Reproductive Behavior 251

Further To 111ate, the queen bee leaves the hive am/

performs a mating flight in an area where
PHENOMENA drones are congregated. The fastest drone
is thefirst to copulate with the queen. Copu­

for Fertility lation is au i11-jfight event that lasts from

I to 3 seconds. When the copulating bees
separate, the entire male genitalia is ripped
Oue day President am/ Mrs. Coolidge were from the 111a/e and stays with the queen. The
,,isiting a government far111. S0011 after male soon dies am/ another male will then
their arrival they were taken off on sepa­ mate with the queen. Up to 17 111ati11gs in
rate tours. Wizen Mrs. Coolidge passed one 111ati11gflight have been observed.
the chicken pens, she paused to ask the
man ill charge if the rooster copulated Females of some species are quite choosy
111ore than once each day. "Dozens of about who gets to fertilize their eggs. 111
times," was the reply. "Please tell that to these cases, mate choice is determined by
the President," Mrs. Coolidge requested. nuptial gifts presented by the male. The
Wl1en the President passed the pens am/ female black-tipped hangfly accepts nuptial
was toltl about tlte rooster, he asked, "Same gifts in the form of food in exchange for
hen eve,y day?'' "OJ, no, Mr. President, copulation. Wl1e11 edible food is presented
a different one eac/, time." The President by the male, the duration of copulation is
nodded slowly and the11 said, "Please tell dependent 011 the size of the gift. If the gift
that to Mrs. Coolidge." is small and can be consumed in 5 minutes
or less, the female will not allow mating. If
The praying mantis has 111111s1,al reproduc­ the gift is large (cannot be consumed in 20
tive hehavi01: As soon as the male mounts minutes), the female will allow mating to
the female a11d accomplishes intromission, take place. If the gift provides a meal of
the female bites his head off. She imme­ only 12 minutes site will leave the gift-giver
diately eats the top half of !tis body while prematurely and seek another gift-giver as
intromissio11 is still taking place. The rea­ a mate.
sou for this behavior is because ejaculation
is permanently inhibited in the male and Satbt bowerbirds build their nests only with
can take place only after the head has been blue objects. Males gather blueflowers, pen
removed. It is not known whether the slang caps, berries and ribbons and arrange them
phrase "bite-your-head-off' was derived under bushes or ill other cozy spots. If a
from this behavior. female "likes" what she sees, she will choose
the nest's decorator as her mate.
Roman snails shoot love darts at one an­
other before copulation to determine ifthey A male newt begins his courtship by jump­
are both 111embers of the sa111e species. ing on the back of the female and rubbing
his jaw against her snout. This releases
S0111e male insects (certainflies and 111os­ a scent that drives the female newt "crazy
quitoes) have evolved 111111sua/ adaptations with desire."
to insure that their genetics will be passed
011. Males have a sharp, specialized penis Wlte11 female rhinoceri are ill heat they will
that can enter a pupa. The 111ale insemi­ run away from a male, then suddenly turn
nates the unborn female. and fight him horn-to-horn, sometimes for
longer titan a day. Only ifhe is.fit enough to
When a grey squirrel comes into estrus, up pursue will she submit. There are no "wimp
to a dozen 1110/es noisily chase /,er through genes" in the rhinocerous gene pool.
the trees. This chase is necessary, because
the female will not ovulate without it.
 252 Reproductive Behavior

During courtship the female balloon fly

will eat tlte male if given the chance. To
achieve copulation and keep from getting
eaten, the male will present the female with
a balloon-shaped cocoon as a ''present".
Unwrapping this ''present" keeps the female
occupied long enough for tlte male to mate
lter and fly off.

Wizen box turtles copulate, the male mounts

the female and remains in an upright posi­
tion in order to facilitate insemination. The
pair may remain in this position for hours
to ensure adequate insemination. At the
conclusion of the event the female will sud­
denly move away, sometimes causing the
male to fall precariously on his hack where
he may remain until his death if he can't
right himself.

Most frogs and toads copulate in the dark.

They are often so eager to mate that the
male will try to mount anything that passes
by. They have been observed keeping afirm
grip on strange objects and even other small

11
animals in the hope that they might turn out
to he females.

The long neck of the giraffe plays an im­

portant role in their reproductive behavior.
First the male samples the female's urine to
ascertain whether she is in estrus. I/so, the
two giraffes then indulge in a form of sexual
preparation by entwining and rubbing their
necks together. Physiologically, this behav­
ior is like a false-mount and 110 doubt causes
the release of oxytocin that moves sperm
in the distal tail of the epididymis into an
ejaculatory position.

The pressure within the penis of the hull at

the time of ejaculation is equivalent to 10
times the pressure within a normal vehicle
tire.
 Reproductive Behavior 253

Key References

Albright, J.L., and C. W. Arave. 1997. The Behaviour

of'Cattle. CAB lntemational, Wellingford, UK. ISBN
0-85199-196-3.
Craig, J.V. 1981. Domestic Animal Behavior: causes
and implications (or animal care and management.
Prentice-Hall, Inc. New Jersey. ISBN 0-13-2 l 8339-
0.
Evans, H.E. 1993. Miller�- Anatomv of' the Dog. 3rd
Edition. W.B.Saunders Co. Philadelphia. ISBN 0-7216-
3200-9.

Grandage, J. 1972. "The erect dog penis: a paradox of

flexible rigidity." Vet Rec: 91: 141-147.

Hart, Benjamin L. 1985. The Behavior o(Domestic

Animals. W.I-I. Freeman and Co., New York. ISBN
0-7167-1595-3.
Houpt, K.A. 1998. Domestic Animal Behavior for
Veterinarians and Animal Scientists. 3rd Edition. Iowa
State University Press, ISBN 0-8 I 38-1061-2.

Katz, L.S. and T.J. McDonald. 1992. "Sexual Behavior

of fann animals" in Repoduction in Farm Animals:
Science, Application and Models. Theriogenology
38:240-254.

Korenman, S.G. 1998. "New insights into erectile dys­

function: a practical approach." Am. J. Med. 105:135-
144.

Signoret, J.P. and J. Balthazart. 1993 "Sexual behavior"

in Reproduction in Mammals and Man. C. Thibault,
M.C. Levasseur and R.H.F. Hunder, eds. Ellipses, Paris.
ISBN 2-7298-9354-7.

Tibary, A. and A. Anouassi. 1997. Theriogenology in

Camelidae. United Arab Emirates. Ministry of Cul­
ture and Information. Publication authorization No.
3849/1/16. ISBN 9981-801-32-1.
The Puerperium & Lactation

Parturition

Fetal Attachment & Gestation

Early Embryogenesis &

Maternal Recognition of Pregnancy

Cyclicity Spermatogenesis

Regulation of Regulation of
Reproduction Reproduction

Tract Function Tract Function

Puberty Puberty

Prenatal Prenatal
Development Development
 Take Home Message
Following insemination, viable spermatozoa that are retained in the female repro­
ductive tract must: 1) transverse the cervix, 2) be transported through the uterus to the
oviduct, 3) undergo capacitation, 4) bind to the oocyte, 5) undergo the acrosome reaction
and 6) penetrate the zona pellucida and fuse with the oocyte plasma membrane. After fu­
sion with the plasma membrane, the fertilizing spermatozoon enters the oocyte cytoplasm
and its nucleus decondenses. The male pronucleus is formed. This signifies successful
fertilization.

Following deposition of semen during the female tract. The remaining spermatozoa must
copulation, spermatozoa are exposed to a series traverse the cervix, enter and traverse the uterus
of different environments that significantly al­ and enter the oviduct. They must undergo ca­
ter their numbers and their function. After their pacitation before they can fertilize the oocyte.
deposition, spermatozoa are lost from the fe­ When sperm encounter the egg they undergo the
male reproductive tract by retrograde transport acrosome reaction and fertilization takes place.
and many are phagocytized by leukocytes within This series of events is summarized in Figure 12- 1.

Figure 12-1. Major Sequence of Events Following Deposition

•
of Spermatozoa in Female Tract

•
Fertilization
• acrosome reaction
• spermatozoon penetrates
oocyte
• male and female pronuclei form
Immediate Transport
• retrograde loss
• phagocytosis
• entrance into cervix/uterus

•
Oviduct
• docking to oviductal cells
,

• '

•
• capacitation completed Cervix
• hyperactive motility • "privileged path-
ways"
• removal of non-
motile sperm
• removal of some
Uterus abnormalities
• capacitation initiated )
• phagocytosis
 256 Sperm in the Female Tract

In some animals (cow, sheep, rabbit, primates, prevent spemiatozoa from undergoing retrograde flow
dog and cat), the male ejaculates the semen into the to the exterior. Female rodents (mice and rats) have a
cranial vagina. In others, (pigs, horses and camelids) relatively solid vaginal plug that is externally visible
semen is either deposited directly into the cervix (pig) following copulation. The presence of the vaginal
or is squirted through the cervical lumen during copula­ plug can be used to detem1ine when mating occurred.
tion (horse). In the dog, pig and the horse most of the Domestic animals do not have a conspicuous vaginal
ejaculate gains entrance into the uterine lumen. plug.
The stallion ejaculates in a series of "jets" in
which a sperm-rich fraction is ejaculated first in 3-4
high pressure squirts. This fraction contains about 80% Spermatozoa are lost ji·om the
of the spennatozoa. The last 5 to 8 "jets" are of lower
female tract by:
pressure and contain fewer spem1. The seminal plasma
in the final "jets" is highly viscous and may serve to • phagocytosis by neutrophils
minimize retrograde spenn loss from the mare's tract. • retrograde transport
Because of the large volume (200 to 400 ml)
of boar ejaculate, most of the semen flows from the
cervix into the uterine lumen. As in the stallion, the
boar ejaculates a series of seminal fractions with dif­ When the female reproductive tract is under
ferent characteristics as ejaculation progresses. The the influence of estradiol during estrus, neutrophils
first fraction consists of accessory fluids and gelatinous (powerful phagocytic white blood cells) sequester in the
coagulum. This fraction contains few sperm. The mucosa of the tract, especially in the vagina and uterus.
second fraction is rich in spennatozoa and this spem1- These neutrophils are poised to attack foreign materials
rich fraction is followed by a final fraction that fonns a that are introduced into the female reproductive tract at
gelatinous coagulum that resembles rice pudding. This insemination. lt should be recognized that, in addition
coagulum reduces retrograde spenn loss. Immediately to spermatozoa, microorganisms are introduced into the
after insemination, semen undergoes va1ying degrees tract during copulation. Thus, the neutrophil population
of retrograde transport (from the cervix towards the is important in preventing these microorganisms from
vulva). colonizing the female tract. From an immunologic

I
In the dog semen is ejaculated in three frac­ perspective, spermatozoa are foreign to the female. As
tions. The first, is a pre-sperm fraction that is thought a result, neutrophils actively phagocytize spermatozoa.

I
to originate from the prostate. The volume of the pre­ They do not discriminate between live and dead spenn.
spenn fraction is usually small but can range from 0.5 In fact, a single neutrophil is capable of engulfing sev­
to 5ml. This pre-spem1 fraction (clear and acellular) is eral motile spem1atozoa (See Figure 12-2).
ejaculated in conjunction with pelvic thrusting by the Srudies have shown that within 6 to 12 hours
male during "first stage coih1s." The second, a spenn after the introduction of spennatozoa into the uterus,
rich fraction, is between 1 and 4 ml and is opalescent there is a large migration of neutrophils from the uter­
in color and contains between 300 million and 2 billion ine mucosa into the uterine lumen (See Figure 12-2).
sperm. The final fraction originating from the prostate While leukocyte infiltration is an important contributor
ranges in volume from I to 80ml. The first two frac­ to post-insemination spermatozoa! losses, this infiltra­
tions are ejaculated without visible force. However, the tion is important for the prevention of reproductive
third fraction is ejaculated in surges of prostatic fluid tract infection.
that squirt into the vagina of the bitch during "second
stage coihls." Because of the "tie" (See Chapter 11)
most of this fraction is forced cranially into the uterus Spermatozoa/ transport consists of a
and is believed to "push" the sperm-rich fraction ahead rapid phase and a sustained phase.
of it into the uterus.
Ejaculate volumes in the tom hlrkey average
only 0.2 to 0.3ml with a range of 0.1 to 0.7ml and it
is therefore difficult to evaluate whether the ejaculate Among the least understood phenomena in
consists of multiple fractions. reproductive physiology are factors that regulate loss
The degree to which spem1atozoa are lost from of spermatozoa from the female tract. The ability of the
the female tract depends upon the physical nah1re of female to retain viable spermatozoa may influence the
the ejaculate and the site of seminal deposition. In fertility of a given mating. Transport of spem1atozoa
some species, the seminal plasma contains coagulat­ following copulation can be divided into two phases.
ing protein(s) that form a conspicuous vaginal plug to These are the rapid transport phase and the sustained
 Sperm in the Female Tract 257
transport phase. Within a few minutes after copula­ The more important component of transport
tion, spem1atozoa can be found in the oviducts. The is the sustained phase in which spermatozoa are trans­
rapid phase of transport was once considered to be ported to the oviducts in a "trickle-like" effect from
important because it delivered spemrntozoa to the site so-called reservoirs in the cervix and the uterotubal
of the fertilization very shortly after copulation, where junction. During the sustained transport phase, sperm
they "postured" themselves for the atTival of oocytes. move into the isthmus and attach to the oviductal epi­
However, further research has shown that spermatozoa thelium. Spenu can attach to the epithelium along the
an-iving in the oviducts within minutes after copulation entire oviduct. However, sperm tempora1ily "dock" to
were not viable. The functional importance of the rapid the epithelium of the lower isthmus near the uterotubal
phase of spe1m transport is not obvious. It may simply junction because this is the first oviductal region they
represent a burst of transport activity brought about by encounter. Spem1 "docking" is crucial to spenn survival
contraction of the muscularis of the female tract in because it elicits a signal cascade in the sperm that pro­
conjunction with copulation. motes viability. Without "docking", spem1 die within
6-10 hours after insemination.
Figure 12-2. Leukocyte
Infiltration Helps Prevent Rapid transport of spermatozoa is primarily
Reproductive Tract Infections the result of elevated tone and motility of
Within 6-12 hours af­ the muscularis of the female tract
ter the introduction of
sperm into the uterus,
.t:J C
E CIJ there is a large infiltra­ As you already know, estradiol is high during
�E the follicular phase when insemination occw-s. Estra­
z�
_...J
tion of neutrophils from
the uterine mucosa into diol stimulates contractions of the muscularis, particu­
·- CIJ
"E..c the uterine lumen. larly the myometrium. Also, prostaglandins in semen
o'i:
I. CIJ (PGF2a and PGE 1 ) cause increased tone and motility of
.IJ -1,,1
�:) the uterus and/or the oviduct. Intermittent contractions
Z.5 of the muscularis propel spermatozoa in both a cranial
CIJ .J:
>-� and a caudal direction. Fluids secreted into the lumen
·..:; � of the female tract also serve as a vehicle for transport.
Control of directionality, while not understood, is prob­
ably under the collective influence of muscular contrac­
12 24 36 48
Insemination tions and fluid distribution and characteristics.
Time - (Hrs) In addition to alteration of tract motility,
seminal plasma from boars has been shown by Ger­
man researchers to advance the time of ovulation in
gilts. For example, when seminal plasma was infused
into the right uterine horn, ovulation occun-ed about 11 12
0 hours earlier in the right ovary than in the left ovary.
SH 0
ST' The left uterine horn did not receive seminal plasma.

1
The specific material in boar seminal plasma inducing
early ovulation has not been identified, but it appears
to be a protein. Identification of these factors could
provide an avenue to control more precisely the time
of ovulation in swine. A similar phenomenon occurs in
camelids where seminal plasma components have been
shown to cause ovulation.

Three leukocytes (A,B and C) phagocytizing The cervix is a major barrier to sperma­
sperm. Sperm heads (SH) can be observed tozoa/ transport and it can also serve as
in the cytoplasm of the leukocytes. A sperm
tail (ST) can also be seen protruding from the a reservoir for spermatozoa.
leukocyte (Micrograph courtesy of R.G. Saacke, Virginia
Polytechnic Institute and State University, Blacksburg)
\.
 258 Sperm in the Female Tract

Figure 12-3. Spermatozoa Travel Through "Privileged Pathways"

in the Cow

During estrus secretion of sulfo­

muci ns from the apical portion
of the cervical mucosa produces
sheets of viscous mucus. Se­
cretion is toward the lumen and
flows in a caudal direction. Less
viscous sialomucins are produced
in the basal crypts of the cervix.
Spermatozoa found in the basal
regions are orientated in the same
direction and traverse the cervix
toward the uterus through these
"privileged pathways" (PP) of low
viscosity sialomucin. (Modified from
Mullins and Saacke 1989, Anat. Rec. 225:106)

To vagina

II
12
 Sperm in the Female Tract 259
Following copulation in the cow and ewe and, vix in spermatozoa! transport and/or retention awaits
to some degree, the mare, spennatozoa must negotiate further clarification in the sow and the mare, where a
the highly convoluted system of grooves within the high proportion of spermatozoa are ejaculated into the
cervix (See Figure 12-3). During estrus, the cervix uterus.
produces mucus. In the cow cervical mucus consists of
two types. One type is a sialomucin, a mucus of low
viscosity. It is produced by cells in the basal areas of Spermatozoa must reside ill the
the cervical c1ypts (See Figure 12-3). A second type, female tract before they acquire
sulfomucin is produced in the apical portions of the maxim11111 fertility.
cervical epithelium covering the tips of the cervical
folds. This type of mucus is quite viscous. The pro­
duction of two types of mucus (one of low viscosity
and one of high viscosity) creates two distinct environ­ As you recall from Chapter 3, spermatozoa
ments within the cervix. Spermatozoa encountering acquire maturity during epididymal transit. However,
the viscous sulfomucin are washed out of the tract. the maturational changes that occur in the epididymis
Those that encounter the low viscosity sialomucin in do not render spermatozoa completely fertile. For
the environment of the crypts of the cervix swim into maximum fertility to be achieved, spermatozoa must
it. Thus, the low viscosity environment of the deeper reside in the female reproductive tract for a minimum
cervical crypts creates "privileged pathways" through period of time. During the time in the female repro­
which spermatozoa can move. ductive tract, some spermatozoa will undergo changes
The ability of spermatozoa to traverse these that allow them to become fertile. These changes are
"privileged pathways" is thought to depend on their referred to as spennatozoal capacitation (See Figure
ability to swim through the basal channels (crypts) of 12-4). The site for capacitation varies among species.
the cervix and the associated low viscosity mucus. In In species where spermatozoa are deposited in the cra­
this context, the cervix may be a filter that eliminates nial vagina, capacitation may begin as spenn ascend
non-motile spermatozoa. The specific role of the cer- and pass through the cervix. In species where semen is

Figure 12-4. Conceptual Version of Mammalian Capacitation

Epididymal Ejaculated Capacitated

Seminal Female
12
�
plasma + tract �
When sperm are ex­
r �
The plasma mem- posed to the female
brane of epididymal tract e n v ironment,
spermatozoa con­ these seminal plasma
tains a complement coatings, along with
of surface molecules some of the surface
(proteins and carbo­ molecules, are re­
hydrates) illustrated moved, thus exposing
here as yellow T's. portions of the mol­
'" ecules that can bind
to the zona pellucida
of the oocyte.
rThe surface molecules in epididymal sperm become
coated with seminal plasma proteins (orange halos)
that mask portions of the membrane molecules.
'" �
 260 Sperm in the Female Tract

deposited into the mid-cervix (sow) or caudal cervix Figure 12-5. Postcapacitation
(mare) and immediately enters the uterus, capacitation Sequence of Events Leading to
is probably initiated within the uterus and completed in
the isthmus of the oviduct as is the case with all spe­ Fertilization
cies. All spermatozoa are not capacitated at the same

•
rate. Instead, they are capacitated over a relatively Hyperactive motility
long period of time (several hours).
Capacitation can occur in fluids other than

•
Binding to
those found in the luminal compartment of the female zona pellucida
reproductive tract. For example, in vitro capacitation

•
has been accomplished in a wide variety of species
using blood serum, a variety of commercial tissue Acrosomal reaction
culture media, Krebs Ringer solution and Tyrodes

•
solution. No single in vitro environment will support Penetration of
capacitation for all species. zona pellucida
There is little doubt that the plasma mem­

•
brane of the sperm (particularly the head) un­ Sperm-oocyte
dergoes marked biochemical changes during ca­ membrane fusion

•
pacitation. During mixing of sperm with seminal
plasma the sperm become coated with various Sperm engulfed
proteins. The coating of seminal plasma proteins is

•
"stripped" away by the female tract environment. The Decondensation of
exact nature of the "stripping process" of capacitation sperm nucleus
is not understood.
An important concept with regard to capacita­ Formation of
tion is that the process can be reversed by returning male pronudeus
capacitated spem1atozoa to seminal plasma. For exam­
ple, when capacitated spermatozoa are removed from
the female reproductive tract and returned to seminal
plasma, they become decapacitated and require ad­ in a small area (like dancers in a disco). Hyperactive
ditional capacitation time in the female reproductive motility occurs throughout the oviduct and is thought
tract before they can regain their fertility. It appears to be brought about by specific molecules produced by
that the seminal plasma components coat the plasma the epithelium there. Hyperactive motility is thought
membrane with surface substances that prevent or to facilitate sperm-oocyte contact.
inhibit interaction of spermatozoa with the egg.
Binding to the zona pe/lucida requires
12 Fertilization is a Complex Process and
Involves a Cascade of Events
specific zona-binding proteins on the
spermatozoa/ membrane.
The process of fertilization involves a series
of specific interactions between spennatozoa and the
oocyte. These are outlined in Figure 12-5. Spermatozoa are known to contain specific
proteins on their plasma membrane surfaces overlying
the acrosome that bind specifically to zona pellucida
Acquisition of hyperactive motility proteins. These zona binding proteins on the plasma
occurs in the oviduct. membrane must be exposed during the capacitation
process before binding to the zona pellucida can occur.
Before zona binding can be understood fully, the mo­
lecular makeup of the zona must be described.
In the oviduct, as capacitation is completed, The zona pellucida of the oocyte consists of
the motility patterns of spennatozoa become hyperac­ three glycoproteins. These glycoproteins have been
tive. The motility pattern changes from a progressive, named zona proteins 1, 2 and 3 (ZPl, ZP2 and ZP3).
linear motility in which they swim in a relatively Zona proteins 1 and 2 are structural proteins providing
straight line (like an Olympic swinuner), into a fren­ the structural integrity of the zona. Zona protein 3 is
zied, dancing motion that is not linear and is localized much like a receptor for a honnone. It binds to proteins

.!..
 Sperm in the Female Tract 261
on the spem1atozoal membrane. Binding of spennato­
zoa to the zona pellucida is believed to require between The acrosomal reaction is an orderly
I 0,000 and 50,000 ZP3 molecules. The current under­ fusion of the spermatozoa/ plasma
standing is that the sperm plasma membrane contains membrane and the outer acrosomal
two zona binding sites. The first binding site, referred membrane.
to as the primary zona binding region is responsible
for adherence of spermatozoa to the zona pellucida.
The second binding site on the spennatozoal plasma The purpose of the acrosomal reaction is
membrane is believed to be acrosome reaction promot­ twofold. First, the reaction enables spennatozoa to
ing ligand. When binding occurs between this region penetrate the zona pellucida. Second, it modifies the
and the ZP3 molecule, a signal transduction occurs. equatorial segment so that it can later fuse with the
This is much like a typical hormone-receptor binding plasma membrane of the oocyte.
complex. Binding initiates the acrosomal reaction. The The acrosomal reaction begins when the plasma
relationship between ZP3 and the spennatozoal plasma membrane of the spermatozoon forms multiple fusion
membrane during binding is illustrated in Figure 12-6. sites with the outer acrosomal membrane. When the
two membranes fuse, many small vesicles are formed
(See Figure 12-7) and this process is called vesicula­
tion. After vesiculation has occurred, the acrosomal
contents are dispersed and the sperm nucleus is left

Figure 12-6. Zona Binding by Sperm and Initiation

of the Acrosomal Reaction

r 'I

Proposed mode l for

zona b i nding an d
the initiation of the
a c r o s o m a l reac-
tion in mammalian
spermatozoa. The
sperm plasma mem-
brane overlying the
acrosome c on tain s

12
two receptor-like re-
gions. The first, called
the zona binding re-
gion (ZBR), reacts
with ZP3 to cause
physical attachment of
the sperm to the zona Sperm plasma --t---­
membrane
pellucida. A second
me mbrane region,
the acrosome reac-
tion promoting region
(ARPR), also binds to Surface of --­
ARPR = Acrosome Reaction zona pellucida
ZP3 and initiates the
Promoting Region
acrosome reaction
by causing the sperm
1AM = Inner Acrosomal
Membrane
p lasma membrane
OAM = Outer Acrosomal
to fuse (arrows) to
Membrane
the outer acrosomal
ZBR = Zona Binding Region
membrane.
OAM 1AM
'- �
 262 Sperm in the Female Tract

Figure 12-7. Schematic Illustration of the Acrosomal Reaction

.. �J' •/ ..
Acrosomal --��
contents
Outer acrosomal
membrane
Inner acrosomal
membrane lt===H-- Fusion
protein
Post nuclear cap

Plasma membrane

Before acrosome During acrosome After acrosome

reaction reaction reaction

,, Before Acrosomal "'

' /
After Acrosomal
..,
During Acrosomal Reaction
Reaction During the reaction, the plas­ Reaction
Before the reaction ma membrane overlying the After the reaction,
begins, all mem­ acrosomal membrane begins the vesicles are
branes of the head to fuse with the outer acrosom­ sloughed, leaving
are intact. al membrane. The fusion of the inner acrosom-
the two membranes leads to al membrane, the
vesiculation that creates pores equatorial seg-
through which the acrosomal ment and the post
enzymes can pass. This al­ nuclear cap intact.
lows the sperm to penetrate \ ,,
through the zona pellucida.

with the inner acrosomal membrane surrounding it. probably takes no more than a few minutes. Following
12 Vesiculation characterizes the acrosomal reaction
and morphologically distinguishes it from a damaged
attachment to the zona pellucida, the acrosome reaction
allows the release of a variety of enzymes. Acrosin is
acrosome. Damage to the acrosome membrane and one enzyme that is released from spermatozoa during the
plasma membrane is irreversible. Damage to these acrosomal reaction. It hydrolyzes zona proteins as well
membranes is brought about by changes in osmotic as enhances the sperm's ability to bind to the zona. In
pressure, sudden cooling, sudden heating or marked the inactive form, acrosin is known as proacrosin which
changes in pH. Damage to the membranes causes has a strong affinity for the zona. Thus, proacrosin aids
premature loss of acrosomal contents and such sperm in binding the spermatozoon to the zona as the acrosomal
cannot accomplish fertilization. reaction proceeds. As proacrosin is converted to acrosin,
the sperm begins to penetrate and make its way through
Release of acl'osomal enzymes allows the zona pellucida. The mechanical force generated by
the flagellar action of the tail may be sufficient to maintain
the spel'matozoon to digest its way spe1111 head contact with the zona pellucida. It is important
thl'ough the zona pellucida. to note that the acrosomal reaction allows the sperma­
tozoon to digest a small hole through the zona through
which it can pass. Placing a hot marble on the surface
The penetration of the zona pellucida by a of a block of chilled butter would be an appropriate anal­
spermatozoon is believed to be a rapid process and ogy. The hot marble would move through the butter in a
 Sperm in the Female Tract 263
small regional hole, but the butter in most of the block When the spermatozoon completely penetrates
would be unchanged. This small regional dissolution the zona and reaches the perivitelline space (the space
leaves the zona predominately intact. Maintenance of between the zona and the oocyte plasma membrane),
an intact zona pellucida is important because it prevents it settles into a bed of microvilli formed from the oo­
blastomeres in the early embryo from separating during cyte plasma membrane. The plasma membrane of
embryogenesis. the oocyte fuses with the membrane of the equatorial
segment and the fertilizing spemmtozoon is engulfed.
The actual fusion of the oocyte plasma membrane with
Fertilization requires fusion of the the equatorial segment is believed to be brought about
equatorial segment and the oocyte by a so-called fusion protein located on this portion
plasma membrane. of the membrane. Prior to the acrosome reaction,
this fusion protein is inactive. After vesiculation and
release of the acrosomal contents, the fusion protein is

Figure 12-8. Illustration of Sperm-Oocyte Fusion

When the spermatozoon
completely penetrates
Post nuclear
the zona and reaches the
cap perivitelline space, it set­
Before
tles into a bed of microvilli
membrane formed b y the oocyte
fusion plasma membrane. The
cortical granules have
migrated to the periphery
of the oocyte.

Cortical granules
The plasma membrane
� of the oocyte fuses with
the equatorial segment
and the fertilizing sper­
matozoon is engulfed.
The cortical granule

12
During
membrane membrane fuses with
fusion the oocyte plasma mem­
Equatorial brane and the cortical
segment contents are released
into to perivitelline space
rtical granules by exocytosis.
� �
"
After the fusion between
the membrane of the
equatorial segment and
the oocyte plasma mem­
After brane occurs, the nucle­
membrane us of the spermatozoon
fusion is within the cytoplasm.
Decondensing
The sperm nuclear mem­
i
sperm
nucleus brane disappears and
Oocyte plasma
membrane the nucleus of the sperm
decondenses.
�
 264 Sperm in the Female Tract

activated, enabling the spenn membrane to fuse or bind chromosomes may pair up with the chromosomes of the
with the oocyte membrane. This process is illustrated female pronucleus. The decondensation of the sperm
in Figure 12-8. nucleus requires the reduction of the many disulfide
cross-links. In the cytoplasm of the oocyte, disulfide
The cortical reaction prevents cross-links in the spem1 nucleus are reduced quickly.
The primary reducing agent is glutathione. When
penetration by additional spermatozoa. disulfide bond reduction occurs, the sperm nucleus
decondenses and the nuclear material is available for
interaction with the female nuclear material. The final
After membrane fusion, the oocyte undergoes a step of fe1tilization is the fusion of the male and female
series of changes that prepare it for early embryogenesis. pronuclei. This fusion is refeITed to as syngamy. Fol­
The most easily recognizable is the cortical reaction. lowing syngamy, the zygote enters the first stages of
During the first and second meiotic divisions of oogen­ embryogenesis that are described in Chapter 13.
esis, small, dense granules called cortical granules move
to the periphery of the oocyte cytoplasm. The contents The Fertile Period Varies Significantly Among
of the cortical granules consist of mucopolysaccharides, Mammalian Females
proteases, plasminogen activator, acid phosphatase and
peroxidase. After membrane fusion between the oocyte The fertile life-span of spem1 after deposition in
and spem1atozoon, the cortical granules undergo exocy­ the female reproductive tract varies immensely among
tosis and their contents are released into the perivitelline species. For example, fertility of spennatozoa is re­
space (See Figure 12-8). Exocytosis of the cortical tained for four to five years in certain reptiles. Among
granules results in the zona block, a process whereby mammals, bat spem1atozoa remain viable after insemi­
the zona pellucida undergoes biochemical changes so nation in the female tract for up to 4-5 months before
that further spem1 cannot penetrate it. Polyspermy is the female ovulates. In general, retention of fertilizing
prevented by the zona block. capacity among domestic animals and humans lasts
Polyspermy is the fertilization of an oocyte by only a few days. Values in Table 12-1 document the
more than one spermatozoon which results in embryo variation in fertilizing ability in the female tract among
death. In addition to alteration of the zona pellucida, various domestic species and women.
the cortical reaction is believed to reduce the ability of In most domestic species the period of estrus is
the oocyte plasma membrane to fuse with additional less than 24 hours. In other words, copulation must take
spermatozoa, thus causing the vitelline block, another place within a time-period that is close to ovulation. In
mechanism that prevents polyspermy. Some species contrast, sperm can remain viable for as long as 5 to 6
have both a zona block as well as a vitelline block, while days before ovulation in women. Another example of
others have either a zona or a vitelline block. a sustained fertile period is the bitch. Ovulation takes
place over about a three day period after the onset of
sexual receptivity. Fertilization can be accomplished as
Pronucleiformation allows the male
12 and female DNA to form a single
long as six days after the onset of sexual receptivity. It
should be pointed out that in a multiparous species like
nucleus.

Table 12-1. Maxima l Duration of

After the sperm nucleus has entered the cy­ Fertilizing Ability of Sperm Within the Female
toplasm of the egg, it becomes the male pronucleus. Reproductive Tract of Various Species
Before the pronucleus can be formed, however, the
nucleus of the sperm must undergo marked changes
within the oocyte cytoplasm. As you will recall, one of
the maturational changes that occurs in the epididymis Species Fertile Life (days)
is the acquisition of large numbers of disulfide cross­ Bitch 9-11
links in the sperm nucleus. Thus, the nucleus of the
mammalian spem1 is almost inert. The keratinoid-like Camelids (camel, llama, alpaca) 4-5
quality of insolubility is considered to be important Cow 1.5-2
during exposure to the female tract environment, sperm Mare 4�
transport and penetration through the zona pellucida. Woman 5-6
After the fertilizing spermatozoon enters the oocyte cy­
toplasm the nucleus must "decondense" so that the male
 Sperm in the Female Tract 265
the dog, several males can sire offspring because the If your favorite baseball player had a batting average of
bitch may be bred by several males during her relatively 0.333 for the season, he had a one in three chance to get
long estrus. Spemmtozoa from all males are eligible to a base-hit during each at-bat. Each at-bat is equivalent
fertilize oocytes. This phenomenon is called superfe­ to the fertile period of an estrous or menstrual cycle.
cundation. Thus, it is not uncommon to observe litters On average, your favorite hitter needs 3 at-bats to get
that have different breeds of puppies. a hit (a pregnancy). It makes no difference how many
It should be emphasized that the long fertile times the batter swings (number of copulations) during
period in women coupled with a high frequency of each "at-bat," his batting average will still be 0.333.
copulation predisposes humans to unwanted pregnan­ Similarly, assuming a threshold number of sperm are
cies and a high global birth rate. Since the woman deposited during the first copulation, the number of
does not have a definite period of sexual receptivity, copulations during each fertile period (an "at bat") will
copulation taking place within 5-6 days of ovulation can not influence the probability of pregnancy because the
result in a pregnancy. Where a poor understanding of first copulation fills the oviductal reservoir and will not
the cycle exists, the probability of pregnancy becomes allow more spem1 to populate the reservoir.
quite high because almost 20% of the menstrual cycle
has the potential to generate a pregnancy.
The question is often asked as to whether the Batting Averages and Pregnancies
number of copulations can influence the chance of preg­ are Similar:
nancy within a given mating period. In spontaneous
ovulators the answer is "probably not". In induced ovu­ • Each "at-bat"= 1 opportunity to achieve
lators (especially in felids), there appears to be a thresh­ pregnancy
old number of copulations required to optimize GnRH • The batting average = probability of
and LH release. The chance of ovulation and therefore becoming pregnant
pregnancies are related to copulation frequency. In
humans, the probability of conception (pregnancy) is • A swing = 1 mating
about 0.33 per cycle. This means if mating takes place
among fertile individuals there is a one-in-three chance • A good "at-bat"= many swings (but
that the woman will become pregnant every cycle (if depletes extragonadal reserves)
sexual intercourse takes place within 2 days of ovula­
tion as Figure 12-9 shows). It is like a batting average.

Figure 12-9. Probability of Conception When Copulation Occurred on

Specific Days Relative of Ovulation in Women
(From Wilcox et al. 1995. NEJM 333:1517)

.4
r Conception can
-�
:E
o c cur wi thin a RI >,
u .3
.Q C
6-day window prior 0 RI
,._ C
to ovulation. At 5 Q. 1),0
days prior to ovula­ "tJ .2
(IJ ,._
tion, the probability .. Q.
RI -
of conception was 0
.I
0.11 and the prob­ ·.p
ability increases w
to about 0.33 two
days before ovu­
lation. -6 -5 -4 -3 -2 -I 0

Day of Copulation
Relative to Ovulation
 266 Sperm in the Female Tract

Delivery of Semen to the Proper Anatomical Artificial Insemination Techniques in

Region of the Female Tract is Required for Domestic Species
Successful Artificial Insemination
Artificial insemination technique requires
It had been erroneously assumed for years that spermatozoa be deposited in the reproductive
that most spermatozoa ascend toward the oviduct soon tract of the female by artificial means. In general,
after they are deposited in the cow uterus by artificial semen is delivered using a pipette to penetrate and
insemination. However, recent studies have shown that bypass the cervix (See Figure 12-11 ). This type
a high proportion of spermatozoa deposited in the uterus of insemination is referred to as transcervical in­
of the cow or ewe are lost from the tract by retrograde semination. In the sow, the insemination pipette is
transport. In most cows, over 60% of spermatozoa positioned within the cervix and semen is delivered
artificially inseminated into the uterus are lost to the into the cranial half of the cervix and flows directly
exterior of the tract within 12 hours after deposition. into the uterine horns. This type of insemination is
Given these findings, a logical interpretation would be referred to as intracervical insemination (See Figure
that artificial insemination of spermatozoa deep into 12-12). In dogs and cats semen is deposited in the cra­
the uterus would result in reduced retrograde loss. nial vagina. This type of insemination is referred to as
This assumption is not true because when spenn are intravaginal insemination (See Figure 12-12).
deposited deep into both uterine horns (as opposed to In cases where sperm are in very limited supply,
the uterine body) the degree of spenn recovered from surgical insemination can be performed by exterioriz­
the vagina (an indication of retrograde loss) is quite ing the reproductive tract and injecting spern1 directly
similar between the two sites of deposition (See Figure into the uterus or uterotubal junction region. Also, use
12-I 0). However, when sperm are deposited in the mid­ of laparoscopy enables insemination to be performed
cervix, a significantly higher degree of retrograde loss without laparotomy (an abdominal incision). In bulls,
of spemiatozoa is encountered (See Figure 12-10). X-Y sorted semen are in short-supply. Therefore, a
Spermatozoa deposited into only one uterine technique has been developed to "thread" the tip of an
horn ofthe cow experience intercornual transpo1i. That insemination pipette through the cervix to the uterohtbal
is, when spennatozoa are deposited into one uterine junction. Such a technique has been reported to gener­
horn (either right or left), they subsequently are redis­ ate excellent results.
tributed so that both uterine horns evenhially contain
substantial numbers of spermatozoa. This phenomenon
also occurs in swine. In cows, fertility is not compro­
mised and in some studies is enhanced when spenn are
deposited within the uterine body or in the right and
left uterine horns.
The important message from the above discus­
sion is that when artificial insemination is perfonned in

12 the cow and semen is deposited into the cervix, a greater

proportion of spermatozoa are lost to the exterior than
when deposition is in the uterus. Thus, when the in­
semination procedure involves cervical deposition (a
serious technique error), fertility may be compromised
because of greater spermatozoa! loss.
 Sperm in the Female Tract 267
Figure 12-10. Insemination into the Uterine Horns
Can Reduce Sperm Loss

22

"'C

18
>
Q)

u Cumulative percentage of sperm recovered

ex::
Q)
14 from the vagina of heifers during an 8 hour
E period after insemination. In one group of
heifers (green bar), sperm was deposited in
Q)
0.
10
the uterine body. In the second group (bur­
V)

�
0
gundy bar), sperm were deposited deep into
-�>
ltS
6 each uterine horn. The cumulative percent
"S of sperm recovered from the vagina did not
E differ between the two treatment groups.
u
::J
2
(Modified from Gallahger and Senger, 1989, J. Reprod. Fert.
86:19)
0 2 3 4 5 6 7 8

Hours After Insemination

"'C
r
12
60 -
� __,.:::.:.,
..... Cumulative percentage of sperm recovered
from the vagina of heifers during an 8 hour
� 50 - period after insemination. In one group of
u
� heifers (blue bar) sperm were deposited in
E
L.
40 - the cervix, while in the second group (bur­
Q)
0. gundy bar) sperm were deposited in the
V)
uterine horns. A significantly higher number
� 30-
- of sperm were found in the vagina of the
g!
-� 20 - animals that were inseminated at midcervix
"S indicating retrograde sperm transport.
§ (Modified from Gallagher and Senger, 1989, J. Reprod. Fert.

r 1
IO­
U 86:19)

_n
0 _..............................._._
.........
....._
.......
_.__.....................................__..__,__
0 2 3 4 s 6 7 8

Hours After Insemination

 268 Sperm in the Female Tract

Figure 12-11. Artificial Insemination Technique in the Cow and Mare

Semen
Inseminating pipette
Hand grasping cervix

r
The radiographs above are from extirpated cow reproductive tracts (dorsal view). In cornual
insemination, one-half of the semen is deposited in each uterine horn. In both examples, the in­
seminant volume is 0.5-ml. Cornual insemination minimizes the possibility of cervical deposition
that results in significant retrograde loss of spermatozoa (See Figure 12-3). RUL= Right Uterine
Lumen; LUL= Left Uterine Lumen; RO= right ovary; LO= left ovary; S= semen; AIS= artificial in­
semination syringe; CX= cervix

Mare ]

12

Uterine
Vagina body

In the mare, the gloved lubricated hand is inserted directly into the vagina and the index finger is
used to guide the insemination pipette into the cervical lumen. A marker (arrow) is used to gauge
the depth of insemination.
\. .)
 Sperm in the Female Tract 269
Figure 12-12 Artificial Insemination Technique in the Sow and Bitch

r Radiographs of an extirpated sow reproductive tracts (dorsal view). An artificial insemination pipette
(AIP) consists of a spiral tip (ST) that is designed so that it can snugly penetrate the interdigitating
prominences (IDP) of the cervix (CX). In the photograph to the right, about 80-ml of radiopaque
contrast medium was infused into the reproductive tract to mimic the inseminant (I). Notice that the
semen becomes distributed within both uterine horns. High volumes (about 80-ml) are necessary
to maximize pregnancies in sows. The vagina (V) and the urinary bladder (UB) can be visualized.
LUL= Left Uterine Lumen; RUL= Right Uterine Lumen.
\.

Bitch

12

'/
Pipette Cervix

Uterine
body

..,
The vulva is elevated manually so that the ventral "tilt" of the vestibule is removed. This allows
the insemination pipette to be inserted with relative ease. The hindquarters of the bitch should be
elevated for about 5 minutes after deposition of the semen to allow pooling in the cranial vagina
and caudal cervix.
\.
 270 Sperm in the Female Tract

Further A Spermatowon Race

PHENOMENA
by Cheryl A. Dudley

Halffrenzied, thick and slicl,

for Fertility and treacherous, through vast dark tmmels,
as motile and penetratingly
zo11a-bo1111d as any race ever,
Some species have delayed fertilization. Tltis 11011e other is so victi111-/ade11,
is a process whereby tlte male inseminates so masked by drunken seizures
tlte female and spermatozoa remain viable or pleasures of full-bodied assaults,
in tlte female tract for a sustained period the tadpoles' mad dash
is like an escaped madman,
of time. When a rooster inseminates a hen
she can lay fertile eggs for over 20 days. a drowner driven to o::i..yge11,
Sperm are stored in special utero-vaginal thejoumey a seas-width heat
glands. Some bats mate in the autumn to life or death
before hibernation. The female does not
ovulate until spl'ing. Sperm are stored in Whe11 theyjolted over the barrier
her tract during tlte winte,: The fertilizing she did11 't realize a race was 011,
life of bat sperm is reported to range from yet in her own primordial way
68 to 198 days depending 011 tlte species of she cheered for them, provided secret
bat. Snakes are reported to store sperm that privileged pathways through crypts
are fertile for up to 6 years. too difficult for most, whose dead,
ffat-floating bodies cluttered the way.
The bifurcation of the glam penis of the The lone victor slithered through, sensed
the trophy ahead-tlte zo11a se1/uci11g him to dip
opossum led to the widespread Appalachian
folk belief that opossums mated through in her warm waters, melt into her soft globe.
the nose, with one fork of the glans penis (The courtship was only long e11011glt for him
to work his way through her pellucida.)
penetrating each nostril. Little scientific
consideration was given to the issues of
sperm transport. A quivering union formed
pri111itive cords that proliferated
Male mammals deliver sperm to the fe­ time and time a11d time again,
male in seminal plasma. However, many swelling tofill the primed pear-palmed

12 lower forms of animals make use of special wo111b where tlte victor celebrated,
packages for delivering spermatozoa to the And a ge11esis bega11.
female reproductive tracl These packages
are called spermatoplwres. Tltese sper­ Cheryl Ditdley typed the 1st Editio11 of Path­
matoplwres are produced within the male wavs to Preg11a11cv and Parturitio11 fro111 the
reproductive tract and are stored there until author� dictation. Size has since graduated
copulation. bi some cephalopods (octopus Cum /cmde in E11glislt from the U11iversity of
am/ squid) tlte male deposits the spermato­ Ida/to and is 110111 a graduate st11de11t in the
phore in the female tract or into the buccal Department of English at that university.
cavity (cheek pouch), from which it can
be conveniently transferred to the female Motility of trout spermatozoa is induced by
tract. In some annelids, spermatophores are the fresh water into wltich it is ejaculated.
"injected" subcutaneously, after which tlte Motility lasts for only about 30 seconds.
spermatozoa spread throughout the female -s During this time the sperm must locate a
body before contacting eggs. single tiny hole in the egg (called a micro­
pyle) through which it enters before fertiliza­
tion can occm: All this happens while being
swept about by moving wate,: