Fertilisation

rermisation is the penetration ofa mature ovum by the spermatozoon and the fus ion of the respective
,rJemale ronuclei. The penetration ofa spermatozoon into ovum activates the egg to start the
derelopen, while the fusion of their pronuclei (amphimixis) results in restoring the diploid (2n) or
Reception cone Haploid
Ovum Sperm sperm nucleus Sperm nucleus
Centrosome
Asters

Haploid nucleus Nucleus of ovum Nucleus of ovum

Paternal nucieus Paternal chromosomes Chromosomes
from spem
Spindle

Asters

Maternal nucleus Maternal chromosomes Chromosomes

from ovum
Daughtercells

Constriction
Fig. 8.1 Diagrammatic representation of process of fertilisation.

|99|
 100 Chordate Embryology
Leeuwenhoek (1683),
main idcn of Fertilisation was enunciated by impregnation
The
parental chromosome number. ovum ruitful, (/.e., to devclop into the embryo) its
to make
who emphasized that in order
(the spem) is essential.
animalculc
with seminal
DUAL ASPECT OF FERTILISATION
Fertilisation :
independent nspecls of
There are (wo cssentially cortex and
of lhc sperm initiates 4 series of changes in the egg
1. Activyation : Penetration cnhancing its mctabolic
activitics.
activates the cgg cytoplasm by pronuclei is known as amphimixis. It is not
fusion of thcmale and female pronucleus may
2. Amphimixis : The cven the single set ofchromosomes of female
that essential for the
development becausc
Fertilisation restores the original diploid chromosome
haploid cmbryo. maternal hereditary
control the development ofa about the intermingling of paternal and
this brings
number of the paretnts. Morcover, without
material.
crustaceans and certain insects) develop regularly
invertebrates (rotifers,
The eggsofcertain development is called
parthenogenesis.
Fertilisation. The method of
sITE OF FERTILISATION
body-ffuid
whether it is sea-water, freshwater or
the aquatic medium, ova are shec
Fertilisation always occurs in and lower chordates, the sperm and
In many marine
invertebrates the body of
of the maternal organism. type of Fertilisation where gametes fuse outside organs ere
Fertilisation occurs. The forms special
in water, where In a large number of terrestrial copulatory
parent is known as
external Fertilisation.
genital tract of femnale. These organs are called the
sperm in the Fertilisation completed inside
developed in male for depositing female gametes. The
Fertilisation of ovun
fusion of male and advanced forms, the
organs. This facilitates the Fertilisation. In still famale. The
termed as internal inside the uterus of the
bod of the femaleis embryonic and foetai development occurs
eggs are called
and the subsequent are called viviparous, while those that lay
young ones
individuals that give birth to
oviparous.
MECHANISM OF FERTILISATION

separatcd into the following five steps

:
The process of Fertilisation can be
spermatoz0a to ova.
1. Approach of
contact.
2. Capacitation and penetration.
3. Acrosonal reaction and
4. Actívatíon of ovum.
5. Amphimixis.

Approach of Spermatozoa to Ova

The male and fenale ganetes are produced either in separate parents or inThe separate organs. 1e
(sperm) is motile, fertilizability and
female gamete (ovum) is innobile, while the male gamete hours. Eggs otaquatie animals
viability of ganetes are often limited varying lroma lew minutes to a fewcase ot aniunals with interna1
with external Fertilisation survive only for a few miues and in humai
Fertilisation for afew Ihours and in case of man hunan cgL survives tor24 hours, Lie span of
Spern is 24 hours, cod's spern 2weeks, bat's spern 3lo+months and honey bee's sperm many months.
Moreover, the energy in the spern is limited. Consequently, the nnajor problenn in sexual reproduction 1S
bringing together the ripe ova and lhe spern.
 Fertilisation 101
1. Chemical Attraction or Trapping of Sperm by Chemotaxis
Achemical substance, the fertillzin, secreted from the cortical region of egg cytoplsm of
sea-urchin egg was presumed to attract the sperm towards the egg. But an actual evidence for
straction is lacking. The encounter bctwcen sperm and ova of sca-urchin is brought about t by chemical
movemets of. spermatoz0a. Their movements are at random and spermatozoa collide with swimming,
the egg as a
matter of pure chance. This applies for most animals. However, in certain cases the chemical
fom eggs serves to trap spennatozoa, that nccidentally enter their field of influence and aresecretion
towands the eggs. For example, intwo species of colonial hydroid, Campanularia, the cells ofguided theca
present nearthe gonangium produce sperm-attracting reagent.. Similarly, a chemical, sperm attractant, is
produced by fish eggs.
2. Mechanical Juxtaposition of Gametes
To facilitate an casy encounter of the spertm and ovum special devices are seen in
animals with
extemal as well as internal fertilisation.

Ova
Funnel

Path of swimming
spermatoza

Fig. 8.2 Atheca of coelenterate, Campanularea.

(a) In forms with external fertilisation: In marine animals (like sea-urchin) a large number of
sexually mature males and females discharge their eggs and sperm in sea-water in close vicinity
and at the same time. Consequently, clouds of sperm and eggs are forned to
Fertilisation. Since, the life span of gametes discharged in sea-water is just a few ensure mass
minutes to a
-few hours, the discharge of ova and sperm at the sametimeis essential..
In fishes, amphibians and freshwater invertebrates, the life span of
sperm is reduced to at
minutes only. Therefore, the sperm are discharged directly on the ova. These animals exND
pseudocopulation or amplexus.
(b) In forms with internal fertilisation: The intermal fertilisation is
foundtract
forms. In them the spermatic fluid (semen) is delivered in the genital mostly nt e
of female by thee
insertion of aspecialized sexual appendage that acts as an intromittent organ. This organ
known as penis and the insemination. The
phenomenon of
insemination occurs during sexual embrace, sperm is called
called copulation or coitus, which is exhibitedi
deposition
reptiles, birds, mammals and insects. In some amphibians, gastropods,turbellarians, trematodo
and cestodes, the:
sperm are received in a specialized 1receptacle after copulation.
 103
Fertilisation
102 Chordate Embryology
In majority of fons, the encounter of cggand spem occurs in upper part ofthe oviduct. However, in
some viviparous fishes (Gmbusta afinis and /ieteraniria formAsa) and certain eutherian mammals, it is
accomplished inside the ovarian tollictes, The spem are deposited in the vagina and from here are
transported to utenis by thc contraction ofuerine wal.
3. Special Devices Eag wth jalty
Aggluttnsted eperm
To bring together the cggs and syemmatoz00, nmber of other devices hanve been encountered in
animals. In spiders and cnistaceans a foOrelimb or a pal in male is modificd for transferTing semen into
Froo 8wimming sporm
genital opcning of female. In cehaloods, one of the arns is modified for holding spermatophores and Nuclaus
transfeing them to the femalc. Sueh a specialized arm is known is heterocotylized arm or
heterocotylus. In Per|Natoysis. the spematophores are applicd to any part ofthe body. The spermatozoa Egg jetty
are released on contact. penctrate the bodywalland make thcir way through the body tissue to reach the
OVa. -Egg Jolly dissotved
Spermatophore Sperm extract
-Sperm Ovum with egg jeily
Fertilizin-antfertilizin reaction.
Cuticle Fig. 8.4
Blood capillary glycoprOtein
or
(1919). It is
a
Ectoderm 3. Fertilizin and Antifertilizin Reaction Lillie Its protein
by F.R.
8 2 , 0 0 0 - 3 0 0 , 0 0 0 .
O00oDdCE Connective Structure and functions of fertilizin were studied of about comprises
molecules
tissue molecular weight are
mucopolysaccharide. Its macromolecules have a
component m o n o s a c c h a r i d e s
pollysaccharide
The
acids, while the
moity is formed of anumber of amino glucose, or
galactose.
fertilizin may
have
fructose, fucose molecuie of
Muscle
of one or more monosaccharides like formula. Each attached to
more than one
Jesterified by sulphuric acid as is shown in the fertilizin mnolecuie becomes
layers following
fertilizin varies
c o m p o s i t i o n of
one
more than one 'active group'. Therefore,
agglutination of : s p e r m a t o z o a .
The
spermatozoon. This brings about the
of antigens.
Fig. 8.3 Spermatophore attached to the body wall of Peripatopsis. from species to species as in the case H
In sorme animals spermatozoa are released in small packets, called spermatozeugmata. In them the
H
sperm are held together by some mutual attraction, the enveloping sheath is absent.
H,OH
OH
H
Capacitation and Contact -o,so
1. Capacitation
species. This is
The egg of a particular species is fertilised only by the sperm of the same
monosaccharide.
sulphonated
Structural fomula of a
fertilizin is
accomplished by a chemical mechanism. A species specific chemical substance, called
Fig. 8.5
secreted by the Both
either in the plasnna membrane
or
produced by the ripe egg, while another chemica! substance antifertilizinislock like an antigen-antibody ripe unfertilised eggs.It remains is produced
fertilizin and antifertilizin are species-specific and form a sort of chemical the egg of The fertilizin is produced by thesea urchin eggs, the jelly consists wholly of fertilizin and
reaction. The fertilizin acts as a receptor for antifertilizin and makes them capable of fertilizing in thejelly, surrounding the egg. In
the cells of zona pellucida
produce fertilizin.
the same species. This process is known as capacitation. by the follicle cells. In mammals,
Capacitation has been studied in sca-urchin., where it occurs in sea-water. In mammals the sarme
S Functions: Fertilizin performs following functions
that they adhere into clumps,
facilitated inside the genital ract of the female. Capacitation is essential for spernm penetration. " makes the sperm sticky so fertilisation,
surface of egg and ensures
makes spern to adhere to the the egg to check
2. Adhesion out tlhe mumber of spermatozoa around
of fertilizin molecules " clumping or agglutination of sperm thins
Tne adnesion o' spermatoz0onto the egg surlace occurs due to the presence the entry of any additional sperm into the egg.
on the surface layer of egg cytoplasm, which receive antifertilizin molecules present on the surtace ol " leads to the capacitation of sperm,
enabling it to penetrate the egg surface.
spermatozoa.
 104 Chordate Embryology
Fertilisation 10S
Jetly
Micropyte
Polar bodies
Fertillzin Micropye Gorrninal vesicle
Spern hoad -Pronucleus Polar hodies
Egg
Egg cytoplasm
shell
Egg
mentbrane
B
Fertilizin
A
Fig. S.6 Clumping of spermatozoa due to their
binding with fertilizin molecules (After Cephaiopod, (From Austin, Fertilization).
Balinsky, 1976). Fig. 8.7 Few eggs with micropyle, A. Egg of Musca; B. Egg of a
Antifertilizin is found on the surface membrane of the the egg is
called corona radiata. It is
Antifertilizin contains acid proteins (b) Hyaluronidase : In mammals the covering around together by an adhesive substances the
with molecular weight of spermatozoa. It is also species specific cells. cemented which dissolve=
of sperm fit into the fertilizin molecuies of egg in the same wayabout 10.000. The antifertilizin molecu Tormed ofa single laver of follicular enzyme,
hyaluronidase,
as a key fits into sperm produce way for the sperm.
Sea-water in hyaluronic acid. The mammalian cells of corona radiata and makes
which
egg water. Sperm added to
eggs have been allowed to stand for some time contains the lock. iyaluronic acid, separates the follicle carbon dioxide that
egg water are fertilizin and is termed a acidification of sea-water by
dissolved by the
spontaneously, but their fertilising agglutinated, adhering by their head. These, later on, separ (C) ln sea-urchin, the egg ielly is
reduced capacity for fertilisation. power is greatly diminished. Eggs treated produced by the spermatozoa.
with antifertilizin also ha radiata shortly after ovuiatic
sheep), the eggs lose their corona
Acrosomal Reaction and (0) In some mammals (horse, cow, direct access to zona pelluciáa.
Penetration enabling the spermatozoa to gain undergone capacitatior
Almost all the animal eggs, except lysin or hyaluronidase till it has
those of The acrosome is not able to release sperm in the sperm acrosome.
membranes or gelatinous porifers and cnidarians, are enveloped by one or mort activation. This involves some specific change
the egg surface has to layers external to the plasma membrane. A spermatozoon
penetrate these membranes so as to reach the egg after attachment 2. Acrosome Reaction
reaching the egg plasma membrane,
undergoes a number of cha
1. Sperm Entry by Way of plasma membrane. The activated spermatozoon on changes are collectively described under
acrosome reaction.
Inthe eggs of nemertines,
Micropyles n its acrosomal region. A!ll these
gastropods, Acrosomal granule
membranes surrounding
the site at Which the egg h¡ve pores,pelecypods, echinoderms,
called micropyles. insects, cephalopods and fish, t Nucleus
the egg
the sperrn enters through was origirally atached to the ovary. In theWhen micropyTe is single, it represents Nuclear
one of the the single micropyle. eggs of insects, fish and envelope
spermatozoa may enter cephalopods
pores of the egg If pores are many, the
surface. The sperm motility Periacrosomal
lysins: The spermalozoa increased when these approach the throughn an
(a) Egg membrane is material
lysins are produced fromproduce Iytic substances which are micropyle.¢
nature. These sperm
Acrosomal
membrane
specific. The the enzymatie
clear the spern lysins dissolve the acrosome of the Egg envelope
in the path for spern to reaclh the membranes present aroundspermatozoa
vitelline plasna
and are
the egg in a local spe
imembranc of tlic cgg of
membrane of the egg.
Actual arca Sperm
membrane
Mytilus caused by the entry of holes have been s
sperm. Fig. 8.8 Electron micrographic structure of acrosomal region of spermatozoon of
Saccogloss
 106 Chordate Embryology
107
according to the structure of
details of the process are common to many animals with minor differences
Fertilisation
acrosome and egg membrancs.
In Saccogiossus and also in other animals an ncrosomal veslcle is present in the sperm head anterior
to the nucleus. It is bounded by an acrnvomal menbvnne. Inside the ncrosomal vesicle is nlarge dense Plasma memttane
acrosomal granule which is suunumted by tine, gramlar material, except in the apical region where an
apical space lies betwven the cnsomal grmule anl ncrosome membrane.
Flhrotn
During the acnsonal ractin, the events ocer in the following sequence : material
(a) the dissolutiv of plasma membrane and acrosomal membrane in the apical region forming an
apical one through nhieh contents ofacrUsomal vesicle nre passed on to the egg.
(b) the union ofthe plasma nennbrnc and acrosomal menbrane around thc margin of the opening.
(c) elease of contents of the acrosomal vesicle, i.c., the acrosomal granule that contains lytic
cnzyes, nvted for spenn penetration, Acrosornal
(d) decpening of the shallow depression of acrosoumal membrane close to the nucleus and formation tubule
of an acnosnal ulbule in the form of a projcction from the inner surface of acrosomal Nucleus Acrosomal
membrane,
rod
Nucleus
Sperm plasma
membrane
Periacrosomal
material Acrosomal Periacrosomal E
membrane material lait
Mitochondria D
Acrosomal A
granule Acrosomal
Nuclear Tubule
membrane
Acrosome Acrosome rod
Periacrosomal
material
Axial rod
Acgosome tubule
Nucleus
Mitochon YFold of acrosome
-drion
Egg
membrane
C o r t i c a l ) cytoplasm Piasma
membrane
Acrosomal
Acrosomal
B Tubule C D granules E
granuie
Fig. 8.9 Acrosomal changes during fertilisation of Saccoglossus (After Torey, 1971).
(e) lengthening of the acrosomal tubule by the eversion of the remaining acrosomal membrane and Distal
centriole
Proximal C.
its addition to-the tubule at its base, centriole B
A
tubule:
(f) liberation of the content of the acrosomal tubule, with long acrosomal
Acrosome reaction.
A. In sea-urchin rod.
Fig. 8.10 acrosomal
(g) penetration of acrosomal tubule gradually through the egg envelopes to reach the egg plasmae B. In Nereis
with an
membrane, projections.
tubule is short with many
(h) fusion of the membrane of acrosomal tubule with egg plasma membrane and activation of egg hydroids), the acrosomal
0) In other forms (e.g, tills the core of tlhe
tubule.
A number of modifications are seen in the pattern discussed above. For example, dense acrosomal rod acrosomal
) In Nereis, an electron membrane of egg and
(a) In animais, whose eygs have athick jelly coat, the acrosomal tubule is very long and thin and acrosonmal tubule is not
fomed. The plasma
V) In mammals the contact and spernatozoon
is drawn into the
helps to penetrate through the jelly-coat as in sea-urchin. dissolve at the point of
membrane of spermatozoon
 108 Chordate Embryology
Activation of Ovum forns a
p r o j e c t i o n
p r o j e c t i o n
and
When acrosomal tubule in J Formation of Fertilisation Cone fusion
a
conical
sequence of events in egg. Thesc spennntozoon contacts tlhe plasma membranc
c y t o p l a s r n i c
form of
arc described ns
nctivatlon of ovum. These are:
of egg. t ngges Theecytoplasm
cy of the egg bulges forward in the region ofmayhe in the the
sperm
ofbyaline cytoplas1n. This is known as Fertilisntlon cone. It
processcs. drawing
periacrosomal
pscudopeodia-like subsides
the
racytoplas1mic cyl1nder or may consist of several cgt.
grnduzily
while
Nucleus p e r m a t o o o n .
the
The fertilisation cone grad1nlly engulfs t#he sper
graule
never
eters
In
mammals
ithin the egg. During this process the acrosonial p interior
of cgg. the vitelline
outside
most
yalerial activates the cgg. into the remains
c y t o p l a s m .

In
Spermatozoon
tail
Variations are noticed in the cntry of he egg
spern
Acrosome enter the
In cchinoderms,
onplete spematozoon cnters cgg cytoplasn.
centriole
proximal
the cgg-
Egg membrane, while in Nereis only the spern hcad andtheir
entry into which
envelope nals, however, spcrn nucleus and mid-picce get physiochemical

changes,
F e r t i l i s a t i o n
A B of of
C Cortical Reactions formation
may
undergocs a number
lead to
the spermatozoa

which
Soon. the cortical region of egg cytoplasm general,
these
of other
Acrosomal
collectively called cortical reactions. In
blocks the entry
tubules
embrane outside the ege plasma membrane. This
and
grike the egg later. Egg studied in
sea-urchin
Egg plasma in
Sea-urchin
have been
Membrane
membrane Formation of Fertilisation cortical
reactions
The formation of fertilisation menbrane and
Fertilisation membrane
Marfish. (800A"))
Egg
Hyaline layer
cytoplasm
Vitelline membrane
D E F Cortical granule
(150-200A)
Sperm plasma (0.5- 1.3u)
membrane
-Sperm nucleus
Centriole
Egg plasma membrane
Egg cytoplasm Periviteline space D
A
layer in sea-urchin egg.
Sperm nucleus membrane and hyaline
Fig. 8.12 Formation of fertilisation
membrane andan inner
plasma
G iscovered with an outer vitelline invoBve
The unfertilised eggofsea-urchin graules. The corical reactions
Fig. 8.11 -Sperm-egg association- during fertilisation-in-a polychaete, membrane is alayer of cortical
(After Austin). membrane. Beneath the plasma
Hydroides hexagonus
he following changes : and forns the outer
separates fronn the plasna membrane, expands
1. The vitelline membrane space Tormed between the tvwo
layers is the perivitelline
1. Cytoplasmic Fusion ayer of the Fertilisation mennbrune. The
and the tw
membrane space. cortical granules. Their
The plasma membrane of acrosomal tubule fuses with the egg plasma
with tha 2. The contact of acrosome with egg surface is lollowedl by tlie rupture of
membranes at the region of contact dissolve. Thus, the egg cytoplasm becomes
continuous
reak down begins initially at the point of spern cntty and spreads through the cortex in all directions.
the sperm. The granules first swell up by imbibing wuler and the mucopolysaccharide contained in them liquifies
components :
he breakdown of granules releases thrce
 111
110 Chordate EmbryologY Fertilisation
Vitellino membrane layer
nd relcases
Plasma membrane jelly-like forms a thick of
the vitelline substance through the The jelly fragments
yutside membrane.
-Homisphere membrane, pores
membrane
dical alveoli fuse to form a newthat forms the
af Vitclline
membranc.
The
-Cortical granule
membrane fertilisatio
of the cg
Eloctron
trnnsparent portion Sperm
Electron opnque Joily coat
Alveol
A Imalarlal
Vitellino membtane
Piasma mombrane
-Hemisphere
-Cortical
membrane
Electron
Aransparent portion Egg cortex Egg
A C
Electron opaque material B
Nereis egg-
Vitelline membrane in
membrane
Fig. 8.14 Cortical reaction and Iformation of
fertilisation
Hemispheric C) In Bony Fishes and Frogs space. The
Fertilisation
fluid.
perivitelline
globule The unfertilised eggs of fishes, and frogs have a narrow amount of
perivitelline
bony
membrane is not formed but the increases the
rupture of cortical granules
D) In Mammalian Eggs egg plasma
C pellucida and
between zona in the
mammalian eggs, the perivitelline space develops releasetheir content
only,
around the egg
Fertilisation membrane embrane. The cortical granules that are present in some mamnalsother membrane is formned
Perivitelline space grivitelline space. Neither the fertilisation membrane nor any does not
Hyaline layer
cyoplasm. amphibians and insects the
cortical reaction
In the eeggs of certain animals such as urodele
in
Cytoplasmic projection
cur at the time of fertilisation.
pole (the blastodisc) but at the vegetal pole
accumulates at the animal Normally, the ectoplasmic
D rsees, the ectoplasn1 outside the poles in Amphibia.
ylea and annelids and at a point The formation
Pcumulatiorn occurs at the point of sperm entry.
Fig. 8.13 Changes in the cortical region of egg cytoplasm and formation of fertilisation me.nbrane in Fertilisation Membrane:
and Formation of penetration of
sea-urchin egg under high magnification. Ogniticance of Cortical Reaction mechanisms against the
reaction are defensive
Erilisation membrane and cortical plasma membrane.
established contact with the egg
The lamellar parts of the ruptured cortical granules appear as dark bodies or folded lamellat aaltionat Spermatozoa after one has
These fuse with the vitelline membrane contributing to its thickening, Metabolic Activation degeneration. The
fertilised, it undergoes
egg pla quiescent stage and ifit is not adeuyl cyclase becomes
The released globules fuse together and form a hyaline layer just outside the Ine ripe unfertilised egg is in metabolic activities. An enzyme AMP
membrane. It adheres to the egg surface and keeps the blastomeres together during
cleavage. number of formation of3-5 cyclic
em penetration introduces a entry. This cnzyne starts the
luid that is
The liquid component of cortical granules forns the perivitelline Ctivated in the plasma membrane by s p e ithe fertilised egg. Thechanges are :
hich activates most of the metabolic plasma membrane becomes
perivitelline space. It contains polymucosacclharides and water. Permeability of Plasma
Membraue : Tlhe egg
ions. Its electric
" Change in the glycol phosphate and K
entry of water, etlhylene,
(B) Formation of Fertilisation Membrane in Noreis Egg The cortical reg more permeable to tlhe positive and then ncgative.
In unfertilisedlegg of Nereis, the vitellinc nembraneiis traversed by microvilli. potential changes first lo calcium chanoes in
sodium, potassiun and of
plasma nem conceniralion
has several layers of cortical alveoli. The ruplure of cortical alveoli destroys the egg " lonic change: The
ionic
fertilised egg
 112 Chordate Embryology Fertilisation

113
NAD-kinase occurs in an innet..
Change in coenzyme : In B8 he enzyme
ate. It is activated durin,
immediately after fertilisation.
The activatcd NAD-kinase directs the interconversion contains
of coenzyme NAD into NADP and NADPI. DNA synthesis begins with sperm penetration. Its ratc increasses
The
unfert1lised
egg
matozoon,
Spern erntry initiates niclear division by introducing its
centriole.
of spert is
NAD kinase
alone. Withthe entrynuclear d i v i s i o n
NAD+ATP NADP +ADP only one centriole and is unable to form mitotic Cspindic and
the egg receives sccond ccntriole (the centriole of
s p e r r n a t o z o o n )
NADP and NADPH are responsible lior tlhe oxidatio-reduction reactions taking place during cell initiated.
respiration and synthesis ofenergy. Tus, inerensed NADP and NADPH contents initiate many synthetic
pathways. and
Migration of Pronuclei and Amphimixis or its head
spernatozOon
events of
" Respiration rate: he ovygen consunption increnses in the fertilised egg indicating an
increase in the espiration rate. llowever, it docs nol changc appreciably in starfish and
1. Future of Spcrmatozoon In tlhe Egg: Whether the whole
ultimatcly
participate in
futurc
amphibians, and it actually deercases aller Fertilisation in cggs of mollusc (Cumingia) middle picce enter the cgg, only the sperm nuclcus and ccntriole it
and fertilisation cone,
annelid(Chactoterus). development. inward from
2. Bchaviour of Pronuclei : head moves assume the forward
Increase in the activity of proteolytic enzymes in the rate of protein synthesis : The middie piece
spermatozoon
ns he centrioles of its
cytoplasn of arie unfertiliscd cyg contains complete machinery for protein synthesis but little rotates thrOugh 180°, so that the mitochondria and This is known
as
or no protein synthesis takes place. Most probably, some inhibitor or repressor proteins are position. Both nucleus and centriole change in appearance. appears
granular.
synthesised in phase of oogenesis, which inactivate the chromosomal genes, mRNA " The nucleus swells up, and its chromatin
becomes vesicular
molecules and ribosomes. During Fertilisation, some proteolytic enzymes remove these male pronucleus.
inhibitorS and activate protein synthesis. The centriole gets surrounded by astral rays and forins
the aster.
cytoplasm tothe site
where
Initiation of nuclear division : The eggs are ready for fertilisation at different stages of through the egg holoblastic
in
pronucleus andi centriole move
centre
meiosis in different animals. At one extreme, the eggs are laid with germinal vesicle intact and Simultaneously, the male site is near the
of sperm in
place. This The movement
do not complete meiosis until fertilisation. In them the germinal vesicle breaks down when union of male and female pronuclei is to take in telolecithal
eggs. cytoplasm. In
mierolecithal eggs and Iin the centre of active cytoplasm subcortical
of corticai and sperm nucleus is marked
spem enters the egg and ineiotic division is completed as in eggs of Ascaris and Nereis. the movement
the egg cytoplasm is usualy accompanied bypigmented taken by
Table 8.1 Time of second maturation division in the oocyte of different animals. and the path
frog egg thesubcorticall cytoplasm is heavilyThis is known as penetratton P
cytoplasm inthe form
Young Primary Fully Grown Female D' Pigment granules trailing along its path. in the peripheral swells up to
Just after the second maturation divisiontnthe egg nucleus lies
First Metaphase Second
pronucleus, which
Ooctye Primary Oocyle Metaphase Pronucleus female its from
of several vesicles, called karyomeres. These fuse to form
pronucleus, the female
pronucleus
Branchiostoma Ascaris, Dog, Fox Chaetopterus Amphioxus Coelenterates
become vesicular. Following the migration of male
Grantia Dentalium Most mammals Echinoids
peripheral position migrates towards the predetermined site of fusion.
Dinophyllus Nereis
Many insects Siredon (sea-urchin)
Original position of female
Hisriobdella Cerebratulus pronucleus Path of female
Peniopatosis Thalassema pronucieus
Saccocimus Nuclear
fusion
First polar body Polar bodies
Path of male
pronucleus
B Entrance of sperm
nucleus
Fig. 8.15 Diagram to show the maturation stage of ovum at the time of egg laying.
Althe other extrene, the cggs ripen only after the completion ofboth meiotic divisions as in urchns
and coclenterates. However, in majority of cases, tlhe eggs are laid at metaphase of the first or seeo D
meiotic division. Nuclear division is delayed until it is initiated by the sperm entry. Thus, the ine
initiation of nuclear division coincides witlh time of fertilisation in majorty of animals. The time Fia, 8.16 Tracks of male and female pronuclei of sea-urchin in their movement towards
second maturation division in different auipals is shown in Table 8.1. the sile of nuclear fusion (After Berrill).
 114 Chordate Embryology
115
the site of fertilisation, has not
What torce goverms the movement of male and female ronueBei to or the attraction exerlcd by the
F e r t i l i s a t i o n
vcen properly ascertained. It may be an attraction force between the two al of either of lhe
astral rays. But both these explanations are inadequate as the experimental removal these
pronuclei does not prevent the other one from following the normal path. CHEMISTRY OF FERTILISATION Since,
3. Amphimixis (union ofnale and lemale gmetes) : The details ofthe fusion of male and female
f e r t i l i s a t i o n .
for gamones.
Bcth egg as well nsspcm contain ncccssary
known as
gametes difter in dieent animals : certain
chemicals are prod1uced by tie gametes and clhemicals that are these are
hornones, are
termed as
nre of the nnnitire of found in
the egg I
" ln sea-urehin axd vetebrates the (wo prouelei nctually iuse together, Their nuclear membranes The hormones of the spem are callcd as
a n d r o g a m o n e
break at the oint of contact and their contents get surrounded a comnon nuclcar membrane Rynogamones. Two types of androgamones
androgamoues and thscThese arc known gynogamone
I
foming the zvyote aucleus. Atthe time of first cleavage tthe nuclear membrane dissolves, nuclear are found in lie spern. namely,
and androgamone lI. Similarly, egg also contains two types of
gynogamones
syindle is fomed ani the chromosomes ofmaternal and paternal origin get arranged on the equator and gynogamone lI.
The sperm
contains
of'spindic. activity. limited
The honone androgamone Iis spern's
beginning its
" In Asaris, some molluscs and annelids the malc and female pronuclei do not fuse. Their limitedaount of encrgy. If it
responsible forConscrving
could frorn
the very
reach the ovum. By
chnosones get arnged on the cquator of the spindle. In these animals, the zygote nucleus is were to
amount ofcnergy would be utilized in tooswim as rapidiy as it be able to
may not
increased.
n fertilisation is
not formod. The patenal and malen1al chromosomes get enclosed in the common nuclear short :a period and sperinprobability
conserving the activity until the spem approaches of for
indispensible
menbrane onty afer the completion of first cleavage in the nuclei of two blastomeres. Androgamone II dissolves the gelatinous
the egg. theegg and is, therefore,
In few animals (eg., Copepods-Crelops) the paternal and maternal nuclear contents reman coating around the
fertilisation. sperm comes
in
seprate even after clcavage. Therefore, cach blastomere contains two nuclei lying side by side. androgarmone I when
the sperm
Gynogamone l which is produced in the ovum neutralizes the II makes
CHANGES IN THE ORGANIZATION OF EGG CYTOPLASM contact with the egg and thereby increasces the activity of the sperm.
Gynogamone
androgamone il an
allows the
AFTER FERTILISATION head sticky and enables the sperm to stick to the egg surface and
In addition to the activation ofthe egg cytoplasm and the restoration of diploid chromosome number, opportunity to dissolve the gelatinous coating.
the fertilisation causes reorganization of egg cytoplasm and also changes the symmetry of the egg. Due lo Table 8.3 Summary of Chemistry
of Fertiisation
the runare of cortical granules, the outer egg surface is replaced by the inner cytoplasm lying below ne Function
cortical grannules. Gamone Gamete
1 Conserves sperm activity. sperm
SITE OF SPERM ENTRY Androgamone Sperm membrane, allowing
2 Dissotves vitelline
Androgamone l Sperm entrance into the egg
The sperm may eniter the egg at any point, usually the sperm avoids the yolk-laden pole of the egg. In androgamone I thereby
increasing
fsh, mollusc and insect eggs the sperm entry is througha definite, funnel-shaped aperture, the micropyle. Gynogamone I Ovum Neutralizes
nactivity
sperm attachment of
sticky to facilitate
TIME OF FERTILISATION Ovum Makes sperm head
Gynogamone l sperm to egg surface.
The eggs are fertilised at difierent stages of maturation in different species of animals as shown in the
table given below THEORIES OF FERTILISATION
Table 8.2 Time of fertiisation and maturation of ovum in different animals. (1887 and 1895)) suggested that the egg
loses its power of mitotic
1. Boveri's Theorv: Boveri the division centre
At metaphase of At metaphase of Female centre degenerates. The sperm introduces
Young primary Fuily grown first meiotic second meiotic pronucleus division during maturation as its division divide.
00cyte primary oocyte division division into the egg during fertilisation and makes the egg ifterent types of
Branchinstorma Amphioxus Coeienterates
---2. boeb's Theory Loeb (1913Lsuggested that speni inttoduces two (b) the
substances bring about cortical cytolysis and
+Ascaris Chaetopterus
Peripatopsis Grantia Cerebratulus Siredon Echinoids
substances in the egg cytoplasm. (a) The lytic
Saccocrus Petinaria Most mammals corrective factor regulates the oxidation.
tlereis
suggested that lvtic substances act as activators
Octornesostorma Aphryotrocha Lovtrup (1974) has supported LLoeb's view and has
Dicyerna membrane, dissolution of cortical granules and
Histriobdella Myzostorna Dentaliunm by causing partial hydrolysis and perforation of plasna
Spirula Many insects subsequent swelling of perivitelline space.
Thalessna 3. Bimolecular Theory of Activation : R.S. Lillie (194Dproposed that the entrance of spermin
of two substances B and S.
Dog and Frog egg leads to the fornation of activating substance A. it is lormed by the uniou egg. Its production is
The substance B is produced by tlhe iycdrolylie process taking place in the
the synthetic processes that are stimulated
stimulated by acid and heat. The substace S is lomedoflrom
these substanees
producu
by hvpertonicity. Entry of sperm liereases
 117
116 Chordate Embryology Fertilisation
4. Bataillion's Theory: Bataillion (1910 and 1916) proposed that the unfertiliscdegg
from dividing due to the
nccumulation of metabolic products. The entry of sperm leads tote thewas
release of these
inhioited suhstances fiom the cortexcintothe perivitelline space and thereby releasing the egg from
In addition to the above cited aspects, fertilisation lea to : division. fertilisation

track.
the inhibition.
5. Sensitizatton to Calclum : The role of calcium in the activation of egg has been emphasized " Liberation of secondary oocyte to complete its second
maturation
mark
the
to cgg-
The theory 'sensitization of calcium' was proposed by Dlacq, Induces movement ofeytoplasnie in the
particles and pigncnt
Ranules
and
by wo of embryologists.
goups(1936)
Braclhet and the theory of calehum release-protoplasmie clotting' was put forward ,Pasteelsby " Induccs centriole which forns the nuclear spindle and initiates
cleavage
of other
sperns.
the following obscrvations: the entry
inside.
Heibrunn (1930-40). The theoriCs ane bascd on changes in the periphery of ezg and this egy
Induces preclute
rotation
of
" Isotonic calcimchloridessolutions or solutions rich in calcium ions effectivcly activate the egg Causes the separation of vitelline icnnbrane which allows the
of calcium jons.
. Actiation of otherchemical agcits is dependent on the presence " Enhances metalolic activities.
calcium-sensitive. . Establishes a ncw plane of cgg axis.
"Eggs deived of calcin1 become
6. Heilbrunn's Theory: 1.V. Heilbrunn (1915, 28 and 43) believedithat an increase in viscosity MONOSPERMY AND POLYSPERMT
initiation of division. This is almost similar to the coagulation o
each
of the egg ctoplasm is the cause for the
surround
activation. At the ti What
biod. Caleium brings about this effect and is, therefore, directly involved in egg
sperm
Monospermy of fertilisation.
that the
myriads
cortex.
of fertilistion spermm liberates calciun that is bound to the egg
laboratory.m o n o s p e r m i c
When fertilisation is accomplished in naturc or in the as been
suggested
But
h.
7. Runnstrom's Theory : J. Runnstrom (1949) pointed out that an inhibitor is present in
sperms.
egg, vet only one enters and fertilises the egg. This is
known
refertilised?
It has of other
removal
of
released when
corten 2nd the vitelline menbrane. This inhibitor is bound to a kinase. The kinase is
passage
prevents the entry of other spems in the ovum? Can an egg beegg blocksthe After the
or
sixteen
stage.
protein substances of spen unite with the inhibitor. This enzyme initiates the development. formation ofthe fertilisation membrane aroundthe fertilised late as eight
by the
two-cell
even at
as covered
the egg of some species of sea-urchin can be refertilised
Occur
8. Repressor Theory of Activation : Monray and Tyler (1965) have suggested that durin refertilisation can gradually
fertilisation membrane from Saccoglossus eggs of egg is egg.
maruraion of egg its energy yielding system is blocked, which stops all the energy requiing reactiong blastomere stage. The sperm receptor, fertilizin,
present on
the surtace
extra
sperm to
the
How such inhibitions oceur and what is the mechanism of activation still remains to be discovered. hyaline layer after fertilisation. This blocks the
attachment of
some
According to Moaroy and Tyler (1963), in unfertilised egg either mRNA or ribosomes or both ar eggs of egg nucleus. This
birds,
reptiles,
is
inactivated in some way. Some proteins must be responsible for this inactivation because the ribosome Polyspermy inthe large yolky the
enters an eggg but fuses with
isolaied from unfertilised sea-urchin eggs and treated with trypsin for 30 minutes become active for As a general rule, only one spern only one
types : nuclei and
.However,
enter the egg.
prozein synthesis. It means mRNA is attached to ribosomes even in the unfertilised egg but this mRNA is amphibians and insects many spermPolyspermy can be of following two one sperm the
fertilisation. more than participate in
masked by a protein. During fertilisation this protein is dissolved by proteolytic enzyme. There is a Known as polyspermic polyspermy
nucleus
In physiological centriole and
increase in the proteolytic activity in the egg immediately after penetration of the sperm. 1. Physiological Polyspermy :
cytoplasm, but only one
sperm
amphibians arnd
centrioles enter the egg urodela
fertilisation and initiation of
cleavage. eggs (eggs of
SIGNIFICANCE OF FERTILISATION moderately yolk ladenelasmobranch fishes.
polyspermy is found in reptiles, birds and nucleus enter the
Fertilisation accomplishes following significant results : Physiological
very large yolky eggs
of
than one sperm
centriole and
some insects) and this case more
Polyspermy : In
I. The gametes or the gen cells constitute the only physical bridge between the suceessive 2. Pathological leads to
pathological
development.
abnormality and
leads to
become active. This Over ripeness is
an
generations. The gametespossess haploid number (n) of chromosomes. As aresult of fusion of egg cytoplasm and than one sperm. in the eggs of
been studiedsuspension
male and a female gameie, the diploid chromosomal coniguration similar to parents îs restored Over-tipe eggs
permit entry of more p a t h o t o g i c a t p o l y s p e r m y - t h a s
of
This fertilisation maintains the characteristic chromosome number. known as eggs in a dense
development. This is placing
can be induced by(such as nicotine, alkaloids and
chloral
pathological
mammals. Polyspermy substances polyandry (fusion of
2 terilisation brings together the genes and chromosomes from two difierent lines of ancestry
and results in new genetic recombinations, which ensure better adaptations to the ihagun
sea-urchin and certain
spermatoz0a, by
treating eggs with
toxic or narcotic Polyspemy may lead to
before insemination. lethal.
environment. hvdrate) or with heat
and cold just pronucleus). The polyandry is
nucleus with one female
D. Tne entry of spermn awakens the quiescent ovum and initiates the egg to divide the developmen more than one sperm Surplus
Spermatozoa
to proceed. or Exclusion of contact the egg surfac
Blocks to
Polyspermy the other sperm that
eBL Cytoplasm,exclusion notice.
The division Tnay be initiated by artilicial its Diferententry into lie mechanisns of surface sperm have been
Once asnerm has madeget of
-tonic or neutral salt solutions, acids, bases,means either exposing the unfertilised egg to
alkaloids. bea, cold, etc. by radiations or by' hypoto not allowed
in.
to
fine needle. puneu later on, are
animals :
in different
 118 Chordate Embryology
1. Elevation of Fertilisatlon Menbrane : In the eggs Fertilisation 119
Branchiostoma and fngs, a of annelids, sea-urchins, starfu
membrane begins fm the fertilisntion membranc is formed around the egg. Formation of fertilii
becomes alout S00 A thick oint of sperm contnet nd is propagatcd over the cgg su face. Finall
and
fomed by the nytun ofalvoi pesent impmcable to spermntozoa. In Nereis,
the fertilisation membrane a number
of
additional
egg. This
in the corteN. Disposal of Supernumerary Male Pronuclei few
insects,
in the
2, Popertv of and aad d i t i o n a l pronuclei female
on one snomatoz Mticropyle: In stugeon anl teleost cggs
birds with the
fuses
at a inne the In the eggs of elas1nobranchs, urodels, reptiles,
form pronucleus eggs, these
evtoplasm, a gelatinous sulbstanee is able to uss micropylar
througlh it. WVhen onc canal is so narrow tha spernatozoa enter the cgg. These surplus
pernatozoa
When one
spcrm
or
rejectcd. In bird division in
cell
membrane is tough and thick bloeks the caal spermatozoon has entered Ge phenomenon is known as physiologienl pollyspermy. citlcr where some ofthem induce
suppresscd
and ustupassable lblocking the cntry of supplementary pronuclcus, the supernumerary male prontelei arc blastodisc,
3. Changc in by thc sperm. sperm. The e rejected pronuclei are scen onthe periphery ofthe
srematoz03 ate theZona Pellucida : In
contact with first mammals the zona pellucida becomes the carly cleavage stages of enbryonic development.
Accessory cleavages
eranules disarrar pcnetration of spcmatozoon. impermeable Zygotic nuclaus
ater the This is known as zona G Pronucleus Supernumerary Primary
spernm
cntry of additionalreaction.
/ Pronuclel
and occlude the The corti cleavag
furrow
sperms in the ovu
QPronucleus C
B
2 Fertilisation A blastodisc of pigeon's egg
Viteline membrane cleavages induced by
them in the
membrane 3
nuclei and
(150-200A") (800A°) 3 l o ACCessory (From C.R. Austin).
Cortica!
granule
Hyaline layer
N Study Questions
diameter
(0.5 to 1.3u) fertilisation in the life cycle
of living beings.
Plasma the significance of process of fertilisation.
4 membrane 1. What is Fertilisation? Discuss that occur in the
diagram the major events
Fig. 8.17 Z. Describe with well-illustrated with internal fertilisation.
Diagram to show the Briefly describe the process
of fertilisation in forms
though lots ofmechanism
fertilisation.
that 3. in the ovum during
them may stick prevents entry
to egg surface.of additional sperms, Give an account of the
metabolic changes occurring process of fertilisation?
4.
fertilizin-antifertilizin reaction. What
is its significance in the
I1 is now believed 5. Discuss in brief the
that cortical reaction of of acrosome.
mpertect protection egg and formation of fertilisation membrane 6. Describe the role
events of cortical reaction and
their significance.
Sea-urchin eggs i takesbecause cortical
about 20
reaction begins at one point and a wavé is -7, What is cortical reaction? DiscusS
yaious
ach vther to maintain
species-specificity-in externally fertilizing
different regions of the eyg seconds and, therefore, cannot check the entrytravels around the egg In
of additional sperms it
8. How do sperm and ova recognize
Ihe two possible surface. animals?
state, fertilisation brings the egg
back into activity. Explain the
9. Eog within the ovary is in quiescent
1. Diaphasie explanations believed to help in sperm
in the Egg Cortex: The first exclusion are :
staternent.
surface within I or 2
Changes 10. Differentiate between
(b) External and internal fertilisation.
2. seconds and confers partial reaction is fast and covers the entire egg
(a) Fertilizin and
antifertilizin,
Fertilisation (d) Vitelline membrane and fertilisation membrane.
fertilisation potential ispotential is establislhed byprotcction.
reaction.
(c) Capacitation and acrosome
evidenced Spern entry and avoids polyspermy. 11. Write short
notes on:
selachians, urodels, rentiles and
Srom those animal eroups which have
The role o
(a) Fertilizin
(b) Cortical reaction
pronucleus birds. I them extra spernatozoa degenerate yolky eggs as in mollusc
atter one fuses with the eg
(c) Capacitation
(d) Hyaluronidase
(e) Spern lysins