Fertilization, Maternal recognition of pregnancy and

Implantation

• Prof G N Purohit
Head Department of Veterinary Gynecology and Obstetrics, College
of Veterinary and Animal Science, Bikaner, Rajasthan, India.
 Some definitions
• Obstetrics Science of midwifery (obstare "stand opposite to)
• A branch of medicine that deals with care of pregnant women
during pregnancy, at child birth and afterwards

• Veterinary Obstetrics:
• A branch of Veterinary Medicine that deals with
• care of animals during pregnancy, at parturition
• and during the postpartum period
• Embryo: from fertilization to the differentiation stage (45 days in
cattle)
• Fetus: from differentiation till parturition
• Conceptus: Fetus with its fetal membranes.
• Nullipara: An animal which has not become pregnant and not
parturated.
• Primipara: An animal which has become pregnant and would
parturate for the first time
• Monotocous(Uniparous): Animals giving birth to single young ones
• Polytocous : Animals giving birth to many young ones
• Embryology: Science of physiological development of the fetus
• Teratology: Embryology + pathology dealing with abnormal fetal
development
•
Carazol: a beta-blocker given to sows to reduce the expulsive time of labor
in sows.
Clenbuterol: A beta-adrenergic that quietens the uterus and inhibits labor.
Psuedopregnancy: False pregnancy occurs in the bitch and the goat and in
many other species like mare.
Feathering: Tapping of the vagina of a whelping bitch with finger leads to
oxytocin release and delivery of a pup.
Single pup syndrome: The presence of a single pup in a bitch leading to
problem in initiation of parturition and dvelopment of an extra large
puppy.
Spalding sign: Overlapping of the fetal skull bones seen on radiography
of a bitch with dead fetuses.
Fergusons reflex: The release of huge quantities of oxytocin in cows
when the fetal legs touches thee pelvis during parturition.
Theilers disease: Infectious hepatitis in mare developing due to
injection of an eCG.
PREGNANCY IN DOMESTIC ANIMALS
• The period of intrauterine development (from fertilization to
parturition) in mammals is
termed pregnancy or gestation.

• Ovum + sperm fertilization Embryo

Completion of gestation Growth in uterus
(Pregnancy) Parturition
 FERTILIZATION
• External Amphibians
• Internal Mammals
• Fertilization in mammals
– Gamete transport
– Changes in gamete structure
– Cleavage and blastocyst formation
– Maternal recognition of pregnancy
 Gamete transport
• Synchronous transport of male and female gametes to oviduct
ampulla should occur

• Semen deposited in vagina (human,

• cattle, buffalo, sheep, goats, rabbits).
• Semen deposited in the uterus
• (in dogs, horses, pigs).
 Oocyte transport
• Except mares the oocyte is picked by the fimbriae and enters
the ostium

• Propelled towards the ampulla by ciliated cells

Fertile life of gametes in domestic animals
Day of
Fertile life
Fertile life entry of Blastocyst
Species ovum
sperm (hrs) embryo in formation
(hrs)
uterus
Cattle 24-48 20-24 3-3.5 7-8
Horse 72-120 6-8 4-5 6
Sheep 30-48 16-24 3 6-7
Swine 24-72 8-10 2 5-6
Dog 120-168 48-72 5 7-8
Buffalo 24-32 6-12 4 5-6
 Changes in gamete structure

• Sperm capacitation, sperm oocyte interaction, acrosome

reaction, oocyte activation, zona reaction and intermingling of
male and female gametes

• Capacitation occurs in female tract

Epididymal
Effect of Capacitation
Ejaculated Capacitated

Seminal
Plasma
and/or Female
+ + Tract
Epididymal
Secretions
Zona Pellucida

Oocyte

Ca2+
Ca2+

Ca2+

Ca2+

Perivitelline
Space
Zona Pellucida

Oocyte

Ca2+

Cortical
Zona Block to
Granules Polyspermy

Perivitelline
Space
Sperm acrosomal Perivitelline Initiates completion of
region binds to sperm Meiosis II and release
Space
receptor on zona of 2nd polar body
Vitelline 1st Polar Body
Membrane Zona Initiates -
Cortical Pellucida Zona &
ZP3
Granule Vitelline
Binding Fusion Reaction

Cortical
Granule
Release
Zona
Initiation of Reaction
Acrosome
Reaction

Continuation of Excluded Sperm

Acrosome-Reacted
Reaction Penetration
Sperm
Zona and Vitelline
Sperm penetrates
zona Reactions block
Redrawn from polyspermy
Wasserman, P.M. 1988 Scientific American. p78
 Blastulation Gap Junctions

Inner Cell Mass Zona Pellucida

Trophectoderm
Na+ [Na+]

H2O

Tight Junctions
 Hatching
• Bovine
– 9 - 11 days
• Equine, Ovine
– 7 - 8 days
• Porcine
– 6 days
Buffalo
6.5 days
 Cow
Conceptus Growth • Day 15, 1-2 mm
Occurs in cow, pig and sheep • Day 18-19, 10-20 cm

Mare remains
Spherical spherical! Embryonic Area

Tubular Elongating
Trophoblast

Filamentous
Elongated Day 15 Porcine Conceptus
Uterine Location of Elongating Ruminant Blastocyst

Corpus
Luteum
Bovine and Ovine
 Pig Intrauterine Migration

Day 5
 Pig Intrauterine Migration

Day 7
 Pig Intrauterine Migration

Day 12

Embryos become fixed

Transuterine migration is rare in cow
and ewe!
 Trans-uterine Migration in the Mare
Begins Day 10

Fixation can occur in either Fixation on day 15

horn! - 16

Corpus
Luteum
Inner Cell Mass
Gastrulation Trophectoderm

Formation of Endoderm
Germ Layers

Blastocoele
Cavity
Gastrulation

Endoderm
Endoderm

Yolk Sack
Gastrulation

Endoderm
Endoderm

Yolk Sack
 Gastrula
Ectoderm Mesoderm
Extraembryonic
Coelom

Yolk Sack

Endoderm Trophectoderm (Chorion)

 Germ Layers
• Ectoderm • Mesoderm  Endoderm
– CNS – Circulatory » Digestive
» Liver
– Sense organs – Skeletal » Lungs
– Mammary glands – Muscle » Pancreas
– Sweat glands – Reproductive tracts » Thyroid gland
» Other glands
– Skin – Kidneys
– Hair – Urinary ducts
– hooves
 Embryonic
Fetal membrane Formation
Amniotic Folds
Ectoderm

Extraembryonic Ceolom

Yolk Sack

Trophectoderm
Chorion
Mesoderm
Endoderm
Amniotic Fold
Amniotic Cavity

Allantois

Extraembryonic Ceolom
Yolk Sack

Chorion
 Amnionic Cavity
Amnion
Uterine
Lumen

Allantois
Yolk Sack

Chorion

Extraembryonic Ceolom Carnuncle

 Amniotic Cavity
Chorion
Amnion

Allantois Cavity

Yolk
Sack

Allantochorion Cotyledon
(Chorioallantois)
 Fetal (Placental) Membranes
• Yolk Sack
– In birds to nourish embryo
– In mammal atrophies but source of blood cells and primordial germ
cells
• Amnion
– Non-vascular, fluid filled
– Fluid produced by fetus
– Protective cushion
– Ruptures at birth (not breaking of water)
 Placental Membranes
• Allantois
– Blood vessels
– Fuses with chorion
• Allantochorion or chorioallantois
• Brings blood vessels to chorion
• Chorion
– Outermost membrane
– Attachment to mother
Maternal Recognition of Pregnancy
 Luteal Regression

PGF2a
(Bovine, Ovine,
Porcine) PGF2a
(Equine)

No Uterine Factor
(human)
 CL Status
CA
Mechanisms

CL Regression
•Luteolytic
•No CL support
(+)
Uterine PGF2a CL Maintenance
CL •Antiluteolytic
» Block PGF2a effects
•Luteotrophic
» Stimulate progesterone production by CL
Filamentous Embryo
(+)

Maintained
 Recognition of Pregnancy in Bovine and Ovine

Conceptus Secretions
• Interferons
•oIFN-t
Mechanism
•bIFN-t
• Antiluteolytic
• Inhibit oxytocin receptor synthesis
• Inhibit PGF2a synthesis

Critical Days
• Bovine - 16 to 17
• Ovine - 12 to 13
 Recognition of Pregnancy in the Porcine
Conceptus Secretions
• Estradiol
E2
E2

Critical Days
PGF2a
E2 11 to 12
PGF2a
PGF2a
E2

E2

E2

PGF2a Mechanism
•Antiluteolytic
» Redirect PGF2a
» Requires 2 embryos per horn
 Recognition of Pregnancy in the Mare

PGF2a PGF2a

Conceptus Secretion Mechanism

• ? • antiluteolytic

Critical Days
10 to 14
Recognition of Pregnancy in the Human
Conceptus Secretes
• HCG

Mechanism
• luteotrophic

HCG

Critical Days
•8 to 12
 Dog and Cat
• CL does not regress if not pregnant
– no uterine effect
• Psuedopregnancy
– CL last similar length as in pregnancy
Implantation
 Time of Implantation
Attachment to Uterus

Species Beginning Completion

pig 13 24
cow 22 40
sheep 15 28
mare 24 - 40 95 - 100
human 7-8 30
The time from ovulation, during which implantation occurs
in different species.

Species Time (days )

Cattle 17 to 35
Sheep 14 to 18
Pigs 12 to 16
Horses 17 to 56
Dogs 14 to 18
Cats 12 to 14
Human 6 to 7
 Type of Implantation
• Centric
– Chorion lies opposed to uterine wall
– Farm animals
• Eccentric
– Chorionic sack in a uterine fold
– Rodent
• Interstitial
– Embryo digests part of uterine wall
– human
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