159

Investigation of the Effects of a1-Adrenoceptor Antagonism and

L-Type Calcium Channel Blockade on Ejaculation and Vas
Deferens and Seminal Vesicle Contractility In Vitro jsm_2410 159..168

Luiz Ricardo de Almeida Kiguti, MSc and André Sampaio Pupo, PhD
Universidade Estadual Paulista (UNESP), Department of Pharmacology, Botucatu-SP, Brazil

DOI: 10.1111/j.1743-6109.2011.02410.x

ABSTRACT

Introduction. Premature ejaculation is one of the most common male sexual dysfunctions. Current pharmacological
treatments involve reduction in penile sensitivity by local anesthetics or increase of ejaculatory threshold by selective
serotonin reuptake inhibitors. a1-Adrenoceptors (a1-ARs) and L-type calcium channels are expressed in the smooth
muscles of the male reproductive tract, and their activations play an important role in the physiological events
involved in the seminal emission phase of ejaculation.
Aim. To evaluate if the inhibition of the contractility of the vas deferens and seminal vesicle by a1-AR antagonism
or the L-type calcium channel blockade can delay ejaculation.
Methods. The effects of the a1-AR antagonist tamsulosin and of the L-type calcium channel blockers, nifedipine and
(S)-(+)-niguldipine, on contractions induced by norepinephrine in the rat vas deferens and seminal vesicles in vitro
and on the ejaculation latency of male rats in behavioral mating tests were evaluated.
Main Outcome Measure. Tension development of vas deferens and seminal vesicles in response to norepinephrine
in vitro and behavioral mating parameters were quantified.
Results. Tension development of vas deferens and seminal vesicle to a1-AR activation was significantly inhibited by
tamsulosin, nifedipine, and (S)-(+)-niguldipine. Tamsulosin displayed insurmountable antagonism of contractions
induced by norepinephrine in the rat vas deferens and seminal vesicle. Ejaculation latency of male rats was not
modified by tamsulosin, nifedipine, or (S)-(+)-niguldipine; however, both the number and weight of the seminal
plugs recovered from female rats mated with male rats treated with tamsulosin were significantly reduced.
Conclusion. Seminal emission impairment by inhibition of vas deferens or seminal vesicle contractility by L-type
calcium channel blockade or a1-AR antagonism is not able to delay the ejaculation. de Almeida Kiguti LR and
Pupo AS. Investigation of the effects of a1-adrenoceptor antagonism and L-type calcium channel blockade
on ejaculation and vas deferens and seminal vesicle contractility in vitro. J Sex Med 2012;9:159–168.
Key Words. Premature Ejaculation; Vas Deferens; Seminal Vesicle; Ejaculatory Latency; a1-Adrenoceptors; L-Type
Calcium Channels

Introduction about 1 minute of vaginal penetration and the

inability to delay ejaculation on all or nearly all

P remature ejaculation is one of the most

common male sexual dysfunctions affecting
5–40% of sexually active men [1–3]. According to
the Committee for Definition of Premature Ejacu-
lation of the International Society for Sexual Medi-
cine, lifelong premature ejaculation is a male sexual
dysfunction characterized by ejaculation that
always or nearly always occurs before or within
vaginal penetrations, accompanied by negative per-
sonal consequences, such as distress, bother, frus-
tration, and/or the avoidance of sexual intimacy [4].
The pharmacological treatment of premature
ejaculation targets the lengthening of the intravagi-
nal ejaculatory latency time, and the current
approaches involve the reduction in the penile sen-
sitivity by topic local anesthetic application or the

© 2011 International Society for Sexual Medicine J Sex Med 2012;9:159–168

160
modulation of ejaculatory threshold by selective
serotonin reuptake inhibitors (SSRIs) [5–8]. As a
result of the importance of serotonin in the control
of the ejaculation, the SSRIs present the best results
in the management of premature ejaculation [9,10].
Ejaculation is a complex reflex encompassing
sympathetic, parasympathetic, and somatic outputs
orchestrated by a spinal ejaculation generator at
lumbosacral spinal cord [11]. The viscero-somatic
events involved in ejaculation can be divided into
two consecutive phases, the seminal emission and
expulsion. During the seminal emission phase of
ejaculation, the sperm stored in the cauda epididy-
mis and secretions from sexual accessory glands are
propelled into the urethra by contractions of the
smooth muscles of cauda epididymis, vas deferens,
seminal vesicle, and prostate. Then, in the expul-
sion phase, the semen deposited in the prostatic
urethra is ejected through the urethral meatus as a
result of rhythmic contractions of striated muscles
of the pelvic floor partially triggered by the disten-
tion of the urethra [12].
The sympathetic nervous system plays a key
role in the seminal emission by releasing norepi-
nephrine, which controls the contractility of the
cauda epididymis, vas deferens, prostate, and
seminal vesicle through a1-adrenoceptor (a1-AR)
activation [13]. The importance of a1-ARs in the
seminal emission is demonstrated by the ejacula-
tion impairment resulting from knocking-out the
three genes encoding a1-ARs in mice and by ejacu-
latory dysfunctions observed during therapeutic
management of benign prostatic hyperplasia with
a1-AR antagonists [14,15]. In addition, the con-
tractions of vas deferens, seminal vesicle, and pros-
tate from different species to several different
pharmacological stimuli, including norepineph-
rine, are dependent on extracellular calcium influx
through L-type voltage-dependent calcium chan-
nels [16–19]. L-Type calcium channels are
members of voltage-dependent calcium channel
family, which are pharmacologically recognized by
their high sensitivity to blockade by a heteroge-
neous class of organic substances collectively
known as calcium channel blockers [20,21].
Because of the involvement of a1-ARs and
L-type calcium channels in the contractions of the
vas deferens and seminal vesicle in response to
norepinephrine and the importance of these con-
tractions in the physiological events that lead to
the emission phase of ejaculation, we hypothesized
that the pharmacological antagonism of a1-ARs
and L-type calcium channels would delay ejacula-
tion by reducing the contractions of these organs,
J Sex Med 2012;9:159–168
Kiguti and Pupo
representing a potentially new therapeutic strategy
for the treatment of premature ejaculation.
Aims
The aims of this study were to determine the effects
of tamsulosin, a selective a1-AR antagonist largely
used in the management of low urinary tract symp-
toms associated with prostatic hyperplasia, and of
the dihydropyridine-calcium channel blockers,
nifedipine and (S)-(+)-niguldipine, in the contrac-
tions of the rat vas deferens and seminal vesicle to
norepinephrine in vitro and on the ejaculation
latency of male rats in behavioral mating tests.
These drugs were particularly chosen to be inves-
tigated because it is known that tamsulosin presents
an insurmountable-like antagonism of vas deferens
contractions induced by norepinephrine in vitro,
whereas it is known that (S)-(+)-niguldipine also
presents high affinity for a1-ARs in addition to
interact with L-type calcium channels.
Materials and Methods
Animals
The experimental procedures were approved by
the local Ethics Committee for the Use of Experi-
mental Animals and are in accordance with the
Guide for the Care and Use of Laboratory
Animals (National Institutes of Health).
Adult male (90–150 days old, 380–450 g) and
female (90–120 days old, 200–220 g) Wistar rats
were used in this study. Animals were maintained
under controlled conditions (25°C, 30% humidity,
12/12-h light/dark cycle) with food and water
available ad libitum.
In Vitro Contraction Studies
Adult male rats were killed by decapitation, and
seminal vesicles or vasa deferentia were isolated,
cleaned of adherent tissues, and mounted in
10-mL organ baths to record isometric tension
development. Tissues were maintained under
9.8-mN resting tension in a nutrient solution with
the following composition (mM): NaCl, 138; KCl,
5.7; CaCl2, 1.8; NaH2PO4, 0.36; NaHCO3, 15;
and dextrose, 5.5, prepared in distilled water and
maintained at 30°C, pH 7.4, constantly bubbled
with 95% O2/5% CO2. This nutrient solution is a
modified Tyrode’s solution optimized to minimize
erratic “spontaneous” contractions and, along with
the organ bath temperature, improves the contrac-
tions of these two smooth muscles (Zuleika P.
Picarelli, personal communication).
a1-ARs, L-Type Calcium Channels, and Ejaculation
After a 30-minute stabilization period, the tissues
were repeatedly challenged with 80-mM KCl to
evaluate tissue viability and maximal response sta-
bilization. Then, a cumulative concentration–
response curve to norepinephrine was constructed
and taken as control curve. Then, tamsulosin (0.1–
100 nM), nifedipine (3.0–1,000 nM), or (S)-(+)-
niguldipine (3.0–1,000 nM) were added to the
organ baths and incubated with the tissues for 45
minutes when a new concentration–response curve
to norepinephrine was obtained in the presence of
these different drugs. To minimize the influence of
norepinephrine removal process and of activation
of receptors that result in relaxation of the smooth
muscle, all concentration–response curves to nore-
pinephrine were constructed in the presence of
cocaine (6 mM), corticosterone (10 mM), and pro-
pranolol (0.1 mM) to block of neuronal and extra-
neuronal norepinephrine uptake and b-ARs,
respectively [22].
Pharmacological Parameters Evaluated
The maximal tension developed (in mN) and the
potency of norepinephrine in inducing contrac-
tions (expressed as pEC50, the –log of concentra-
tion of norepinephrine inducing 50% of maximal
response) were derived by nonlinear curve fitting
with GraphPad Prism software (GraphPad Inc.,
San Diego, CA, USA). The inhibitory potencies of
nifedipine, (S)-(+)-niguldipine, and tamsulosin on
the maximal tension developed to norepinephrine
in the vas deferens and seminal vesicle were
expressed as pIC50 (the –log of concentration of
drug inducing 50% of its maximal inhibitory
effect).
Mating Behavior Analysis
Sexually experienced male rats were anesthetized
with ketamine + xylazine (70 mg/kg + 15 mg/kg,
i.p.), and the right jugular vein was catheterized
with polyethylene (PE) tubing (PE10 connected to
PE50) to assist drug administration. Catheters
were exteriorized and fixed in the rat dorsum with
cotton silk. A 3-day recovering period after
surgery was allowed before evaluation of mating
behavior.
Mating behavior analysis was performed 4 hours
after establishment of dark phase of the cycle under
red dim light illumination. Male rats received
vehicle (0.2 mL/rat, i.v.), tamsulosin (0.3, 1.0, or
3.0 mg/kg, i.v.), nifedipine (10, 50, or 100 mg/kg,
i.v.), or (S)-(+)-niguldipine (10, 50, or 100 mg/kg,
i.v.) and were placed individually in the observation
cages for habituation. After 15 minutes of habitua-
161
tion, a naturally receptive female was introduced
into the observation cage, and the copulatory
behavior was taped for 30 minutes after first mount
or intromission. The following parameters were
recorded: latency to the first mount or intromission
(time elapsed from presentation of receptive female
until first mount or intromission), latency to the
first intromission (time elapsed from presentation
of receptive female until first intromission),
number of intromissions until ejaculation (number
of mounts with pelvic thrusting), ejaculation late-
ncy (interval between first intromission until ejacu-
lation), and post-ejaculatory refractory time (PR,
time between ejaculation until the first mount or
intromission of next ejaculatory series). In addition,
the wet weights of the seminal plugs recovered at
the end of each copulatory analysis were recorded.
Statistical Analysis
Data are presented as mean ⫾ standard error of
mean or median and interquartile range. Statistical
comparisons were performed by Student’s t-test or
by analysis of variance followed by Dunnet a pos-
teriori for parametric data and by Kruskal-Wallis
followed by Dunn’s test for non-parametric data.
The values of P < 0.05 were considered significant.
Drugs and Solutions
Drugs were obtained from the following sources:
tamsulosin hydrochloride from IFFECT (Hong
Kong); (S)-(+)-niguldipine hydrochloride from
Tocris (Ellisville, MO, USA); nifedipine, norepi-
nephrine ([L]-[-]-norepinephrine bitartrate salt
monohydrate), corticosterone, and dimethyl sul-
phoxide (DMSO) from Sigma (St. Louis, MO,
USA); prazosin hydrochloride and propranolol
([+/-]-propranolol hydrochloride) from RBI
(Natick, MA, USA); and cocaine (Cocainum hydro-
chloricum puriss.) from Boehringer-Ingelheim
(Ingelheim, Germany).
Stock solutions for in vitro contraction studies
were prepared in distilled water (tamsulosin, pra-
zosin, propranolol, cocaine, and norepinephrine),
DMSO (100%) (corticosterone and [S]-[+]-
niguldipine), or ethanol (100%) (nifedipine).
Solutions administrated to male rats during
mating behavior analysis were prepared in 0.9%
NaCl (tamsulosin) or in 50% ethanol v/v in 0.9%
NaCl (nifedipine and [S]-[+]-niguldipine).
Results
In Vitro Contraction Studies
Figure 1A shows typical recordings of the contrac-
tions induced by cumulative addition of norepi-
J Sex Med 2012;9:159–168
162 Kiguti and Pupo

Figure 1 Contractions induced by norepinephrine in the rat vas deferens and seminal vesicle. (A) Representative trace
recordings showing tension development in the rat vas deferens and seminal vesicle in response to cumulative addition of
norepinephrine to the organ baths. Black dots represent the time points of norepinephrine addition and numbers under the
dots represent the respective norepinephrine concentrations. (B) Mean concentration–response curves to norepinephrine in
the vas deferens and seminal vesicle. Symbols represent mean and vertical bars, when larger than the symbols, represent
the standard error of mean for 17 and 18 different tissues.

nephrine to the rat vas deferens and seminal vesicle
in vitro, and Figure 1B shows the respective
concentration–response curves. Norepinephrine
was more potent in inducing contractions of the
vas deferens than of the seminal vesicle (pEC50
values of 6.90 ⫾ 0.05 N = 17 in the vas deferens vs.
6.05 ⫾ 0.06 N = 18 in the seminal vesicle;
P < 0.0001, Student’s t-test).
Increasing concentrations of the dihydropyri-
dine-calcium channel blockers, nifedipine and (S)-
(+)-niguldipine, reduced the contractions induced
by norepinephrine in the vas deferens (Figure 2A,
B) and seminal vesicle (Figure 2D, E). The con-
tractions of the vas deferens in response to nore-
pinephrine were nearly abolished by the maximal
concentrations of nifedipine and (S)-(+)-
niguldipine, whereas in the seminal vesicle,
approximately 30% of the contractions to norepi-
nephrine were resistant to the maximal concentra-
tions of (S)-(+)-niguldipine and nifedipine. In
addition, the inhibitory potency of (S)-(+)-
niguldipine in both tissues was significantly
greater than that of nifedipine (Figure 2C, F,
Table 1). Further, as evidenced by the rightward
displacements of the concentration–response
curves to norepinephrine, (S)-(+)-niguldipine, but
not nifedipine, reduced the potency of norepi-
nephrine in inducing contractions in the vas def-
erens and seminal vesicle.
The effects of the a1-AR antagonist tamsulosin
on the contractions of the vas deferens and seminal
vesicle to norepinephrine were determined. Tam-
sulosin presented a complex insurmountable
antagonism in both tissues, characterized by con-
comitant reductions of the maximal tension devel-
oped and of the potency of norepinephrine in
inducing the contractions (Figure 3A, D). To
check if the insurmountable antagonism of nore-
pinephrine contractions presented by tamsulosin
would be related to the experimental conditions
used rather than to a specific pharmacological
characteristic of this drug, the effects of the selec-
tive a1-AR competitive antagonist prazosin were
also determined. In contrast to what was observed
with tamsulosin, prazosin showed a classical com-
petitive antagonism inducing rightward displace-
ments of concentration–response curves to
norepinephrine with no significant alterations in
the maximal contractions attained in the vas def-
erens or seminal vesicle (Figure 3B, E). Table 1
presents a summary of the inhibitory effects of
nifedipine, (S)-(+)-niguldipine, and tamsulosin on
the contractions induced by norepinephrine in the
rat vas deferens and seminal vesicle.
Mating Tests
The effects of nifedipine, (S)-(+)-niguldipine, and
tamsulosin on copulatory behavior of sexually
experienced male rats were evaluated. Male rats
treated with vehicle, nifedipine, (S)-(+)-
niguldipine, or tamsulosin presented normal
exploratory and precopulatory behavior. All

J Sex Med 2012;9:159–168

a1-ARs, L-Type Calcium Channels, and Ejaculation 163

Figure 2 Inhibitory effects of L-type calcium channel blockers on the contractions induced by norepinephrine in the rat vas
deferens and seminal vesicle. Concentration–response curves to norepinephrine in the absence and presence of increasing
concentrations of nifedipine (A and D) and (S)-(+)-niguldipine (B and E). Maximal contractions induced by norepinephrine in
the vas deferens (C) and seminal vesicle (F) in the presence of increasing concentrations of nifedipine and (S)-(+)-
niguldipine. Symbols represent mean and vertical bars, when larger than the symbols, represent the standard error of mean
for three to six different tissues.

animals treated with vehicle, nifedipine, (S)-(+)-
niguldipine, or tamsulosin ejaculated during copu-
latory tests.
Table 2 presents the recorded parameters of
copulatory behavior of male rats from the different
treatment groups. Nifedipine 10 mg/kg reduced the
PR, and this effect was not dose-dependent as the
PR of animals treated with nifedipine 50 or
100 mg/kg was not different from the PR of vehicle-
treated males. No other parameter of copulatory

Table 1 Inhibitory effects of dihydropyridine-calcium channel blockers, nifedipine and (S)-(+)-niguldipine, and of the
a1-adrenoceptor antagonist tamsulosin on the contractions induced by norepinephrine in the vas deferens and seminal
vesicle
Parameter evaluated
Rightward displacement in
norepinephrine concentration–
Maximal response response curve (fold over
inhibition (%) pIC50† control curve)‡
Nifedipine
Vas deferens 100 ⫾ 0.00 7.60 ⫾ 0.07 No effect
Seminal vesicle 65.66 ⫾ 3.57 7.63 ⫾ 0.08 No effect
(S)-(+)-niguldipine
Vas deferens 94.78 ⫾ 0.33 7.90 ⫾ 0.05* 549.5
Seminal vesicle 71.00 ⫾ 1.86 8.09 ⫾ 0.12* 21.4
Tamsulosin
Vas deferens 85.69 ⫾ 1.85 9.08 ⫾ 0.09 3,499
Seminal vesicle 95.28 ⫾ 0.51 8.81 ⫾ 0.12 173.8

Data are mean ⫾ standard error of mean of four to six different tissues.
*Different from the inhibitory potency of nifedipine in the same organ (P < 0.05, Student’s t-test).
†-log IC
50 concentration effective at 50% of its inhibitory effect.
‡Effect of the maximal concentration of antagonist used.

J Sex Med 2012;9:159–168

164 Kiguti and Pupo

Figure 3 Inhibitory effects of a1-adrenoceptor antagonists, prazosin and tamsulosin, on the contractions induced by
norepinephrine in the rat vas deferens and seminal vesicle. Concentration–response curves to norepinephrine in the presence
of different concentrations of tamsulosin (A, D) and prazosin (B, E). Maximal tension attained in the vas deferens (C) and
seminal vesicle (F) in the presence of prazosin and tamsulosin as percent of those attained in the absence of drugs. Symbols
represent mean and vertical bars, when larger than the symbols, represent the standard error of mean for four different tissues.

behavior was modified by nifedipine. Treatment of treated with nifedipine, (S)-(+)-niguldipine, or

male rats with (S)-(+)-niguldipine or tamsulosin did tamsulosin was not modified at any dose.
not modify any parameter of copulatory behavior of During the mating tests, each ejaculation of
male rats. The ejaculation latency of male rats vehicle-treated male rats resulted in a seminal plug

Table 2 Parameters of copulatory behavior of sexually experienced male rats treated with nifedipine, (S)-(+)-niguldipine,
or tamsulosin
Nifedipine (mg/kg)
Vehicle(ethanol, N = 11) 10.0 (N = 8) 50.0 (N = 7) 100.0 (N = 7)
LMI (s) 14.2 ⫾ 2.0 14.0 ⫾ 3.6 9.7 ⫾ 1.7 10.0 ⫾ 2.6
LI (s) 28.7 ⫾ 7.1 29.6 ⫾ 10.8 23.2 ⫾ 4.0 14.3 ⫾ 3.1
NI 9.0 (6.0–10.0) 10.0 (9.0–11.0) 9.0 (5.0–15.0) 10.0 (8.0–11.0)
EL (s) 274.5 ⫾ 30.3 308.0 ⫾ 47.8 201.6 ⫾ 29.6 260.0 ⫾ 43.4
PR (s) 330.2 ⫾ 9.0 268.3 ⫾ 15.4* 327.9 ⫾ 16.5 318.1 ⫾ 17.9
(S)-(+)-Niguldipine (mg/kg)
Vehicle(ethanol, N = 11) 10.0 (N = 7) 50.0 (N = 7) 100.0 (N = 6)
LMI (s) 14.2 ⫾ 2.0 14.2 ⫾ 3.3 12.1 ⫾ 2.2 9.2 ⫾ 1.3
LI (s) 28.7 ⫾ 7.1 36.2 ⫾ 12.8 20.1 ⫾ 3.4 17.3 ⫾ 5.4
NI 9.0 (6.0–10.0) 10.0 (5.0–13.0) 7.0 (5.0–10.0) 10.0 (9.0–11.0)
EL (s) 274.5 ⫾ 30.3 272.1 ⫾ 50.0 340.7 ⫾ 44.4 288.7 ⫾ 15.1
PR (s) 330.2 ⫾ 9.0 357.0 ⫾ 21.6 320.5 ⫾ 17.8 315.0 ⫾ 37.8
Tamsulosin (mg/kg)
Vehicle(saline, N = 10) 0.3 (N = 9) 1.0 (N = 6) 3.0 (N = 6)
LMI (s) 10.90 ⫾ 1.96 14.44 ⫾ 3.04 14.40 ⫾ 5.27 12.17 ⫾ 2.18
LI (s) 12.6 ⫾ 2.3 21.4 ⫾ 5.0 22.0 ⫾ 9.2 14.3 ⫾ 2.6
NI 8.0 (7.0–15.0) 9.0 (7.0–12.0) 11.0 (9.0–11.0) 9.0 (8.0–12.0)
EL (s) 347.70 ⫾ 61.90 270.30 ⫾ 42.13 399.70 ⫾ 55.83 240.30 ⫾ 52.72
PR (s) 342.8 ⫾ 13.1 354.8 ⫾ 29.1 327.2 ⫾ 14.3 326.3 ⫾ 26.9

Data are mean ⫾ standard error of mean (LMI, LI, EL, and PR) or median and interquartile range (NI).
*P < 0.05 vs vehicle (analysis of variance followed by Dunnet).
LMI = latency for the first mount or intromission; LI = latency for the first intromission; N = number of animals; NI = number of intromissions until ejaculation;
EL = ejaculation latency; PR = post-ejaculatory refractory time.

J Sex Med 2012;9:159–168

a1-ARs, L-Type Calcium Channels, and Ejaculation 165

formation in the female vagina. The seminal plugs

presented as white, waxy appearance masses dis-
lodged from female vagina by successive intromis-
sions of subsequent ejaculatory series. Treatment
of male rats with nifedipine or (S)-(+)-niguldipine
had no effect in the weight of seminal plugs recov-
ered during copulatory tests. In contrast, the
weights of seminal plugs of male rats treated with
tamsulosin (0.3 and 1.0 mg/kg) were reduced by
50% (Figure 4). In addition, seminal plugs of only
33% (2/6) of the animals treated with tamsulosin
(1.0 mg/kg) could be recovered, whereas no
seminal plugs were recovered from female rats
mated with male rats treated with tamsulosin
(3.0 mg/kg).

Discussion
The present study investigates if the inhibition of
the contractility of the vasa deferentia and seminal
vesicles by L-type calcium channel blockers and/or
a1-AR antagonists can delay ejaculation in view of
the importance of the contractions of these organs
in the sequential events involved in the ejaculatory
reflex in males. To investigate this hypothesis, the
effects of the dihydropyridine-calcium channel
blockers, nifedipine and (S)-(+)-niguldipine, and
of the a1-AR antagonist tamsulosin on the con-
tractility of the vas deferens and seminal vesicle in
vitro and on the ejaculation latency of male rats in
behavioral mating tests were determined.
The contractions of the vas deferens and seminal
vesicle in response to norepinephrine were potently
inhibited by the dihydropyridine-calcium channel
blockers, nifedipine and (S)-(+)-niguldipine, indi-
cating the participation of calcium influx through
L-type calcium channels in the contractions result-
ing from a1-AR activation. In fact, electrophysi-
ological and radioligand binding assays have shown
the presence of L-type calcium channels in the
smooth muscle cells from rat and human vas defer-
ens and seminal vesicle [16,23–28].
Interestingly, the contractions induced by nore-
pinephrine in the vas deferens were completely
abolished by nifedipine and (S)-(+)-niguldipine,
whereas in the rat seminal vesicle, approximately Figure 4 Seminal plug weight of male rats treated with
vehicle, nifedipine, (S)-(+)-niguldipine, or tamsulosin.
30% of the contraction in response to norepineph- Seminal plugs dislodged by successive intromissions during
rine remained in the presence of these two copulatory behavior were recovered at the end of behavioral
dihydropyridine-calcium channel blockers. Simi- analysis and weighted. *P < 0.05 vs. vehicle, **P < 0.001
larly to what was found in the present study, vs. vehicle (analysis of variance + Dunnet a posteriori).
a significant part of the contractions of guinea Data are mean ⫾ standard error of mean of 26 seminal
plugs recovered from 6 to 11 animals. Data for tamsulosin
pig and human seminal vesicles in response (1.0 mg/kg) represent mean of four seminal plugs recovered
to norepinephrine is reported to be resistant from two different males (see text). NR = no seminal plug
to dihydropyridine-calcium channel blockers, recovered.

J Sex Med 2012;9:159–168

166
indicating that calcium mobilization from intrac-
ellular stores and/or extracellular calcium influx
through non-L-type calcium channels contribute
to the contractions induced by a1-AR activation in
this tissue [29,30].
The inhibitory profiles displayed by (S)-(+)-
niguldipine and nifedipine in the concentration–
response curves for norepinephrine in the seminal
vesicle and vas deferens were qualitatively different
as (S)-(+)-niguldipine also decreased the potency
of norepinephrine, in addition, to reduce the
maximal contractions of these organs. This finding
is in agreement with the high affinity of (S)-(+)-
niguldipine for a1-ARs, at which it behaves as a
competitive antagonist [31].
Although the contractions induced by norepi-
nephrine in the vas deferens and seminal vesicle
were effectively inhibited by either nifedipine or
(S)-(+)-niguldipine in vitro, the treatment of male
rats with these two drugs was unable to affect most
of the parameters evaluated in behavioral mating
tests, including the ejaculation latencies. This
would suggest that the rats were treated with doses
of nifedipine or (S)-(+)-niguldipine not large
enough to affect contraction, but similar doses of
these two calcium channel blockers were shown to
significantly reduce the mean arterial pressure in
conscious rats and to impair the spontaneous and
carbachol-induced contractions of the rat urinary
bladder in vivo, indicating that the absence of
effect of nifedipine and (S)-(+)-niguldipine on
ejaculation latency is not because of ineffective
doses [32,33]. Furthermore, supporting the notion
that effective doses were used in this study is the
fact that male rats treated with nifedipine pre-
sented reduced PR; this may be related to the
ability of L-type calcium channel blockers to
modify both dopaminergic and serotonergic neu-
rotransmissions in the central nervous system as
these neurotransmitters were shown to modulate
the PR in rats [34–38].
The a1-AR antagonist tamsulosin presented
high potency in antagonizing contractions of rat
vas deferens and seminal vesicle to norepinephrine
in vitro. However, the maximal contractions
induced by norepinephrine in both the vas defer-
ens and seminal vesicle were concentration-
dependently reduced by tamsulosin; this behavior
is not consistent with simple competitive antago-
nism, such as that displayed by prazosin for which
no significant effect on the maximal contractions
induced by norepinephrine was observed. Similar
insurmountable antagonism has been described
for tamsulosin in the contractions of rat and
J Sex Med 2012;9:159–168
Kiguti and Pupo
human vas deferens and in the rabbit bladder neck
and femoral vein to a1-AR activation [39–42].
Seminal emission was strikingly impaired in
male rats treated with tamsulosin as the weight and
number of the seminal plugs recovered during
mating tests were significantly reduced. The
seminal plugs are formed by the clotting of
seminal vesicle secretions by coagulating gland
fluid, and their final weight is determined by the
amount of seminal vesicle contends emitted during
seminal emission [43,44]. The lower doses of tam-
sulosin (0.3 and 1.0 mg/kg) reduced the weight of
seminal plugs by 50%, and at 3.0 mg/kg, there was
a complete inhibition of seminal plug formation.
As judged from the results of the in vitro contrac-
tion studies, this effect might be related to the
inhibition of the contraction of the seminal
vesicles. In accordance to the idea that the reduced
number and weight of seminal plugs recovered
from rats treated with tamsulosin are related to a
lower contractility of the seminal vesicles, studies
by Kim et al. and Giuliano et al. have shown that
similar doses of tamsulosin impair the increase in
intraluminal seminal vesicle pressure induced by
hypogastric nerve stimulation in rats [45,46].
These pressure responses are physiologically rel-
evant as similar events were observed during ejacu-
lation in copulating male rats [47]. However,
although tamsulosin has impaired the seminal
emission, the ejaculation latency of male rats in
mating tests was not modified.
Interestingly, the reduction in the weight of the
seminal plugs recovered from male rats treated
with tamsulosin correlates well with the profile of
ejaculatory dysfunctions observed in men taking
tamsulosin for the management of benign pros-
tatic hyperplasia, as the incidence and severity of
abnormal ejaculation are dose-dependent [48].
Men taking tamsulosin (0.4 mg) experience sig-
nificant reductions in the ejaculate volume, and at
0.8 mg, the incidence of anejaculation is very high
(35%) [49,50]. In the human male, the secretions
from the seminal vesicle amount for 50–80% of
the ejaculate volume, and conditions of reduced
seminal vesicle secretion output are related to
diminished ejaculate volume [51,52]. In addition,
the transrectal color Doppler ultrasonography
studies in healthy men treated with tamsulosin
have shown that the contraction of the seminal
vesicles during ejaculation is inhibited [50]. The
reduction in the maximal tension developed and
probably in the transport efficiency of vas deferens
and seminal vesicle due to insurmountable antago-
nism would be responsible for reductions in the
a1-ARs, L-Type Calcium Channels, and Ejaculation
sperm count and ejaculate volume in men taking
tamsulosin [49,53].
Conclusions
The physiological events related to the seminal
emission phase of ejaculation are significantly
impaired by a1-AR antagonism or L-type calcium
channel blockade in vitro and at least to a1-AR
antagonism by tamsulosin in vivo. The present
results show that seminal emission impairment by
a1-AR antagonism or L-type calcium channel
blockade is not able to delay the ejaculation. The
insurmountable antagonism of norepinephrine-
induced contractions displayed by tamsulosin in
the seminal vesicle and vas deferens may contrib-
ute to the high incidence of reduced ejaculate
volume and sperm output observed in the men
treated with tamsulosin for relief of the symptoms
of benign prostatic hyperplasia.
Acknowledgments
The authors would like to thank Professor Oduvaldo
Câmara Marques Pereira, Dr. Renata Carolina Piffer,
and Carina Leonelli for the scientific guidance in the
behavioral analysis and to Dr. Flávia Hebeler Barbosa
Trovão for the critical review of the article.
Corresponding Author: Luiz Ricardo Kiguti, MSc,
Pharmacology, UNESP, Department of Pharmacology,
Instituto de Biociências, Botucatu, São Paulo 18618-
970, Brazil. Tel: (55) 1438116253; Fax: (55)
1438153744; E-mail: luizkiguti@ibb.unesp.br
Conflict of Interest: None.
Statement of Authorship
Category 1
(a) Conception and Design
Luiz Ricardo de Almeida Kiguti; André Sampaio
Pupo
(b) Acquisition of Data
Luiz Ricardo de Almeida Kiguti
(c) Analysis and Interpretation of Data
Luiz Ricardo de Almeida Kiguti; André Sampaio
Pupo
Category 2
(a) Drafting the Article
Luiz Ricardo de Almeida Kiguti; André Sampaio
Pupo
(b) Revising It for Intellectual Content
Luiz Ricardo de Almeida Kiguti; André Sampaio
Pupo
167
Category 3
(a) Final Approval of the Completed Article
Luiz Ricardo de Almeida Kiguti; André Sampaio
Pupo
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