15th lecture
SPERM TRANSPORT IN ♀
REPRODUCTIVE TRACT
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2
Continued
• After insemination the spermatozoa are exposed
to a different environment in which it passes
through different processes & make it capable to
fertilize the oocyte.
• After deposition, most of the spermatozoa are lost
in ♀ RT by the retrograde movement or transport
& many of these are phygocytized by leukocytes
within the ♀ RT.
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Continued
• The remaining spermatozoa in the ♀ RT must
1) Travels/pass through the cervix.
2) Entered & be transported thorough the uterus &
oviduct, must undergo through capacitation before
they can fertilize the oocytes
When a sperm encounter/bind the oocyte/egg, they
undergo acrosome reaction & penetrate the zona
pellucida & fuse with the oocyte plasma membrane.
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Continued
• After fusion with the plasma membrane the fertilizing
spermatozoon enter the oocyte cytoplasm & its
nucleus decondenses.
• Male & ♀ pronuclei are formed & this mean
fertilization take place.
• Semen is deposited in cranial vagina (cow, sheep,
primate, dog, cat etc.). In some animal like pig, mare
& camelid semen is squirted through the cervical
lumen during copulation (& is directly deposited in
cervix). 5
Continued
• Stallion: Ejaculate in series of jets/movements &
the 1st fraction of the ejaculate is sperm rich
having 80% of spermatozoa & is ejaculated in 3-4
squirts of high pressure but the last few
squirts/jets having less spermatozoa & the final
jet/squirt is highly viscous that prevent retrograde
movement of the sperms in mares.
6
Continued
• Dog: semen is ejaculated in 3 fractions. The 1st
pre-sperm fraction & is thought to originate from
prostate. Its volume is 0.5-5ml & is clear &
acellular.
• The 2nd, a sperm rich fraction, volume 1-4 ml &
opalescent in color, sperm 300 million to 2 billion
sperm. The final faction originate from prostate,
volume 1-80 ml & is squirted into the vagina &
because of copulatory tie, this fraction is forced
cranially into the uterus & is thought to push the
sperm rich fraction into the uterus.
7
Continued
• So mostly the loss of spermatozoa in ♀ tract
depend on the physical nature & site of the
deposition.
• In some specie the seminal plasma contain
coagulating protein (coagulum) that form a
prominent vaginal plug, that prevent retrograde
movement of the sperms (♀ rodent have a solid
vaginal plug that is visible but in domestic animals
no vaginal plug is observed).
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Continued
• During the estrus when ♀ RT is under the
influence of E2 as there is more blood supply so
the Neutrophils (powerful phagocytic WBC)
isolates in the mucosa of the tract, especially in
the vagina/uterus
• These Neutrophils are supposed to attack foreign
bodies that are introduced into the ♀ RT during
insemination i.e. microorganism which later on
colonize the RT.
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Continued
• From immunological perspective, sperm are
foreign bodies to the ♀.
• so as result, neutrophils potently phagocytise the
spermatozoa.
• They don’t discriminate between live & dead
sperms.
• A single Neutrophils can engulf several motile
sperms (after 6-12 hrs of AI a large number of
Neutrophils infiltrated from uterine mucosa to
uterine lumen & play an important role in
spermatozoal losses & also prevent RT infections).
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Spermatozoal Transport
• After insemination, its transport can be divided into
two phases.
1) Rapid transport phase: (15 - 30 min)
After copulation, within a few minutes,
spermatozoa may be found in the oviduct, initially this
transport was considered important because
spermatozoa were made available in the site of
fertilization where they wait/postured for the oocyte.
Later on research revealed that, its availability just
after copulation in the oviduct is of no use. It simply
tells us burst of transport activity achieved
through contraction of muscularis. 12
Continued
2) Sustained transport phase: (6-12 hr.) An important
phase in which the sperm are transported to the site
of fertilization in the trickle-like effect from the so
called reservoir in the cervix & utero-tubal junction.
• This phase delivered spermatozoa to the ampulla in a
more uniform manner over a sustained period of
time.
• After semen deposition, most of the sperms moves
towards the oviduct but recent research tells us that
most of the sperm are lost from the ♀ RT by
retrograde transport. In cow 60% or more are lost
within 12 hrs after deposition(AI). 13
Continued
• Sperm deposited deep in uterus or uterine body,
the retrograde sperm recovered from vagina are
same, however deposited in cervix the retrograde
sperm recovered are high.
• When deep in one horn, a sufficient number of
spermatozoa may be found in the other horn too,
because of retrograde movement & fertility is not
compromised.
• In some studies fertility is enhanced when in
uterine body.
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15
Continued
• At the time of insemination estradiol is high that
stimulate contraction of the myometrium.
• Similarly PGF2α & PGE1 in semen causes, tone &
motility of uterus/oviduct.
• So this intermittent contraction propel sperms,
cranially & caudally.
• Fluid secretion into lumen of ♀ RT serve as vehicle
for transport.
• Control of directionality is not understood.
16
Continued
• To minimize the retrograde transport, we may
treat the animal with some pharmaceutical
products to change the smooth muscle motility &
this alter sperm transport.
• So small number of sperm in straw can be used
with acceptable fertility.
• E.g. in one test a rabbit injected with
phenylephrine/Ergonovine (both smooth muscle
stimulant) sperm reaching to oviduct was as
compared to non-treated rabbit.
17
Continued
• In doe inseminated with low no of sperm (90k)
+phenylephrine or Ergonovine caused
fertilization rate as compared with non-treated
animals.
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Cervix is major barrier to sperm transport
• After natural service, the sperm must pass the
highly convoluted system in the cervix.
• As you know during estrus , cervix produce a
mucus. In cow this mucus is of two types.
1. Sialomucin: mucus of low viscosity, that is produced
by the cells in the basal area of cervical crypts.
2. Sulfomucin: quite viscous mucus & is produced by
the cells present in the apical portion of the cervical
folds.
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Continued
• Now two types of environment to the spermatozoa
in the cervix.
• Spermatozoa encounter the viscous sulfomucin,
are washed out as they can’t swim in it.
• Those encounter the sialomucin in the crypt of
cervix, swim into it & creates privileged pathway
through which spermatozoa can move.
• The ability of sperm to traverse this pathway
depends on their ability to swim through the basal
channels (crypt ) of the cervix, so the non-motile
sperm are eliminated.
21
Capacitation
• Refers to the physiological changes spermatozoa
must undergo in order to have the ability to
fertilize the ovum.
• As you know the spermatozoa acquire maturity
during epididymal transit & these changes
occurred in epididymis but don’t make them
capable to fertilize.
• Spermatozoa must stay in the female tract for
sometime before they acquire the ability to
fertilize, these changes are referred to as
capacitation.
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Continued
• Site for capacitation varies among species. All
spermatozoa are not capacitated at same rate.
Instead they are capacitated over long period of
time & this reflect individual sperm differences &
as well as location in the tract.
• Capacitation can take place in fluid other than the
fluid found in the luminal compartments of ♀ RT,
e.g. in-vitro capacitation has been done in a variety
of species, using blood serum, Krebs's ringer
solution & Tyrodes solutions.
24
Continued
• There is doubt that the plasma membrane of
sperm undergo marked biochemical changes
during capacitation.
• However during mixing of sperm with seminal
plasma the sperm become coated with various
protein & the coating of this seminal plasma
protein is stripped away.
25
General information
• Rooster inseminate a hen, lay fertile eggs for 20
days(semen is stored in utero-vaginal gland).
• Some bats mate in autumn before hibernation &
♀ doesn’t ovulate until spring(sperm store in RT &
its fertilizable life is 68-198 days depending on
specie).
• Snakes are reported to store sperm that are fertile
for 6 years. 26
