BIOCHEMICAL
Vol. 83, No. 3, 1978
AND BIOPHYSICAL RESEARCH COMMUNICATIONS Pages
August14,1978
GLUTATHIONE S-TRANSFERASE
ACTIVITIES
1093-1098
IN RAT AND MOUSE SPERM AND HUMAN SEMEN
Insu P. Lee, and John R. Bend
Hasan Mukhtar,
Health Laboratory of Pharmacology, National Institute of Environmental Park, NC 27709 Sciences, P. 0. Box 12233, Research Triangle
Received
June 26,197s
SUMMARY: Glutathione S-transferase activity was found in sperm of the rat and DBA/ZJ and C57 BL/bJ mi-de. In rat sperm activities with benzo(a)pyrene 4,5-oxide, styrene 7,8-oxide, and 1-chloro-2,4-dinitrobenzene were 0.88, 1.07, and 26.1 nmoles/min/mg protein, respectively. A5-3-Ketosteroid isomerase activity of rat sperm was 4.9 nmoles/min/mg protein. These specific glutathione S-transferase and A5-3-ketosteroid isomerase activities in sperm represent 0:4-4.1% of rat liver cytosol values. Human semen also contained significant glutathione z-transferase activity. It is postulated that these enzymes could function in the metabolism and detoxification of certain electrophilic xenobiotics, if present in sperm. INTRODUCTION: The glutathione important
role
Enzymic
reactions
trophiles widely
involving
distributed
(l-5)
species
Glutathione
organisms, including
ferase
B (9,
lo),
thione
transferases, metabolites,
including hormones,
present
ligandin,
bind
dyes,
with
many organic
and most
isomerase glutathione (11).
activity
The gluta-
anions
noncovalently,
of
S-trans-
including and covalently
Abbreviations used: SO, styrene 7,8-oxide, 4,5-BPO, benzo(a)pyrene, CDNB, l-chloro-2,4-dinitrobenzene; HEPES, N-2-hydroxypiperazine-N'-2-ethane sulfonic acid; PBS (0.01 M sodium phosphate, 0.145 M sodium chloride, CTAB, cetyltrimethylammonium bromide.
1093
is
and serum (8)
as ligandin
and drugs
an
of elec-
activity
(7),
As-3-ketosteroid
identified
play
a variety
in hepatic
testes
associated
was previously
with
z-transferase
(61,
Recently,
was shown to be principally
bilirubin,
being ovary
of rat liver
which
enzymes,
of many xenobiotics.
of glutathione
(l-5).
studied.
of cytosolic
and detoxification
conjugation
in living
tissues
mammalian
a family
in the biotransformation
have been described
extrahepatic all
S-transferases,
4,5-oxide; pH 7.2);
0006-291X/78/0833-1093$01.00/0 Copyright 0 1978 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol. 83, No. 3, 1978
BIOCHEMICAL
bind
and/or
several
S-transferases
carcinogens
hydrocarbons,
epoxides
has recently
biotransformation
in hepatic
aromatic been
metabolites.
To our
amines,
of alkene
knowledge
have not
been previously
glutathione
S-transferase
of carcinogenic
reviewed
and extrahepatic
MATERIALS
their
in the detoxification
aromatic
the
AND BIOPHYSICAL
(5,
12).
and arene
tissues
enzymes
and other
reported activities
oxides
in the
in sperm. in rat
The role aminoazo
of the
glutathione
dyes,
polycyclic
electrophiles
Our laboratory
of mammalian
involved
RESEARCH COMMUNICATIONS
including is
by glutathione and aquatic
in
S-transferases species.
biotransformation Here we describe
interested
of xenobiotics the presence
of
and mouse sperm and human semen.
AND METHODS:
Vasa deferentia were dissected from male Sprague-Dawley rats (300-350 g body weight), OBA/ZJ, and C57 BL/6J mice (30-40 g body weight). Sperms in the vas deferens of rats were gently flushed out by washing with ice-cold PBS with a syringe fitted with a 23-gauge needle. For collection of sperm from mice, vas deferens were squeezed gently with the help of a fine forceps. The collected sperms were washed twice with PBS by centrifugation at 1OOOxg for 15 min at 4*C. To remove the small amount of red blood cell contamination, the sperm preparation was subjected to hypotonic shock for 20 set, using distilled water. The osmolarity of the sperm suspension was immediately readjusted to 300 mOsm using 0.45 M NaCl. The sperm preparation was then repeatedly washed by resuspension (three times) and sedimentation at 1500x9 for 15 min to remove cellular contaminants. Finally the sperms were resuspended in PBS containing 0.1% CTAB and sonicated for 15 set with a Branson 20 KHz sonicator equipped with a microprobe (setting of 40). The sonicated sperm suspensions were used as the enzyme source. Identical volumes of 0.1% CTAB in PBS were added to blank assay tubes and to tubes used to determine nonenzymatic reaction rates. Samples of human semen were obtained from normal healthy volunteers (29-38 years of age). Glutathione S-transferase activities with SO, 4,5-BPO, or CDNB as substrate were assayed according to procedures described earlier (6, 8). As-3-Ketosteroid isomerase activity was determined spectrophotometrically (9). The cuvette (in a fina? volume of 3.0 ml) contained: 24 mM Tris base, 12.5 mM K HP0 sufficient H PO to attain a pH of 8.5, 100 pM GSH, 68 pM A5-androstene3:17-%;one (in 20 p130f4methanol) and enzyme source (200 pl) which was used to initiate the reaction. The absorbance of the sample cuvette was recorded at 25'C in a UV vis Gilford spectrophotometer at 248 nm. One unit of enzyme is defined as the amount causing the isomeration of 1 nmol of As-androstene-3,17dione per minute I.? A4:Tndrostene-3,17-dione. The activities were calculated from EM = 16000 M cm . All reaction rates reported have been corrected for the nonenzymatic contribution. All chemicals used were of the highest purity available commercially were obtained from the sources described previously (8). As-Androstene-3,17dione (m.p. 155-157O) was prepared by Dr. J. H. Maguire according to the cedure of Jones and Gordon (13).
and pro-
RESULTS AND DISCUSSION: Specific
rat
sperm
glutathione
$-transferase
1094
activities
with
4,5-BPO,
a K-
BIOCHEMICAL
Vol. 83, No. 3, 1978
region
arene
oxide,
nmoles/min/mg aryl
protein,
halide
sperm
and with
of specific ratios
S-transferase
hepatic
in rat
sperm may differ
and extrahepatic
11) earlier
demonstrated
ferases
in rat
liver
partially
purified
was shown
in our
S-transferases activity
4.1% of the
BL/6J
sperm were
slightly
activities
were
four 47-112
mice
activity
ketosteroid the mature
retained
CDNB than
(1,
S-trans-
Similarly, liver
with
and lung, for
all
it
glutathione isomerase
protein,
with 2.
those
which
4,5-BP0
repre-
observed
in rat
sperm,
in Table
In the
although Gluta-
3.
activity
nmoles/min/ml
benzo(a)pyrene
in mouse
BP0 as substrate.
S-transferase
as substrates.
and CDNB in C57
The activities
and mice with
and 1.0-3.2
P-450,
associates
1).
had glutathione
cytochrome
those
A5-3-ketosteroid
in human semen is given
ejaculate
from
glutathione
nmoles/min/mg
in Table
in rats
in these
Semen from
ranging
ejaculate,
respec-
sperm preparations
hydroxylase,
from
we were
or epoxide
activity.
The origin
the
similar
and his
rabbit
activities
given
with
CDNB and 4,5-BP0
to detect
hydrase
sperm are
individuals
with
unable
S-transferase
These
and 1.4 percent
specificities. from
(Table
1).
quantitatively
different
The specific
used
S-transferases
SO was not a substrate
was.
activity
higher very
that
sperm was 2.46
hepatic
nmoles/min/ml
tively,
from
4,5-BP0 rat
S-transferase healthy
S-transferases (14)
glutathione
and DBA/W
thione
substrate
laboratory
whereas
Specific
overlapping
glutathione
in sonicated
sents
of seven
0.4,
glutathione
by Jakoby
the presence
and 1.07
The differences
and/or
Studies
0.88
(Table
1.9,
rat.
(total)
qualitatively
tissues.
with
protein
in the
the various
were
the more commonly
represent
activities
since
RESEARCH COMMUNICATIONS
oxide,
with
nmoles/min/mg
activities
cytosolic
of hepatic
alkene
Activity
CONB was 26.1
may be relevant
present
SO, a mutagenic
respectively.
substrate
glutathione
AND BIOPHYSICAL
and functional
isomerase spermatozoa, reproductive during
role
activity
in sperm
obtained tract
spermatogenic
of glutathione
from
after cell
cell
are
the
uncertain.
vas deferens,
differentiation
differentiation.
1095
S-transferase It
and As-3-
is
not
acquired or if
these
The presence
known whether these
enzymes
enzymes were of gluta-
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Vol. 83, No. 3, 1978
TABLE 1
SPECIFIC GLUTATHIONE z-TRANSFERASE AND A5-KETOSTEROID ISOMERASE ACTIVITIES IN RAT SPERM
Substrate
nmoles
Benzo(a)pyrene Styrene
product/min/mg
4,5-oxide
0.88
f 0.27
(45.6)
1.07
+ 0.32
(275)
26.05
+ 2.54
(1832)
2.46
+ 0.48
(58.9)
7,8-oxide
trobenzene
I-chloro-2,4-dini A5-androstene-3,17-dione
Data
represent
values
are
mean f S.D.
given
of three
in parentheses
determinations.
for
protein
Corresponding
hepatic
comparison.
TABLE 2
SPECIFIC GLUTATHIONE z-TRANSFERASE
AND DBA/W MICE
FROM C57 BL/6J
nmoles product/min/mg 4,5-BP0
Mouse Strain
0.97,0.71
74,52
DBA/25
0.92,0.64
58,39
values
S-transferase
detoxification It cluding hydantoin
protein CDNB
C57 BL/6J
Individual thione
ACTIVITIES IN SPERM
is
represent
activity
methadone (21,
22)
metabolites
to note (18, are
19>,
pooled
from
in human semen suggest
of electrophilic interesting
sperm
that
excreted
(20),
in high
that
it
of environmental
relatively
thalidomide
25 mice.
large
and phenytoin
concentrations
1096
amounts
in the
may assist
in the
chemicals. of some drugs (5,5-diphenylsemen of rabbit
in-
BIOCHEMICAL
Vol. 83, No. 3, 1978
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
TABLE 3 SPECIFIC GLUTATHIONE z-TRANSFERASE ACTIVITIES IN HUMAN SEMEN WITH BENZO(a)PYRENE 4,5-OXIDE (4,5-BPD) OR l-CHLORO-2,4-DINITROBENZENE (CDNB) AS SUBSTRATE
Subject
Semen Volume ml
Age (Years)
nmoEes product/mi Per ml CDNB
4,5-BP0
n Per ejaculate 4,5-BP0
CDNB
A
29
4.4
1.0
47
4.4
207
B
32
2.9
1.3
64
3.8
186
C
36
1.8
3.2
112
5.8
202
D
38
5.3
1.4
49
7.4
260
5.4 21.6
214 + 32
Mean f S.D.
and man.
Recently
1.7 + 1 .o
chlordecone
found
in sperm of dosed
solic
fraction
of rat i-transferase
cells
(7).
Failure
sperm
suggests
oxide
biotransformation Human seminal
peptidase is
activity
known to contain
nase,
malate
determine roid
(23).
germ cells than
to detect
epoxide
the
protective
major
in rodent
(15),
a key enzyme enzymes
a similar
in the present
(or
been
pathway
of these
with
phosphatase,
enzymes
However,
it
between
glutathione
S-transferase
isomerase
activities
and the
functionality
of sperm
these
have electrophilic
compounds
chemicals
known to be localized intermediary
1097
(2.5-fold)
in rat
alkene
or arene
glutathione. y-glutamyl Seminal lactate
in sperm
relationship
with
a cyto-
transfluid
dehydroge-
17).
the
been pretreated
for
synthesis.
(16,
investigation.
P-450)
shown to contain
such as acid
that
of interstitial
cytochrome
in glutathione
was also
higher
fraction
conjugation
and transaminases
significance
demonstrated
enzymatic
sperm is
has previously
pesticide,
had significantly
hydrase
fluid
other
a polychlorinated
We have earlier
activity
dehydrogenase,
The biological not assessed
rats
testicular
glutathione
that
(Kepone),
68.0 + 30.3
and human semen was
may be of interest and As-3-ketostein animals
in sperm,
metabolites.
to
that
have
particularly
if
Vol. 83, No. 3, 1978
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
REFERENCES 1. Z: 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. :54: 16. 17. 18. 19. 20. 21. 22. 23.
Habig, W. H., Pabst, M. J., and Jakoby, W. B. (1974) J. Biol. Chem., 249, 7130-7139. Mukhtar, H. and Bresnick, E. (1976) Biochem. Pharmac., 25, 1081-1084. James, M. O., Fouts, J. R., and Bend, J. R. (1976) Biochem. Pharmac., 25, 187-193. Hayakawa, T., Udenfriend, S., Yagi, H., and Jerina, D. M. (1975) Arch. Biochem. Biophys., 170, 438-451. Jerina, D. M. and Bend, J. R. (1977) Glutathione S-Transferases. In Biological Reactive Intermediates (Jollow, Cl., Kocsis, J. J., Snyder, R., and Vainio, H., eds.) pp. 207-236, Plenum Press, New York. Mukhtar, H., Philpot, R. M., and Bend, J. R. (1978) Biochem. Biophys. Res. Commun., 81, 89-98. Mukhtar, H., Lee, I. P., Foureman, G. L., and Bend, J. R., Chem.-Biol. Interact. In press. Mukhtar, H. and Bend, J. R. (1977) Life Sci., 21, 1277-1286. Benson, A. M. and Talalay, P. (1976) Biochem. Biophys. Res. Commun., 69, 1073-1079. Benson, A. M., Talalay, P., Keen, J. H., and Jakoby, W. H. (1977). Proc. Natl. Acad. Sci., U.S.A., 74, 158-162. Habig, W. H., Pabst, M. J., Fleischner, G., Gatmaitan, Z., Arias, I. M., and Jakoby, W. B. (1974) Proc. Natl. Acad. Sci. U.S.A., 71, 3879-3882. G. (1977) Cancer Res., 37, 8-14. Smith, G. H., Ohl, V. S., and Litwack, Jones, J. B. and Gordon, K. D. (1972) Can. J. Chem., 50, 2712-2718. Maguire, J. H., Fouts, J. R. and Bend, J. R. (1977) Fed. Proc., 36, 951. Krishnaswamy, P. R., Tate, S. S., and Meister, A. (1977) Life Sci., 20, 681-686. Cannon, D. C. (1974) In Clinical Diagnosis by Laboratory Methods, pp. 13001306, W. B. Saunders,Thiladelphia. Eliasson, R. (1973) -In Human Reproduction, pp. 39-51, Harper and Row, New York. Gerber,.N. and Lynn, R. K. (1976) Life Sci., 19, 787-792. Swanson, B. N. and Gerber, N. (1977) Proc. West. Pharmacol. Sot., 20, 477-482. H. (1967) Nature, 214, 1018-1020. Lutwak-Mann, C., Schmid, K. and Keberle, N. (1977) Proc. West. Pharmacol. Leger, R. M., Swanson, B. N., and Gerber, sot., 20, 69-72. Swanson, 6. N., Leger, R. M., Gordon, W. P., Lynn, R. K., and Gerber, N. (1978) Drug Metab. Dispos., 6, 70-74. Egle Jr., J. L., Fernandez, B. S., Guzelian, P. S., and Borzelleca, J. F. (1978) Drug Metab. Dispos., 6, 91-95.
1098
Vol. 83, No. 3, 1978
AND BIOPHYSICAL RESEARCH COMMUNICATIONS Pages
August14,1978
GLUTATHIONE S-TRANSFERASE
ACTIVITIES
1093-1098
IN RAT AND MOUSE SPERM AND HUMAN SEMEN
Insu P. Lee, and John R. Bend
Hasan Mukhtar,
Health Laboratory of Pharmacology, National Institute of Environmental Park, NC 27709 Sciences, P. 0. Box 12233, Research Triangle
Received
June 26,197s
SUMMARY: Glutathione S-transferase activity was found in sperm of the rat and DBA/ZJ and C57 BL/bJ mi-de. In rat sperm activities with benzo(a)pyrene 4,5-oxide, styrene 7,8-oxide, and 1-chloro-2,4-dinitrobenzene were 0.88, 1.07, and 26.1 nmoles/min/mg protein, respectively. A5-3-Ketosteroid isomerase activity of rat sperm was 4.9 nmoles/min/mg protein. These specific glutathione S-transferase and A5-3-ketosteroid isomerase activities in sperm represent 0:4-4.1% of rat liver cytosol values. Human semen also contained significant glutathione z-transferase activity. It is postulated that these enzymes could function in the metabolism and detoxification of certain electrophilic xenobiotics, if present in sperm. INTRODUCTION: The glutathione important
role
Enzymic
reactions
trophiles widely
involving
distributed
(l-5)
species
Glutathione
organisms, including
ferase
B (9,
lo),
thione
transferases, metabolites,
including hormones,
present
ligandin,
bind
dyes,
with
many organic
and most
isomerase glutathione (11).
activity
The gluta-
anions
noncovalently,
of
S-trans-
including and covalently
Abbreviations used: SO, styrene 7,8-oxide, 4,5-BPO, benzo(a)pyrene, CDNB, l-chloro-2,4-dinitrobenzene; HEPES, N-2-hydroxypiperazine-N'-2-ethane sulfonic acid; PBS (0.01 M sodium phosphate, 0.145 M sodium chloride, CTAB, cetyltrimethylammonium bromide.
1093
is
and serum (8)
as ligandin
and drugs
an
of elec-
activity
(7),
As-3-ketosteroid
identified
play
a variety
in hepatic
testes
associated
was previously
with
z-transferase
(61,
Recently,
was shown to be principally
bilirubin,
being ovary
of rat liver
which
enzymes,
of many xenobiotics.
of glutathione
(l-5).
studied.
of cytosolic
and detoxification
conjugation
in living
tissues
mammalian
a family
in the biotransformation
have been described
extrahepatic all
S-transferases,
4,5-oxide; pH 7.2);
0006-291X/78/0833-1093$01.00/0 Copyright 0 1978 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol. 83, No. 3, 1978
BIOCHEMICAL
bind
and/or
several
S-transferases
carcinogens
hydrocarbons,
epoxides
has recently
biotransformation
in hepatic
aromatic been
metabolites.
To our
amines,
of alkene
knowledge
have not
been previously
glutathione
S-transferase
of carcinogenic
reviewed
and extrahepatic
MATERIALS
their
in the detoxification
aromatic
the
AND BIOPHYSICAL
(5,
12).
and arene
tissues
enzymes
and other
reported activities
oxides
in the
in sperm. in rat
The role aminoazo
of the
glutathione
dyes,
polycyclic
electrophiles
Our laboratory
of mammalian
involved
RESEARCH COMMUNICATIONS
including is
by glutathione and aquatic
in
S-transferases species.
biotransformation Here we describe
interested
of xenobiotics the presence
of
and mouse sperm and human semen.
AND METHODS:
Vasa deferentia were dissected from male Sprague-Dawley rats (300-350 g body weight), OBA/ZJ, and C57 BL/6J mice (30-40 g body weight). Sperms in the vas deferens of rats were gently flushed out by washing with ice-cold PBS with a syringe fitted with a 23-gauge needle. For collection of sperm from mice, vas deferens were squeezed gently with the help of a fine forceps. The collected sperms were washed twice with PBS by centrifugation at 1OOOxg for 15 min at 4*C. To remove the small amount of red blood cell contamination, the sperm preparation was subjected to hypotonic shock for 20 set, using distilled water. The osmolarity of the sperm suspension was immediately readjusted to 300 mOsm using 0.45 M NaCl. The sperm preparation was then repeatedly washed by resuspension (three times) and sedimentation at 1500x9 for 15 min to remove cellular contaminants. Finally the sperms were resuspended in PBS containing 0.1% CTAB and sonicated for 15 set with a Branson 20 KHz sonicator equipped with a microprobe (setting of 40). The sonicated sperm suspensions were used as the enzyme source. Identical volumes of 0.1% CTAB in PBS were added to blank assay tubes and to tubes used to determine nonenzymatic reaction rates. Samples of human semen were obtained from normal healthy volunteers (29-38 years of age). Glutathione S-transferase activities with SO, 4,5-BPO, or CDNB as substrate were assayed according to procedures described earlier (6, 8). As-3-Ketosteroid isomerase activity was determined spectrophotometrically (9). The cuvette (in a fina? volume of 3.0 ml) contained: 24 mM Tris base, 12.5 mM K HP0 sufficient H PO to attain a pH of 8.5, 100 pM GSH, 68 pM A5-androstene3:17-%;one (in 20 p130f4methanol) and enzyme source (200 pl) which was used to initiate the reaction. The absorbance of the sample cuvette was recorded at 25'C in a UV vis Gilford spectrophotometer at 248 nm. One unit of enzyme is defined as the amount causing the isomeration of 1 nmol of As-androstene-3,17dione per minute I.? A4:Tndrostene-3,17-dione. The activities were calculated from EM = 16000 M cm . All reaction rates reported have been corrected for the nonenzymatic contribution. All chemicals used were of the highest purity available commercially were obtained from the sources described previously (8). As-Androstene-3,17dione (m.p. 155-157O) was prepared by Dr. J. H. Maguire according to the cedure of Jones and Gordon (13).
and pro-
RESULTS AND DISCUSSION: Specific
rat
sperm
glutathione
$-transferase
1094
activities
with
4,5-BPO,
a K-
BIOCHEMICAL
Vol. 83, No. 3, 1978
region
arene
oxide,
nmoles/min/mg aryl
protein,
halide
sperm
and with
of specific ratios
S-transferase
hepatic
in rat
sperm may differ
and extrahepatic
11) earlier
demonstrated
ferases
in rat
liver
partially
purified
was shown
in our
S-transferases activity
4.1% of the
BL/6J
sperm were
slightly
activities
were
four 47-112
mice
activity
ketosteroid the mature
retained
CDNB than
(1,
S-trans-
Similarly, liver
with
and lung, for
all
it
glutathione isomerase
protein,
with 2.
those
which
4,5-BP0
repre-
observed
in rat
sperm,
in Table
In the
although Gluta-
3.
activity
nmoles/min/ml
benzo(a)pyrene
in mouse
BP0 as substrate.
S-transferase
as substrates.
and CDNB in C57
The activities
and mice with
and 1.0-3.2
P-450,
associates
1).
had glutathione
cytochrome
those
A5-3-ketosteroid
in human semen is given
ejaculate
from
glutathione
nmoles/min/mg
in Table
in rats
in these
Semen from
ranging
ejaculate,
respec-
sperm preparations
hydroxylase,
from
we were
or epoxide
activity.
The origin
the
similar
and his
rabbit
activities
given
with
CDNB and 4,5-BP0
to detect
hydrase
sperm are
individuals
with
unable
S-transferase
These
and 1.4 percent
specificities. from
(Table
1).
quantitatively
different
The specific
used
S-transferases
SO was not a substrate
was.
activity
higher very
that
sperm was 2.46
hepatic
nmoles/min/ml
tively,
from
4,5-BP0 rat
S-transferase healthy
S-transferases (14)
glutathione
and DBA/W
thione
substrate
laboratory
whereas
Specific
overlapping
glutathione
in sonicated
sents
of seven
0.4,
glutathione
by Jakoby
the presence
and 1.07
The differences
and/or
Studies
0.88
(Table
1.9,
rat.
(total)
qualitatively
tissues.
with
protein
in the
the various
were
the more commonly
represent
activities
since
RESEARCH COMMUNICATIONS
oxide,
with
nmoles/min/mg
activities
cytosolic
of hepatic
alkene
Activity
CONB was 26.1
may be relevant
present
SO, a mutagenic
respectively.
substrate
glutathione
AND BIOPHYSICAL
and functional
isomerase spermatozoa, reproductive during
role
activity
in sperm
obtained tract
spermatogenic
of glutathione
from
after cell
cell
are
the
uncertain.
vas deferens,
differentiation
differentiation.
1095
S-transferase It
and As-3-
is
not
acquired or if
these
The presence
known whether these
enzymes
enzymes were of gluta-
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Vol. 83, No. 3, 1978
TABLE 1
SPECIFIC GLUTATHIONE z-TRANSFERASE AND A5-KETOSTEROID ISOMERASE ACTIVITIES IN RAT SPERM
Substrate
nmoles
Benzo(a)pyrene Styrene
product/min/mg
4,5-oxide
0.88
f 0.27
(45.6)
1.07
+ 0.32
(275)
26.05
+ 2.54
(1832)
2.46
+ 0.48
(58.9)
7,8-oxide
trobenzene
I-chloro-2,4-dini A5-androstene-3,17-dione
Data
represent
values
are
mean f S.D.
given
of three
in parentheses
determinations.
for
protein
Corresponding
hepatic
comparison.
TABLE 2
SPECIFIC GLUTATHIONE z-TRANSFERASE
AND DBA/W MICE
FROM C57 BL/6J
nmoles product/min/mg 4,5-BP0
Mouse Strain
0.97,0.71
74,52
DBA/25
0.92,0.64
58,39
values
S-transferase
detoxification It cluding hydantoin
protein CDNB
C57 BL/6J
Individual thione
ACTIVITIES IN SPERM
is
represent
activity
methadone (21,
22)
metabolites
to note (18, are
19>,
pooled
from
in human semen suggest
of electrophilic interesting
sperm
that
excreted
(20),
in high
that
it
of environmental
relatively
thalidomide
25 mice.
large
and phenytoin
concentrations
1096
amounts
in the
may assist
in the
chemicals. of some drugs (5,5-diphenylsemen of rabbit
in-
BIOCHEMICAL
Vol. 83, No. 3, 1978
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
TABLE 3 SPECIFIC GLUTATHIONE z-TRANSFERASE ACTIVITIES IN HUMAN SEMEN WITH BENZO(a)PYRENE 4,5-OXIDE (4,5-BPD) OR l-CHLORO-2,4-DINITROBENZENE (CDNB) AS SUBSTRATE
Subject
Semen Volume ml
Age (Years)
nmoEes product/mi Per ml CDNB
4,5-BP0
n Per ejaculate 4,5-BP0
CDNB
A
29
4.4
1.0
47
4.4
207
B
32
2.9
1.3
64
3.8
186
C
36
1.8
3.2
112
5.8
202
D
38
5.3
1.4
49
7.4
260
5.4 21.6
214 + 32
Mean f S.D.
and man.
Recently
1.7 + 1 .o
chlordecone
found
in sperm of dosed
solic
fraction
of rat i-transferase
cells
(7).
Failure
sperm
suggests
oxide
biotransformation Human seminal
peptidase is
activity
known to contain
nase,
malate
determine roid
(23).
germ cells than
to detect
epoxide
the
protective
major
in rodent
(15),
a key enzyme enzymes
a similar
in the present
(or
been
pathway
of these
with
phosphatase,
enzymes
However,
it
between
glutathione
S-transferase
isomerase
activities
and the
functionality
of sperm
these
have electrophilic
compounds
chemicals
known to be localized intermediary
1097
(2.5-fold)
in rat
alkene
or arene
glutathione. y-glutamyl Seminal lactate
in sperm
relationship
with
a cyto-
transfluid
dehydroge-
17).
the
been pretreated
for
synthesis.
(16,
investigation.
P-450)
shown to contain
such as acid
that
of interstitial
cytochrome
in glutathione
was also
higher
fraction
conjugation
and transaminases
significance
demonstrated
enzymatic
sperm is
has previously
pesticide,
had significantly
hydrase
fluid
other
a polychlorinated
We have earlier
activity
dehydrogenase,
The biological not assessed
rats
testicular
glutathione
that
(Kepone),
68.0 + 30.3
and human semen was
may be of interest and As-3-ketostein animals
in sperm,
metabolites.
to
that
have
particularly
if
Vol. 83, No. 3, 1978
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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