- Email: [email protected]
Effect of α-chlorohydrin on in situ PH in rat testis and epididymis
CONTRACEPTION
EFFECr
3F a-CHL3RCHY3HIN Carlton
ON -IV
SITU -__-
R. Cafli:;ch
Renal-Electrolyte Physiology Department of Internal Yedicine, Gal veston, Texas 77550
and
pE LY RXT TESTIS Thomas
Q.
l~bW3LOr‘jr, IJniversity
DuRos?,
AN3 EPI3IDYMIS Jr.
Div;sion of Texas
of Nephrglogy, Medi?71 Sranch,
ABST-tAACT l’qe effect seminiferous
of
the male coniraccptive, tubules and e2ididymd.l.
a-ch:orohydrin, on -in situ pH ii -duct of tne rat has been studied employing microelectrode techniques. 4fter eight days of low-dose -in vivo _(l%lg/Kg/day), a significant increase in cr-chlorohydrin administration acidity of luminal fluid in seminiferous tubules, proximal caput, middle caput , and proxi;nal cauda epididymidis WIs observed. Increased dcidi ty in t he testis and epididymis mdy play an important. role in t’le ant.ifertility effect of a-:hlorohyl!‘in.
INTRODUCT iON T7e actions propanediol)
of
tne have
male been
antifertility studied
agent, a-chlorohydrin extens:vely, especially
There are two knswn effects (1,2,3). tract of the m.-lle rat. The first, produces a characteristic lesion in occlusion epithel.ium
(3-chl.oro-1,2 in the rat
of
tnis compound on the reproductive elicited by ?I single high dose, the cdput cpididymidis le&Jing to an
of
the cpididymal duct, and subsequent deg,?neration of germinal which can result in prolonged or even permanent sterility The second effect, produced by low doses of ry-chlorohydrin, (4,5). 1 ndllces en,3yme .lctivity c’hanges in the of the testis and inhibits secretion of accessory sex organs (5) and rendtArs mitwe sperm in the ICiUsing ci uda opididymidis incapable of fertiLiz+tion, without morphological changes in t’he epididymis or in sperm (‘i,d,Yj. Recent studies acidification in arterial blood. JS
much
as
0.5
normal significant in the rat hdvr? demo?r+rated a. II seminiferous tubules and epididymal duct below that Furthermore, fluid pH along the epididymis can vary ptl
unit
(10).
T?e
pF!
profile
along
the
lnngth
of
of
by tne
cpididymis mdy be important for maturation, the development of sperm motility (11) and the maintenance of sperm quiescence (12). The functional significance of an acid milieu in seminiferous tubul.?s hds yet to be determined.
Submitted Accepted
for publication August for publication October
FEBRUARY
1990 VOL. 41 NO. 2
31, 1989
11,
1989
207
CONTRACEPTION The aim of the present study was to determine directly luminal fluid pH in seminiferous tubules and three sections of the rat epididymis before It was assumed and after administration of low doses of a-chlorohydrin. that any change in activity of Sertoli cells or epididymal cells or any leakage of constituents from spermatozoa might be reflected in the composition of the luminal fluid in which spermatozoa are suspended.
Adult male Sprague-Dawley rats 280-350 g body weight were obtained from Harlan Laboratories (Indianapolis, IN) and acclimatized for several days pri3r to experimental use. They were then divided at random into two groups: 8-day sham and 8-day treated. Eacn animal in the treated group (N=13) received a daily intraperitoneal injection of a-chlorohydrin (Aldrich Chemical Company, Milwaukee, WI) at a dose of 15mg/kg body weight in 0.1 ml of normal saline. Each animal in the sham group (N=‘7) received Animals were allowed free a daily injection of 0.1 ml of normal saline. access to food and water until the morning of the experiment. At the end of the treatment period, the rats were anesthetized with intraperitoneal Inactin (Byk Gulden, Konstanz, FRG) lOOmg/kg body weight snd placed on a thermostatically controlled table that maintained body Surgical preparation of animals for testis and temperature at 37%. epididymis micropuncture was accomplished as reported previously (10). Femoral artery pressure was monitored and :a maintenance solution of Ringers bicarbonate was administered at a rate of 1% body weight/hour. The scrotal sac was incised and the left or right testis and epididymis exposed, placed in a Lucite cup, stabilized with 3% agar and b;lthed continuously with paraffin oil (water equilibrated) maintained at 33-34°C. The tunica albuginea and epididymal capsule were slit to expose seminiferous tubules and segments of the proximal chput, middle caput, and proximal cauda epididymidis as described previously (10). All animals were maintained on a volume regulated rodent ventilator (Harvard Apparatus Inc., Willis, MA) and arterial blood gases monitored frequently (30-45 minutes). For both groups the inspired gas was 40% oxygen (balance nitrogen) . The -in situ pH of luminal fluid i? seminiferous tubules and epididymal -duct was determined directly with double br?eled glass membrane pH microelectrodes of 20-30 micrometer tip diameter exactly as reported previously (10). Construction and testing of double barreled electrodes followed the procedure of Pucacco and Carter (13). Each pH microelectrode was calibrated in standard phosphate buffer solutions of pii 6.00, 6.84 and 7.40 prior to and after --in situ use. Recalibration was performed after three to five pun-tures at each site. The pH sensitivity of double barreled electrodes ranged from 58-61 mV/pH uni-t at 33-34OC.. Electrodes having a sensitivity of less than 57 mV/pH 1ulj.t were not used. 411 calibrations and determinations for pE were carried out on a standard micropuncture table totally enclosed in a Faraday aage that was connected to earth ground.
208
FEBRUARY 1990 VOL. 41 NO. 2
CONTRACEPTION Results are expressed as mean values + SEM for each group. Analysis of variance and Student’s lrtll test were used for determination of the statistical significance of the differences between values. A p value of 0.05 or less was considered to constitute statistical significance.
RESULTS Group
I: _- sham
The rats used in these experiments (N=7) were in normal acid-base status. The mean arterial blood pH was 7.39tO.01, PCOz 38.8ko.4 mmHg and bicarbonate concentration 22.7tO.3 mM. is presenLed in theTable (Sham). A summary of the micropunzture findings The mean -in situ pH vallles in seminiferous tubu?as (ST), proximal zaput -(PCP), middle caput (MCP), and proximal cauda epididymidis (PCD) were the prevailing systemic arterial blood pk! significantly more acid than The pH values obtained in PCP and MCP were identical to each (p
EfPect of a-chlyohydrin TABLE: tubules and epididymal due',
on -i? situ pH in rat seminiferous
ST
PCP
MCP
Sham
%.97 kO.01
b4.60 to.01
b6.57 kO.02
‘6.84 k3.01
a-Chlorohydrin
“6.89” kO.01
bb. 52* kO.01
b6.bg*” +0.02
%. 74” +O.Dl
Val ues are meansGEM. Within rows, mean vai ues with significantly different (p
‘The -in situ pH vslues determined in -duct of sndm-treated animals in this determined in control animals of a The specific function of an (10). present Nun&?fined bl>t, may play an
FEBRUARY 1990 VOL. 41 NO. 2
no
letter different
PCD
in
common
from
control
(.a ,b ,c) values
are in
seminiferous tubules and epididymal study rlere almost identical to values previous study from this laboratory acid milieu in the cpididymis is at important, role in control of sperm
209
CONTRACEPTION cytosolic pH and thus development of sperm motility during epididymdl Acidic luminal fluid in the epididymis transit (11). may also be important for the maintenance of sperm quiescence during apididymal Tne function of an acid miliell in seminiferous tubules is storage (12). yet to be determined. The present studies provide the first evidence that a-chl,orohydrin, or a can significantly reduce luminal pH in metabolite of a-chlorohyd*~in, seminiferous tubules, proximal caput, middle caput, and proximal ,osuda rate knotin to induce epididymidis when administered at a low-dose sterility but not morphological changes in male rats (7). a-Chlorohydrin is known to rapidly enter the corpus and nauda npididymidis of the fat after low-dose intravenous administration (8). Tna drug also enters the testis and cauda epididymidis after intraperitoneal administration at a high dose (14). It is not yet fully understood how this agent enters the if a-chlorohydrin acts directly on the epididymal male reproductive tract, spermatozoa, or through its action on the function of the epididymal epithelium, affects sperm viability secondarily. Studies by Bdck et al. (15) have demonstrated an increase in the enzyme content of cauda epididymidis fluid suggesting a-chlorohydrin may have a direct action of other studies hdVf? shown thdt this lumi nal spermatozoa. Conversely, compound cdn significantly reduce sugar transport across rat caput epithelium (16) and sodium and water reabsorption in the cduda epididymidis (17). a-Chlorohydrin, in a dose similar to that used in the present study, has also been shown to induce changes in the biochemical composition of the testis and accessary reproductive organs in male rats The authors suggest that the observed changes could be due to the (6). antiandrogeniz nature of a-chlorohydrin. Preliminary studies from our laboratory have shown that administration of the nonsteroidal antiandrogen flutamide associated with significant (25mg/ kg/ day/ 15days ) was alkalinization of luminal fluid in caput and cauda epididymidis but not in seminiferous tubules of the rat (unpublished observations). These findings suggest that lurninal acidification in the cpididymis is under androgen control. The present results, therefore, are not compatible with an antiandrogenic effect of a-chlorohydrin. In conclusion, the present study has demonstrated for the first time that low-dose administration of a-chiorohydrin can reduce significantly in situ -pH in seminiferous tubules and epididymal duct of tne rat. This effect may be the result of action of a-chlorohydrin on the function of seminiferous tubule, proximal caput, middle cdput, and proximal %uda epididymidis duct epithelium and/or a direct effect on luminal spermatozoa causing leakage of cellular contents. Increased acidity of luminal fluid in the testis and epididymis may play an important role in the antifertility effect of u-chlorohydrin, therefore.
The authors acknowledge the excellent technical .assistance of Galen Bevel. The secretarial assistance of Gail Pollard is also appreciated. This research ROl HD 19988.
210
was supported
in part by National
Institutes
of Health grant
FEBRUARY 1990 VOL. 41 NO. 2
CONTRACEPTION
REFERENCES 1.
Jones AR. The antifertility Life Sci 1978;23:1625-1646.
2.
Jones AR. J Biol Sci
Antifertility 1983;36:333-350.
3.
Lob1 .JJ. antifertility Regulation
a-Chlorohydrin: Review of a model post-testicular agent. In: Cunninham CR, Schill WB, Hofez SF, eds. The Hague: Martinus Nyhof, 1980:109of male fertility.
actions
actions
of
of
a-chlorohydrin
a-chlorohydrin
in
in
the
the
male.
male.
Aust
122.
4.
5.
Hoffer AP, Hamilton tne fat epididymis Effects of a single Brown-Woodman epididymidis Contraception
DW,
after high
pathology of Fawcett DW. The ultrastructural administration of a-chlorohydrin (lJ-5897): dose. Anat Ret 1373;175:203-208.
PDC, White IG. and spermatozoa 1974;ll
of
Effect the
Hundril RS, Mangat HK. Effect composition of testis and accessory Indian J Exp Biol 1978;16:1278-1279.
7.
Samgjlik propanediol
8.
Crabo B, and rats.
9.
Vickery BH, Erickson GI, Bennett action of low doses of cc-chlorohydrin 1974;38:1-lo.
Chang (U-5897)
cr-chlorohydrin and general
of cc;uda physiology.
:69-7'7.
6.
E,
of rat
of a-chlorohydrin on biochemical reproductive organs on adult rats.
MC. Antifertility on male rats. BioI
Applegren LE. Distribution of J Reprod Fert 197?;30:161-163. JP.
attivity Reprod :14C]
of 3-chloro-1 1970;2:299-334.
a-chlorohydrin
Nechanism of in the male rat.
in
,2-
mice
antifertility J Reprod Fert
10.
Direct evaluation of acidification Caflisch CH, DuBose TD, Jr. Am J testis and epididymis. Role of carboni:: anhydrase. (Endocrinology and Metabolism ) 1989;in press.
11.
Vijayaraghavan S, Critchlow Evidence for a role for LM, Hoskins DD. cellular alkalinization in the cyclic adenosine 3’,5’-monophosphatemediated initiation of motility in bovine caput spermatozoa. Biol Reprod 1985;32:489-500.
12.
Carr DW, Ussclman MC, Acott TS. viscoelastic drag of sperm motility: Aeprod ;985;33:588-595.
13.
14.
Pucacco
B ioc’hein
Carter NW. 1976;30:427-434.
LR,
A glass
Effects of A species
membrane
by rat Physiol
pH, lactate, comparison.
pl! mi croelectrode
and Rio1
.
The entry of a-chlorohydrin Edwards FM, Jones AR, Waites GM. body fluids of male rats and the effect upon the incorporation 1975;43:225-232. glycerol into lipids. J Reprod Fert
FEBRUARY 1990 VOL. 41 NO. 2
Anal
into of
211
CONTRACEPTION
15.
Back DJ, Slaver TD, Shenton JC, Boyd GP. The cf fecLs of achlorohydrin on the composition of rat and rabbit epididymdl plasma: 4 possible explanation of species difference. J Heprod Flert 1975;45:117-128.
agents u16. Hinton BT, Hernandez H, Howards SS. The male antifertility chl orohydr i n , 5-thio-D-glucose, and 6-chloro-6-deoxy-D-glucose interference with sugar transport across the epithelium OF rat caput epi di dymi di s. J Androl 1983 ;‘I :216-221 . 17. Wong PYD, Yeung CH, Ngai F1K. Effect in perfused rat ca uda processes 1977;16:637-544.
212
of a-cholrohydrin epididymidis.
on transport Contraception
FEBRUARY 1990 VOL. 41 NO. 2
EFFECr
3F a-CHL3RCHY3HIN Carlton
ON -IV
SITU -__-
R. Cafli:;ch
Renal-Electrolyte Physiology Department of Internal Yedicine, Gal veston, Texas 77550
and
pE LY RXT TESTIS Thomas
Q.
l~bW3LOr‘jr, IJniversity
DuRos?,
AN3 EPI3IDYMIS Jr.
Div;sion of Texas
of Nephrglogy, Medi?71 Sranch,
ABST-tAACT l’qe effect seminiferous
of
the male coniraccptive, tubules and e2ididymd.l.
a-ch:orohydrin, on -in situ pH ii -duct of tne rat has been studied employing microelectrode techniques. 4fter eight days of low-dose -in vivo _(l%lg/Kg/day), a significant increase in cr-chlorohydrin administration acidity of luminal fluid in seminiferous tubules, proximal caput, middle caput , and proxi;nal cauda epididymidis WIs observed. Increased dcidi ty in t he testis and epididymis mdy play an important. role in t’le ant.ifertility effect of a-:hlorohyl!‘in.
INTRODUCT iON T7e actions propanediol)
of
tne have
male been
antifertility studied
agent, a-chlorohydrin extens:vely, especially
There are two knswn effects (1,2,3). tract of the m.-lle rat. The first, produces a characteristic lesion in occlusion epithel.ium
(3-chl.oro-1,2 in the rat
of
tnis compound on the reproductive elicited by ?I single high dose, the cdput cpididymidis le&Jing to an
of
the cpididymal duct, and subsequent deg,?neration of germinal which can result in prolonged or even permanent sterility The second effect, produced by low doses of ry-chlorohydrin, (4,5). 1 ndllces en,3yme .lctivity c’hanges in the of the testis and inhibits secretion of accessory sex organs (5) and rendtArs mitwe sperm in the ICiUsing ci uda opididymidis incapable of fertiLiz+tion, without morphological changes in t’he epididymis or in sperm (‘i,d,Yj. Recent studies acidification in arterial blood. JS
much
as
0.5
normal significant in the rat hdvr? demo?r+rated a. II seminiferous tubules and epididymal duct below that Furthermore, fluid pH along the epididymis can vary ptl
unit
(10).
T?e
pF!
profile
along
the
lnngth
of
of
by tne
cpididymis mdy be important for maturation, the development of sperm motility (11) and the maintenance of sperm quiescence (12). The functional significance of an acid milieu in seminiferous tubul.?s hds yet to be determined.
Submitted Accepted
for publication August for publication October
FEBRUARY
1990 VOL. 41 NO. 2
31, 1989
11,
1989
207
CONTRACEPTION The aim of the present study was to determine directly luminal fluid pH in seminiferous tubules and three sections of the rat epididymis before It was assumed and after administration of low doses of a-chlorohydrin. that any change in activity of Sertoli cells or epididymal cells or any leakage of constituents from spermatozoa might be reflected in the composition of the luminal fluid in which spermatozoa are suspended.
Adult male Sprague-Dawley rats 280-350 g body weight were obtained from Harlan Laboratories (Indianapolis, IN) and acclimatized for several days pri3r to experimental use. They were then divided at random into two groups: 8-day sham and 8-day treated. Eacn animal in the treated group (N=13) received a daily intraperitoneal injection of a-chlorohydrin (Aldrich Chemical Company, Milwaukee, WI) at a dose of 15mg/kg body weight in 0.1 ml of normal saline. Each animal in the sham group (N=‘7) received Animals were allowed free a daily injection of 0.1 ml of normal saline. access to food and water until the morning of the experiment. At the end of the treatment period, the rats were anesthetized with intraperitoneal Inactin (Byk Gulden, Konstanz, FRG) lOOmg/kg body weight snd placed on a thermostatically controlled table that maintained body Surgical preparation of animals for testis and temperature at 37%. epididymis micropuncture was accomplished as reported previously (10). Femoral artery pressure was monitored and :a maintenance solution of Ringers bicarbonate was administered at a rate of 1% body weight/hour. The scrotal sac was incised and the left or right testis and epididymis exposed, placed in a Lucite cup, stabilized with 3% agar and b;lthed continuously with paraffin oil (water equilibrated) maintained at 33-34°C. The tunica albuginea and epididymal capsule were slit to expose seminiferous tubules and segments of the proximal chput, middle caput, and proximal cauda epididymidis as described previously (10). All animals were maintained on a volume regulated rodent ventilator (Harvard Apparatus Inc., Willis, MA) and arterial blood gases monitored frequently (30-45 minutes). For both groups the inspired gas was 40% oxygen (balance nitrogen) . The -in situ pH of luminal fluid i? seminiferous tubules and epididymal -duct was determined directly with double br?eled glass membrane pH microelectrodes of 20-30 micrometer tip diameter exactly as reported previously (10). Construction and testing of double barreled electrodes followed the procedure of Pucacco and Carter (13). Each pH microelectrode was calibrated in standard phosphate buffer solutions of pii 6.00, 6.84 and 7.40 prior to and after --in situ use. Recalibration was performed after three to five pun-tures at each site. The pH sensitivity of double barreled electrodes ranged from 58-61 mV/pH uni-t at 33-34OC.. Electrodes having a sensitivity of less than 57 mV/pH 1ulj.t were not used. 411 calibrations and determinations for pE were carried out on a standard micropuncture table totally enclosed in a Faraday aage that was connected to earth ground.
208
FEBRUARY 1990 VOL. 41 NO. 2
CONTRACEPTION Results are expressed as mean values + SEM for each group. Analysis of variance and Student’s lrtll test were used for determination of the statistical significance of the differences between values. A p value of 0.05 or less was considered to constitute statistical significance.
RESULTS Group
I: _- sham
The rats used in these experiments (N=7) were in normal acid-base status. The mean arterial blood pH was 7.39tO.01, PCOz 38.8ko.4 mmHg and bicarbonate concentration 22.7tO.3 mM. is presenLed in theTable (Sham). A summary of the micropunzture findings The mean -in situ pH vallles in seminiferous tubu?as (ST), proximal zaput -(PCP), middle caput (MCP), and proximal cauda epididymidis (PCD) were the prevailing systemic arterial blood pk! significantly more acid than The pH values obtained in PCP and MCP were identical to each (p
EfPect of a-chlyohydrin TABLE: tubules and epididymal due',
on -i? situ pH in rat seminiferous
ST
PCP
MCP
Sham
%.97 kO.01
b4.60 to.01
b6.57 kO.02
‘6.84 k3.01
a-Chlorohydrin
“6.89” kO.01
bb. 52* kO.01
b6.bg*” +0.02
%. 74” +O.Dl
Val ues are meansGEM. Within rows, mean vai ues with significantly different (p
‘The -in situ pH vslues determined in -duct of sndm-treated animals in this determined in control animals of a The specific function of an (10). present Nun&?fined bl>t, may play an
FEBRUARY 1990 VOL. 41 NO. 2
no
letter different
PCD
in
common
from
control
(.a ,b ,c) values
are in
seminiferous tubules and epididymal study rlere almost identical to values previous study from this laboratory acid milieu in the cpididymis is at important, role in control of sperm
209
CONTRACEPTION cytosolic pH and thus development of sperm motility during epididymdl Acidic luminal fluid in the epididymis transit (11). may also be important for the maintenance of sperm quiescence during apididymal Tne function of an acid miliell in seminiferous tubules is storage (12). yet to be determined. The present studies provide the first evidence that a-chl,orohydrin, or a can significantly reduce luminal pH in metabolite of a-chlorohyd*~in, seminiferous tubules, proximal caput, middle caput, and proximal ,osuda rate knotin to induce epididymidis when administered at a low-dose sterility but not morphological changes in male rats (7). a-Chlorohydrin is known to rapidly enter the corpus and nauda npididymidis of the fat after low-dose intravenous administration (8). Tna drug also enters the testis and cauda epididymidis after intraperitoneal administration at a high dose (14). It is not yet fully understood how this agent enters the if a-chlorohydrin acts directly on the epididymal male reproductive tract, spermatozoa, or through its action on the function of the epididymal epithelium, affects sperm viability secondarily. Studies by Bdck et al. (15) have demonstrated an increase in the enzyme content of cauda epididymidis fluid suggesting a-chlorohydrin may have a direct action of other studies hdVf? shown thdt this lumi nal spermatozoa. Conversely, compound cdn significantly reduce sugar transport across rat caput epithelium (16) and sodium and water reabsorption in the cduda epididymidis (17). a-Chlorohydrin, in a dose similar to that used in the present study, has also been shown to induce changes in the biochemical composition of the testis and accessary reproductive organs in male rats The authors suggest that the observed changes could be due to the (6). antiandrogeniz nature of a-chlorohydrin. Preliminary studies from our laboratory have shown that administration of the nonsteroidal antiandrogen flutamide associated with significant (25mg/ kg/ day/ 15days ) was alkalinization of luminal fluid in caput and cauda epididymidis but not in seminiferous tubules of the rat (unpublished observations). These findings suggest that lurninal acidification in the cpididymis is under androgen control. The present results, therefore, are not compatible with an antiandrogenic effect of a-chlorohydrin. In conclusion, the present study has demonstrated for the first time that low-dose administration of a-chiorohydrin can reduce significantly in situ -pH in seminiferous tubules and epididymal duct of tne rat. This effect may be the result of action of a-chlorohydrin on the function of seminiferous tubule, proximal caput, middle cdput, and proximal %uda epididymidis duct epithelium and/or a direct effect on luminal spermatozoa causing leakage of cellular contents. Increased acidity of luminal fluid in the testis and epididymis may play an important role in the antifertility effect of u-chlorohydrin, therefore.
The authors acknowledge the excellent technical .assistance of Galen Bevel. The secretarial assistance of Gail Pollard is also appreciated. This research ROl HD 19988.
210
was supported
in part by National
Institutes
of Health grant
FEBRUARY 1990 VOL. 41 NO. 2
CONTRACEPTION
REFERENCES 1.
Jones AR. The antifertility Life Sci 1978;23:1625-1646.
2.
Jones AR. J Biol Sci
Antifertility 1983;36:333-350.
3.
Lob1 .JJ. antifertility Regulation
a-Chlorohydrin: Review of a model post-testicular agent. In: Cunninham CR, Schill WB, Hofez SF, eds. The Hague: Martinus Nyhof, 1980:109of male fertility.
actions
actions
of
of
a-chlorohydrin
a-chlorohydrin
in
in
the
the
male.
male.
Aust
122.
4.
5.
Hoffer AP, Hamilton tne fat epididymis Effects of a single Brown-Woodman epididymidis Contraception
DW,
after high
pathology of Fawcett DW. The ultrastructural administration of a-chlorohydrin (lJ-5897): dose. Anat Ret 1373;175:203-208.
PDC, White IG. and spermatozoa 1974;ll
of
Effect the
Hundril RS, Mangat HK. Effect composition of testis and accessory Indian J Exp Biol 1978;16:1278-1279.
7.
Samgjlik propanediol
8.
Crabo B, and rats.
9.
Vickery BH, Erickson GI, Bennett action of low doses of cc-chlorohydrin 1974;38:1-lo.
Chang (U-5897)
cr-chlorohydrin and general
of cc;uda physiology.
:69-7'7.
6.
E,
of rat
of a-chlorohydrin on biochemical reproductive organs on adult rats.
MC. Antifertility on male rats. BioI
Applegren LE. Distribution of J Reprod Fert 197?;30:161-163. JP.
attivity Reprod :14C]
of 3-chloro-1 1970;2:299-334.
a-chlorohydrin
Nechanism of in the male rat.
in
,2-
mice
antifertility J Reprod Fert
10.
Direct evaluation of acidification Caflisch CH, DuBose TD, Jr. Am J testis and epididymis. Role of carboni:: anhydrase. (Endocrinology and Metabolism ) 1989;in press.
11.
Vijayaraghavan S, Critchlow Evidence for a role for LM, Hoskins DD. cellular alkalinization in the cyclic adenosine 3’,5’-monophosphatemediated initiation of motility in bovine caput spermatozoa. Biol Reprod 1985;32:489-500.
12.
Carr DW, Ussclman MC, Acott TS. viscoelastic drag of sperm motility: Aeprod ;985;33:588-595.
13.
14.
Pucacco
B ioc’hein
Carter NW. 1976;30:427-434.
LR,
A glass
Effects of A species
membrane
by rat Physiol
pH, lactate, comparison.
pl! mi croelectrode
and Rio1
.
The entry of a-chlorohydrin Edwards FM, Jones AR, Waites GM. body fluids of male rats and the effect upon the incorporation 1975;43:225-232. glycerol into lipids. J Reprod Fert
FEBRUARY 1990 VOL. 41 NO. 2
Anal
into of
211
CONTRACEPTION
15.
Back DJ, Slaver TD, Shenton JC, Boyd GP. The cf fecLs of achlorohydrin on the composition of rat and rabbit epididymdl plasma: 4 possible explanation of species difference. J Heprod Flert 1975;45:117-128.
agents u16. Hinton BT, Hernandez H, Howards SS. The male antifertility chl orohydr i n , 5-thio-D-glucose, and 6-chloro-6-deoxy-D-glucose interference with sugar transport across the epithelium OF rat caput epi di dymi di s. J Androl 1983 ;‘I :216-221 . 17. Wong PYD, Yeung CH, Ngai F1K. Effect in perfused rat ca uda processes 1977;16:637-544.
212
of a-cholrohydrin epididymidis.
on transport Contraception
FEBRUARY 1990 VOL. 41 NO. 2