fisiologi reproduksi.

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

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

All the available (or acceptable) curves obtained

during validation, with a minimum of a 3 should

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 84.341.56

8000 pg/ml 53549 56791 42013 52995 44232 44345 35294 45046 82.60 78.08 84.01

85.00 82.423.06

4000 pg/ml 25711 28903 20876 26894 21698 24346 17745 23675 84.39 84.23 85.00

88.83 85.411.78

2000 pg/ml 12894 13764 11143 13547 10521 11230 8971 11693 81.59 81.59 80.51

86.31 82.502.59

1000 pg/ml 6690 6902 5621 6854 5429 5885 4525 5713 81.15 85.27 80.50

83.35 82.572.18

500 pg/ml 3112 3487 2876 3269 2471 2959 2386 2879 85.83 84.86 82.96

88.07 85.432.13

250 pg/ml 1615 1775 1313 1656 1265 1473 1089 1334 78.33 82.99 82.94

80.56 81.202.23

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 2}nd _{run 3}rd _{run 4}th _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

1.660.05 0.850.04 0.420.02 0.210.01 0.1210.004 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

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

These relationship employ many of

regulatory

mechanisms

used by other body systems for

maintaining homeostasis, such as

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)

In humans, attraction the opposite sex not only

influenced by secondary sexual characteristic

but also

strongly affected by the complexities

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

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

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

Granulosa cells produce one or more factors

that act on primary oocytes

to maintain them

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,

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

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

The externally visible portions of reproductive

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 mother and fetus): uterus

5. Giving birth to the baby (parturition)

Overview of Female Reproductive

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, only

Chronology of Reproductive Function

Sequential changes in reproductive development or function:

Sex determination

Genetic inheritance sets the gender of individual which is established at the moment of fertilization

Sex differentiation

Multiple process in which development of reproductive system occurs in fetus

Maturation of system at puberty

Definition

Sex determination is concerned with the regulation of the development of the primary of gonadal sex, while sex differentiation encompasses the events

subsequent to gonadal organogenesis

The processes are regulated by at least 70 different that are located on the sex chromosomes and

autosomes and that act through a variety of

mechanisms including organizing factors, gonadal

Sex Determining Region

An important point: early embryos of both sexes

possess indifferent common primordial that have an inherent tendency to feminize, unless in the

interference by masculinizing factors

Sex-determining region Y (SRY) gene is found to be the primary sex determinant that induce the indifferent

gonad into testes

SRY is expressed in XY gonads in Sertoli cell progenitors at the stage of sex cord formation

Translation of Genetic Sex

The SRY protein is detected at an early age of gonadal

differentiation in XY embryos, where it induces Sertoli cell differentiation (in adult, it is present in both Sertoli and germ cells)

In embryonic and fetal life, SRY gene product regulates gene expression in a cell autonomous manner

The precise molecular mechanisms by which SRY triggers testis development are unknown, nor is it yet known how SRY is regulated

Differentiation of Reproductive Tract

Although male and female external genitalia develop from the same embryonic tissue, but reproductive tract develop from the different system

2 primitive duct systems: Mullerian ducts and Wolfian ducts develop in all embryos, so that the early embryo has potential to develop either a male or a female

reproductive tract

Development of reproductive tract is determined by the presence of 2 hormones secreted by fetal testes: testosterone and Mullerian-inhibiting factor

In female fetus: reproductive tract develops from

Differentiation of Gonads

The genes directly determine only whether the individual will have testes or ovaries

Sex differentiation depends upon the presence or absence of substance produced by the genetically gonads, in particular, testes

Difference between male and female exist at 3 levels: - genetic sex: depends on combination of sex chr

- gonadal sex: determined by genetic sex , the presence of

a Y hr → SRY → H-Y antigen

Differe tiatio of Go ads…..

The reproductive organs are embryogically developed from the intermediate mesoderm

The male and female gonads derive from an area called

urogenital ridge, a condensation tissue near adrenal gland The gonad develops a cortex and a medulla

Until the 6th week of uterine life, primordial gonads are

u differe tiated → ide ti al i oth se es

A gene on Y chromosome (SRY gene) is expressed at this time in urogenital ridge cells and triggers the development of testes

Differe tiatio of Go ads…..

In genetic males, during the 7th and 8th weeks, the medulla develops into a testis, and the cortex regresses

Leydig a d Sertoli ells appear → testostero e a d MIS are

secreted

In genetic females, at about 11 weeks, the cortex develops into ovary and the medulla regresses

Embryonic ovary does not secrete hormones

Hormonal treatment of the mother has no effect on gonadal differentiation in humans

Whe the e ryo has fu tio al testes → ale i ter al

Differentiation of Genitalia

So far as its internal duct system and external genitalia are concerned, fetus is capable of developing into

either gender

Before the functional of fetal gonads, primitive reproductive tract includes a double genital duct system: Wolfian ducts and Mullerian ducts and a

Differentiation of Genitalia

…..

Normally, most of reproductive tract develops from only one of the duct systems

In female fetus: Mullerian ducts persist and Wolfian ducts regress

External genitalia and outer part of vagina do not develop from the duct system, but from other

structures at body surface

Ovaries (unlike testes), do not play a role in the development processes of genitalia

In other words, female development occurs

Differentiation of Internal Genitalia

The

internal genitalia

are bi-potential until gonads

undergo differentiation

In the

7

th

week

of gestation, the embryo has both

male and female internal genitalia

Internal genitalia

develop from the

different

system

Two primitive duct systems:

paramesonephric

ducts (Mullerian duct)

and

mesonephric ducts

(Wolffian duct)

develop in all embryos, so that the

early embryo has potential to develop either a

Differe tiatio of I ter al Ge italia…..

Development of internal genitalia is determined

by the presence of 2 hormones secreted by fetal

testes: testosterone and Mullerian-inhibiting

factor

In normal

male fetus

, the Wolffian duct system

develops into epididymis ducts and vas deferens,

and

Mullerian ducts degenerate

In normal

female fetus

, the Mullerian duct system

Differentiation of External Genitalia

Male and female external genitalia develop from the same embryonic tissue

Undifferentiated external genitals consist of:

- Genital tubercle which rise to exquisitely sensitive erotic tissue

- Urethral folds surrounding a urethral groove - Genital (labioscrotal) swellings

The external genitalia are bi-potential until the 8th

week of gestation

Thereafter, the urogenital slit disappears → male

genitalia form, or the urogenital slit remains open →

Differe tiatio of E ter al Ge italia…..

In normal

female fetus

Genital tubercle

Develop into clitoris without penetration by urethral opening

Paired urethral folds

Does not fuse and become labia minora

Urogenital slit

Urogenital slit remains open and become vagina

Genital (labioscrotal) swellings

DEVELOPMENT OF

Development of Ovaries

Descend to brim of pelvis during third month of development

During fetal life, the outer surface of ovary is covered by a germinal epithelium, which embryologically is derived from epithelium of germinal ridges

As the female fetus develops, cells that give rise to ova arise from endoderm of yolk sac and migrate to

ovaries during embryonic development

Primordial (primitive) germ cells migrate from

Develop e t of Ovaries…..

Primordial ova differentiate and migrate into the substance of ovarian cortex

In the 3rd _{month of prenatal development: oogonia} divided mitotically into primary oocytes (2n) until 20-24 weeks

± 7 month after conception, fetal oogonia cease dividing

Develop e t of Ovaries…..

Each ovum collects around it a layer of spindle cells from ovarian stoma and causes them to take on epithelioid characteristics; then called granulosa cells

Ovum surrounded by a single layer of granulosa cells is called primordial follicle with ovum at stage of primary oocyte

Primary oocytes enter prophase of reduction division (meiosis I), but do not complete

All the eggs present at birth are primary oocytes containing 46 chromosomes, each with two sister chromatids

Components of Female Reproductive Tract

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

Components of Female Reproductive

Tra t…..

Oviduct s (Fallopian tubes)

- in close association with ovaries,

- pick up ova on ovulation and serve as

fertilization site

Uterus

-

thick-walled hollow

- responsible for:

Co po e ts of Fe ale Reprodu tive Tra t…..

Vagina

- expandable tube

- connects uterus to external environment

Cervix

- lowest portion of uterus

- projects into vagina

Cervical canal

- small opening of cervix

Co po e ts of Fe ale Reprodu tive Tra t…..

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

- erectile tissue (analog with penis)

Development of Brain

At least in some species, the development of the brain as well as the external genitalia is affected by

androgens early in life.

In rats, a brief exposure to androgens during the first few days of life causes the male pattern of sexual

behavior and the male pattern of hypothalamic

control of gonadotropin secretion to develop after

puberty. In the absence of androgens, female pattern develop

Develop e t of Brai …..

In humans, early exposure of female fetuses to

androgens also appears to cause subtle but significant masculinizing effects on behavior

However, women with adrenogenital syndrome due to congenital adrenocortical enzyme deficiency develop normal menstrual cycles when treated with cortisol. Thus, the human, like the monkey, appears to retain

Tugas

SIKLUS OVARIUM

Kuliah 6

Rahmatina B. Herman

Bagian Fisiologi

Female Monthly Se