THIS CLASS IS IN BRIEF FOR MSc STUDENTS

1. MENOPAUSE
CRYOPRESERVATION
OESTROUS CYCLE

5. Neuroprotective and CNS effects- insomnia, fattigue
Tibolone-synthetic norsteroid derivative-estrogenic,
progestrogenic, and weak androgenic properties
Dose-2.5mg OD
ERT in primary ovarian failure-
Estrogens in cyclical pattern-
Ethinyl estradiol-0.01mg- 3 wk for 4 m
-0.02mg- 3wk for 1yr

6. Dysfunctional Uretine Bleeding -Mainly progestins
Dysmenorrhoea- mainly NSAIDS, cyclic estrogens
Acne and hirsuitism-cyclic estrogens
ERT in primary ovarian failure-
Estrogens in cyclical pattern-
Ethinyl estradiol-0.01mg- 3 wk for 4 m
-0.02mg- 3wk for 1yr
Estrogen adverse effects
Gynacomastia, Feminisation, Prostatic Ca,
Migraine, Breast tenderness, Amenorrhea,
Gall stones, hepatic dysfunction
C/I- diabetes, hepatic failure, thromboembolic disorder

7. 1. Oral contraceptives
2. HRT to limit estrogen’s effects on the endometrium
3. Uterine Bleeding disorders -DUB
4. Premature labor (decrease uterine contractions)
5. Stimulate Appetite in AIDS or cancer patients
6. Endometriosis-danazol(non-progestrogenic, non-
estrogenic, but exhibits hypoestrogenic,
hyperandrogenic, cause atropy of endometrium)
7. Premenstrual syndrome
8. Threatened abortion
9. Endometrial Carcinoma

9. Progressive accomplishment of alterations in genital tract
characteristic of estrus, metestrus, diestrus and
proestrus produced by changes in ovarian hormones
Occurs in non-primate females
Estrous period or estrus Period of sexual receptivity in
female Commonly called "heat".

10. Animals Length of cycle Length of heat
Sheep 16-17 days 24-36 hours
goat 21 days 32-40 hours
pig 19-21 days 48-72 hours
cow 20-22 days 18-24 hours
horse 19-25 days 4-7 days

11. Proestrus (FOLLICULAR PHASE)
Days 17 to 21 of estrous cycle of cow
Ovarian follicles are growing rapidly
Increase in Estrogen secretion by growing follicles
Decrease in Progesterone from corpus luteum (CL)
Mucosal layers of vagina and uterus multiply
Estrus
Day 1 of estrous cycle of cow
Period of sexual receptivity
Standing Heat
Surges of FSH and LH from anterior pituitary gland
pig, sheep and horse ovulate during this phase

12. Metestrus Days 2 to 4 of estrous cycle of cow
Short transitional phase after ovulation with
decreasing Estrogen and increasing Progesterone
concentrations Ruptured follicle is reorganizing into a
Corpus Leuteum(CL)Cow ovulates during this phase
Diestrus (LUTEAL PHASE)
Days 5 to 16 of estrous cycle of cow  Continuation of
luteal phase in species with long estrous cycles
CL becomes fully developed and Progesterone exerts a
dominant influence on oviducts, uterus, vagina,
pituitary and hypothalamus

13. 1. 3.
2.
ENDOCRINE REGULATION OF ESTROUS CYCLE

14. I. Introduction
- The term menopause is derived from Greek Meno
(months) and pause (cessation). The word means
cessation of menstruation.
- Cliamacteric which is by dictionary definition is
period of life when fertility and sexual activity decline.
It is a wide term leading to:
Pre Menopause
Peri Menopause
Post Menopause

15. - It is 3-5 years period before menopause with
increase frequent irregular anovulatory bleeding
followed by episodes of ammenorrhea and
intermittent menopausal symptoms.
Menopause:
- The point in time at which menstrual cycles
permanently cease. It is a retrospective diagnosis
after 12 months of ammenorrhea women classified
as being menopause.
- Mean age – 51 years.

16. The number of primordial follicle decline even before birth
but dramatic just before menopause.
Increase FSH, LH from about 10 years before menopause.
Close to menopause: There will be -anovulation
-inadequate Leuteal phase →
decrease progesterone but not astrogen level → lead to
DUB and endometrial Hyperplasia
- at menopause dramatic decrease of Estrogen→
menstruation ceases and symptoms of menopause started.
But still ovarian stroma produce →small androstenedione
and testosterone but, main postmenopausal estrogen is
estrone produced by Peripheral fat from adrenal androgen.

17. Hot flushes cutaneous vasodilation
- occurs in 75% of women
- more severe after surgical menopause
- continue for 1 year
- 25% continue more than 5 years
Urinary Symptoms
- urgency
- frequency
- nocturia
Psychological changes decreased level of central
neurotransmitters
- Depression, Irritability, Anxiety
- Insomia
- lose of concentration

18. Cryo is Greek word. (krayos – frost)
It literally means preservation in “frozen state.”
The principle - to bring plant cells or tissue to a
zero metabolism and non dividing state by
reducing the temperature in the presence of
cryoprotectant.
It can be done :
Over solid carbon dioxide (at -79 degree)
Low temperature deep freezer (at -80 degree )
In vapor phase nitrogen (at -150 degree)
In liquid nitrogen (at -196 degree)
Cryopreservation

19. Once the material is sucessfully conserved to particular
temperature it can be preserved indefinately.
Once in storage no chance of new contamination of fungus
or bacteria.
Minimal space required.
Minimal labour required.
Cryopreservation

20. The cryopreservation technique followed by the
regeneration. It involves following steps :
Selection of material.
Addition of cryoprotectant.
Freezing.
Storage in liquid nitrogen.
Thawing.
Washing and reculturing.
Measurement of viability.
Regeneration
Mechanism of cryopreservation

21. Genotypic drift
Senescence leading to extinction of cell line
Transformation to tumor related properties
Contamination
Distribution to others
Saving reagents, time
Equipment failure such as incubator
Cross-contamination by other cell lines

22. Sex determined in pre-implantation embryos
Approach- either invasive or non invasive
Splitting of sexed embryos
First successful embryo sexing done by Gardner,
1968 in rabbits by cytological method (Barr body
observation)

23. Non invasive Methods--
Immunological assay of HY antigen
Quantification of X-linked enzyme
Differential growth of male & female embryo
Invasive Methods--
Cytogenetic analysis
observing Barr bodies
chromosome analysis
Y-specific DNA probe
Y-specific DNA primer & PCR

24. Barr body forms after certain stage in embryo depending
on species
Performed at blastocyst stage
Affected normal embryonic development in some case
outdated now
Not applicable in domestic species
Gardner, 1968

25. Sexing from Trophoblast biopsy, day 12 -- 15
Accuracy 58.5--68%
sexed embryo at 6-7 days but low efficiency & low
survival rate reported
Slides prepared at metaphase stage
Depends on how many cells at metaphase stage
Takes an expert 5 hours to process 12 - 15 embryo

26. H-Y detection used for murine, bovine, porcine embryo.
Detection as early as 8 cell stage.
Can have two approaches cytotoxic or immunofluorscent.
In pigs detected only after removal of zona pellucida.
Accuracy
84% in cattle, 85% in goat, 81% in pigs.
Immunofluorescent Assay
detection stage specific
Monoclonal and polyclonal antibodies has equal effect
Poor quality embryo show fluorescence unrelated to
presence of antigen

27. Revolutionized the technique of
embryo sexing
Reduced time requirement
Increased efficiency
Embryos have been successfully
sexed in a number of farm
animals by using this technique

29. Embryo sexing done for cattle, sheep, pigs, horses, goats,
buffalo & humped cattle
Differential growth rate not applicable to choose embryos
Single cell is sufficient for sexing
Rapid sexing within 2 hours by using multiplex PCR they
used BOV-97M & bovine 1.715 satellite DNA sequence
Non Electrophoretic method for PCR sexing reduced time
requirement to less than 2 hours

30. Altering the male & female sex ratio in farm animals
Increase in milk & meat production
Getting quality bulls
Conservation of rare breeds of farm animals