BIOLOG\
OF
12, 393-395
REPRODUCTION
Phospholipids
(1975)
of Goat
Spermatozoa
Y.
C.
JAIN
National
Dairy
Phospholipids
of goat
dimensional
thin
percent
Research
spermatozoa
choline,
sphingomyelin.
0.9
2.8
lysophosphatidyl
percent
choline.
glycerol
(cardiolipin)
plasma
phospholipids
and
percent
4.3
percent percent
were
18.3,
plasma
(choline
1.8 percent The
4.1,
phosphatidyl
5.5.
by
two
of
25.0
5.0
phasmalogen), 8.0
percent
9.6,
and
4.4
per-
percent
diphosphatidyl
values 2.0
percent 11.8
inositol.
corresponding
3.5.
quantified comprised
plasmalogen).
ethanolamine. acid.
15.0,
and were
(ethanolamine
lysophosphatidyl 3.0,
separated
phospholipids choline
serine,
9.1.
Plasma
India
were
ethanolamine
phosphatidic
19.1.
Seminal
5, 1974
phosphatidal
phosphatidyl
0.5
Karnal,
spermatozoal
phosphatidal
the
ANAND
Institute,
seminal
The
27.8
ethanolamine,
R.
November
and
chromatography.
phosphatidyl
phosphatidyl cent
layer
S.
AND
Accepted
and
for
the
seminal
0.7
percent,
respec-
the
thin
layer
was
exposed
tively.
The
phospholipid
of a number and man Hartree,
composition
of
semen
at..
have 1962;
been Pursel
studied (Masaki and Graham,
The
is available
present
conducted
investigation
Semen
was
and
16000
Lipids
collected
g
and
the
for
30
were of
stored
ammonia
seminal
mm
two
silica
and
gel
al.,
by
1969).
paring
using
(1957)
and
percent.
of
the
and
use
of
run
Entenman,
of
under
by
on 0.5 mm
in the
first
second were
spots
with
identical Different
spots
chromatogram
with
by charring
at
estimated by the method (1969) and the plasmalogens
of
aration-reaction-separation
technique
reagents were
by of in
com-
180#{176}C.Phosphorus Ahovcova and were quantitated (Viswanathan
visible
rights
plasmalocalculated
±
content seminal
layer
phospholopid lipid was plasma
of
seminal plasma of spermatozoa distribution
based
is shown
spermatozoa Aldehydogenic
percent
of
the
as
tide
et
and
15.3
percent
on
in Table
well lipids
phospholipids
of
57.5 41.7
chromatographic
I. The
in Fig.
of the plasma
0.1 g/l00 g of wet 0.003 g/lOO ml of
average
matozoa and 22.1 percent lipids of seminal plasma. 52.7 percent of the total
was Odavic by sep-
of Reproduction.
±
phospholipids, content,
hr
by 25.
which
28.7
H2SO4
393 All
the
that
of the plasma.
addition
(Skidmore made
The
is shown
one
developed separated
DISCUSSION
to be 1.2 and 0.057
thin
for
was
value
AND
HCI with
content
phatidal choline, phosphatidyl sphingomyelin were the major
authentic
50 percent
1975 by The Society for the Study of reproduction in any form reserved.
Copyright ©
the phosphorus
in the to
represented
phospholipid
The
phorus
(Rouser
identified
conditions spray
(60:
direction those
analogues
The
dried and then lyso forms
pattern of phospholipids was very similar to different
direction
acid-water
the
semi-specific 1962).
the
followed
in
Phospholipids
mobilities
by spraying (w/w)
vol.)
achieved
chloroform-methanol-25
by vol.)
was
of the spermatozoal and that of seminal
an atmosphere
chromatography
(65:25:5,
plate of nitrogen The 2-acyl
plate
neutralization
plasma.
content percent
was
di-acyl
its
seminal pro-
with
to
Sloane-Stanley
phospholipids
layer
G plates
the
compounds to
content.
was found spermatozoa
into
fractions
chloroform-acetone-methanol-acetic
70:20:20:10. et
their
gen
the
two
10#{176}C under
of thin
from
temperature.
according
The
by
The total phospholipid goat spermatozoa and the
by centrifugation
the
v/v)
and
at
NH4OH
percent
from
Lees
Separation
dimensional
thick
from
track
followed
RESULTS
fractionated
refrigerated
mm
by multiplying
semen.
vagina
and
4
lipid
vapours.
therefore,
artificial
plasma
at
(2:1.
Folch,
an
developing the
under the atmosphere in the second direction.
METHODS
pooled
extracted
in chloroform
nitrogen.
by
bucks,
chloroform-methanol cedure
was,
AND
Alpine
spermatozoa at
goat
for
to fill this gap.
MATERIALS Beetal
about
After
direction,
fumes
and 1967;
Johnson et a!., 1969: Poulos el a!., 1972, 1973: Scott et a!., 1967; Quinn and White, 1967: Poulos and White, 1973) but no such information
1968).
first
of species, viz,, bull, boar, ram
the
of phos-
I. Phos-
choline and phospholipids as seminal constituted of
sper-
of the phosphoIn spermatozoa choline phosphathe total ethanol-
394
JAIN
AND
ANAND
amine
phosphatide
logen whereas corresponding
LPI!.
amine
plasmalogens
percent
and
6
resembles 2
(2)
C-A-N
I. Two
pattern
of goat
technique origin
as
PA,
phosphatidyl
erol;
EP,
malogen:
X,
Separation
serine:
P1,
PC,
ethanolamine
inositol;
lipids
choline;
PE,
diphosphatidyl
plasmalogen:
less polar
man
lysophos-
phosphatidyl
CP.
plas-
(Poulos
TABLE OF
GOAT
plasand re-
that
of the plasma
of
the
ram
and White, 1967), 1962: Pursel and and Anand,
than that of boar (JohnPoulos et a!., 1972) or
and
in
White,
boar and phosphatidyl
ethanolamine
triglyceride
etc.).
COMPOSITION
closely
data) 1969:
1973). As
in the
SPERMATOZOA
human semen ethanolamine
plasmalogens
comparatively
PHO5PHOLIPID
96.8
total
composition the seminal
and
1967) or buffalo (Jam
whereas gomyelin,
glyc-
choline
(cholesterol,
the
goat, the choline plasmalogens are the major phospholipids of spermatozoa and the seminal plasma of the ram, bull and buffalo
ethanolamine;
phosphatidyl DPG,
0,
uncharacterized LPC,
constituted of
1967: Quinn and Hartree,
unpublished son et a!.,
Abbreviations:
acid:
ethanolamine; LPL,
text.
lysophosphatidyl
sphingomyelin;
phosphatidyl
the
phosphatidic LPE.
Graham,
chromatographic
phospholipids. in
compounds):
choline:
SPH,
layer
more
(Scott et a!., bull (Masaki
(eo:7o-.20:20:IO)
thin
spermatozoa
(non-migrating
phatidyl
1120
described
components; PS,
-NM-
dimensional
and percent
The phospholipid goat spermatozoa
PS,
FIG.
86.4
spectively.
“I
LPE .,,-.-l.PC
_
the 24.8
malogen content of the spermatozoa the seminal plasma phospholipids,
N
P1”
as plasma-
percent, respectively. Choline plasmalogens were more predominant than ethanol-
t ci
present
were
in the seminal plasma values were 51.1 and
larger
are
sphinand
present
amounts.
The
in phos-
I
AND
THE
SEMINAL
PLASMA
(As
PERCENT
OF
THE
TOTAL
PHOSPHORUS
Spermat
Component Phosphatidylcholine(PC)
Phosphatidal
choline
(Choline
plasmalogen)
(CP) Phosphatidyl Phosphatidal
ethanolamine ethanolamine
plasmalogen) Sphingomyelin
‘
(PE) (Ethanolamine
(EP) (SPH)
Phosphatidyl
serine
Phosphatidyl
inositol(PI)
Lysophosphatidyl
(PS) choline
(LPC)
Lysophosphatidylethanolamine(LPE) Diphosphatidyl
glycerol
Phosphatidic Non-migrating Values each
time.
(Cardiolipin)
(DPG)
acid (PA) ±
are based
Uncharacterized(0.X) on six analyses
of spermatozoal
lipid
%
ozoa
Seminal
p lasma
%
Range
Average
21.6-35.3
25.0
15.2-22.0
18.3
25.3-30.3
27.8
15.7-22.4
19.1
Range
Average
1.9-9.0
5.0
1.1-15.6
9.1
0.4-2.6
0.9
1.6-4.3
3.0
9.8-15.3
11.8
0.8-4.0
2.8
10.6-
1.1-5.9
19.8
15.0 4.1
0.6-2.7
1.8
1.5-6.0
3.5
1.4-8.7
4.4
3.2-9.8
5.5
1.9-7.0
4.3
4.3-14.2
9.6
1.6-12.7
8.0
0.3-5.0
2.0
0.0-1.1
0.5
0.2-1.4
0.7
5.2-10.5
7.7
5.6-16.4
and
five of seminal
plasma
pooled
from
at least
10.1 30 ejaculates
phatidyl
serine,
GOAT
SPERM
constitutes
The
phospholipids
Aust.
Biochem.
normally
than ten percent of the human and seminal plasma phospholipids, is pres-
more boar ent
which
in
much
pholipids
smaller
of the
amounts
bull,
in the
buffalo,
ram
and
A.,
POULOS.
(1973) passage
through
goat.
Biophys.
Acta
nal The
authors
Sundaresan, cil
of
are
Agricultural
lowship
grateful
to
the
for encouragement
and
Research
to one of them
for
Director, to the
Dr.
Indian
providing
D.
Coun-
UNDP
fel-
(YCJ).
J.
method
for
separated matog. (1957)
the by
cation
of
total
Chem.
226, L.
JOHNSON,
seminal
frequency
chem. POULOS,
A
simple
of phospholipids
chromatography.
J.
M.
AND
method
for
lipids
the
from
A.,
isolation
animal
R.
GERRITS,
analysis
plasma AND
and
purifi-
J.
tissues.
E.
Biol.
F.
of
bull
E.
spermatozoa
cold
shock.
phorus
Fert.
DARIN.
P.
by the
19, 95.
(1962)
Distribution
and
hyaluronidase
spermatozoa.
C.
AND
WHITE,
I. G.
F.
phosphoand
(1967)
and
semi-
Phospho-
seminal
I.
and
seminal
J. Biol.
of polar
G.
(1967)
plasma.
Phospholipid
of epididymal
lipids
phosphorus
and
plasma
Sci.
20,
in
ejaculated relation
S.
AND
YAMAMOTO,
layer
chromatographic
and
determination
analysis
to
1205.
thin
FLEISCHER.
T.
SCOTT,
W.,
ticular
VOGLMAYR,
Lipid and
J.
composition
ejaculated
SKIDMORE,
W.
dimensional phosphatides.
of
A. of phos-
spots.
K.
AND
and
Lipids
5.
ram
B.
SETCHELI..
metabolism
in
spermatozoa.
tes-
Biochem.
W.
0.
(1972)
D.
(1968)
plasmalogens.
AND
C.
ENTENMAN,
thin layer chromatography J. Lipid Res. 3, 471. C.
VISWANATHAN.
Bio-
chromatography A.
E.
WHITF.
dimensional
by
The
14, 203.
Aust.
G..
(1973)
35, 265.
GRAHAM,
content
Two
Biochim.
spermatozoa
spermatozoa
AND
ram
of the bull.
J. 102. 456.
J. 84. 347. A.,
J.
P.
during
494.
spermatozoa
as affected
phospholipid tails
YOUNG,
porcine
J. Reprod.
HARTREE.
and
J. AND of
phospholipids
activity, heads
P.
cholesterol
(1969)
H.
AND
and
P. (1967)
of ejaculation. J.
G.
SLOANE-STANLEY,
Fert.
Fert.
ROUSER.
Chro-
497.
of metabolic between
analysis
layer
Quantitative
MASAKI,
(1969)
J. Reprod.
J. Reprod.
B.
SETCHELL,
spermatozoa
I. G. human
G.
AND
in tract
of
bovine
Proc.
194.
WHITE,
of
separation
LEES, simple
and
R.
ODAVIC.
quantitative thin
A
(1969)
AND
V.
PURSEL,
K.
the genital 306,
AND
plasma.
QUINN,
40, 90. J.,
FOLCH,
K.
J. changes
composition
lipids
REFERENCES AHOVCOVA.
A.
lipid
spermatozoa.
5, 28.
VOGLMAYR.
phos-
ENTS
of mammalian Soc.
Phospholipid
PouLos.
ACKNOWLEDGM
395
PHOSPHOLIPIDS:
V.,
Two
PHILLIPS,
F.
dimensional in
the
J. Chromatog.
of AND
reaction
analysis 35, 66.
(1962)
of
rat
Two liver
LUNDBERG,
thin
layer
phosphatide
OF
12, 393-395
REPRODUCTION
Phospholipids
(1975)
of Goat
Spermatozoa
Y.
C.
JAIN
National
Dairy
Phospholipids
of goat
dimensional
thin
percent
Research
spermatozoa
choline,
sphingomyelin.
0.9
2.8
lysophosphatidyl
percent
choline.
glycerol
(cardiolipin)
plasma
phospholipids
and
percent
4.3
percent percent
were
18.3,
plasma
(choline
1.8 percent The
4.1,
phosphatidyl
5.5.
by
two
of
25.0
5.0
phasmalogen), 8.0
percent
9.6,
and
4.4
per-
percent
diphosphatidyl
values 2.0
percent 11.8
inositol.
corresponding
3.5.
quantified comprised
plasmalogen).
ethanolamine. acid.
15.0,
and were
(ethanolamine
lysophosphatidyl 3.0,
separated
phospholipids choline
serine,
9.1.
Plasma
India
were
ethanolamine
phosphatidic
19.1.
Seminal
5, 1974
phosphatidal
phosphatidyl
0.5
Karnal,
spermatozoal
phosphatidal
the
ANAND
Institute,
seminal
The
27.8
ethanolamine,
R.
November
and
chromatography.
phosphatidyl
phosphatidyl cent
layer
S.
AND
Accepted
and
for
the
seminal
0.7
percent,
respec-
the
thin
layer
was
exposed
tively.
The
phospholipid
of a number and man Hartree,
composition
of
semen
at..
have 1962;
been Pursel
studied (Masaki and Graham,
The
is available
present
conducted
investigation
Semen
was
and
16000
Lipids
collected
g
and
the
for
30
were of
stored
ammonia
seminal
mm
two
silica
and
gel
al.,
by
1969).
paring
using
(1957)
and
percent.
of
the
and
use
of
run
Entenman,
of
under
by
on 0.5 mm
in the
first
second were
spots
with
identical Different
spots
chromatogram
with
by charring
at
estimated by the method (1969) and the plasmalogens
of
aration-reaction-separation
technique
reagents were
by of in
com-
180#{176}C.Phosphorus Ahovcova and were quantitated (Viswanathan
visible
rights
plasmalocalculated
±
content seminal
layer
phospholopid lipid was plasma
of
seminal plasma of spermatozoa distribution
based
is shown
spermatozoa Aldehydogenic
percent
of
the
as
tide
et
and
15.3
percent
on
in Table
well lipids
phospholipids
of
57.5 41.7
chromatographic
I. The
in Fig.
of the plasma
0.1 g/l00 g of wet 0.003 g/lOO ml of
average
matozoa and 22.1 percent lipids of seminal plasma. 52.7 percent of the total
was Odavic by sep-
of Reproduction.
±
phospholipids, content,
hr
by 25.
which
28.7
H2SO4
393 All
the
that
of the plasma.
addition
(Skidmore made
The
is shown
one
developed separated
DISCUSSION
to be 1.2 and 0.057
thin
for
was
value
AND
HCI with
content
phatidal choline, phosphatidyl sphingomyelin were the major
authentic
50 percent
1975 by The Society for the Study of reproduction in any form reserved.
Copyright ©
the phosphorus
in the to
represented
phospholipid
The
phorus
(Rouser
identified
conditions spray
(60:
direction those
analogues
The
dried and then lyso forms
pattern of phospholipids was very similar to different
direction
acid-water
the
semi-specific 1962).
the
followed
in
Phospholipids
mobilities
by spraying (w/w)
vol.)
achieved
chloroform-methanol-25
by vol.)
was
of the spermatozoal and that of seminal
an atmosphere
chromatography
(65:25:5,
plate of nitrogen The 2-acyl
plate
neutralization
plasma.
content percent
was
di-acyl
its
seminal pro-
with
to
Sloane-Stanley
phospholipids
layer
G plates
the
compounds to
content.
was found spermatozoa
into
fractions
chloroform-acetone-methanol-acetic
70:20:20:10. et
their
gen
the
two
10#{176}C under
of thin
from
temperature.
according
The
by
The total phospholipid goat spermatozoa and the
by centrifugation
the
v/v)
and
at
NH4OH
percent
from
Lees
Separation
dimensional
thick
from
track
followed
RESULTS
fractionated
refrigerated
mm
by multiplying
semen.
vagina
and
4
lipid
vapours.
therefore,
artificial
plasma
at
(2:1.
Folch,
an
developing the
under the atmosphere in the second direction.
METHODS
pooled
extracted
in chloroform
nitrogen.
by
bucks,
chloroform-methanol cedure
was,
AND
Alpine
spermatozoa at
goat
for
to fill this gap.
MATERIALS Beetal
about
After
direction,
fumes
and 1967;
Johnson et a!., 1969: Poulos el a!., 1972, 1973: Scott et a!., 1967; Quinn and White, 1967: Poulos and White, 1973) but no such information
1968).
first
of species, viz,, bull, boar, ram
the
of phos-
I. Phos-
choline and phospholipids as seminal constituted of
sper-
of the phosphoIn spermatozoa choline phosphathe total ethanol-
394
JAIN
AND
ANAND
amine
phosphatide
logen whereas corresponding
LPI!.
amine
plasmalogens
percent
and
6
resembles 2
(2)
C-A-N
I. Two
pattern
of goat
technique origin
as
PA,
phosphatidyl
erol;
EP,
malogen:
X,
Separation
serine:
P1,
PC,
ethanolamine
inositol;
lipids
choline;
PE,
diphosphatidyl
plasmalogen:
less polar
man
lysophos-
phosphatidyl
CP.
plas-
(Poulos
TABLE OF
GOAT
plasand re-
that
of the plasma
of
the
ram
and White, 1967), 1962: Pursel and and Anand,
than that of boar (JohnPoulos et a!., 1972) or
and
in
White,
boar and phosphatidyl
ethanolamine
triglyceride
etc.).
COMPOSITION
closely
data) 1969:
1973). As
in the
SPERMATOZOA
human semen ethanolamine
plasmalogens
comparatively
PHO5PHOLIPID
96.8
total
composition the seminal
and
1967) or buffalo (Jam
whereas gomyelin,
glyc-
choline
(cholesterol,
the
goat, the choline plasmalogens are the major phospholipids of spermatozoa and the seminal plasma of the ram, bull and buffalo
ethanolamine;
phosphatidyl DPG,
0,
uncharacterized LPC,
constituted of
1967: Quinn and Hartree,
unpublished son et a!.,
Abbreviations:
acid:
ethanolamine; LPL,
text.
lysophosphatidyl
sphingomyelin;
phosphatidyl
the
phosphatidic LPE.
Graham,
chromatographic
phospholipids. in
compounds):
choline:
SPH,
layer
more
(Scott et a!., bull (Masaki
(eo:7o-.20:20:IO)
thin
spermatozoa
(non-migrating
phatidyl
1120
described
components; PS,
-NM-
dimensional
and percent
The phospholipid goat spermatozoa
PS,
FIG.
86.4
spectively.
“I
LPE .,,-.-l.PC
_
the 24.8
malogen content of the spermatozoa the seminal plasma phospholipids,
N
P1”
as plasma-
percent, respectively. Choline plasmalogens were more predominant than ethanol-
t ci
present
were
in the seminal plasma values were 51.1 and
larger
are
sphinand
present
amounts.
The
in phos-
I
AND
THE
SEMINAL
PLASMA
(As
PERCENT
OF
THE
TOTAL
PHOSPHORUS
Spermat
Component Phosphatidylcholine(PC)
Phosphatidal
choline
(Choline
plasmalogen)
(CP) Phosphatidyl Phosphatidal
ethanolamine ethanolamine
plasmalogen) Sphingomyelin
‘
(PE) (Ethanolamine
(EP) (SPH)
Phosphatidyl
serine
Phosphatidyl
inositol(PI)
Lysophosphatidyl
(PS) choline
(LPC)
Lysophosphatidylethanolamine(LPE) Diphosphatidyl
glycerol
Phosphatidic Non-migrating Values each
time.
(Cardiolipin)
(DPG)
acid (PA) ±
are based
Uncharacterized(0.X) on six analyses
of spermatozoal
lipid
%
ozoa
Seminal
p lasma
%
Range
Average
21.6-35.3
25.0
15.2-22.0
18.3
25.3-30.3
27.8
15.7-22.4
19.1
Range
Average
1.9-9.0
5.0
1.1-15.6
9.1
0.4-2.6
0.9
1.6-4.3
3.0
9.8-15.3
11.8
0.8-4.0
2.8
10.6-
1.1-5.9
19.8
15.0 4.1
0.6-2.7
1.8
1.5-6.0
3.5
1.4-8.7
4.4
3.2-9.8
5.5
1.9-7.0
4.3
4.3-14.2
9.6
1.6-12.7
8.0
0.3-5.0
2.0
0.0-1.1
0.5
0.2-1.4
0.7
5.2-10.5
7.7
5.6-16.4
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