BIOLOGY
OF
REPRODUCTION
Regional
19,
Histology
(1978)
1063-1069
and HistoChemistry of the Ductus Rhesus Monkey (Macaca mulatta)’ J. ALSUM
DONALD
and
Department
0/Animal
University
St.
ALAN
Epididymis
in the
G. HUNTER Science,
0/Minnesota,
Paul,
Minnesota
55108
ABSTRACT Examination of epididymides from 6 sexually mature male rhesus monkeys revealed epididymal epithelial and stereocilial height was maximal in the capur region and minimal at the cauda. Luminal diameter was just the reverse. Strong acid phosphatase activity was localized principally in the supranuclear area of the columnar cells in the distal caput and corpus epithelia. Stereocilia were practically devoid of this activity. Moderate alkaline phosphatase activity was localized in the basal cells of the corpus and proximal cauda epididymidis, while activity in the principal cells was slight. Luminal contents of ductuli efferentes and epididymis had much more alkaline phosphatase activity than tissues. PAS positive materials were most evident in the apical epithelium of the ductuli efferentes and distal caput and in the luminal contents beyond the proximal caput. Location of the cytoplasmic droplet on the sperm midpiece changed from proximal to distal between distal caput and proximal corpus. Progressive sperm motility changed dramatically between distal corpus (15%) and proximal cauda (75%). It was hypothesized that some rhesus sperm within the distal corpus (second portion of the middle segment) would have acquired fertilizing ability.
INTRODUCTION
Growth
and
of the androgens
differentiation
rhesus during
1972a)
and
monkey both
adult
period
transit
(Dinakar
mean
rhesus days
epididymal segments for the caput, corpus (Amann
sperm
acquire
fertilizing
involves
factors
extrinsic 1975;
al.,
is
region
where
sperm
Therefore, currently
of
most
intrinsic
to
within
the
first
acquire only
controlled breeding to determine the
based on means
epididymides
in various
‘Paper Minnesota
been
other
studies
laboratory reported
ability. data
available
to on the
and
domestic
(Nicander,
1957,
June 29, 1978. January 24, 1978. No. 10,339 Agricultural
of the Experiment
the
and
Journal Series of the Station.
1063
studies
(Ramos
in the
and
Dym,
to
relate,
undertaken
The
specific
objectives
the
histometrically
where
phosphatase
positive rhesus
type
spermatozoa location
MATERIALS
epididymis
fertilizing
within
degree the
was
determine
alkaline
(PAS)
and
Parallel rare
the regional epididymal epitheactivity to that of post-testicular
to
Schiff
in
Bedford, et al.,
monkey
study
describe
epididymis;
as
are
maturation.
to
tase,
well
and Nicander, 1971; 1975; Orgebin-Crist
unlikely that will be performed
were:
species as
Our
spermatozoal
the and 5.6 respec-
maturation
sperm
1975).
primate
where possible, hal and luminal
epididymal
This
Hamilton,
1977b).
1974).
mammalian
this occurs. and histochemical
Accepted Received
al.,
through
During
capacity.
hypotheses are the
have
et
are 1.1, 3.8 and cauda,
predict where Histological animals
epididymis
sperm
1976).
of
(Bedford Hamilton,
1976). It experiments exact
times
et
transit,
of the
are dependent upon the prenatal (Alexander,
The
tively
1958; subhuman
and
substances
epididymis;
of potential from of
acid
each their
AND
periodic
rhesus
phosphaacid-
are localized to determine the motility
epididymal cytoplasmic
inherent region droplets.
METHODS
Six adult macaque monkeys (Macaca mulatta) were castrated and the excurrent ducts dissected from the testes. Sexual normality was established by normal testicular histology and histochernistry and by the presence of normal, motile sperm in the cauda epididymidis. Tissues representing testis and 7 regions (Fig. 1) from 1 epididymis of each monkey were frozen in isopentane and tissues from the contralateral epididymis were used to provide sperm samples. Cryostat sections were cut at 8 tom, air dried and stained with hematoxylin and eosin (Cooper, 1963) and for acid phosphatase (I3arka and Anderson, 1962) using 30 and 60 mm staining times, for alkaline phosphatase (Thompson, 1966) using 2 and 4 mm staining times and for periodic acid-Schiff (PAS) positive materials (Humason, 1967) using 30 and 60 mm staining times. Staining intensity was scored
1064
ALSUM
AND
HUNTER
from height to volume and the percent packed sperm cells calculated. To determine the location of the cytoplasmic droplet, fluid from 2 males was aspirated into 0.5 ml formal saline, placed on a slide and evaluated under phase contrast. Position of the cytoplasmic droplet on the midpiece was determined on 200 sperm per region. Positions were classified as being proximal, mid, distal or absent.
if firent
RESULTS
Histometric
via
The and
luminal
and
the
various in
corpus
of
rhesus
epididymal
regions.
1), caput (regions (regions 6-7).
2-3),
Table
ter. Measurements were made with a light microscope equipped with a calibrated eye piece micrometer such that 3 m differences could be resolved. Contents from the contralateral epididymides were collected by making an incision across each specific epididymal region and aspirating the accumulated fluid. Testicular fluid was collected similarly. For motility studies, fluid from 2 males was aspirated into 0.5 ml TES NaK yolk buffer (Graham et al., 1972) at room temperature, placed on a microscope slide, warmed to 37#{176}Cand evaluated microscopically by 3
For relative sperm concentration studies, fluid from 2 males was aspirated into a 400 MI Beckman teflon tube and centrifuged in a Beckman Model 152 Microfuge for 5 mm. Seminal columns and sperm columns were measured with sliding calipers, converted
I. Measurements
Epididymal
of rhesus
region
Ductuliefferentes,
excurrent
Lumin
77.1
al diameter
2, proximal
161.2
Caput Corpus
3, distal 4, proximal
145.7 218.7
5.5 8.5 6.4 9.6
5, distal
227.3 328.7
7.9 14.2
618.0
31.4
6, proximal 7, distal
Cauda Cauda
a Values *Not
are measured.
in microns
and
represent
the
efferentes
regions
are
Epididymal
was
fixation
cauda.
in the
Luminal
both
studies
stereocihial and tubular
than Both
narrower
(1977b). ter
enlarges
populations
between
the
from
close,
about were
of show
of
heights
relatively
those studies
report-
were smaller and Dym to different
epithehial
were
and was
were
and/or
lengths were diameters
and
caput
diameter
(Table 1) by Ramos partially due
Epididymal
summa-
epithehial
maximal
procedures
monkeys.
stereocihia
ductuli
but
3-fold about
Ramos and the luminal ductuli
our
shorter 1.5-fold Dym
diame-
efferentes
and proximal caput and then narrows in the distal caput. However, in the bonnet monkey (Macaca radiata), the height of the epididymal epithehium in
and
the
caput
(Flechon
et
Ramos ductuhi ily sional
lumen region
al.,
and
Dym
maximal
at
(1977a)
in monkeys
ciliated
basal
were
minimal
the
cauda
1976).
efferentes
of
diameter and
and cells,
nonciliated macrophages
the
reported
consisted
primar-
cells and
and
occa-
intraepithe-
ducts.a
Caput
Corpus
1
at
the
of
the
just the reverse. Identical findings ed by Ramos and Dym (1977b).
people.
TABLE
1.
Our measurements than those reported (1977b). This was
independently by 3 people, averaged for individual monkeys and means and standard errors were determined on a group basis. Five random transverse histological measurements per region were made for epithelial height, stereocilia length and luminal diame-
length
in
epididymal
height
minimal
(region cauda
height, diameter
stereocihial
1. Diagram efferentes (regions 4-5),
DISCUSSION
Observation epithehial
rized
FIG. Ductuli
AND
means
±
SEM
Epith dial
21.5 69.6 78.4 64.0
1.0 2.0 2.3 2.4
52.1 49.9 40.2
for 6 animals.
height
Stereocilia
height
S
18.4 16.2 7.5
1.4 1.2 0.7
2.0
10.3
0.6
2.2 2.3
7.5
0.6
4.0
0.5
MONKEY
hal
lymphocytes.
presence
of
basal
pseudostratified Vasectomy
causes to
diameter
the
enlarge
and
than
size
were
body
1972b).
region
7. These
ductuhi
four
We
in the
in
agree
that
with
no
unique
head
may
or
function
division
regions
was
such and
epididymis. initial
of
Regions
the
4
and
the
5,
characterized
by
supranuclear and
an
(Table
2).
Regions
ment,
were a
very
Acid
Phosphatase The
data
7,
similar
The 0.01 Moderate
segsperm.
Acid
testes
controls activity was area of epidid-
the
activity
rhesus
Riar
et
luminal
rhesus
acid
epididymides.
of acid
phosphatase (Hunter et and Risley,
Moniem
and
Glover,
monkey
most
closely
the
ram
(Moniem
and
Glover,
rat
(Nikkanen
and
phosphatases
of
are
and
the
Vanha-Perttula,
strong
acid
apically
in
(regions
3-5)
vation
epididymal
the
that
compounds
and
the
that
and
tract, corpus
corpus
activity
was but
primarily
regions.
was
in
suggested occurring The
spermatozoal
region
The
localized epithehia
luminal
epididymides, materials
luminal
in that
under 1963).
activity,
weak
rhesus
caput
increased
in obser-
density
in agreement
with
concept. Acid
caput
the
absorption
(deDuve,
phosphatase the distal caput and
enzymes
with
of
conditions
rhesus monkey absorption of
throughout the distal
lysosomal
associated
breakdown acidic
the that
are
usually
slightly
increased distal
of
The
devoid
through
comparison
Epididymal region
2.
incuba-
practically
phosphatase
Regional
Fig.
both
in the supranuclear in all regions of were
from
in for
M NaF-treated to strong
2).
rhesus
and
Acid
epithehium with
testes
from
minimal
1952;
Montagna,
which
terminal
packed
and
acid
1977).
a wider
lower
efferentes
varies with species 1969; Martan
The
enzyme
detectable (Fig.
in
Martan,
1972b). 1972b)
concentration
the
1972;
resembled
were
epithehium,
by
was
Stereocihia
2.
stereocihia. stereocihia,
sperm
al.,
the
decreased
distribution
epididymis
1963; by
segment,
summarized
localized principally the columnar cells
TABLE
the
that
found
regional
in the
showing
epididymidis
also
cauda
biochemical
ductuhi
activity
The
Activity are
periods. negative.
activity.
the
of
lumen
of activity
gradually
in
6 and
wide
portion
regular
characterized
and
pattern
middle
area
major
long
more
vacuoles
lumen
of
characterized
extremely
with
(1973)
with
barely
activity caput
corpus
to distal
caput . region with little 1972). The luminal contents
tubules,
phosphatase
ductuli
segment
3, the
were
al.
into
was
1
initial
2 and
Nicander,
tissue
exhibited the
proximal
conflict
the
seminiferous
phosphatase
to
epididymis
Region
segment,
epithelium
and
the
that
part
according
(Glover of
Regions
the
divided
be
in
gradually
rhesus
epididymides through
Our
efferentes
on
strong
results
was mainly in beyond (Riaretal., of
other
epididymal
region
studies
times
characteristics
present
1971).
ymis.
as
(1977b)
epididymis
morphology
tion were
columnar.
regions.
The
high
3, remained 4 and decreased
the
rhesus
much
histological
region
simple
adult
as
Dym
distinguishing
the
1065
EPIDIDYMIS
epithehium
confirmed
of
(Alexander,
Ramos
of
results making
instead
efferentes
and
Our cells
DUCTUS
phosphatase
catabolism
sperma
of
may
substances
concentration,
Droplets
Droplets
Droplets
Spermatocrit
proximal
distal
absent
10b
92 85 87 2 1 0 0
be
involved
absorbed
anatomy
and
in
from
motility
the
the
patterns.
Motility
(%) Ductuli Caput Caput Corpus Corpus
Cauda Cauda
efferentes, 2, proximal 3, distal 4, proximal 5, distal 6, proximal 7, distal
aSamples
were
of everything on bRete testis-
cNO
tested.
1
.
49 69 69 61 73 obtained
from
6 monkeys.
each monkey. Therefore, Ductuli efferentes.
0 6 5 89 92 78 51 However, samples
from
8 9
7 9 7 22 49
None 1% circular + 5% stationary 10% progressive + 5-25% stationary 5-15% full progressive + 60% stationary 5-15% full progressive + 60% stationary 70-75% full progressive + 10% stationary 75% full progressive
epididymal
sample
2 monkeys
were
volumes used
for
were each
insufficient determination.
for
analysis
1066
ALSUM
4
Acmd
AND
HUNTER
contain
Pase
this
A
of
C3
droplet
I
this
enzyme
suspending (Dott
the
ram,
associated
T Yl
and
Dingle,
found
that
levels
I
2
VE
3
4
Caput
Alk.
5
6
Corpus
7
VD
Pose
apical
dyl in
Cauda
-
A C3
lumen
transit
through
largest
change
took
corpus,
activity
..‘‘#{149}.4...
....
T
vi
was
(Riar
Alkaline
Phosphatase
The
0 yE
2
3
5
4
Coput
6
Corpus
7
VD
The
testes,
was
C
caput of
in
secretory
I
vi
The
0 T
VE
3
2
4
Coput
5
6
Corpus
7
VD
ductuli
Caudo
the
luminal
than
the
from
ductuli 5
through
cauda.
Our
and
lumen
(Moniem
Nicander,
1965,
1966)
testicular
origin.
was
activity mass
indicating
that
epididymal this
Slight
observed of
in
the
rhesus
the
enzymes
sperm. might
1972b).
and
such
be However,
acid the
were the
artifact cytoplasmic
1972b;
proteins
lumen
spermato-
epithelium.
also
present was
(Moniem droplets
droplet not
could
and
sperm plasma.
of
rabbit
not
They
high
the
on al.,
and
ram
levels
rhesus In
activity in
sperm.
addi-
in the
Glover
in the it
were
whole
than
hypothesized
regional
1972).
more
demonstrate
despite
corpus constant
on
phosphatase and
increased distal
phosphatase
et
Moniem
alkaline
and
essentially
(Riar
epididymis
The
activity
activity
data
the
of
epitheli-
activity.
enzyme
data
illustrated
caput
efferentes
alkaline
data
a
alkaline
columnar
remained
tissue
our
localized
epididymidis
tissue
tion,
for
through
biochemical
epididymal
of
phosphatase
luminal caput
Since an
Glover,
as
to
proximal
Luminal
of
similar
and negative
more
histochemical
of the
cells
attributed
ductuli
efferentes
region
and (1970)
basal
slight
much
tissues.
basal
corpus
cells.
apical
of
had
greatest Bavdek
the
showed
contents
distribution
epididymal
the
regions and efferentes
He
virtually
corpus
epididymis FIG. 2. Acid and alkaline phosphatase activity and periodic acid-Schiff staining activity in apical portions of rhesus monkey excurrent duct epithelium and luminal content. Mean ± SEM. Activity: 0 = no reaction, I = slight, 2 = moderate, 3 = strong, 4 = intense; apical, is.. = lumen.
in
All
media activity
the
4-6).
these
while
of
The in
efferentes
were
phosphatase, urn
cells.
activity
to
2.
epididymides
epididymidis.
activity
epithelia
rhesus
Fig.
phosphatase
(regions
caput
and
the
deficient
substrate
detected
strong
rabbit
Glover,
that
in
basal
cauda
observed
fixed,
proximal
efferentes,
was
proximal
on
the than
alkaline
the
in
activity
is
phosphatase
summarized
ductuli
strong
cell
zoal
activity
acid
and ductus deferens varied between within tissues. Activity in the ductuli
4
caput
Activity
are
negative. the
The the
1973).
incubated
were in
Cauda
et al.,
data
controls
in
lipid
epididymis.
phosphatase
higher
monkey
total
progressively
between
luminal
strongest
significantly
total
phosphatiand
the
the
be
(1975)
protein,
decreased
acid
man,
al.
et
acid,
place
where
In
to
maturation
choline,
sperm
of
ceases its
sphingomyelin
monkey
the
extent
Arora
sialic
of release
a lesser
during
1968). of
during
was
T
sperm
ethanolamine,
high.
to
phosphatidyl
rhesus
and
4
and
and
phosphatase
the
(Dott
1968)
bull
acid
phospholipid,
0
the
with
disruption
Dingle,
In
of osmolarity
alteration
causes
and
enzyme.
in
and
fluid
the
lining (1972a)
cytoplasmic
However,
in
hamster
in
the
they or
epididymal that
alkaline
rat
MONKEY
phosphatase cell by
played a role dephosphorylation
phosphate and
groups
the
in the mis.
sperm
epididymal
and
would
thus
following
of sperm phosphate
compounds
are
in
the
distal
early
maturation
The more
phospholipid,
dyl
lower
1975).
Also,
luminal cauda.
decreased with
alkaline
phosphatase
between
distal
change
in the
droplet
the
and
position
from
seems tered
in
caput
proximal
disrupted
during
reactions
are
Acid-Schiff data
difference periods. incomplete significant
to
strong
the
apical
tes the
ductuli
efferentes
rat
and
(1955)
in the
epididymis.
which
He
was
Stereocilia
most and
from moderate (region 3) to (region
6).
The
PAS was
out
the
the duct,
basal were The
tubules
reaction
of the in
the
epididymis,
lamina strongly luminal through
Apical in the (region
3).
the
the
contents corpus
from
2).
bonnet Sperm
1975).
efferentes
location
and
of
the
midpiece,
and
droplet
midpiece.
cyto-
occurred
proximal
changed
corpus
from
The
appreciable
proximal
droplet
numbers
testes
Microscopic
had
been
of cauda
sperm.
during
transit
that
testis and
the
increased
to corpus
epididymis
observations
histological suggested
stallions
was
being
caput
Sperm to
of
slides verified most of the as
(Nicander,
corpus
caput.
concentration
rete
1965).
cycle
(region Apical the
epididymal region.
collagenous
surrounding PAS positive.
the
the
this fluid
finding. arising in
the
reported
in
bulls,
1958)
and
was
high
2).
epididymal
absorbed
density
distal
(Table
This from efferent
rams,
boars
(Crabo,
from
proximal
cauda.
diminished
cauda
basal
for
atocrit
ducts
PAS
distal
was observed. deferens was
from
Sperm
efferen-
distal caput epididymidis in the proximal cauda distal
in the caput
The
on the
distal
Sperm
in the slight the
were
(Table
in
a secretory the
sperm
ductuli
on the sperm
distal 2).
shed
prevalent
activity
rhesus
epididymis
reported and Hafez,
change
droplet
between to
PAS
In
faintest
results.
only slightly intensities. of moderate
throughout
in
the
testes,
tail forward
Droplet
A rapid
were
apical
increased activity hum of the ductus cells
epithelia
active
rete
showing finally,
6 (75%).
region
(Table
caput
et al., Bower,
proximal caput were not progressively motile while about 10% of the distal caput sperm were
materials
considerable
proposed
the
plasmic
ductuli
distal
increases
(Hunter and
sperm and
data were (Flechon
from
Little
epididymis. was strongest
noted
material
2.
our
testis,
the
Similar monkey
of circular
transported
2 incubation diffusion nor
were
Similar
boar (Malone
PAS vary
progressively motile. The greatest change in progresstve motility occurred between distal corpus region 5 (15%) and proximal cauda
the epididymis. In general, moderately to strongly PAS
throughout PAS activity
reactive
PAS
the
positive epithelial Maneely
It
lumen.
Fig.
the
intensity
and throughout stereocilia were
2).
percentage
Cytoplasmic
in
influencing
of
a
stronger
0.1% maltdiastase the tissue PAS staining PAS positive materials epithelia
of
the
in
staining
the
Activity
of
factors
Incubation reduced Diffuse
as
along
is sequesand as they in
between neither
reactivity
increased
was
maturation,
summarized
was seen Therefore,
motility,
to
(Table
observed
(PAS)
are
then
cytoplasmic
distal
the
of not
Motility.
The
contents
phosphatase droplets
positive
beating,
corpus
sperm
to
become
The
luminal
of the
point.
et al., in the
increase
proximal
likely that alkaline into the cytoplasmic
Periodic
the
that
reported in the hamster
Sperm
caput
the
beyond been and
amounts which did
1962).
phosphati-
from
Concomitant
have 1972)
increasing (Fig. 2)
total
regions: choline,
acquired materials
from
sphingomyelin (Arora levels of phospholipid
the
fluid
sperm
epididymal
phosphatidyl
ethanolamine,
involved
in the epididycontaining
rhesus
1067
markedly
plasma be
EPIDIDYMIS
gradually positive
in the function of the and transport of
between cell
DUCTUS
the
least
Acquisition
of Fertilizing
Regions
7),
tes
and
1-3
constitute
initial
region
epithe-
Fertile
sperm
have
active.
there
in every
species
columnar
the
Through-
location
fibers
of
epididymal
of
seminiferous
epididymal
region
poor
sperm
of
Capacity
the
the of
the
not
been
motility
sperm
to
have
originate
In addition,
and
the
droplet not
efferenepididymis.
found
cytoplasmic
from those regions are fertility. Region 4 (proximal
studied,
ductules rhesus
so far studied.
the sperm concentrated segment of the epididymis. 4,
Hypothesis
proximal
on
portion of the In the species to
sperm
compatible with corpus) constituted
pass
through
middle so far such
a
1068
ALSUM
zone
prior
to
acquiring
greatest
potential
developed
between
and
proximal
that
upon
cauda
sperm
ability
and
(region
(regions
probably
is gradually
the
corpus
(region
5)
6). It is hypothesized
distal
corpus
distal
6,
part
7).
5),
of
region
4
capacity
by
rhesus
and
in
N. J. (1972a).
ductus
epididymis
effects
of
119-133. Alexander,
Prenatal
fetal
R. Pickett,
duction, transit
R.,
monkey.
Biol.
N.
and
changes contents
Prasad,
in of
M.R.N.
the
for
different
1, 128-131. Bedford, J. M. (1975).
of
the
System. Amer. 17.
Crabo,
(D.
W. Hamilton
Physiol.
Soc.,
B. (1965). Studies on epididymal content in bulls Scand. 6 Suppl. 5, 1-94. deDuve, C. (1963). In: Lysosomes.
the and
and
of Vet.
of testosterone on the glands of the castrated
ion
and
A.
G.,
the
freezing Sci.
J. Dairy
function efferentes,
of the ductus
and
ducnili
deferens in the rat. In: Sect. 7, Endocrinology. R. 0. Greep, eds.). The
Society,
Washington,
D.
and of
Alteration epididymal
Biol. Reprod. Bower, M. polysaccharide
phosphatase Amer.
pp.
D. A.,
(1972). during
Abstr.
Technique.
Tissue Francisco.
San
Schellpfeffer,
T. E. localization
Maneely,
on
I. Bull.
Animal
Co.,
Barker, L. seminal antigens
Abstr.
buffers
Structure
the
G. L. (1967).
and
Zool.
in
C.
W. H.
301-308.
Johnson, W. L. of porcine transit
and
7, 133. (1962). Regional complexes and
hamster
epididymis.
2, 427-428.
R. B. (1955).
The
distribution
of polysac-
charide complexes and of alkaline glycerophosphatase in the epididymis of the rat. Acts Anat. 24, 314-329.
fate
Washing-
composition boars. Acts
of
and
259-301.
alkaline
(de Reuck, A.V.S. M.P., eds.). J. and A. Churchill p. 1. R. and Prasad, M.R.N. (1974).
Effects of micro-quantities epididymis and accessory
Hunter,
Cytochem.
and
zwitter
of spermatozoa.
Physiological
pp.
Freeman
Malone,
hexazonium
Bedford, J. M. and Nicander, L. (1971) Ultrastructural changes in sperm membranes and the acrosome during maturation of spermatozoa in testes and epididymis of rabbit and monkey. J. Anat. 108, 527-543. Cooper, A. R. (1963). Instructions for pathologists and technicians at Peter Bent Brigham Hospital. In: Cryostat Frozen Sections in Surgical Pathology. (Cited in Microtome-Cryostat Handbook, 2nd ed., 1964). International Equipment Co., Needham Heights, Massachusetts. pp. 15-16.
Cameron, Ltd., London. Dinaker, N., Arora,
American
maturation.
Histochemical
transport,
some
epididymis, and ductus Handbook of Physiology. D. W. Hamilton and
Mulat-
in the caput epididymis Aspects of Andrology
Maturation,
J. T. (1968). Distribution in the spermatozoa of bull and ram. Exp.
55, 372-378. Hamilton, D. W. (1975). epithelium lining
and
in the epididymis. In: Handbook Sec. 7, Endocrinology. Vol. V.
Male Reproductive R. 0. Greep, eds.). ton, D.C. pp. 303-3
and
of
storage
Humason,
(1975).
regions Macaca
acid phosphatase using as coupler. J. Histochem.
10, 741-753. Bavdek, S. (1970) Basal cells of rabbits. Morphological
of spermatozoa of Physiology.
and
15,
spermatozoa
epididymis of the rhesus monkey, ta. Contraception 11, 689-700. Barka, T. and Anderson, P. J. (1962). methods pararosaniin
The 135,
Reprod.
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Flechon, J. E., Bustos-Obregon, E., Steger, R. W. and Hafez, E.S.E. (1976). llltrastructure of testes and excurrent ducts in the bonnet monkey (Macaca radiata). J. Med. Primatol. 5, 321-335. Glover, T. D.. and Nicander, L. (1971). Some aspects of structure and function in the mammalian epididymis. J. Reprod. Fert. Suppl. 13, 39-50. Graham, E. F., Crabo, B. G. and Brown, K. 1. (1972).
D.C.
Dinakar,
Biochemical luminal
of the
monkey. J. Anat.
P., Johnson, L., Thompson, D. L. and B. W. (1976). Daily spermatozoal proepididymal spermasozoal reserves and time of spermatozoa through the epididy-
rhesus
Dingle, enzymes
mulatta.
12 19-1227. 5.
Vasectomylong-term monkey. J. Reprod. Fert.
N.
mis of the 586-592. Arora,
rhesus Am.
Macaca
droplets Cell Res. 52, 523-540 Flechon, J. E. and Hafez, E.S.E. (1975). Spermiation and epididymal maturation of spermatozoa in the bonnet macaque (Macaca radiata) as viewed by scanning electron microscopy. Fertil. Steril. 26,
Effect
J. (1972b). effects in the rhesus 31, 399-406.
Amann,
development
in the castration.
monkey,
lysosomal
sperm region
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Alexander,
rhesus
60, 399-408. Dott, A. M. and cytoplasmic
(region
Fertilizing
acquired
HUNTER
The
motility
have mostly acquired fertilizing would then be stored in the terminal
segment in
distal
the
ability.
progressive
the
leaving
rhesus
fertilizing for
AND
J.
Martan,
(1969).
physiology.
Epididymal Biol.
histochemistry
Reprod. Suppl. P. L. (1963). The
and
1, 134-154. epididymis
J. and Risley, of mated and unmated rats. J. Morphol. 133, 1-16. Moniem, K. A. and Glover, T. D. (1972a). Alkaline phosphatase in the cytoplasmic droplet of mammalian spermatozoa. J. Reprod. Fert. 29, 65-69. Moniem, K. A. and Glover, T. D. (1972b). Comparative histochemical localization of lysosomal Martan,
enzymes
in mammalian
epididymides.
J. Mat.
111,437-452. W. (1952). Some cytochemical on human testes and epididymides. Acad. Sci. 55, 629-642.
Montagna,
Nicander, L. (1957). cytochemistry rabbits. Acts
On the regional of the ductus Morph. Neer-scand.
Nicander, L. (1958). and cytochemistry stallions, rams scand. 1, 337-362. Nicander, L. (1965).
of absorbing
cells
observations Ann. N. Y. histology and epididymidis in 1, 99-108.
Studies on the regional histology of the ductus epididymides in and bulls. Acts Morph. NeerAn
electron
in the
microscopical
posterior
caput
study
epididy-
MONKEY
mis Nicander,
of
rabbits.
L. (1966).
DUCTUS
Z. Zellforsch. 66, 829-847. A study of multivesicular bodies
Re-examination
of
the
dependence
of
epididymal sperm viability on epididymal environment. J. Reprod. Fert. Suppl. 24, 115-128. Ramos, A. S. and Dym, M. (1977a). Ultrastructure of the ductuli efferentes in monkeys. Biol. Reprod.
Riar,
1069
17,
339-349. A. S. and
the 501-
monkey 53 1.
Ramos,
in the epididymis. (Abstr.) J. Ultrastruct. Res. 14, 424-425. Nikkanen, V. and Vanha-Perttula, T. (1977). Acid phosphatases of the rat epididymis. II. Biochemical characteristics, subcellular distribution and histochemical localization. Andrologia 9, 115-132. Orgebin-Crist, M. C., Danzo, B. J. and Cooper, T. G.
(1976).
EPIDIDYMIS
Current Riar,
S. S..
B. S. and
composition
Science Setty,
M. (1977b).
epididymis.
S. S., Setty, chemical
Dym,
Fine structure
Kar, of
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A. B. (1972).
Bio-
41, 453-455. B. S. and Kar,
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A. B. (1973).
Studies
on the physiology and biochemistry of mammalian epididymis: Biochemical composition of epididymis. A comparative study. Fertil. and Steril. 24, 355-363. Thompson, S. W. (1966). Selected Histochemical and Histopathological Methods. C. C. Thomas, Springfield, Ill.
OF
REPRODUCTION
Regional
19,
Histology
(1978)
1063-1069
and HistoChemistry of the Ductus Rhesus Monkey (Macaca mulatta)’ J. ALSUM
DONALD
and
Department
0/Animal
University
St.
ALAN
Epididymis
in the
G. HUNTER Science,
0/Minnesota,
Paul,
Minnesota
55108
ABSTRACT Examination of epididymides from 6 sexually mature male rhesus monkeys revealed epididymal epithelial and stereocilial height was maximal in the capur region and minimal at the cauda. Luminal diameter was just the reverse. Strong acid phosphatase activity was localized principally in the supranuclear area of the columnar cells in the distal caput and corpus epithelia. Stereocilia were practically devoid of this activity. Moderate alkaline phosphatase activity was localized in the basal cells of the corpus and proximal cauda epididymidis, while activity in the principal cells was slight. Luminal contents of ductuli efferentes and epididymis had much more alkaline phosphatase activity than tissues. PAS positive materials were most evident in the apical epithelium of the ductuli efferentes and distal caput and in the luminal contents beyond the proximal caput. Location of the cytoplasmic droplet on the sperm midpiece changed from proximal to distal between distal caput and proximal corpus. Progressive sperm motility changed dramatically between distal corpus (15%) and proximal cauda (75%). It was hypothesized that some rhesus sperm within the distal corpus (second portion of the middle segment) would have acquired fertilizing ability.
INTRODUCTION
Growth
and
of the androgens
differentiation
rhesus during
1972a)
and
monkey both
adult
period
transit
(Dinakar
mean
rhesus days
epididymal segments for the caput, corpus (Amann
sperm
acquire
fertilizing
involves
factors
extrinsic 1975;
al.,
is
region
where
sperm
Therefore, currently
of
most
intrinsic
to
within
the
first
acquire only
controlled breeding to determine the
based on means
epididymides
in various
‘Paper Minnesota
been
other
studies
laboratory reported
ability. data
available
to on the
and
domestic
(Nicander,
1957,
June 29, 1978. January 24, 1978. No. 10,339 Agricultural
of the Experiment
the
and
Journal Series of the Station.
1063
studies
(Ramos
in the
and
Dym,
to
relate,
undertaken
The
specific
objectives
the
histometrically
where
phosphatase
positive rhesus
type
spermatozoa location
MATERIALS
epididymis
fertilizing
within
degree the
was
determine
alkaline
(PAS)
and
Parallel rare
the regional epididymal epitheactivity to that of post-testicular
to
Schiff
in
Bedford, et al.,
monkey
study
describe
epididymis;
as
are
maturation.
to
tase,
well
and Nicander, 1971; 1975; Orgebin-Crist
unlikely that will be performed
were:
species as
Our
spermatozoal
the and 5.6 respec-
maturation
sperm
1975).
primate
where possible, hal and luminal
epididymal
This
Hamilton,
1977b).
1974).
mammalian
this occurs. and histochemical
Accepted Received
al.,
through
During
capacity.
hypotheses are the
have
et
are 1.1, 3.8 and cauda,
predict where Histological animals
epididymis
sperm
1976).
of
(Bedford Hamilton,
1976). It experiments exact
times
et
transit,
of the
are dependent upon the prenatal (Alexander,
The
tively
1958; subhuman
and
substances
epididymis;
of potential from of
acid
each their
AND
periodic
rhesus
phosphaacid-
are localized to determine the motility
epididymal cytoplasmic
inherent region droplets.
METHODS
Six adult macaque monkeys (Macaca mulatta) were castrated and the excurrent ducts dissected from the testes. Sexual normality was established by normal testicular histology and histochernistry and by the presence of normal, motile sperm in the cauda epididymidis. Tissues representing testis and 7 regions (Fig. 1) from 1 epididymis of each monkey were frozen in isopentane and tissues from the contralateral epididymis were used to provide sperm samples. Cryostat sections were cut at 8 tom, air dried and stained with hematoxylin and eosin (Cooper, 1963) and for acid phosphatase (I3arka and Anderson, 1962) using 30 and 60 mm staining times, for alkaline phosphatase (Thompson, 1966) using 2 and 4 mm staining times and for periodic acid-Schiff (PAS) positive materials (Humason, 1967) using 30 and 60 mm staining times. Staining intensity was scored
1064
ALSUM
AND
HUNTER
from height to volume and the percent packed sperm cells calculated. To determine the location of the cytoplasmic droplet, fluid from 2 males was aspirated into 0.5 ml formal saline, placed on a slide and evaluated under phase contrast. Position of the cytoplasmic droplet on the midpiece was determined on 200 sperm per region. Positions were classified as being proximal, mid, distal or absent.
if firent
RESULTS
Histometric
via
The and
luminal
and
the
various in
corpus
of
rhesus
epididymal
regions.
1), caput (regions (regions 6-7).
2-3),
Table
ter. Measurements were made with a light microscope equipped with a calibrated eye piece micrometer such that 3 m differences could be resolved. Contents from the contralateral epididymides were collected by making an incision across each specific epididymal region and aspirating the accumulated fluid. Testicular fluid was collected similarly. For motility studies, fluid from 2 males was aspirated into 0.5 ml TES NaK yolk buffer (Graham et al., 1972) at room temperature, placed on a microscope slide, warmed to 37#{176}Cand evaluated microscopically by 3
For relative sperm concentration studies, fluid from 2 males was aspirated into a 400 MI Beckman teflon tube and centrifuged in a Beckman Model 152 Microfuge for 5 mm. Seminal columns and sperm columns were measured with sliding calipers, converted
I. Measurements
Epididymal
of rhesus
region
Ductuliefferentes,
excurrent
Lumin
77.1
al diameter
2, proximal
161.2
Caput Corpus
3, distal 4, proximal
145.7 218.7
5.5 8.5 6.4 9.6
5, distal
227.3 328.7
7.9 14.2
618.0
31.4
6, proximal 7, distal
Cauda Cauda
a Values *Not
are measured.
in microns
and
represent
the
efferentes
regions
are
Epididymal
was
fixation
cauda.
in the
Luminal
both
studies
stereocihial and tubular
than Both
narrower
(1977b). ter
enlarges
populations
between
the
from
close,
about were
of show
of
heights
relatively
those studies
report-
were smaller and Dym to different
epithehial
were
and was
were
and/or
lengths were diameters
and
caput
diameter
(Table 1) by Ramos partially due
Epididymal
summa-
epithehial
maximal
procedures
monkeys.
stereocihia
ductuli
but
3-fold about
Ramos and the luminal ductuli
our
shorter 1.5-fold Dym
diame-
efferentes
and proximal caput and then narrows in the distal caput. However, in the bonnet monkey (Macaca radiata), the height of the epididymal epithehium in
and
the
caput
(Flechon
et
Ramos ductuhi ily sional
lumen region
al.,
and
Dym
maximal
at
(1977a)
in monkeys
ciliated
basal
were
minimal
the
cauda
1976).
efferentes
of
diameter and
and cells,
nonciliated macrophages
the
reported
consisted
primar-
cells and
and
occa-
intraepithe-
ducts.a
Caput
Corpus
1
at
the
of
the
just the reverse. Identical findings ed by Ramos and Dym (1977b).
people.
TABLE
1.
Our measurements than those reported (1977b). This was
independently by 3 people, averaged for individual monkeys and means and standard errors were determined on a group basis. Five random transverse histological measurements per region were made for epithelial height, stereocilia length and luminal diame-
length
in
epididymal
height
minimal
(region cauda
height, diameter
stereocihial
1. Diagram efferentes (regions 4-5),
DISCUSSION
Observation epithehial
rized
FIG. Ductuli
AND
means
±
SEM
Epith dial
21.5 69.6 78.4 64.0
1.0 2.0 2.3 2.4
52.1 49.9 40.2
for 6 animals.
height
Stereocilia
height
S
18.4 16.2 7.5
1.4 1.2 0.7
2.0
10.3
0.6
2.2 2.3
7.5
0.6
4.0
0.5
MONKEY
hal
lymphocytes.
presence
of
basal
pseudostratified Vasectomy
causes to
diameter
the
enlarge
and
than
size
were
body
1972b).
region
7. These
ductuhi
four
We
in the
in
agree
that
with
no
unique
head
may
or
function
division
regions
was
such and
epididymis. initial
of
Regions
the
4
and
the
5,
characterized
by
supranuclear and
an
(Table
2).
Regions
ment,
were a
very
Acid
Phosphatase The
data
7,
similar
The 0.01 Moderate
segsperm.
Acid
testes
controls activity was area of epidid-
the
activity
rhesus
Riar
et
luminal
rhesus
acid
epididymides.
of acid
phosphatase (Hunter et and Risley,
Moniem
and
Glover,
monkey
most
closely
the
ram
(Moniem
and
Glover,
rat
(Nikkanen
and
phosphatases
of
are
and
the
Vanha-Perttula,
strong
acid
apically
in
(regions
3-5)
vation
epididymal
the
that
compounds
and
the
that
and
tract, corpus
corpus
activity
was but
primarily
regions.
was
in
suggested occurring The
spermatozoal
region
The
localized epithehia
luminal
epididymides, materials
luminal
in that
under 1963).
activity,
weak
rhesus
caput
increased
in obser-
density
in agreement
with
concept. Acid
caput
the
absorption
(deDuve,
phosphatase the distal caput and
enzymes
with
of
conditions
rhesus monkey absorption of
throughout the distal
lysosomal
associated
breakdown acidic
the that
are
usually
slightly
increased distal
of
The
devoid
through
comparison
Epididymal region
2.
incuba-
practically
phosphatase
Regional
Fig.
both
in the supranuclear in all regions of were
from
in for
M NaF-treated to strong
2).
rhesus
and
Acid
epithehium with
testes
from
minimal
1952;
Montagna,
which
terminal
packed
and
acid
1977).
a wider
lower
efferentes
varies with species 1969; Martan
The
enzyme
detectable (Fig.
in
Martan,
1972b). 1972b)
concentration
the
1972;
resembled
were
epithehium,
by
was
Stereocihia
2.
stereocihia. stereocihia,
sperm
al.,
the
decreased
distribution
epididymis
1963; by
segment,
summarized
localized principally the columnar cells
TABLE
the
that
found
regional
in the
showing
epididymidis
also
cauda
biochemical
ductuhi
activity
The
Activity are
periods. negative.
activity.
the
of
lumen
of activity
gradually
in
6 and
wide
portion
regular
characterized
and
pattern
middle
area
major
long
more
vacuoles
lumen
of
characterized
extremely
with
(1973)
with
barely
activity caput
corpus
to distal
caput . region with little 1972). The luminal contents
tubules,
phosphatase
ductuli
segment
3, the
were
al.
into
was
1
initial
2 and
Nicander,
tissue
exhibited the
proximal
conflict
the
seminiferous
phosphatase
to
epididymis
Region
segment,
epithelium
and
the
that
part
according
(Glover of
Regions
the
divided
be
in
gradually
rhesus
epididymides through
Our
efferentes
on
strong
results
was mainly in beyond (Riaretal., of
other
epididymal
region
studies
times
characteristics
present
1971).
ymis.
as
(1977b)
epididymis
morphology
tion were
columnar.
regions.
The
high
3, remained 4 and decreased
the
rhesus
much
histological
region
simple
adult
as
Dym
distinguishing
the
1065
EPIDIDYMIS
epithehium
confirmed
of
(Alexander,
Ramos
of
results making
instead
efferentes
and
Our cells
DUCTUS
phosphatase
catabolism
sperma
of
may
substances
concentration,
Droplets
Droplets
Droplets
Spermatocrit
proximal
distal
absent
10b
92 85 87 2 1 0 0
be
involved
absorbed
anatomy
and
in
from
motility
the
the
patterns.
Motility
(%) Ductuli Caput Caput Corpus Corpus
Cauda Cauda
efferentes, 2, proximal 3, distal 4, proximal 5, distal 6, proximal 7, distal
aSamples
were
of everything on bRete testis-
cNO
tested.
1
.
49 69 69 61 73 obtained
from
6 monkeys.
each monkey. Therefore, Ductuli efferentes.
0 6 5 89 92 78 51 However, samples
from
8 9
7 9 7 22 49
None 1% circular + 5% stationary 10% progressive + 5-25% stationary 5-15% full progressive + 60% stationary 5-15% full progressive + 60% stationary 70-75% full progressive + 10% stationary 75% full progressive
epididymal
sample
2 monkeys
were
volumes used
for
were each
insufficient determination.
for
analysis
1066
ALSUM
4
Acmd
AND
HUNTER
contain
Pase
this
A
of
C3
droplet
I
this
enzyme
suspending (Dott
the
ram,
associated
T Yl
and
Dingle,
found
that
levels
I
2
VE
3
4
Caput
Alk.
5
6
Corpus
7
VD
Pose
apical
dyl in
Cauda
-
A C3
lumen
transit
through
largest
change
took
corpus,
activity
..‘‘#{149}.4...
....
T
vi
was
(Riar
Alkaline
Phosphatase
The
0 yE
2
3
5
4
Coput
6
Corpus
7
VD
The
testes,
was
C
caput of
in
secretory
I
vi
The
0 T
VE
3
2
4
Coput
5
6
Corpus
7
VD
ductuli
Caudo
the
luminal
than
the
from
ductuli 5
through
cauda.
Our
and
lumen
(Moniem
Nicander,
1965,
1966)
testicular
origin.
was
activity mass
indicating
that
epididymal this
Slight
observed of
in
the
rhesus
the
enzymes
sperm. might
1972b).
and
such
be However,
acid the
were the
artifact cytoplasmic
1972b;
proteins
lumen
spermato-
epithelium.
also
present was
(Moniem droplets
droplet not
could
and
sperm plasma.
of
rabbit
not
They
high
the
on al.,
and
ram
levels
rhesus In
activity in
sperm.
addi-
in the
Glover
in the it
were
whole
than
hypothesized
regional
1972).
more
demonstrate
despite
corpus constant
on
phosphatase and
increased distal
phosphatase
et
Moniem
alkaline
and
essentially
(Riar
epididymis
The
activity
activity
data
the
of
epitheli-
activity.
enzyme
data
illustrated
caput
efferentes
alkaline
data
a
alkaline
columnar
remained
tissue
our
localized
epididymidis
tissue
tion,
for
through
biochemical
epididymal
of
phosphatase
luminal caput
Since an
Glover,
as
to
proximal
Luminal
of
similar
and negative
more
histochemical
of the
cells
attributed
ductuli
efferentes
region
and (1970)
basal
slight
much
tissues.
basal
corpus
cells.
apical
of
had
greatest Bavdek
the
showed
contents
distribution
epididymal
the
regions and efferentes
He
virtually
corpus
epididymis FIG. 2. Acid and alkaline phosphatase activity and periodic acid-Schiff staining activity in apical portions of rhesus monkey excurrent duct epithelium and luminal content. Mean ± SEM. Activity: 0 = no reaction, I = slight, 2 = moderate, 3 = strong, 4 = intense; apical, is.. = lumen.
in
All
media activity
the
4-6).
these
while
of
The in
efferentes
were
phosphatase, urn
cells.
activity
to
2.
epididymides
epididymidis.
activity
epithelia
rhesus
Fig.
phosphatase
(regions
caput
and
the
deficient
substrate
detected
strong
rabbit
Glover,
that
in
basal
cauda
observed
fixed,
proximal
efferentes,
was
proximal
on
the than
alkaline
the
in
activity
is
phosphatase
summarized
ductuli
strong
cell
zoal
activity
acid
and ductus deferens varied between within tissues. Activity in the ductuli
4
caput
Activity
are
negative. the
The the
1973).
incubated
were in
Cauda
et al.,
data
controls
in
lipid
epididymis.
phosphatase
higher
monkey
total
progressively
between
luminal
strongest
significantly
total
phosphatiand
the
the
be
(1975)
protein,
decreased
acid
man,
al.
et
acid,
place
where
In
to
maturation
choline,
sperm
of
ceases its
sphingomyelin
monkey
the
extent
Arora
sialic
of release
a lesser
during
1968). of
during
was
T
sperm
ethanolamine,
high.
to
phosphatidyl
rhesus
and
4
and
and
phosphatase
the
(Dott
1968)
bull
acid
phospholipid,
0
the
with
disruption
Dingle,
In
of osmolarity
alteration
causes
and
enzyme.
in
and
fluid
the
lining (1972a)
cytoplasmic
However,
in
hamster
in
the
they or
epididymal that
alkaline
rat
MONKEY
phosphatase cell by
played a role dephosphorylation
phosphate and
groups
the
in the mis.
sperm
epididymal
and
would
thus
following
of sperm phosphate
compounds
are
in
the
distal
early
maturation
The more
phospholipid,
dyl
lower
1975).
Also,
luminal cauda.
decreased with
alkaline
phosphatase
between
distal
change
in the
droplet
the
and
position
from
seems tered
in
caput
proximal
disrupted
during
reactions
are
Acid-Schiff data
difference periods. incomplete significant
to
strong
the
apical
tes the
ductuli
efferentes
rat
and
(1955)
in the
epididymis.
which
He
was
Stereocilia
most and
from moderate (region 3) to (region
6).
The
PAS was
out
the
the duct,
basal were The
tubules
reaction
of the in
the
epididymis,
lamina strongly luminal through
Apical in the (region
3).
the
the
contents corpus
from
2).
bonnet Sperm
1975).
efferentes
location
and
of
the
midpiece,
and
droplet
midpiece.
cyto-
occurred
proximal
changed
corpus
from
The
appreciable
proximal
droplet
numbers
testes
Microscopic
had
been
of cauda
sperm.
during
transit
that
testis and
the
increased
to corpus
epididymis
observations
histological suggested
stallions
was
being
caput
Sperm to
of
slides verified most of the as
(Nicander,
corpus
caput.
concentration
rete
1965).
cycle
(region Apical the
epididymal region.
collagenous
surrounding PAS positive.
the
the
this fluid
finding. arising in
the
reported
in
bulls,
1958)
and
was
high
2).
epididymal
absorbed
density
distal
(Table
This from efferent
rams,
boars
(Crabo,
from
proximal
cauda.
diminished
cauda
basal
for
atocrit
ducts
PAS
distal
was observed. deferens was
from
Sperm
efferen-
distal caput epididymidis in the proximal cauda distal
in the caput
The
on the
distal
Sperm
in the slight the
were
(Table
in
a secretory the
sperm
ductuli
on the sperm
distal 2).
shed
prevalent
activity
rhesus
epididymis
reported and Hafez,
change
droplet
between to
PAS
In
faintest
results.
only slightly intensities. of moderate
throughout
in
the
testes,
tail forward
Droplet
A rapid
were
apical
increased activity hum of the ductus cells
epithelia
active
rete
showing finally,
6 (75%).
region
(Table
caput
et al., Bower,
proximal caput were not progressively motile while about 10% of the distal caput sperm were
materials
considerable
proposed
the
plasmic
ductuli
distal
increases
(Hunter and
sperm and
data were (Flechon
from
Little
epididymis. was strongest
noted
material
2.
our
testis,
the
Similar monkey
of circular
transported
2 incubation diffusion nor
were
Similar
boar (Malone
PAS vary
progressively motile. The greatest change in progresstve motility occurred between distal corpus region 5 (15%) and proximal cauda
the epididymis. In general, moderately to strongly PAS
throughout PAS activity
reactive
PAS
the
positive epithelial Maneely
It
lumen.
Fig.
the
intensity
and throughout stereocilia were
2).
percentage
Cytoplasmic
in
influencing
of
a
stronger
0.1% maltdiastase the tissue PAS staining PAS positive materials epithelia
of
the
in
staining
the
Activity
of
factors
Incubation reduced Diffuse
as
along
is sequesand as they in
between neither
reactivity
increased
was
maturation,
summarized
was seen Therefore,
motility,
to
(Table
observed
(PAS)
are
then
cytoplasmic
distal
the
of not
Motility.
The
contents
phosphatase droplets
positive
beating,
corpus
sperm
to
become
The
luminal
of the
point.
et al., in the
increase
proximal
likely that alkaline into the cytoplasmic
Periodic
the
that
reported in the hamster
Sperm
caput
the
beyond been and
amounts which did
1962).
phosphati-
from
Concomitant
have 1972)
increasing (Fig. 2)
total
regions: choline,
acquired materials
from
sphingomyelin (Arora levels of phospholipid
the
fluid
sperm
epididymal
phosphatidyl
ethanolamine,
involved
in the epididycontaining
rhesus
1067
markedly
plasma be
EPIDIDYMIS
gradually positive
in the function of the and transport of
between cell
DUCTUS
the
least
Acquisition
of Fertilizing
Regions
7),
tes
and
1-3
constitute
initial
region
epithe-
Fertile
sperm
have
active.
there
in every
species
columnar
the
Through-
location
fibers
of
epididymal
of
seminiferous
epididymal
region
poor
sperm
of
Capacity
the
the of
the
not
been
motility
sperm
to
have
originate
In addition,
and
the
droplet not
efferenepididymis.
found
cytoplasmic
from those regions are fertility. Region 4 (proximal
studied,
ductules rhesus
so far studied.
the sperm concentrated segment of the epididymis. 4,
Hypothesis
proximal
on
portion of the In the species to
sperm
compatible with corpus) constituted
pass
through
middle so far such
a
1068
ALSUM
zone
prior
to
acquiring
greatest
potential
developed
between
and
proximal
that
upon
cauda
sperm
ability
and
(region
(regions
probably
is gradually
the
corpus
(region
5)
6). It is hypothesized
distal
corpus
distal
6,
part
7).
5),
of
region
4
capacity
by
rhesus
and
in
N. J. (1972a).
ductus
epididymis
effects
of
119-133. Alexander,
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R. Pickett,
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