Neurourology and Urodynamics

Cyclophosphamide-Induced Alterations of the Micturition

Reflex in a Novel In Situ Urinary Bladder Model in the
Anesthetized Rat
Patrik Aronsson,* Thomas Carlsson, Michael Winder and Gunnar Tobin
Department of Pharmacology, Institute of Neuroscience and Physiology, Sahlgrenska Academy at the University of
Gothenburg, Gothenburg, Sweden

Aims: Cyclophosphamide-induced cystitis alterations have been reported to occur both at efferent and afferent level in
the micturition reflex arc. In particular, the stretching of the bladder wall causing urothelial release of ATP has been
proposed as one of the pivotal mechanisms causing these alterations. To evaluate functional changes at efferent and
afferent levels of the micturition reflex following cyclophosphamide treatment we have applied a novel in situ half bladder
rat model. Methods: Male Sprague-Dawley rats were treated with either saline or cyclophosphamide (100 mg/kg), and
stretch-, electric-, methacholine-, and ATP-induced responses were thereafter measured at 60–72 hr postinjection under
pentobarbitone anesthesia. In the novel in situ half bladder model, the urinary bladder was prepared via a midline
incision, where the two halves were separated all the way to the urethra as previously described. Results: Following
bladder stretch of 30–80 mN, of the half that was not used for tension measurement, the cyclophosphamide-treated
animals evoked significant two- to threefold larger contractile responses as compared to saline-treated control animals.
A sensitization of the afferent arm was shown in cyclophosphamide-treated animals, since afferent stimulation evoked
similar responses as in control animals despite that the efferent pelvic nerve stimulation displayed a lower contraction-
frequency relationship in cyclophosphamide-treated animals. Atropine reduced the stretch(reflex)-evoked contraction by
up to 50% in control and 75–80% in cyclophosphamide-treated rats. Subsequent addition of PPADS further reduced the
contractions. Conclusion: The micturition reflex response is increased following cyclophosphamide-induced cystitis, as
compared to control. The likely cause is sensitization at mechanosensor level in the micturition arc, which overrides the
decrement of the efferent cholinergic effects. Neurourol. Urodynam. # 2014 Wiley Periodicals, Inc.

Key words: ATP; cystitis; methacholine; pelvic nerve stimulation; stretch

INTRODUCTION impulses in the bladder innervation are conveyed via fibers in

Cystitis is a common adverse effect in patients treated with
the alkylating anti-neoplastic agent cyclophosphamide (CYP).
Since the CYP-induced cystitis shares many features with
bladder pain syndrome/interstitial cystitis (BPS/IC), the CYP-
treated rodent is a commonly used model for the study of this
kind of bladder pathophysiology.1 CYP-induced cystitis is
characterized by frequent micturitions of small volumes and
evokes changes that may affect the urodynamics at most levels
of the micturition reflex arc.2,3 Even though the detrusor
contractile capacity and the efferent signaling in the micturi-
tion reflex are altered, afferent changes have been intensively
discussed as a major mechanism in BPS/IC bladder
dysfunction.4
The afferents inducing the voiding reflex are activated in two
principal ways; either directly via stimulation of mechano-
gated receptors that activate afferent nerve endings or
indirectly by nerve fiber receptors activated by mediators
released from the tissue, for example, by mechanical stretch of
the bladder wall.5 While sacral reflexes enable bladder filling,
the voiding engages a spinobulbospinal reflex.6 At intense
afferent activity, induced by a substantial bladder filling,
signals are transmitted via the spinal cord to the brainstem and
the midbrain. By that, efferent fibers in the periaqueductal gray
area activate the pontine micturition center, which in turn
induces parasympathetic nerve-evoked detrusor contraction.
The sensory input in the myelinated Ad and non-myelinated
C-fibers is conveyed to the spinal cord mainly via the pelvic
nerve, but also to some extent via the hypogastric nerve and
from the bladder neck via the pudendal nerve.5 The efferent
the same nerves as the afferent fibers. While the pelvic nerve
contains the parasympathetic nerve fibers, the hypogastric
nerve and the pudendal nerve contain sympathetic and
somatic fibers, respectively.
The micturition contraction is evoked by the parasympa-
thetic transmitter acetylcholine, which in the rat co-acts with
adenosine 50 -triphosphate (ATP).7 The parasympathetic con-
traction is partly caused by ATP activating P2X1 purinoceptors,
and partly by acetylcholine acting on muscarinic M3 recep-
tors.8,9 The effects of ATP on sensory nerves are mediated via
P2X2, P2X3 and P2X2/3 purinoceptors. However, acetylcholine
may also affect the afferents.5,10
In the rat, CYP-treatment induces increase of the expression
of P2X purinoceptors on sensory neurons.11 Also, the release of
urothelial ATP is increased,12 and both phenomena are likely to
contribute to the afferent sensitization of the micturition reflex
observed in CYP-induced cystitis.2 Furthermore, a hyperexcit-
ability probably occurs in the central nervous system that may
involve glutamate, substance P and calcitonin gene-related
Karl-Erik Andersson led the peer-review process as the Associate Editor
responsible for the paper.
The authors declare that they have no conflict of interest.

Grant sponsor: Vetenskapsradet and Hj€ arnfonden

Correspondence to: Department of Pharmacology, University of Gothenburg, Box
431, SE-405 30 Gothenburg, Sweden.
E-mail: patrik.aronsson@pharm.gu.se
Received 10 October 2013; Accepted 6 January 2014
Published online in Wiley Online Library
(wileyonlinelibrary.com).
DOI 10.1002/nau.22562

# 2014 Wiley Periodicals, Inc.

2 Aronsson et al.
peptide.2 The efferent transmission seems not to be affected per
se. However, both the acetylcholine- and ATP-evoked contrac-
tile effects are changed.13 The major reason for the observed
reduction of the contractile response is a lowered contractile
capacity of the detrusor smooth muscle. Regarding acetylcho-
line, alterations of the muscarinic receptor expression also
contribute to a smaller micturition contraction, partly indirect-
ly via the release of urothelial factors such as nitric oxide.14 All
together, these changes result in that conscious CYP-treated
rats micturate more frequently with smaller volumes at each
occasion in comparison to control rats.15
In order to enable the assessment of effects of exogenous
agonists (methacholine and ATP) at different levels in the
micturition reflex, as well as the impact of inflammatory
changes on afferent and efferent neuronal transmission levels,
we have elaborated an in situ rat model for urinary bladder
studies. In most in vivo studies, bladder pressure is measured
by introducing a catheter into the bladder. Recently, we
presented a novel model well suited for functional bladder
studies aiming for discrimination of afferent and efferent reflex
mechanisms.16,17 In our model, we split the bladder into two
parts—one being used for measuring tension and the other to
induce the reflex. Also, the micturition reflex can be activated
by stretch and the afferent and efferent innervations can be
electrically stimulated separately. Since the CYP-induced
cystitis in the rat shares many features with the cystitis
occurring in patients treated with CYP, as well as with BPS/IC,
the model may also be well suited for determining mechanisms
in several types of bladder disorders.
The aims of the present study were to evaluate CYP-induced
functional changes at efferent and afferent levels of the
micturition reflex. Thus, inflammation was induced by CYP-
treatment and functional cholinergic and purinergic responses
were examined in situ by intravenous agonist administration,
electrical nerve stimulation of the pelvic nerve and by stretch-
activation of the micturition reflex.
MATERIALS AND METHODS
The present study design, in which 36 male rats (340–460 g)
of the Sprague-Dawley strain were used, was approved by the
local ethics committee at the University of Gothenburg. In order
to induce cystitis, CYP (Sigma-Aldrich, St. Louis, MO; 100 mg/kg
i.p.) in combination with the analgesic buprenorphine (Temge-
sic1; Schering-Plough, Brussels, Belgium; 10 mg/kg s.c.) was
administered 60–72 hr prior to sacrifice, ensuring peak inflam-
mation at the time of the experiment.3 Control groups instead
received an injection of saline (0.9%; 1 ml/kg i.p.). On the day of
the experiment the rats were anaesthetized with medetomi-
dine (Domitor1Vet.; Orion Pharma, Espoo, Finland; 0.1 mg/kg i.
p.) and pentobarbitone (Pentobarbitalnatrium vet.; APL, Stock-
holm, Sweden; 30 mg/kg i.p.). All analgesics and anesthetics
were purchased from Apoteket Farmaci (Stockholm, Sweden).
An adequate anesthesia depth was controlled by the absence of
blood pressure- and heart rate changes after strong paw-
pinches. Before each experiment in CYP pre-treated animals, the
bladders were checked for typical signs of inflammation, such
as thickening and reddening of the bladder wall. The surgical
procedures were performed as has previously been reported.16
Briefly, the urethra and the urinary bladder as well as the pelvic
nerve were exposed ventrally after separation of the prostatic
glands. Threads were ligated at the top and sides and below the
entrance of the pelvic nerve on each side of the bladder.
Thereafter the bladder was divided into two parts along the
midline from the top of the bladder all the way to the urethra.
The two parts were completely separated from each other but
Neurourology and Urodynamics DOI 10.1002/nau
with preserved intact ureters and blood and nerve supplies. The
two lower ligatures were fixed to the underlying tissue, while
the upper ligature from one of the bladder parts was connected
to an adjustable isometric force transducer (Linton Instrumen-
tation, Norfolk, UK), and the other, contralateral, bladder half
was used for force application (30, 50, and 80 mN). The blood
pressure was monitored continuously via a catheter placed into
the femoral artery. A cannula placed in the femoral vein was
used for all drug administrations, that is, methacholine
(muscarinic receptor agonist; 2, 5, and 10 mg/kg i.v.), ATP (P2
purinoceptor agonist; 10 and 100 mg/kg i.v.), atropine (musca-
rinic receptor antagonist; 1 mg/kg i.v.) and PPADS (P2 purino-
ceptor antagonist; 2 mg/kg i.v.), all purchased from Sigma-
Aldrich.
The pelvic nerve was exposed either on the side of tension
measurement (electrical stimulation of efferent fibers) or on the
contralateral side (electrical stimulation of afferent fibers) and
placed on a bipolar platinum electrode. The protocol involved
stimulation at 2, 5, 10, 20, and 40 Hz continuously until a
maximum of the response was reached (approximately 30 sec;
8–12 V square wave; 2 msec pulse width). The basal tension of
the bladder half used for measurements (contractile side) was
maintained at 5–10 mN.
Statistics and Calculations
Statistical significance was determined by Student’s t-test for
unpaired data. For multiple comparisons, statistical signifi-
cance was determined by one-way analysis of variance
(ANOVA) or two-way ANOVA followed by the Bonferroni or
Tukey’s multiple comparison test. P values <0.05 were regarded
as statistically significant. Graphs were generated and param-
eters computed using the GraphPad Prism program (GraphPad
Software, Inc., San Diego, CA). Data are presented in the form of
mean  SEM.
RESULTS
Mechanical stretch of the contralateral bladder half induced
force-dependent contractile responses from the ipsilateral half
in both control and CYP-treated rats. Application of a force of
30 mN was close to the threshold in control rats (1.5  0.2 mN),
while application of 50 and 80 mN induced larger contractions
(3.1  0.4 and 3.6  0.7 mN, respectively; n ¼ 8; Fig. 1). In the
CYP-treated rats, the mechanical stretch of the contralateral
half evoked significantly larger responses (4.8  1.3, 7.0  1.1,
and 11.1  2.4 mN at 30, 50, and 80 mN, respectively; n ¼ 8;
P < 0.05, P < 0.01, and P < 0.05; Fig. 1). The reflex-induced
responses always showed some delay after applying the force
in both control and CYP-treated rats (30–60 sec). While the
response in the control rats almost instantaneously vanished
after the removal of the force, the response in the CYP-treated
showed a gradual decline. Furthermore, in the CYP-treated rats,
spontaneous contractions generally persisted for some mi-
nutes, even though it varied intra-individually.
In contrast to stretch-stimulation, the contractile responses
to electrical nerve stimulation of the efferent (ipsilateral) pelvic
nerve were reduced in the CYP-treated rats (overall two-way
ANOVA; P < 0.01). At low stimulation frequencies (5 Hz)
almost no reduction occurred in comparison with control
rats. However, at stimulation frequencies above 5 Hz, the
reduction was more prominent and at 40 Hz the contractile
response was reduced by about 40% in CYP-treated rats; the
responses were 21.5  2.4 mN (n ¼ 5) and 14.5  2.8 mN (n ¼ 4)
in controls and CYP-treated rats, respectively (Fig. 2A). The
stimulation of the contralateral pelvic nerve was performed in
 Afferent and Efferent Functional Changes in Cystitis 3

Fig. 1. Contractions following contralateral bladder half stretch stimulation in control and CYP-treated rats. The ipsilateral contractile response to mechanical
stretch of the contralateral bladder half to 30, 50, and 80 mN (3, 5, and 8 g weights) in control (lower part of A) and CYP-treated animals (lower part of B). Also
shown are the blood pressure recordings (upper parts of A and B) and the duration of stretch-stimulation (thin gray line below recordings). CYP-treated animals
(n ¼ 8) showed a significantly higher contraction to contralateral stretch at all three weights applied, compared to control animals (C; n ¼ 8). Un-paired t tests;

P < 0.05 and   P < 0.01. Vertical bars represent the SEM.

order to activate afferents that via the central nervous system
would induce contractile responses of the bladder half used for
tension measurement. This type of stimulation caused almost
identical responses in the control and the CYP-treated rats; at
40 Hz, 15.1  3.2 (n ¼ 4) and 16.1  3.1 (n ¼ 5) mN, respectively
(Fig. 2B).
The muscarinic receptor agonist methacholine (2, 5, and
10 mg/kg) and the P2 purinoceptor agonist ATP (10 and 100 mg/
kg) were injected intravenously. Methacholine-evoked con-
tractions were reduced in CYP-treated rats in comparison with
control rats (3.4  0.6 mN vs. 5.8  0.5 mN at 10 mg/kg i.v.; n ¼ 9;
P < 0.01; Fig. 3A). The responses to ATP remained unaffected by
the CYP-treatment (at 100 mg/kg i.v., 5.9  2.0 vs. 6.4  1.6 mN in
CYP-treated and control rats, respectively; n ¼ 7, n.s.; Fig. 3B).
The administration of atropine (1 mg/kg i.v.) reduced the ATP
(100 mg/kg i.v.) response by 72.7  2.1% in control rats and by
81.8  3.0% in CYP-treated rats (P < 0.05; n ¼ 4).
In another series of mechanical stretch experiments in
control and CYP-treated rats, muscarinic receptor and P2
purinoceptor antagonists were consecutively administered
after an initial round of contralateral stretch-stimulation (30,
50, and 80 mN) in the absence of antagonists. Atropine (1 mg/kg
i.v.) reduced the responses to the mechanical stretch of the
contralateral side by 35–50% (n ¼ 7–9) in control rats, and after
the additional administration of PPADS (2 mg/kg i.v.) only a
trace contraction remained at stretch with 30 mN (reduced by
95%). After PPADS administration, the reductions seemed to be
less prominent at 50 and 80 mN. Here, the responses were
reduced by 60–70% of the initial contractions in the absence of
antagonists (n ¼ 4–7; Fig. 4A). The stretch-evoked responses to
50 mN were 3.8  0.7, 1.9  0.3, and 1.2  0.3 mN in the absence
of antagonists, presence of atropine and subsequent addition of
PPADS, respectively. In the CYP-treated rats, atropine had even
greater effects and consequently the responses were reduced by
75–80% (P < 0.01; n ¼ 6–8). In the presence of atropine and
PPADS, the responses were reduced by 80–95% compared to the
initial response in the absence of antagonists (n ¼ 4–5; Fig. 4B).
Stretch-evoked responses to 50 mN were 13.1  2.8, 2.9  1.3
and 1.3  0.3 mN in the absence of antagonists, presence of
atropine and subsequent addition of PPADS, respectively
(Fig. 4C–E).
DISCUSSION
In the present study, a novel model has been applied in order
to examine the effect of CYP-induced cystitis at different levels
of the micturition reflex. Furthermore, it confirms that a
contralateral influence occurs in the micturition reflex—stretch
on one side of the urinary bladder evokes contraction of the
other.16 Despite a reduced contractile response to parasympa-
thetic stimulation (i.e., efferent pelvic nerve stimulation), the
stretch-evoked response was markedly enlarged in CYP-treated
rats, most likely by sensitized mechanisms in the afferent part
of the micturition arc. However, a general decrement of the
contractile capacity occurs in CYP-treated rats.18 The currently
performed stimulations in the in situ half bladder preparation
also demonstrated that the detrusor contractile capacity is
reduced after CYP-treatment, partly due to a decrement of the
cholinergic function. The contractile effect of ATP did not seem
to be affected by CYP-treatment, but it may have been masked
by other counteracting factors such as decreased smooth
muscle contractility. This latter fact may also have had an
impact on the contractions evoked by electrical stimulation
of afferents, which were rather similar in control and in CYP-
treated rats.
The data generated agree well with the previously reported
results.16 However, the responses to electrical stimulation of
the afferents within the pelvic nerve were presently larger than
previously reported. This may be explained by refined surgical
procedures and that further work was undertaken to perform
the stimulations within the anesthetic window, in which the
reflexes work while the rat is still anaesthetized. It is important
to consider the use of alternative anesthetics, for instance
urethane, when designing and interpreting in vivo studies. This
has previously been thoroughly discussed concerning the
currently used half bladder model.16
The comparisons of reflex-induced responses in control and
in CYP-treated animals in the current study, revealed the
contractions to be markedly larger in the inflamed bladders.
However, the pattern showed a tendency to vary in the two
groups. Namely, in the CYP-treated rats, spontaneous contrac-
tions occurred for a longer period of time following stretch-
stimulation. In the rat, several studies have shown that both

Neurourology and Urodynamics DOI 10.1002/nau

4 Aronsson et al.

A Efferent nerve
stimulation A Methacholine B ATP
10 10
25 Control Control
Control CYP CYP
8 8

Contraction (mN)

Contraction (mN)
CYP
20
6 6
Contraction (mN)

15 4 ** 4

**
2 2
10 *
0 0
5 2 5 10 10 100
Dose (µg/kg) Dose (µg/kg)
0
Fig. 3. Bladder contractile responses to methacholine and ATP in control and
2 5 10 20 40
CYP-treated rats. The half bladder contractions following i.v. administration
Stimulation frequency (Hz) of methacholine (A; 2, 5, and 10 mg/kg) and ATP (B; 10 and 100 mg/kg).
Significant decreases in contractile responses to methacholine in CYP-treated
B Afferent nerve animals (black columns; n ¼ 9) was reached at 2 and 10 mg/kg, as compared to
stimulation controls (white columns; n ¼ 9). The ATP-evoked response was not
statistically significantly affected by the CYP-treatment (n ¼ 7 in both
groups). Un-paired t tests;  P < 0.05 and   P < 0.01. Vertical bars represent the
25 SEM.
Control
CYP
20 substantial enlargement of the reflex-evoked, and not of the
Contraction (mN)

electrical afferent responses, in the CYP-treated rats indicates

15
10
5 treated rats than in control rats. Tentatively, ATP may activate
0
2 5 10 20 40
Stimulation frequency (Hz)
Fig. 2. Bladder contractile responses to efferent and afferent electric
stimulation in control and CYP-treated rats. Half bladder contractions to
efferent (ipsilateral; A) and afferent (contralateral; B) electric pelvic nerve
stimulation at 2, 5, 10, 20, and 40 Hz (8–12 V). The efferent stimulation
showed an overall significant decrease in contractile response between
CYP-treated (n ¼ 4) and control animals (n ¼ 5). The response to afferent
stimulation on the other hand was not affected by the CYP-treatment.

P ¼ 0.004 (two-way ANOVA F(4,35) ¼ 9.273). Vertical bars represent the
SEM.
CYP-induced cystitis and spinal cord transection can increase
the spontaneous bladder activity, in particular during the filling
phase.16,19 The origin of these spontaneously arisen contrac-
tions has been debated, but several studies have suggested the
urothelium and/or sub-urothelium to be involved.20 Increased
ATP release has been discussed in the context of increased
frequency and amplitude of the spontaneous contractions,20
and must be considered as one tentative mechanism causing
the alterations of reflex-evoked contractions in the CYP-treated
rat.
The electrical stimulation of afferents within the pelvic nerve
evoked almost identical responses in control and CYP-treated
rats. In the perspective of the enlargement of the reflex-evoked
responses in CYP-treated rats, the resemblance of the afferent
responses to electrical stimulation of the pelvic nerve may
seem contradictory, particularly in view of the sensitization of
sensory pathways that occur after CYP-treatments.11 However,
the reduced contractility of the smooth muscle may have
masked the effect of sensitized afferent pathways. But the
that most of the alterations occur at the mechanoreceptor level.
Regarding the response to intravenous administration of ATP, a
substantial part has been shown to be atropine-sensitive.16 In
the current study, the atropine-sensitive part was larger in CYP-
the afferent arm in the micturition reflex and by that inducing
an activation of parasympathetic efferent nerve fibers that
contributes to the contractile ATP response. If so, it speaks in
favor of the assumption made above.
The parasympathetic response in the urinary bladder mainly
involves two transmitter substances—acetylcholine and ATP.21
When the contractile effects of the two transmitters have been
examined in vitro, methacholine (selective to cholinergic
muscarinic receptors) induces much larger responses than
ATP on a molar basis.18 In the current study, ATP and
methacholine induced responses of similar size on a molar
basis. One explanation to this can be that ATP co-acts with
acetylcholine either directly or indirectly via activation of
afferents.16 Nevertheless, while the methacholine-evoked
contraction was reduced after CYP-treatment, the ATP-evoked
response was preserved.
In vitro findings, however, indicate the opposite in that the
response to large concentrations of ATP is reduced during CYP-
induced cystitis.13 As mentioned before, the contractility of the
smooth muscle seems to be hampered and this may possibly
only be noticeable at contractions close to the maximum
response. In vitro, one is able to examine effects close to the
receptor-maximum responses, which is not possible in vivo (e.
g., due to the blood pressure effects). The reports in the
literature according to the impact of inflammation on ATP
contractile responses are inconclusive. In the feline IC model,
P2X1 purinoceptors have been shown to be down-regulated.12
However, contractile responses to ATP have been reported to be
enhanced in BPS/IC.22,23 The present findings in CYP-induced
cystitis in the rat, often regarded as an animal model for BPS/IC,
support this notice of an increased importance of purinergic
signaling.
The interpretation of a specific transmitter contribution
to nerve-evoked responses is often imprecise because of

Neurourology and Urodynamics DOI 10.1002/nau

 Afferent and Efferent Functional Changes in Cystitis 5

Fig. 4. Contractile responses to contralateral bladder half stretch stimulation in the absence and presence of muscarinic and purinergic antagonists. Increasing
mechanical stretch stimulation (30, 50, and 80 mN) in the absence (white columns) and presence of the muscarinic receptor antagonist atropine (gray
columns), and atropine combined with the P2 purinoceptor antagonist PPADS (black columns), in controls (A; n ¼ 4–9) and CYP-treated rats (B; n ¼ 4–8). PPADS
subsequent to atropine almost completely blocked the contractile responses in both groups. Original traces of contraction to 50 mN stretch stimulation in the
CYP-treated animal in the absence of antagonist (C), after atropine (D), and after subsequently administered PPADS (E). Panel A: One-way ANOVAs,
F(2,35) ¼ 25.20, P < 0.0001; F(2,37) ¼ 5.61, P ¼ 0.0077; F(2,18) ¼ 12.81, P ¼ 0.0005, for 30, 50, and 80 mN, respectively. Panel B: One-way ANOVAs, F(2,20) ¼ 18.96,
P < 0.0001; F(2,16) ¼ 11.67, P ¼ 0.001; F(2,15) ¼ 21.88, P < 0.0001, for 30, 50, and 80 mN, respectively.   P < 0.01 and    P < 0.001. Vertical bars represent SEM.

co-transmitter interactions and composite transmitter effects
at different levels in the synapse. When employing an
antagonist for determining a specific transmitter contribution
to a response, the contribution of another transmitter may be
substantially changed. The administration of the muscarinic
receptor antagonist atropine and the P2 purinoceptor blocker
PPADS almost abolished the reflex-evoked responses in both
control and CYP-treated animals. However, atropine seemed to
have a greater effect in the CYP-treated animals than in
controls. This may at first glance seem peculiar, but considering
what has been mentioned previously, namely a hampered
cholinergic and preserved purinergic function in CYP-induced
cystitis, it seems logical to assume that the CYP-treatment
causes sensitization of mechanisms either at the afferent level
in the bladder or in the propagation of the signals. In this way,
any given stimulation, such as stretch of the bladder wall, may
result in more intense parasympathetic activity. One should
bear in mind that a substantial atropine-resistance part of the
parasympathetic response exists. However, other substances
than acetylcholine and ATP may also affect the responsiveness
to a contractile stimulus. In further experiments, local effects on
the detrusor responsiveness need to be scrutinized. The current
study implicates that drugs acting on the mechanosensor/
afferent nerve transmission of the urinary bladder may be
interesting candidates for pharmacological treatment of dis-
eases such as BPS/IC. It is also of great interest to evaluate the
impact of CYP on the central nervous system. In such
experiments, stimulation can be performed at different levels
of the micturition arc, such as pontine or cortical centers.
CONCLUSION
The present study shows that in our novel in situ half bladder
model, CYP-treatment hampers cholinergic responses, has less

Neurourology and Urodynamics DOI 10.1002/nau

6 Aronsson et al.
effect on purinergic contractions and reduces efferent para-
sympathetic nerve-evoked responses. In spite of this, the reflex-
evoked response is enlarged in CYP-treated rats. This is mainly
caused by increases in the mechanoreceptor function, but it
may involve sensitization of sensory pathways as well. Thus, in
CYP-induced cystitis, mechanoreceptor/afferent sensitization
overrides any decrements in efferent cholinergic responses and
smooth muscle contractility.
ACKNOWLEDGEMENTS
 14. Andersson MC, Tobin G, Giglio D. Cholinergic nitric oxide release from the
The authors wish to thank Ra dman och Fru Ernst Collianders
Stiftelse, Kungliga och Hvitfeldtska stiftelsen, Wilhelm och

Martina Lundgrens Vetenskapsfond and Vetenskapsra det for
 micturition pattern in normal and cyclophosphamide pre-treated conscious
financial contribution. T.C. was financed by Vetenskapsra det
and Hja €rnfonden.
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