Diploid fertilized Ovum
Mitosis
Ovum with X sex chromosome
Fertilized by
Sperm with Y sc Sperm with X sc
Embryo with XY sc Genetic sex Embryo with XX sc
Sex-determining region of Y chr (SRY) stimulates Production of H-Y antigen
In plasma membrane of undifferentiated gonad
H-Y antigen directs differentiation of gonads into testes
No Y chr, so no SRY and no H-Y antigen
With no H-Y antigen, undifferentiated gonads
develop into ovaries
Testosterone
Promotes development of undifferentiated external genitalia along male lines
(e.g. penis, scrotum)
Testes secrete hormone and factor
Phenotype sex Mullerian-inhibiting factor Dihydrotestosterone (DHT) Converted to Degeneration of Mullerian ducts
Transforms Wolfian ducts into male reproductive tract
(e.g. epididymis, ductus deferens, ejaculatory duct,
Absence of testosterone
Undifferentiated external genitalia along female lines
(e.g. clitoris. labia)
Ovaries does not secrete hormone and factor
Phenotype sex
Absence of Mullerian- inhibiting factor
Degeneration of Wolfian ducts
Errors in Sexual Differentiation
Genetic sex and phenotype sex are usually
compatible
Occasionally, discrepancies occur
between genetic and anatomic sexes
Errors i Se ual Differe tiatio …..
1. If testes in
a genetic male
fail to properly
differentiate and secrete hormones, the result
is the development of an apparent anatomic
female in a genetic male, who, of course will
be sterile.
Errors i Se ual Differe tiatio …..
2. Testosterone acts on Wolfian ducts to convert
them into a male reproductive tract;
If testosterone derivative dihydrotestosterone
(DHT) that responsible for masculinization of
external genitalia because of genetic deficiency
of the enzyme which converts testosterone
Errors i Se ual Differe tiatio …..
3. Adrenal gland normally secretes a weak
androgen, dehydroepiandrosterone in
insufficient quantities to masculinize females.
If, pathologically excessive secretion of this
hormone in a genetically female fetus during
critical developmental stages imposes
Errors i Se ual Differe tiatio …..
Sometimes, the discrepancies between genetic
sex and apparent sex are not recognized until
puberty, when discovery produces
psychologically traumatic gender identity crisis
For instance: a masculinized genetic female
with ovaries, but with male type external
Errors i Se ual Differe tiatio …….
Less dramatic cases of inappropriate sex
differentiation often appear as sterility
problems
Therefore, important to diagnose any
problems in sexual differentiation in infancy. It
can be reinforced, if necessary, with surgical
and hormonal treatment, so that psychosexual
development can proceed as normally as
Tugas
1. Hubungan sistem limbik (
limbic system
)
dengan pengaturan fungsi seks
DASAR-DASAR
BIOMOLEKULER
REPRODUKSI WANITA
Rahmatina B. Herman
Bagian Fisiologi
Fakultas Kedokteran Universitas Andalas
Sex Determination and Differentiation
Reproductive cells/gamete each contain a half
set of chromosomes (
haploid
)
Gametogenesis is accomplished by
meiosis
The sex of and individual is determined by
combination of sex chromosomes
Sexual differentiation along male or female
lines depends on the presence/absence of
Absence of testosterone
Undifferentiated external genitalia along female lines
(e.g. clitoris. labia)
Ovaries does not secrete hormone and factor
Phenotype sex
Absence of Mullerian- inhibiting factor
Degeneration of Wolfian ducts
Summary of 4 possible defects produced by maternal nondisjunction of sex chromosomes at the time of meiosis
The YO combination is believed to be lethal, the fetus dies in utero
/Ovarian agenesis/Turner syndrome
Female Reproductive System
Reproductive system of women shows regular cyclic
changes that may be regarded as periodic preparation for fertilization and pregnancy
In humans and primate, the cycle is a menstrual cycle and its conspicuous feature is periodic vaginal bleeding that
occurs with the shedding of uterine mucosa (menstruation)
In other mammal: the sexual cycle is called estrous cycle, no episodic vaginal bleeding occurs, but the underlying endocrine events are essentially similar
- in some species: ovulation occurs spontaneously - in other species: ovulation is induced by copulation
Ovaries
Primary female reproductive organs
Perform dual function:
- producing ova (oogenesis)
- secreting female sex hormones:
estrogen and progesterone which act together to: > promote fertilization of ovum
> prepare female reproductive system for pregnancy
Oogenesis
Undergo numerous mitotic divisions
± 7 month after conception, fetal oogonia
cease dividing
From this point on, no new germ cells are
generated
Still in the fetus, all oogonia develop into
Primary Oocyte
Begin a first meiotic division by replicating their DNA However, they do not complete the division in the fetus
Accordingly, all the eggs present at birth are primary
oocytes containing 46 chromosomes, each with two
sister chromatids
Cells are said to be in a state meiotic arrest
State meiotic arrest continues until puberty and the onset of renewed activity in ovaries
Only primary oocytes destined for ovulation will ever
Pri ary Oo yte…..
Each daughter cells receives 23 chromosomes, each with 2 chromatids
One of the two daughter cells, secondary oocytes retains virtually all cytoplasm (other is first polar body)
Thus, the primary oocytes:
- Already as large as the egg will be
- Passes on to be secondary oocyte half of its
Secondary Oocyte
The second meiotic division occurs in a
fallopian tube after ovulation, but
only if the
secondary oocyte is fertilized (penetrated by a
sperm)
Daughter cells each receive 23 chromosomes,
each with a single chromatid
One of the two daughter cells, termed an ovum
retains
nearly all cytoplasm
(other is second
Final Result of Oogenesis
Net result of oogenesis is
that
each primary oocyte
can produce only one
Summary of Oogenesis
Birth
Puberty
Oogonia
Chromosomes Per cell
Chromatids Per cell
46 2
2 46
23
23 1
Summary of
Ooge esis…..
Oogonia: mitotic divisions until ± 7 month after conception
Mitosis of oogo iu → primary oocyte
Meiosis of primary oocyte, but do not complete
(beginning of the 1st eioti divisio → eioti arrest
Primary oocyte at birth containing 46 chromosomes
1st meiotic division is completed just before ovulation
→ secondary oocyte
2nd meiotic division occurs in a fallopian tube after
Comparison of Spermatogenesis and Oogenesis
Spermatogenesis
Three major stages:
1.
Mitotic proliferation
2.
Meiosis
3.
Packaging/ spermiogenesis
:
physically
reshaping/ remodeling
± 64 days, from spermatogonium to mature sperm
Up to
several hundred million sperm may reach
Follicle
From the time of birth, there are many
primordial
follicles
,
each containing 1 primary oocyte
Progression of some primordial follicles to
preantral and early antral stages occurs
- throughout infancy and childhood, and
- then during the entire menstrual cycle
Therefore, although most of follicles in ovaries are
still primordial, there are also always present
a
Menstrual Cycle
At the start of each
menstrual cycle, 10-25 the
follicles begin to develop into larger follicles
In humans, usually one of the larger follicles in one ovary starts to grow rapidly on ± the 6thday, becomes the dominant follicle
The dominant follicle continues to develop, and
others (in both ovaries) regress and become a
degenerative process called
atresia
(an example of
programmed cell death, or
apoptosis
)
Ovulation
Mature follicle (Graafian follicle): ± 1,5 cm in
dia eter, that it alloo s out o ovary’s surfa e
Ovulation occurs when the thin walls of follicle
and ovary at site where they are joined rupture
because of enzymatic digestion
Secondary oocyte surrounded by its tightly
Ovulatio …..
Occasionally, 2 or more follicles reach
maturity and more than 1 egg may be
ovulated
This is the most common of cause of
multiple births
In such cases, siblings are fraternal, not
LH
Follicular steroid hormones (progesterone)
Proteolytic enzymes (collagenase)
Follicular hyperemia and
Prostaglandin secretion
Weakened follicle wall Plasma transudation
into follicle
Degeneration
of stigma Follicle swelling
Follicle rupture
Evagination of ovum
Indicators of Ovulation
A surge in LH secretion triggers ovulation
- Ovulation normally occurs ± 9 h after the peak of LH surge - The ovum lives for ± 72 h after ovulation, but it is
fertilizable for a much shorter time
Research shows:
> Intercourse on the day of ovulation: pregnancy 36% > Intercourse on days after ovulation: pregnancy 0
> Intercourse 1-2 d before ovulation: pregnancy 36%
> A few pregnancies resulted from intercourse 3-5 d before ovulation (8% on day 5 before ovulation)
I di ators of Ovulatio …..
A change (usually rise) in
basal body temperature
caused by secretion of progesterone, since
progesterone is thermogenic
- The rise starts 1-2 d after ovulation
- Obtaining an accurate temperature chart should
use a digital thermometer and take oral/rectal
temperatures in the morning before getting out
of bed
Formation of Corpus Luteum
After mature follicle discharges its antral fluid and egg, it collapses around antrum and undergoes rapid transformation
Granulosa cells enlarge greatly, and entire glandlike structure formed, known as corpus luteum (CL)
CL secretes estrogen, progesterone, inhibin
If the discharged egg (now in a fallopian tube) is not fertilized, CL reaches its maximum development
within ± 10 days.
CL then rapidly degenerates by apoptosis
Granulosa Cell
Primordial follicles surrounded by a single layer of
granulosa cells
Granulosa cells secrete: - estrogen ,
- small amounts of progesterone just before
ovulation
- peptide hormone inhibin
During childhood granulosa cells secrete: - nourishment for ova
Gra ulosa Cell…..
Further development from primordial follicle stage is characterized by
- an increase in size of oocyte
- a proliferation of granulosa cells into multiple layers - separation of oocyte from inner granulosa cells by a
thick of material: zona pellucida
Gra ulosa Cell…..
Inner layer of granulosa cells remains closely
associated with oocyte by means of
cytoplasmic processes
that
traverse zona
pellucida
and form
gap junctions
with oocyte
Nutrients and chemical messengers
are passed
to oocyte through gap junctions
Theca Formation
As follicle grows by mitosis of granulosa cells,
connective tissue cells surrounding granulosa
cells differentiate and form layers known as
theca
Shortly
after theca formation
,
-
Primary oocyte
reaches
full size
(115
m in
diameter)
Process of Atresia
Atresia is not limited to just antral follicles, follicles
can undergo atresia
at any stage
This process is already occurring
in utero
so that
the 2-4 million follicles and eggs are present at
birth represent only a small fraction of those
present at earlier time in the fetus
Atresia then
continues all through pubertal life
so
that only 200,000-400,000 follicles remain when
active reproductive life begins.
Pro ess of Atresia…..
Sites of Synthesis of Ovarian Hormone
Estrogen is synthesized and released into blood:
- during follicular phase mainly by granulosa cells
- after ovulation, by CL
Progesterone is synthesized and released into
blood:
- in very small amounts by granulosa and theca
cells just before ovulation
- major source is CL (after ovulation)
Inhibin is synthesized and released into blood:
- by granulosa cells
Ovarian Cycle
1. The follicular phase:
- ovarian follicle growth
- ovulation
2. The luteal phase:
Uterine Cycle
1. Proliferative phase:
-
estrogen
estrogen phase
-
before ovulation
2. Secretory phase:
-
progesterone
progestational phase
-
after ovulation
3. Menstruation
Cyclical Changes in Cervix
The mucosa of cervix does not undergo cyclical desquamation
There are regular changes in cervical mucus:
- Estrogen makes the mucus thinner, watery, and more
alkali e → pro otes the survival a d tra sport of sper s - At the time of ovulation the mucus is:
> thinnest and fern-like pattern on slide
> its elasti ity i reases → a drop a e stret hed i to a long (8 - ≥ , a d thi thread
- Progesterone makes the mucus thick, tenacious, and cellular
Microscopic of patterns formed of cervical mucus on dried smeared slide. Progesterone makes the mucus thick and cellular.
In anovulatory, no progesterone is present to inhibit fern-like pattern
Estrogen:
fern-like pattern
Estrogen,
no progesterone: Fern-like pattern Progesterone:
Cyclical Changes in Vagina
Under influence of estrogen:
- the vaginal epithelium becomes cornified that
can be identified in the vaginal smear
Under influence of progesterone:
- secretion of thick mucus
- the vaginal epithelium proliferates and
becomes infiltrated with leukocytes
Cyclical Changes in Breast
Although lactation normally does not occur until the end of pregnancy, cyclical changes take place in the breasts during the menstrual cycle
Estrogens cause proliferation of mammary ducts
Progesterone causes: growth of lobules and alveoli
The breast swelling, tenderness, and pain experienced by many women during the 10 day preceding
e struatio ← due to diste tio of the du ts,
hyperemia, and edema of the breast interstitial tissue
Normal Menstruation
Menstrual blood is predominantly arterial, only 25% of the blood being of venous origin
Containing tissue debris, prostaglandins, and relatively large amount of fibrinolysin from endometrial tissue
Fi ri olysi lyses lots → o lots i e strual lood
Usual duration is 3-5 d, but 1-8 d can occur normally The average amount of blood lost is 30 ml (range
normally from light spotting – 80 ml)
The amount of blood affected by various factors,
Anovulatory Cycles
Anovulatory cycles are common for the first 12-18 months after menarche and before the onset of menopause
Whe ovulatio does ot o ur → o CL → effe ts of
progesterone on endometrium are absent
Estrogens continue to cause growth, and proliferative endometrium becomes thick enough to break down and begins to slough
The time it takes for bleeding usually < 28 d from the last menstrual period
BIOSINTESIS DAN
MEKANISME KERJA
HORMON REPRODUKSI WANITA
Kuliah 3
Rahmatina B. Herman
Bagian Fisiologi
Ovarian Sex Hormones
Types of ovarian sex hormones:
1. Estrogens
The most important of estrogens is estradiol
2. Progestins
The most important of progestins is progesterone
Production:
> Non-pregnant: by - ovaries
- adrenal cortices
> Pregnancy: by - ovaries
- placenta
Ovarian
Se Hor o es…..
Transport to target organ:
> Loosely bound with plasma albumin
> More tightly bound with a beta globulin: sex binding globulin
- specific estrogen-binding globulin
- specific progesterone-binding globulin
Mechanism of action:
> Location of receptors: - Cell interior
> Signal transduction mechanism:
- Receptors directly alter gene transcription
Types of Estrogens
Beta-estradiol (17β-estradiol):
- major estrogen
- synthesized by ovaries
- strength: 12x estrone and 80x estriol
• Estrone
- by adrenal cortices, ovaries, and some other tissues
• Estriol
Synthesis
Cholesterol
Androstenedione
Testosterone
Estrone
Estradiol
Secreted by Ovaries
Aromatase
Sy thesis…..
The naturally estrogens (17β-estradiol, estrone, and estradiol) are C18 steroids
The biosynthesis of estrogens depends on the enzyme aromatase (CYP19), which converts testosterone to estradiol and androstenedione to estrone
The reaction of converting androstenedione to estrone also occurs in:
Synthesis by Ovaries
Estrogen is synthesized and released into blood
- during follicular phase mainly by granulosa cells
- after ovulation by corpus luteum
- during pregnancy by placenta
Granulosa cells require help to produce estrogen
because they are deficient in the enzymes required to produce androgens that are the precursors of estrogen
They are aided by theca cells
Synthesis by
Ovaries…..
Ovarian follicle
(Diffusion)
Theca cells
Synthesize androgens
Granulosa cells
Convert androgens to estrogen
Sy thesis y Ovaries…..
Theca interna cells have many LH receptors
LH acts via cAMP to increase conversion of cholesterol to androstenedione
Theca interna cells supply androstenedione to granulosa cells
Estradiol produced by granulosa cells when provided with androgen and secreted into follicular fluid
Granulosa cells have many FSH receptors
FSH facilitates the secretion of estradiol by acting via cAMP to increase aromatase activity
Secretion
Concentration of estradiol in plasma during menstrual cycle is depend on menstrual phase
Almost of this estrogen comes from ovary
Two peaks of secretion occur: one just before ovulation and one during the midluteal phase Estradiol secretion rate is:
- 36 μg/d in the early follicular phase - 380 μg/d just before ovulation
- 250 μg/d during midluteal phase
Transport and Metabolism
Circulating estradiol: 2% is free, 60% is bound to albumin and 38% is bound to sex hormone-binding globulin /gonadal steroid-binding globulin (specific estrogen-binding globulin)
Metabolism by liver:
- conjugated glucoronide and sulfate - excreted: - most in urine
- in the bile (1/5 → e terohepati circulation
3 Basi e ha is s of estroge ’s effe ts o target
organs/ cells
- Promote proliferation and growth specific cell - Development of primary sex characteristic
- Development of most secondary sex characteristics
Non reproductive effects - Promotes fat deposition - Increases bone density - Closes epiphyseal plates
On Sex-specific tissues
Essential for egg maturation and release
Stimulates growth and maintenance of entire female reproductive tract
Stimulates granulosa cell proliferation which lead to follicle maturation
Thins cervical mucus to permit sperm penetration Enhances transport of sperm by stimulating upward contractions of uterus and oviduct
Stimulates growth of endometrium and myometrium
Induces synthesis of endometrial progesterone receptors
Triggers onset of parturition by increasing uterine responsiveness to oxytocin during late gestation through a twofold effect by
- inducing synthesis of myometrial oxytocin receptors
- increasing myometrial gap junctions so that uterus can contract as a coordinated unit in response to oxytocin
Other reproductive effects
• Promotes development of secondary sexual characteristics
• Controls GnRH and gonadotropin secretion: - Low levels: inhibit secretion
- High levels responsible for triggering LH surge
• Stimulates duct development in breasts during gestation
• Inhibits milk secreting action of prolactin during gestation
On uterus & external sex organ
:
- Increase the size
- External genitalia enlarge with deposition of fat
- Change vaginal epithelium from cuboidal stratified - Endometrium changes:
> proliferation of the stroma
> greatly increased development of endometrial glands
On Fallopian tube
:
- Proliferation of glandular tissue
- Increase the number of ciliated epithelial cell - Enhance the activity of the cilia
On breast:
- Development of the stromal tissue
- Growth of an extensive ductile system - Deposition of fat
- Develop lobules and alveoli (initiate growth)
- Characteristic growth and external appearance of the mature female breast
On metabolism:
- Increase metabolic rate (1/3 of testosterone)
On electrolyte balance: Na retention
- Slight and rarely significance, except in pregnancy
On protein deposition:
- Slight increase in total body protein
On fat deposition:
- Increase quantities of fat in subcutaneous tissue decreased specific gravity flotation in water
On skin:
- Develop texture which is soft and smooth - Thicker than children and more vascular
- Increase secretion of axillary sweat gland acne (by adrenal androgen)
On hair distribution:
- No greatly effect (opposite to testosterone)
On skeleton:
- Increase osteoblastic activity growth
- Early uniting of the epiphyses with the shafts of long bone (stronger than testosterone)
Osteoporosis caused by estrogen deficiency: - Diminished osteoblastic activity
- Decreased bone matrix
- Decreased deposition of Ca & Phosphate
On central nervous system
- Increase libido in humans and estrous behavior in
animals by direct effect on neurons in hypothalamus - Increase proliferation of dendrites on neurons in rats
Intracellular function:
- circulate in blood only a few minutes target cells - combine with receptor protein in cytoplasm
- activate specific portions of chromosomal DNA
- initiate transcription process DNA – RNA
> division of cell
> protein formation in a few specific target organs
Synthesis
Cholesterol
Pregnenolone
17-Hydroxy progesterone
Dehydroepi androsterone
Synthesis
The most important type of progestin is
progesterone
Progesterone is C21 steroid
Progesterone is synthesized and released into
blood
> by ovaries:
- major source is CL (after ovulation)
- In very small amounts by granulosa and theca cells just before ovulation
- placenta
Secretion
Concentration of progesterone in plasma during menstrual cycle is depend on menstrual phase
Progesterone secretion rate is 0.9 ng/ml during the follicular phase, and late in follicular phase,
progesterone secretion begins to increase
During luteal phase, corpus luteum produces large
ua tities → plas a o e tratio arkedly
increased to a peak value of approximately 18 ng/ml Stimulating effect of LH is due to activation of adenylyl cyclase and involves subsequent step that is
Transport and Metabolism
Circulating progesterone: 2% is free, 80% is bound
to albumin and 18% is bound to sex
hormone-binding globulin /corticosteroid-hormone-binding globulin
(specific progesterone-binding globulin)
Progesterone has a short half-life
Converted in liver to pregnanediol, which is
conjugated to glucuronic acid
Final preparation of the uterus for pregnancy
- Prepares a suitable environment of a developing embryo/ fetus
- Promotes formation of a thick mucus plug in cervical canal
- Inhibits uterine contractions during gestation
Final preparation of the breast for lactation
- Stimulates alveolar development in breasts during gestation
- Inhibits milk-secreting action of prolactin during gestation
Inhibits hypothalamic GnRH and gonadotropin secretion
On uterus:
- promote secretory change during the latter half of
y le → prepari g for i pla tatio
- decrease frequency and intensity of uterine
o tra tio → preve t e pulsio of i pla ted
ovum
On Fallopian tube:
- pro ote se retory ha ge → utritio for fertilized ovum
On breast:
- Development of lobules and alveoli
- Proliferate, enlarge, to become secretory of alveolar cells
- Cause breast swell
development in lobules and alveoli
Increase fluid in subcutaneous tissue
On electrolyte balance:
- In large quantity: Na retention (less than aldosterone)
- More often: increased Na and water excretion
Competition with aldosterone for binding
with receptor so that effect of aldosterone on Na retention is blocked
with net results increased Na excretion
The effects of progesterone, like those of other
steroids are brought about by an action on DNA to initiate synthesis of RNA
Progesterone receptor is bound to a heat shock protein
The synthetic steroid mifepristone binds to the
receptor but does not release the heat shock protein, and it blocks the binding of progesterone
Mifepristone combined with prostaglandin can be used to produce elective abortions
Tugas
Anatomy-Physiology of Ovaries:
- Immature follicle
- Mature follicle
- Theca cells
- Granulosa cells
PRINSIP FISIOLOGIS
BERBAGAI MACAM
ESAI HORMON
Kuliah 4
Rahmatina B. Herman
Bagian Fisiologi
Hormones
1. Hormones are chemical messengers that
enter the blood, which carries them from
endocrine glands to the target cells
2. Hormone action in the target cells/organ
3. Hormone
metabolism
Hormone Concentrations in Blood
Most hormones are present in blood in
extremely minute quantities, some in
concentrations of pg / ml (1 pg = 1 billionth of
mg)
Almost impossible to measure by usual
chemical means
Extremely sensitive methods:
Hormones Removal From Plasma
Hormones are cleared from plasma in several
ways:
1. Metabolic destruction by tissues
2. Binding with tissues
3. Excretion by liver into bile
4. Excretion by kidneys into urine
A decreased Metabolic Clearance Rate of
Metabolic Clearance Rate of Hormones
Two factors can affect hormone concentration
in blood:
- rate of secretion
- rate of removal which is called metabolic
clearance rate
Hormones Measurement
1. Measurement of hormone
secretion rate
2. Measurement of circulating hormones
concentration
3. Measurement of hormone action
4. Measurement of hormone
metabolite
product
Measurement of Hormone Secretion Rate
A simple method for estimating hormone
secretion is:
- Measuring the concentration of natural
hormone in plasma by means of a
radioimmunoassay procedure (C)
- Measuring metabolic clearance rate (MCR)
- By multiplying C x MCR, one derives a value
that is equal to steady-state of hormone
Measure e t of Hor o e Se retio Rate.….
However, hormone production often increases or decreases rapidly
In such case, one can measure the changing rate of secretions only by:
- collecting samples of arterial blood entering the
gland (AB) and samples of venous blood leaving the gland (VB)
- measuring rate blood flow through the gland (BF) - by multiplying BF x (VB-AB), one can derive the
Measurement of Hormones Concentration
Physiologically variable that fluctuates
each day with a cyclical periodically
Measurements of particular variable are
usually obtained at a single time of day
In certain types of hormonal diseases,
plasma concentration of the hormone
Measure e t of Hor o es Co e tratio …..
Thus, if the hormone in the blood was measured
at only one time of day, the disorder might be
missed
To avoid this problem is to obtain repeated
measurements of the hormone over a 24-hour
period
Ideally, repeated blood measurements could be
drawn to provide as complete a profile as possible
of the minute-to-minute changes in circulating
hormone level.
A simpler method is to obtain a 24-hour
cumulative urine sample
Metabolites of many hormones appear in the
urine as part of the daily process of clearing excess
hormones from the blood
The more hormones in the blood, the more it or its
metabolites appear in the urine
A-24 hour measurement will provide information
on the integrated, or summed, amount of
hormone produced during the day and night.
So that it is a time-averaged mean
In fact that time-averaged means reveal
nothing of the countless small (sometimes
large) fluctuations in circulating hormone
concentration that occurred during that time
It reveals whether or not abnormally low or
high total amounts of hormone were produced
Measurement of Hormones Through Its Action
Different hormone has different effects on
target organ, so that the effects of the
hormones on the target organ may reflect
the hormone secretion or production
For examples: Basal Metabolic Rate (BMR)
Measure e t of hor o es through its a tio …..
Hormones has pharmacological effects on
target organs so that it also may reflect
the
concentration
of that hormone
For examples: measurement of
cardiovascular parameters may reflect
Measurements of
Metabolic Clearance Rate of Hormones
To calculate Metabolic Clearance Rate (MRC),
one makes following 2 measurements:
1. Rate of disappearance of the hormone
from plasma per minute (D)
2. Concentration of the hormone in each
ml plasma (C)
How to Measure
Quantitative:
- Blood samples : hormones
- Urine samples : metabolites product
hormone excretion
Qualitative:
- direct effect on target organ
Ho to Measure..…
Blood samples: hormones
Most hormones are unstable, so that need
appropriate approach:
- before assaying
> drawing samples
> transportation: temperature
> storing: temperature and long life
- during assaying:
> direct assessment
Valid and reliable
on method, tools, competencies
- Intra-assay validation (intra-day
validation)
- Inter-assay validation (inter-day
validation)
- Standard Curve
Guideline on Bioanalytical
Method Validation
Method Validation
The main objective of method validation is to
demonstrate the reliability of a particular method for the determination of an analyte concentration in a specific biological matrix, such as blood, serum,
plasma, urine, or saliva
If an anticoagulant is used, validation should be
performed using the same anticoagulant as for the study samples
Method Validatio …..
Main characteristics of bioanalytical method that are essential to ensure the acceptability of the
performance and the reliability of analytical results are:
- Selectivity
- Lower limit of quantification (LLOQ)
- the response function and calibration range
(calibration curve performance / standard curve) - Accuracy
- Precision
- Matrix effects
- Stability of the analytes in biological matrix
Method Validatio …..
During method validation and analysis of study
sample, a blank biological matrix will be spiked
with the analytes of interest using solutions of
reference standards to prepare
calibration
,
standards quality control samples
and
stability
samples
In addition, suitable
internal standards
(IS) can
be added during sample processing in
Selectivity
The analytical method should be able to differentiate the analytes of interest and internal standard (IS) from
endogenous components in the matrix or other
component in the sample
Selectivity should be proved using at least 6 individual sources of the appropriate blank matrix, which are
individually analysed and evaluated for interference
Normally, absence of interfering components is
Lower Limit of Quantification (LLOQ)
LLOQ is the lowest concentration in a sample which
can be quantified reliably, with an acceptable accuracy LLOQ is considered being the lowest calibration
standard
The analyte signal of LLOQ sample should be at least 5 times the signal of blank sample
LLOQ should be adapted to expected concentrations and to the aim of study: for bioequivalence studies
LLOQ should be not higher than 5% of Cmax
Calibration Curve Performance
Before carrying out the validation of the analytical
method, it should be known what concentration range is expected
The range should be covered by calibration curve range, defined by LLOQ being the lowest calibration standard and the upper limit of quantification (ULOQ) being the highest calibration standard
A minimum of 6 calibration concentration levels should be used, in addition to the blank sample (processed matrix sample without analyte and without IS) and a zero sample
(processed matrix with IS)
Cali ratio Curve Perfor a e…..
The calibration curve parameters should be
reported (slope and intercept in case of linear fit)
The back calculated concentrations of the
calibration standards should be presented
together with the calculated mean accuracy values
Cali ratio Curve Perfor a e…..
The back calculated concentrations should be within
±15% of the nominal value, except for LLOQ for which it should be within ±20%
At least 75% of calibration standards, with a minimum of 6 calibration standard levels, must fulfill this
criterion
Standard Curve
A standard curve is a type of graph used as a quantitative research technique.
A graphic plot of tracer binding versus the known
concentration of test substances in a set of standards usually prepared by serial dilution or incremental
addition.
Multiple samples with known properties are measured and graphed
Accuracy
Within-run accuracy
Determined by analysing in a single run a minimum of 5 samples per level at a minimum 4 concentration levels
which are covering the calibration range.
The mean concentration should be within 15% of nominal values, except for LLOQ should be 20% of the nominal value
Between-run accuracy
For the between-run accuracy, LLOQ, low, medium and high QC samples from at least 3 runs analysed on at least 2
different days should be evaluated
The mean concentration should be within 15% of the
Precision
Within-run precision
For the validation of the within-run precision, there should be a minimum of 5 samples per concentration level at
LLOQ, low, medium and high QC samples in a single run
The within-run CV value should not exceed 15% for QC samples, 20% for LLOQ
Between-run precision
For the validation of the between-in run precision, LLOQ, low, medium and high samples from at least 3 runs
analysed on at least 2 different days should be evaluated The between-run CV value should not exceed 15% for QC
Matrix Effects
Matrix effects should be investigated when using mass spectrometric methods
Using at least 6 lots of blank matrix from individual donors For each analyte and IS, matrix factor should be calculated for each lot of matrix, by calculating the ratio of peak area
in the presence of matrix (measured by analysing blank matrix spiked after extraction with analyte), to the peak area in absence of matrix (pure solution of analyte)
The IS normalized MF should also be calculated by dividing the MF of analyte by the MF of the IS
The CV of IS normalized MF calculated from 6 lots of matrix should not be greater than 15%
This determination should be done at a low and at a high level of concentration (maximum of 3 times LLOQ and
Stability
Evaluation of stability should be carried out to ensure that
every step taken during sample preparation and sample analysis, as well as the storage conditions used do not affect the concentration of analyte
The following stability tests should be evaluated:
- stability of the stock solution and working solutions of the analyte and internal standard (IS)
- freeze and thaw stability of the analyte in the matrix from freezer storage conditions to room temperature or sampling processing temperature
- short term stability of the analyte in matrix at room temperature or sampling processing temperature
- long term stability of analyte in matrix stored in freezer
Percentage recoveries of internal standard amitriptyline
Concentration of amitriptyline
Pure AT
(height in chrom.)
Extracted AT
(height in chrom.)
Recovery
(%)
20000pg/ml 31485 25358
28832 31115 27719 28899 32143 28922 20594 26866 29189 22865 21839 25846 91.86 81.21 93.18 93.81 82.49 75.57 80.41
Percentage recoveries of derivatized NE
Concentration
of NE (height in chrom.) Pure NE (height in chrom.) Extracted NE Recovery (%) Mean SD
16000 pg/ml 103455 109876 83387 104638 89075 90632 69863 88943 86.10 82.49 83.78
85.00 84.341.56
8000 pg/ml 53549 56791 42013 52995 44232 44345 35294 45046 82.60 78.08 84.01
85.00 82.423.06
4000 pg/ml 25711 28903 20876 26894 21698 24346 17745 23675 84.39 84.23 85.00
88.83 85.411.78
2000 pg/ml 12894 13764 11143 13547 10521 11230 8971 11693 81.59 81.59 80.51
86.31 82.502.59
1000 pg/ml 6690 6902 5621 6854 5429 5885 4525 5713 81.15 85.27 80.50
83.35 82.572.18
500 pg/ml 3112 3487 2876 3269 2471 2959 2386 2879 85.83 84.86 82.96
88.07 85.432.13
250 pg/ml 1615 1775 1313 1656 1265 1473 1089 1334 78.33 82.99 82.94
80.56 81.202.23
Above: Scanning chromatogram of derivatized extracted NE (DNE) and extracted amitriptyline (AT)
Peak ratio of derivatized NE and amitriptyline (for standard curve)
Concentra tion of NE
(pg/ml)
Peak ratio
Mean SD
1st run 2nd run 3rd run 4th run
8000 4000 2000 1000 500 100 (LLOQ) 1.59 0.86 0.39 0.21 0.117 0.019 1.64 0.90 0.40 0.22 0.111 0.024 1.72 0.81 0.41 0.19 0.107 0.026 1.67 0.83 0.43 0.20 0.099 0.021
1.65 0.05 0.85 0.04 0.41 0.02 0.21 0.01 0.11 0.008 0.023 0.003
The standard curve based on peak ratio of derivatized NE and internal standard amitriptyline. The concentrations of NE were from 8000 pg/ml to 500 pg/ml and 100 pg/ml, the lower limit of quantitation (LLOQ).
y = 0 ,0 0 0 2 x R2 = 0 ,9 9 9 7
0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8
0 1000 2000 3000 4000 5000 6000 7000 8000 9000
C o nc e nt ra t io n o f N o re pine phrine ( pg/ m l)
Accuracy and precision of the assay for NE with amitriptyline as internal standard (n=4) Conc of NE Intra-assay variation (Intra-day variation) Inter-assay variation (Inter-day variation) Peak ratio
Mean SD
CV (%)
Peak ratio
Mean SD CV
(%) 1st run 2nd run 3rd run 4th run 1st run 2nd run 3rd run 4th run 8000 4000 2000 1000 500 100 (LLOQ) 1.67 0.90 0.41 0.22 0.121 0.024 1.62 0.86 0.42 0.20 0.118 0.028 1.70 0.82 0.44 0.19 0.129 0.026 1.65 0.88 0.40 0.20 0.124 0.021
1.66 0.03 0.87 0.03 0.42 0.02 0.20 0.01 0.123 0.005 0.025 0.003
1.81 3.45 4.76 5.00 4.07 12.00 1.67 0.90 0.41 0.22 0.121 0.024 1.59 0.82 0.40 0.20 0.117 0.026 1.68 0.86 0.43 0.19 0.119 0.027 1.72 0.83 0.44 0.21 0.127 0.020
1.660.05 0.850.04 0.420.02 0.210.01 0.1210.004 0.024 0.03
3.01 4.71 4.76 4.76 3.31 12.50
Radioimmuno-Assay
For hormones assessment
Principle of radioimmuno-assay:
Antibody (globulin) which is specific for the hormone that will be assessed must be produced from the
animal in a great amount (commercially available)
The antibody then mixed with:
- animal serum which contain hormone to be assessed (h)
Radioimmuno-
Assay..…
Antibody and hormone will be bound (ab-h &
ab-hsr)
Hormone to be assessed and hormone labeled
by radioisotope
competitively
binds the
antibody
Concentration
of ab-hsr then measured, soon
after the binding
has reached equilibrium
,
Radioimmuno-
Assay..…
To make
assay highly ua titative ,
radioimuno-assay must also be applied for
sta dard
solution from pure un-labeled
hormone with some levels of
concentration
The results then will be arranged in a
PERKEMBANGAN
ORGAN REPRODUKSI WANITA
DARI JANIN SAMPAI LAHIR
Kuliah 5
Rahmatina B. Herman
Bagian Fisiologi
Overview of The Reproductive System
The organ of male and female may be grouped
by
function
Testes and ovaries (called
gonads
), function in
production of gametes
: sperm cells and ova
Gonads also
secrete hormones
The ducts of reproductive systems transport,
receive, and store gametes
Overview of The Reproductive System
…..
Accessory sex glands
produce materials that
support gametes
In females, the
breasts
are also considered
accessory reproductive organs
The production of gametes and fluid, and their
discharge into ducts classify the gonads as exocrine glands
Overview of Female Reproductive System
Fe ale’s role i reprodu tio is ore o pli ated tha ale’s:
1. Production of ova (oogenesis) by ovaries
2. Reception of sperm: vagina-cervix
3. Reception of sperm and ovum to a common site for union (fertilization or conception): Fallopian tube
4. Maintenance of the developing fetus until it can survive in outside world (gestation or pregnancy), including
formation of placenta (organ exchange between
