BIOLOGY
OF
REPRODUCTION
37, 22 5-234
Distribution
(1987)
and Number of Spermatozoa
in the Oviduct of the Golden
after Natural Mating and Artificial
lnseminatio&
T. TIMOTHY
SMITH,2
FUKASHI
KOYANAGI,
Department
of Anatomy
University
of Hawaii
Honolulu,
and RYUZO
Hamster
YANAGIMACHI
and Reproductive
Biology
School
of Medicine
Hawaii
96822
ABSTRACT
A group
ovulation.
killed
female hamsters
was mated
with
males
of proven
fertility
nother group
of females
was artificially
inseminated
several
at various
times
after
the onset of mating
or artificial
insemination,
of
serially,
and
spermatozoa
Regardless
remained
The
“sperm
reached
reached
females
ascending
considerably
complete
were
of the type
in
ampulla.
and/or
tozoa
tozoa
were
to
the
lower
counted
individually
of insemination,
of the isthmus
segments
lower
segments
of the
reservoir.”
In females
the
the
cephalic
ampulla
mated
the
during
ovulation,
The
than
oviduct,
mated
particularly
or artificially
spermatozoa
number
in naturally
mated
animals,
that
but
In mammals,
it is in the
oviduct
where
spermatozoa
lized
eggs (zygotes)
begin
millions
of spermatozoa
part of the female
genital
the
cervix,
utero-tubal
The
ampullary
region
of the
and eggs meet and fertito develop.
At
coitus,
junction,
and/or
the
lower
reach
the upper
regions
of the tract,
ensuring
only a small number
of spermatozoa
reaches the
ampulla
(for
Bedford,
1970;
1975;
Mortimer,
reviews,
Hunter,
1978,
see
Blandau,
1969,
1973,
1980;
Ahlgren
1983;
Polge,
1978;
1982;Hawk,
1983;Overstreet,
1983).
Before
spermatozoa
become
capable
eggs, they must undergo
the process
of
February
10, 1987.
December
15, 1986.
‘This study was supported
by
2Reprint
requests:
T. Timothy
Reproductive
Biology,
University
West
Road,
Honolulu,
HI 96822.
number
site
where
during
semen
coitus
the
apparently
vary
fertilization
information
capacitation
1980;
large
as a “sieve”
no sperma-
enough
is
from
to
species
is deposited
(e.g.,
many
may begin
enter
the
to ensure
and
species.
“directly”
rodents,
and
many
as a model
com-
In
species
into the
dog, pig),
uterus
sperm
or soon
species
mechanisms
in mammals.
has been
in vitro
Yanagimachi,
others
for the
after the
in which
have
used
analysis
of
Although
obtained
concerning
(for
reference,
see
1981;
Meizel,
1984),
much
sperm
Bavister,
the
ques-
capacitation
occurs
in vivo
As a preliminary
to address-
ing these
questions,
it is necessary
spermatozoa
are distributed
in the
tract
before
and during
fertilization.
to know
how
female
genital
Several
pre-
vious
studies
have reported
the distribution
of hamster spermatozoa
in the female
genital
tract
(Strauss,
1956;
Chang
and
Sheaffer,
1957;
Yamanaka
and
NIH Grant
HD-03402.
Smith,
Department
of Anatomy
of Hawaii
Medical
School,
1960
before
was
initiated
in the uterus
oviduct.
For
tions
of where
and how
remains
to be answered.
Accepted
Received
oviduct
to the
afew
spermathe rule. When
capacitation
Our
laboratory
golden
hamster
1973;
of fertilizing
capacitation.
entered
the
ascending
Although
rather
than
were
sectioned
is deposited
in the vagina
at coitus
(e.g.,
cow, human),
capacitation
may begin
in the
or as spermatozoa
pass through
the cervix.
et al.,
Harper,
and
3 h in the caudal
isthmus
after artificial
insemination
where
may
that
or during
Females
fixed
without
of ovulation.
exception
was
before
appeared
to be acting
to ovulation,
virtually
of about
oviduct
pleted
semen
rabbit,
vagina
or “sieve”
spermatozoa
the
the
capacitation
spermatozoa
are deposited
in the lower
tract (i.e., vagina or uterus),
part of the oviduct
each act as a “barrier”
to sequentially
reduce
the number
of
that
that
low
hours
ovulation.
were
isthmus)
isthmus,
prior
a minimum
entered
this
oviducts
caudal
the caudal
inseminated
spent
of spermatozoa
several
before
in the oviduct.
of spermatozoa
isthmus
or ampulla
until
the commencement
by 1 h after the onset
of mating,
they were
ampulla.
lower
fertilization.
as to their location
the vast majority
(the intramural
and
or time
INTRODUCTION
but
either
hours
A
and
East-
225
226
SMITH
Soderwall,
1960; Yanagimachi
the methods
used in these
ciently
methods
or
in the
contents
of
genital
tract.
female
a
segments
physiologically
various
times
and
spermatozoa
the
several
mating
because
many
crypts
of
(Thibault,
using
tract
have
in
cannot
Another
specified
mating
the
first
used
(Adams,
are
to
1956)
in
can
flushing
Nichol,
1983).
later
fertilized.
transport
Hunter,
main
acts
technique.
at a
was
in
more
rabbits
recently
in the ewe and
1981, 1984; Hunter
focus
of these
enough
spermatozoa
complete
fertilization.
respect
to the number
in the tract,
more
by examining
serial
method,
Moricard
studies
was to
had reached
However,
the
can
sow
and
the
this
number
only
be
zation.
Yanagimachi
and Chang
distribution
of spermatozoa
in
hamster
oviduct,
but
after mating.
Thibault
length
and distribution
direct
results
can
sections
of the tract.
and
Bossu
(1951)
the number
of spermatozoa
of the rat oviduct
around
in the ampullary
the time of fertili-
(1963)
serial
examined
the
sections
of the
limited
their observations
to 1 h
et al. (1975)
serially
sectioned
of the
bovine
oviduct
different
times
after
cow
oviduct,
the
4 cm;
(about
1979)
and follows
Because
of these
hamster
Battalia
oviduct
and
Yanagi-
an extremely
tortuous
path
anatomical
features,
we have
use of the serial section
number
and distribution
of
in the hamster
oviduct
and artificial
insemination.
at various
times
and
counted
AND
METHODS
nimals
Mature
and
golden
hamster
(12-20
weeks
males
Animals
were raised
room under
controlled
1900
h; dark
1900
mature
fourth
the
old)
day
of
ovulation)
the
were
Females
consecutive
vaginal
used
for
in the
study.
checked,
each
for the presence
(Orsini,
1961).
discharge
as Day
1 of
(the
day
the
estrous
together
large
mating
and
of
were
determined,
females
in small
cages (20 X 26 X
estrus
on the evening
of
were
housed
individually
large cages (30 X 35 X 17 cm). Those used for
tion of spermatozoa
for artificial
insemination
raised
old)
by 1800 h of every
0100 and 0300 h of
days,
discharge
designated
cycle.
After
their
cycles
were housed
individually
15 cm). Females
exhibited
Day 4.
Males
weeks
used
were
post-ovulatory
was
(7-10
and kept in an air-conditioned
light conditions
(light 0500 to
to 0500
h). Under
this regime,
day.
morning
for four
of a post-ovulatory
The
females
females
came into estrus
day and ovulated
between
following
each
inferred.
Lastly,
with
of spermatozoa
entire
by
which
transport
employed
the
chosen
to make
practical
technique
to examine
the
A
easily
technique
technique
also has a limitation
in that
and specific
position
of spermatozoa
counted
segment
out
a ligature
This
sperm
and has been
The
this
machi,
in situ.
epithelial
epithelial
crypts
However,
the
the female
geni-
to place
at
MATERIALS
be overcome
using
to
short
spermatozoa
after mating
tract
at various
times
after
this effect
has on the number
examine
determine
when
oviduct
to effect
be obtained
Using
this
distribution
technique
the
medium,
has been
to examine
sperm
(Hunter
et al., 1980;
at
Although
be flushed
be determined
approach
that
solution
this
remain
problem
place
of the
to observe
what
eggs
the
are flushed
insemination
used
cannot
This
detergent
method,
salt
to dislodge
spermatozoa
from
the
(Overstreet
and
Cooper,
1978a,b).
exact location
of spermatozoa
within
tal tract
and
mating.
is relatively
the
number
of spermatozoa
tends
to be underestimated
and
1973).
one
are counted.
spermatozoa
the
In
or artificial
studies
to study
sperma-
of the tract
collected
(e.g.,
Braden,
1953),
present
in the
tract
used
mammalian
balanced
after
excellent
been
have
distribution
of different
with
the
number
Compared
three
transport
tozoa
of
1963),
but
not suffi-
AL.
quantitative.
In general,
the
and Chang,
studies
were
ET
housed
in groups
in
collecwere
of up to six in
cage.
Mating
Each
male
least
estrous
of proven
two days
intromissions
mm after
male’s
mating.
the
female
was
placed
in the
fertility
that had been
before
each experiment.
(which
normally
introduction
of
cage)
was
The female
considered
remained
cage
of
a
rested
for at
The onset of
takes place
the female
within
into
the
to be the
in the male’s
onset
cage
of
for
3 0-60
mm until a vaginal smear
least one ejaculation
had occurred.
1
confirmed
that at
The female
was
the spermatozoa
in the various
segments.
Considering
the length
of the bovine
oviduct
(about
14 cm), this
was an admittedly
laborious
task, but worthwhile
in
then
returned
to its previous
cage. A group
of 18
females
was mated
at 2000 h of Day 4 (5-7
h prior
to ovulation).
Another
group
of 9 females
was mated
that
at 0200
they
obtained
a very
detailed
picture
of
sperm
h of Day
1 (during
the time
of ovulation).
TRANSPORT
SPERM
Artificial
IN THE
last
Insemination
Spermatozoa
were
collected
midis
and suspended
in 0.9%
of approximately
1.7 X 108
females
was
ether,
uteri
suspension
26-gauge
this,
were exposed,
was injected
Examination
between
attached
to a tuberculin
Oviduct
At
various
times
mating
after
or artificial
1962),
prewarmed
to
37#{176}C,was
insemina-
dorsal
peritoneal
cavity
to fix the genital
Ten minutes
later, oviducts
with
ajoining
uteri
fresh
were excised
and re-fixed
A.F.A.
fixative
for 6-24
ture.
After
fixation,
tissue,
the
xylene,
and
was
serially
sectioned
ovarian
end
sections
oviducts
dehydrated
with
to
with
avoid
pm)
alcohols,
of
of
oviduct
beginning
spermatozoa.
Individual
spermatozoa
section
the
and
oviduct
a thin
at
the
subsequent
Sections
wall
than
were
scored
in
and
much
the preceding
counted
according
which
found
in the cumulus
in the
infundibulum
spermatozoa.
indication
they
in
to
were
the
each
segment
found.
Those
oophorus,
on and in eggs, and
were
recorded
as ampullary
when a spermatoof the vitellus,
or a
were
of fertilization.
RESU LTS
Within
experimental
groups,
the
number
of
sper-
matozoa
found
in each oviduct
varied
greatly
from
animal
to animal.
In Tables
1-3,
the range has been
given
to show
this variation
between
animals.
For
each
time
period,
there
was
no significant
(p>O.OS)
between
the number
in right and left oviducts
(paired
cleared
Each
contamination
uterine
them in
temperatrimmed
in paraffin.
(10
the
tract in situ.
ovaries
and
were
in graded
embedded
into
by immersing
h at room
having
and lumen
decondensing
sperm
nucleus
or both
sperm
and egg
pronuclei
was/were
discernible
within
the vitellus.
Resumption
of meiotic
division
was taken as another
ether. Ten to 15
10% formalin,
water;
Orsini,
injected
loops
diameter
An egg was recorded
as fertilized
zoon was seen on/in
the surface
syringe.
tion,
females
were anesthetized
with
ml of A.F.A.
fixative
(25%
ethanol,
10% glacial
acetic
acid,
55% distilled
three
loops.
of
and 0.2 ml of fresh sperm
into
each uterus
through
a
or
outside
serial
2000
of Spermatozoa
in the
excess
epididy-
4 (5 to 7 h prior
to ovulation).
animals
were anesthetized
with
Day
needle
caudae
NaCl at a concentration
cells/ml.
A group
of 21
inseminated
artificially
and 2100
h of
To accomplish
from
two
larger
227
OVIDUCT
In
all
tozoa
duct
three
were
experimental
found
throughout
than half of
of the oviduct
groups,
in the
the
difference
of spermatozoa
t-test).
lower
most
segments
period
of
found
spermaof the
observation.
oviMore
the spermatozoa
in the lower
segments
had formed
small groups
(Fig. 2). The
hydrolyzed
with
1 N HC1 at 60#{176}C
for 6 mm, stained
with
Schiff’s
reagent
and counterstained
with
Fast
Green
F.C.F.,
(Fisher
Scientific,
Los Angeles,
CA),
spermatozoa
in the cephalic
isthmus
were usually
observed singly and were located
towards
the center
of
the lumen.
Spermatozoa
in the ampulla
were found
with
most commonly
complex.
the
mated
exception
prior
hematoxylin
to
of
the
ovulation
and
sections
which
eosin
for
from
were
a female
stained
photography
1 and 2; sperm
tails do not stain
well
Green F.C.F.).
Figure
1 shows
cross sections
of the
with
(see
Figs.
with
Fast
and
distinct
of the
trarily
oviduct
1)
divided
utero-tubal
the
junction
and
segments
were apparpresent
study,
we arbiinto
the
four
segments:
intramural
isthmus,
which
makes
a single
loop
within
the wall
of the
uterus;
2) caudal
isthmus,
the portion
composed
of
the first four or five loops of the oviduct
outside
the
uterine
wall;
3) cephalic
isthmus,
the next three
or
four loops having
slightly
lumen
than the preceding
larger
loops;
outside
diameter
and 4) ampulla,
and
the
1 shows
mating
(1
spermatozoa
of
the
the
oocyte-cumulus
the number
segments
h before
5-7
upper
lower
portions
of the hamster
oviduct.
Although
morphologically,
the oviduct
makes
a more
or less
continuous
transition
from
the utero-tubal
junction
to the infundibulum,
ent. For the purpose
Table
various
within
of
the
ovulation.
oviduct.
By
of spermatozoa
oviduct
At
h post-coitum
was observed
of
1 h after
the
in
mated
onset
of
of sperma-
to the caudal isthmus.
From
percentage
of spermatozoa
isthmus
remained
The
total
number
tozoa
continued
oviduct
the
majority
found
in the caudal
constant
(72-81%).
in the
found
females
[p.c.] ), a large number
of
in the lower two segments
3 h p.c.,
tozoa
(7 3%) had moved
this
time
onward,
the
oophorus
more
or less
of sperma-
to increase
to a maxi-
mum
at 7 h p.c.; after this time,
the total
declined.
In this group,
a single spermatozoon
was found
in the
infundibulum
of one oviduct
at 1 h p.c. No spermatozoa were found
in the periovarian
a single spermatozoon
was found
some
females
at
5 and
7 h p.c.,
space. Although
in the ampullae
of
it was
not
until
9 h
228
SMITH
p.c.
that
males.
spermatozoa
By
11 h p.c.,
spermatozoa
tion
reached
had
was nearing
the
a significant
arrived
at the
completion
ampulla
number
ampulla
(Fig.
ET AL.
Table
in all fe(3-3
3) of
2 shows
oviduct
of
the
females
number
of spermatozoa
mated
during
the
period.
As with
the animals
mated prior
a considerable
number
of spermatozoa
and fertiliza-
3).
in the
ovulatory
to ovulation,
was found
in
Ampulla
I
lnfundlbulum
/
Caudal
Isthmus
Cephalic
let h m us
II
#{149} Caudal
Isthmus
Uterine
Lumen
,2#{149}
Intramural
FIG.
mated
1. A
prior
composite
to ovulation
Isthmus
of the upper
and lower
and killed
1 h after the
regions
of the
onset
of mating.
hamster
Section
oviduct
stained
in cross section
with hemotoxylin
showing
and
the
eosin.
various
X 30.
segments.
Taken
from
a female
SPERM
TRANSPORT
IN THE
229
OVIDUCT
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*
U’.
‘‘
N
-,
Cs
0
N
5Cs...,
.
.0
0’.
N
2.
‘-.0
V0
Cs
Cs’-
C
0
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CE
cE
4-
9.C
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,
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N
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SQ SQ en
N
-
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N
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._.0
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ON
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-
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9
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en
en
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I
en
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0’
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en
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en
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o
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9
Cs
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ii
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N
SQ
N
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ocen
0
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0
en
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en
en
i-
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‘
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E#{176}
N
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o.E
V
H
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4-
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05
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D
Cs
en
*
.-,
en,t’.
‘E
i
N
e,
N
*
en
*
SQ SQ
SQ
-
s
,
Eu
-
2
.0
-
,j
‘0
,,
.9
-
5
0
..
N
.
‘0
.9
<0
VCs
V
0’V
0’0
1-0
VCs
0’V
.
‘0
‘
0’0
230
SMITH
ET
AL.
*
>s
Cs
00
0
0
0
00
000
000
‘4-
0
‘1450
‘1450
.0
8
N*
N
0
N
N
en
SQ
0’
Os
N
en
N
0
8
Cs
9
0
‘.12
V
SQN
I
I
en
‘nQN*
0*O5SQ
NOenN
N
V
VV
*
.-‘ONN
N
en
C
*
9
N
en
-
I
Cs
V
I
I
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0
00-N
9
e
4n,nr’len
00
N
oen,n’1’.
00
-
Cs
I
,-,‘-‘‘
0’.
Cs
I
t-en’nen
‘ON’flSQ
*50
gg
‘0
N
0
N
ooen*0’
SQNen*
N©’0’O
*0s
NV’.
‘no
N
*
*
‘0
I
SQ
I
-
FIG. 2. Cross
section
ster mated
5-7
h prior
mating.
Shown
is a small
hal crypt.
Section
stained
U’.
9-
through
the caudal
isthmus
of a female
hamto ovulation
and killed
1 h after the onset
of
group
of spermatozoa
wedged
into an epithewith hemotoxylin
and eosin.
X500.
12
Cs
15
u
V
.0
.0
‘0
Cs
9
E
0
9-
0
u
.0
‘0
u
0
Cs
.0
0.
U’
V
C
U
_
the
*
IF’.
n’ON
‘0 N
I
I
en
N
00
0
‘150
*
II”
0.0
00
0
SQ
SQ
en
SQ
N
en
N
SQ
*
-en
-
0
V
.0
*
_
V
5
V
.0
*
V
.0
SQ
I
I
p.c.;
early
I’
90
C
N
N
0
N
en.n*’0
N
N
N
if’.
414*414154
U’.
N
N’O0.’.
eno’.in*
*
en*
U’.
Oen
I.
SQU’.
.0
.9
9
‘C
4.b
.9
5
Cs
‘0
.0
Cs
Cs12
0’O
N
U’
‘0
N
I
I
I
I
NU’.NSQ
‘14’OSQN
04-’4-
cf. Table
in estrus
in the infundibulum
more spermatozoa
Os en
*
U’.U’.
SQ
N
en
ON
Nil’.
en
NXN-4
N
*
0s
N
‘0*
en
SQ
OSQON
SQ -
*
05
*
of
the
oviduct
of spermatozoa
at an earlier
time
2) when
compared
(32%
at 1 h p.c.;
.0
A larger
caudal
at 1 h
to animals
mated
cf. Table 1). From
appear
that
spermatozoa
were being
the caudal
isthmus
at a faster rate in
Oen*SQ
of one oviduct
at 1 h p.c. A few
(1-3)
were found
in the ampulla
000SQU’.
U’
Cs
0.0
a 1 h p.c.
was found
in the
in this group
(50%
animals
mated
during
ovulation
as compared
to animals mated
several hours before
ovulation.
In animals
mated
near ovulation,
two spermatozoa
were found
‘OSQNU’.
I
-tell
SQ
U
0.9-
portions
this,
it would
transported
to
Cs
.
lower
proportion
isthmus
en
N
u’.
N
-
N
-o
en
-
Mt
Cv
.9
0.0
V
12
C-
.011
Cs
5en
UE
N
N
eno
Os’0
N
U’.U’.
en
0N
en
0.
Cs
*
en
en*
SQ 0
‘0
U’.
N
-
‘0
N
0s
O’nen’O
SQ *
SQ
en
N
N
N
N’O
V’0
VQ
II
if’.
0
N
en
in
0’-
‘015
.0
U,
‘----4
00
UI
Cd,
II
V
en
00
I
-
en
U’.
0’*
en
0OY
V
s
u
.0
‘0
0
<I
N
<
VCs
0
V
.9
12
0.0’
5
FIG.
3. Fertilization
rates
following
mating
and artificial
insemination
early
in estrus
(5-7
h prior
to ovulation)
and following
mating
near the time
of ovulation.
Abbreviations:
M,, mating
in early
estrus;
M3, mating
during
ovulation;
Al, artificial
insemination
early in estrus.
TRANSPORT
SPERM
at 3 h p.c. The number
had increased
markedly
time,
the
eggs
had
of spermatozoa
(2-243)
by
reached
the
in the ampulla
5 h p.c. At this
lower
ampulla
and
fertilization
was nearly
complete
(Fig.
The results
of artificial
insemination
3).
performed
7 h before
in Table
ovulation
total
number
after
artificial
after
tozoa
natural
in the
nation
and
of
observation
the
of
spermatozoa
insemination
mating.
oviduct
then
entering
was
slowly
declined
period.
When
although
oviduct
was
the
of
number
smaller
the
distribution
two
groups.
of
following
of
The
lower
the
rate
was
of sperm
in the
though
when
ovulation
on, the number
ampulla
increased.
the total
number
relatively
conditions
of
may
play
which
the
spermatozoa
artificial
small,
all
to
eggs
variation
in
ported
have
tered.
into
study
has
might
that
the total
number
of
the hamster
oviduct.
there
is a large
spermatozoa
Two factors
have
for
extended
to the caudal
transmay
been responsible
for the variation
we encounFirstly,
there is a wide range in the number
of
clear
in animals
tion.
From
observed
in
animals
mated
we
directly,
at
to
Sheaffer,
1957).
the
movement
30-
the caudal isthmus.
Yanagimachi
and
number
of
to 60-mm
experiments.
ejaculations
mating
Our
that
period
was
only concern
ejaculation
had occurred.
ber of ejaculations
may
the number
oviduct.
To
experiments,
this
occurred
problem,
during
the
not controlled
in our
was that at least one
Variations
in the total numhave had a marked
effect
on
of spermatozoa
observed
later
in the
eliminate
this large variation
in future
the number
of ejaculations
during
the
mating
period
could
using the behavioral
al. (1977).
be determined
criteria
described
and
by
controlled
Bunnell
et
an
mural
numbers
in
the
lumen
isthmus
at
this
time,
of
of
spermatozoa
before
the
ovulation.
lumen
diameter
caudal
been
of
the
due
intra-
facilitating
through
this
(1963)
in the
the
have
thereby
Al-
diameter
to
could
Chang
spermatozoa
of
the
transport
increase
ovula-
during
ovulaintramural
at a faster rate than in
of ovulation
ejaculated
by different
males and by an
male
on different
occasions
(Chang
and
to
majority
mated
hours
sperm
time
in this
isthmus.
through
measure
faster
the
prior
vast
caudal
isthmus
not
the
instead,
move
is particularly
hours
animals
several
did
the
isthmus
caudal
oviduct,
to remain
the
moved
energy
encounter
after
is the intramural
but,
This
to the
after
containing
appear
several
that
to the
though
not
onward,
had moved
we
for
oviduct
in the hamster
mated
spermatozoa
isthmus
the
periods,
isthmus.
3 h p.c.
spermatozoa
individual
Compounding
entered
do
into
is that
may
not
be the best
motility.
Perhaps
more
that
spermatozoa
utero-tubal
junction
Spermatozoa
contractions,
spermatozoa
if a medium
segment
directly
the
factors
spermatozoa
the
This
sperm
stimu-
under
explanation
suspend
isthmus.
tion,
revealed
to
insemination
We
present
of
possible
insemination.
to support
spermaozoa
DISCUSSION
The
uterine
sources had been used.
During
sperm transport
ampulla
was about
of spermatozoa
inducing
entry
follow3). This
These
the
folless
Coital
absent
insemination.
in
NaC1
first
segment
negotiating
the
factors.
were
artificial
Another
0.9%
to the
to several
facilitate
oviduct.
used
due
into the oviduct
Tables
1 and
components
a role
may
in the
at the
been
semen
similar
It should
be noted
that alof spermatozoa
entering
the
oviduct
in this group
was
were fertilized
(Fig. 3).
have
and
transport
insemimated
to ovula-
into the oviduct
was considerably
than the number
transported
ing natural
mating
(compare
artificial
medium
The
arrived
ber of spermatozoa
transported
lowing
artificial
insemination
lus
the
231
Although
care was taken
to artificially
inseminate
each
female
with
a number
of spermatozoa
comparable
to the
number
ejaculated
during
1 h of
natural
mating
(Chang
and Sheaffer,
1957),
the num-
could
this
in
OVIDUCT
insemination,
tion).
spermatozoon
of
spermatozoa
artificial
was about
the same in artificially
animals
as that
in animals
naturally
the same time
period
(5-7
h prior
5 h after insemination,
begin.
From this time
than
found
following
1), it can be seen
spermatozoa
overall
oviduct
ampulla
nated
during
first
oviduct
for the remainder
the percentage
of
are compared
to percentages
prior
to ovulation
(Table
that,
the
notably
53. The
The total
number
of spermaincreased
until
3 h after insemi-
in the segments
spermatozoa
group
mating
are summarized
IN THE
the
segment
observed
intramural
to
large
and
caudal
isthmus
at 1 h p.c. We found,
in all three groups,
that
the
majority
of spermatozoa
were
in the caudal
isthmus
from
3 h p.c. or 5 h post insemination
onward.
is
It would
acting
as
appear,
then,
a reservoir
for
period
from
mating
is not
unique
to
other
species
to
the
(Hunter,
that
ovulation.
hamster
1973,
the
caudal
spermatozoa
This
and
1980,
has
1984,
isthmus
during
the
phenomenon
been
found
1985).
in
232
SMITH
Of the
large
isthmus,
number
groups
of five
were
generally
that
more
observed
groups
of
living
wall
distance,
for
a time,
then
to
observations).
of
this
At
it
sage
through
the
interact
in
some
region.
This
vivo
the
rat
and swim
again
not
with
may
some
the
nature
epithelium.
that
the
the
for
during
their
pasmay
epithelium
represent
of
part
that
of the
in
process.
(1976)
observed
remained
in the
and were transported
of ovulation.
(Shalgi
and
This
Kraicer,
street
and Cooper,
1978b),
sow (Hunter,
1981),
and
1983).
In agreement
served,
in animals
prior
to ovulation,
has also
1978),
that
guinea
lower
isthmus
to the ampulla
been
reported
rabbit
(Over-
mouse
(Zamboni,
ewe (Hunter
and
1972),
Nichol,
with these findings,
we also obmated
or artificially
inseminated
that spermatozoa
did not advance
to the ampulla
in significant
numbers
until about the
time of ovulation.
For the hamster,
it is not well understood
what
triggers
caudal
the
movement
isthmus
to the
tion.
served
Battalia
and
coordinated
hamster
were
may
oviduct
absent
be that
Yanagimachi
propulsive
during
at other
spermatozoa
until
the coordinated
pels them
towards
consistent
with
our
of spermatozoa
ampulla
at the time
the
periovulatory
times
of
remain
movement
the ampulla.
observation
from
the
of ovula-
(1979)
movements
have
of
period
the estrous
in the lower
obthe
that
cycle.
It
isthmus
of the oviduct
proThis
hypothesis
is
of animals
mated
or
inseminated
5 to 7 h prior
to ovulation.
This hypothesis does not, however,
explain
our observation
that
most spermatozoa
remained
in the caudal isthmus
for
a minimum
ovulation,
propulsive
be explained
of 3 h in animals
mated
at the time of
during
the period
when these coordinated
movements
take place. This could partially
by
our
observation
that
many
sperma-
wedged
and
displaced
many
the
at this
muscular
cient to
movements
transport
ampulla.
not
time.
It
easily
we also
which,
transported
that
to the
coordinated
of the oviduct
alone
spermatozoa
from
the
the
easily
in the lumen,
is possible
Perhaps
of the
been
However,
free
been
crypts
have
contractions.
have
could
epithelial
would
spermatozoa
ampulla
the
into
probably
by muscular
observed
are insuffiisthmus
to
spermatozoa
themselves
must undergo
some physiological
changes (e.g., initiation of hyperactivated
motility)
in the isthmus
before
advancing
to the ampulla.
In females
mated
several
hours
before
ovulation,
until
tozoa
remained
in the
the time
of ovulation
could
have become
lower
isthmus
for
after
the
more
effect
have
from
onset
of mating
hours
before
fertilization
completed
the
onset
of
the
lower
for
the
the
to
must
3-h
period
beginning
addition
isthmus
only
ampulla
spermatozoa
until
in
ovulaby 1 h
there
the
during
mating
Therefore,
reservoir,
to
2). These
capacitation
fertilization.
sperm
and remained
ascending
(Table
7 h,
(Table
1). These spermacapacitated
at any time
during
this 7-h period.
In females
mated during
tion,
spermatozoa
reached
the lower
isthmus
2
and Mahi
were
spermatozoa
How-
spermatozoa
tozoa
presumably,
(unpublished
know
oviduct
isthmus,
way
pig
spermatozoa
before
ovulation
in
speculate
interaction
Yanagimachi
time
do
to
caudal
capacitation
at the
we
the
is tempting
to the oviduct
it was impos-
These groups
of
stuck
to the ovi-
free
stuck
with
groups
of the
through
AL.
oviduct
was of a permanent
occasions,
we have
to break
present,
association
ever,
These
the folds
oviduct.
become
become
caudal
had formed
spermatozoa
wall
of a freshly
excised
spermatozoa
appeared
to
duct
half
in close association
our fixed
material,
tell
if this association
However,
on different
to
nature.
in the
than
or more
spermatozoa.
found
wedged
into
oviduct
wall, lying
epithelium.
From
sible
of spermatozoa
we observed
ET
to
of
acting
as a
also be acting
may
as a site for sperm
capacitation
in vivo, and this 3-h
period
may represent
the minimum
capacitation
time
for
hamster
spermatozoa
in vivo.
Few
spermatozoa
were
found
in the
cephalic
isthmus,
even when fertilization
was in progress
in the
ampulla.
The spermatozoa
leaving
the caudal isthmus
must
be transported
very
rapidly
to the ampulla.
Either
an increase
in forward
progressive
motility
of
spermatozoa
and/or
an increase
in the propulsive
movement
this.
In
of
all
the
oviduct
experimental
spermatozoa
observed
could
be responsible
groups,
in the
the
ampulla
during
stage of fertilization
observed
previously
Yanagimachi,
1982),
was very small.
in the hamster
mouse (Zamboni,
(Shalgi
and
spermatozoa
1978).
We
to arrive
after
Kraicer,
continued
fertilization
was
the
of
early
This has been
(Cummins
and
1972),
and rat
observed
that
in the ampulla
complete.
for
number
These
many
even
late-arriving
spermatozoa
may function
to break up the cumulus
oophorus
to facilitate
the passage of the eggs into the
more
We
constricted
did
find
in infundibulum
mals; however,
rule.
Because
lumen
a very
of the oviductal
small
number
isthmus.
of spermatozoa
at 1 h p.c. in naturally
mated
anithis was the exception
rather
than the
these spermatozoa
were found
in the
SPERM
infundibulum,
we
may represent
spermatozoa
thought
the
1 h preceding
clarify
this,
we
from
1 h p.c.
females
could
curred
have
during
our
examined
first
the
that
p.c. No spermatozoa
space. The spermatozoa
these
IN
been
To
space
sacrificed
of 4
at 1 h
were found
in the periovarian
found
in the infundibulum
at
been
from
sectioning
contamination
or
staining,
that
but
this
oc-
seems
unlikely
since they
were
lumen
of the infundibulum,
these
spermatozoa
being
found
within
the narrow
and the probability
of
deposited
there
by chance
seems
small.
Therefore,
the
found
in the
infundibulum
represent
for
rapidly
rapid
transported
transport
has
oviduct
of
been
Yamanaka
and
recovered
from
very
spermatozoa.
in
Soderwall
spermatozoa
at 1 h p.c.
spermatozoa
reported
the
few
of females
the
past.
(1960),
“ampullary
must
the
hamster
According
spermatozoa
segment”
to
can be
of the ham-
ster genital
tract within
2 mm after
coitus.
However,
upon
careful
reading
of these authors’
description,
it
has become
clear
that
their
term
“ampullary
segment”
per
the
referred
to the
entire
se. Rabbit
spermatozoa
ampulla
during
the
were found
to be dead and
space
before
fertilization
Cooper,
1978a).
Since
material,
it is impossible
the spermatozoa
found
they
to
were
infer
alive,
did
not
the
ampulla
were transported
to
phase”
of transport
cleared
began
to the periovarian
(Overstreet
and
our data are based on fixed
to ascertain
the condition
of
in the infundibulum
at 1 h
p.c.
But,
since
eggs were
reached
the mid
or lower
reasonable
oviduct,
that
“rapid
not fertilized
ampulla,
it
that
not
these
until
would
spermatozoa,
participate
they
seem
even
if
in fertilization.
Since
that
this
have
the
sperm
paper
confirmed
use
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