Prolonged Inhibitory Effects of Repeated Tibial Nerve Stimulation on the Micturition Reflex in Decorticated Rats.

OBJECTIVE: This study aimed to determine whether a short-term repeated stimulation of tibial nerve afferents induces a prolonged modulation effect on the micturition reflex in a decorticated rat model. MATERIAL AND METHODS: Fifteen female Sprague-Dawley rats (250-350 g) were fully decorticated and paralyzed in the study. Tibial nerve stimulation (TNS) was delivered by inserting two pairs of needle electrodes close to the nerves at the level of the medial malleolus. Constant flow cystometries (0.07 mL/min) at approximately ten-minute intervals were performed, and the micturition threshold volume (MTV) was recorded and used as a dependent variable. After four to five stable recordings, the tibial nerves of both sides were stimulated continuously for five minutes at 10 Hz and at an intensity of three times the threshold for α-motor axons. Six same stimulations were applied repeatedly, with an interval of five minutes between each stimulation. Mean MTV was calculated on the basis of several cystometries in each half-hour period before, during, and after the six repeated TNS. RESULTS: During the experiment, all the animals survived in good condition with relatively stable micturition reflexes, and a significant increase in MTV was detected after TNS. The strongest effect (mean = 178%) was observed during the first 30 minutes after six repeated stimulations. This obvious threshold increase remained for at least five hours. CONCLUSIONS: A prolonged poststimulation modulatory effect on the micturition reflex was induced by short-term repeated TNS in decorticated rats. This study provides a theoretical explanation for the clinical benefit of TNS in patients with overactive bladder and suggests decorticated rats as a promising model for further investigation of the neurophysiological mechanisms underlying the bladder inhibitory response induced by TNS.

Repeat periods of electrical stimulation prolong the modulation of the micturition reflex in the rat.

AIMS: The aim of this study was to determine if the duration of the micturition reflex modulation could be prolonged by repeated periods of afferent stimulation in the decorticated rat. METHODS: Eighteen female Sprague-Dawley rats were used for the study, 10 for intravesical electrical stimulation (IVES), and 8 for Ano-genital pudendal afferents stimulation. Repeated constant flow cystometries were performed with body-warm saline (0.06-0.1 mL/min) at about 10 min interval. The selected afferents were stimulated continuously for 5 min at maximal intensity. The same stimulation was repeated six times with a pause of 5 min between the stimulations. The mean threshold volume of cystometries performed during one hour before and each hour after the stimulation were compared. RESULTS: After six periods of IVES, the micturition threshold volume decreased to its lowest value (62% of control) during the first hour and remained at 80% 4 h later (n = 10, P < 0.01). Ano-genital afferent stimulation produced a corresponding increase in the micturition threshold volume. The long-lasting poststimulation effect was again observed for more than 5 h. During the first hour the mean threshold volume increased to 211% of controls and it remained at about this level for the entire observation period (n = 8, P < 0.01). CONCLUSIONS: Repeated short periods of stimulation prolonged the modulatory effect well beyond the stimulation period. The findings provide experimental evidence supporting the clinical application of IVES and ano-genital stimulation for treatment of neurogenic urinary bladder dysfunction.

Cause of residual urine in bladder outlet obstruction: an experimental study in the rat.

PURPOSE: We explored the role of bladder mechanoreceptors in post-void residual urine in rats with bladder outlet obstruction. MATERIALS AND METHODS: Partial bladder outlet obstruction was induced by a urethral ligature in 11 adult female Sprague-Dawley® rats. Nine sham operated rats served as controls. The outcome was evaluated in acute experiments using α-chloralose anesthesia 6 weeks later. Bladders were catheterized for infusion, pressure recording and intravesical electrical stimulation. Bladder efferent activity was recorded from a thin pelvic nerve branch close to the bladder. Micturition contractions were triggered at different bladder volumes by a brief train of electrical stimulation of bladder afferents while monitoring post-stimulus efferent activity and reflex bladder contractions. The degree of obstruction was assessed by bladder wet weight at the end of the experiment. RESULTS: Bladder weight, micturition threshold volume, anatomical bladder capacity and peak contraction force were significantly increased in obstructed rats. In sham operated controls a triggered micturition reflex was sustained by afferent feedback from the bladder until the bladder was empty. In contrast, reflex discharges failed with substantial volume remaining in the bladder in obstructed rats. The minimal micturition reflex volume correlated positively with bladder weight, micturition threshold volume and maximal bladder capacity (r ≥0.74). CONCLUSIONS: In rats with partial bladder outlet obstruction the micturition reflex failed before the bladder was empty due to a decreased afferent drive from bladder mechanoreceptors. Similar changes may contribute to post-void residual urine in humans with bladder outlet obstruction.

Bladder mechanoreceptor changes after artificial bladder outlet obstruction in the anesthetized rat.

AIMS: Experimental animal models of bladder outlet obstruction (BOO) have reproduced several features of BOO in man, i.e., detrusor hypertrophy, instability, frequency, and residual urine. This study was focused on the mechanisms underlying the development of residual urine in patient with benign prostatic hyperplasia (BPH) by examining changes in tension sensitivity of bladder mechanoreceptors in rat model. METHODS: Female adult Sprague-Dawley rats including 12 BOO and 17 sham operated rats were used in this study. Cystometrograms together with the bladder afferent activity were recorded. Tension sensitivity of the afferents was determined by plotting the normalized afferent response against the contraction evoked bladder pressure at different volumes. Degree of obstruction was assessed by the wet weight of the bladder at the end of the experiment. RESULTS: The bladder weight, maximal bladder capacity, micturition threshold volume, peak contraction force, and volume at peak contraction force were all significantly increased in obstructed animals. The threshold volume for afferent activation was increased (mean 0.60 ml compared to 0.15 ml in controls; P < 0.001), positively correlated with the bladder weight (r = 0.74). The tension sensitivity of the bladder mechanoreceptors and the slope of their normalized pressure-response functions were significantly lower at the comparable volumes in the obstructed animals. CONCLUSIONS: Rats with BOO had bladder mechanoreceptors with higher threshold volumes and lower tension sensitivity. Such changes would result in a weaker afferent drive of the micturition reflex. Similar changes may contribute to the development of residual urine and retention in patients with BOO.

Feedforward and recurrent inhibitory receptive fields of principal cells in the cat's dorsal lateral geniculate nucleus.

Principal cells in the dorsal lateral geniculate nucleus receive both feedforward and recurrent inhibition. Despite many years of study, the receptive field structure of these inhibitory mechanisms has not been determined. Here, we have used intracellular recordings in vivo to differentiate between the two types of inhibition and map their respective receptive fields. The feedforward inhibition of a principal cell originates from the same type of retinal ganglion cells as its excitation, while the recurrent inhibition is provided by both on- and off-centre cells. Both inhibitory effects are strongest at the centre of the excitatory receptive field. The diameter of the feedforward inhibitory field is two times larger, and the recurrent two to four times larger than the excitatory field centre. The inhibitory circuitry is similar for X and Y principal cells.

Gating of the micturition reflex by tonic activation of bladder cold receptors in the cat.

AIMS: To determine whether C afferents can modify the gating of the Adelta micturition reflex in order to identify the neuronal site of interaction of the two afferent systems. METHODS: Adult female cats, anaesthetized with alpha-chloralose, had their bladder and urethra catherized through a slit in the proximal urethra. Micturition threshold volume was assessed by cystometry and bladder efferent activity recorded simultaneously. The bladder was filled at a slow rate (1.2-3.5 ml/min) with either body-warm saline (control) or menthol solution (0.06 mM) or by cold saline (4 degrees C). RESULTS: Of 14 trial sessions in 5 animals, the threshold volume of the Adelta micturition reflex was consistently reduced by menthol infusions from a control median (md) value of 16.8 to 10.2 ml (P < 0.01). The threshold pressure was also somewhat decreased from md 0.7 to 0.5 kPa (P < 0.05), while the peak pressure or pressure slope did no differ in two situations. Similar results were obtained with slow cold infusions into the bladder (nine sessions in three animals). The threshold volume decreased from md 19.8 to 17.4 ml (P < 0.05). The bladder reflex response to slow menthol or cold infusions had the typical features of an Adelta micturition reflex in that the efferent activity was largely abolished by the bladder Adelta mechanoreceptor unloading. CONCLUSIONS: Gradual tonic activation of bladder cold receptors lowers the threshold volume of the ordinary Adelta micturition, pointing to a segmental spinal mechanism for the gating of the micturition reflex.

Experimental radiofrequency brain lesions: a volumetric study.

OBJECTIVE: This study describes the production, under strictly standardized and controlled conditions, of radiofrequency lesions with identical neurogenerator settings: in vitro in two different albumin solutions (nongelatinous and gelatinous) and in vivo in the thalamus of the pig. METHODS: The radiofrequency lesions were investigated in vitro by the use of a specially designed video system and in vivo by magnetic resonance imaging. Moreover, the size of the in vivo lesions was estimated with the use of histological sectioning. The statistical analysis included the calculation of a correlation coefficient for the length, width, and volume for each lesion estimation. RESULTS: A high correlation (R = 0.96, P < 0.005; n = 14) was found between clot sizes in the two albumin solutions. Albumin clots generated in gelatinous albumin showed systematically larger volumes. In the pig, two concentric zones were seen in all magnetic resonance images and all histological preparations. The width correlation of the completely coagulated brain tissue (inner zones) was R = 0.94, P < 0.005, and n = 7. The corresponding correlation between magnetic resonance images and gelatinous albumin was R = 0.93, P < 0.005, and n = 7. As a rule, the in vitro clots were smaller than the outer zone but larger than the inner zone of the magnetic resonance imaging-recorded lesions for all of the electrode and temperature combinations tested. In vivo lesions generated with the same electrode and parameter settings showed high reproducibility. CONCLUSION: The value of presurgical electrode tests to validate the electrode function and lesion size in vitro has become evident in this study, which shows a high correlation between the in vitro albumin clots and the in vivo lesions observed on magnetic resonance images.

Bladder cooling reflex and external urethral sphincter activity in the anesthetized and awake guinea pig.

A spinal bladder cooling reflex, triggered by cold receptors of transient receptor potential melastatin type in the bladder wall, has been identified in several mammals, including man. This reflex and its influence on the external urethral sphincter were further characterized in the urethane anesthetized and awake guinea pigs. A total of 214 bladder infusions were performed in the 12 animals. Compared to controls, cold fluid induced a significant decrease in the threshold volume for reflex bladder contractions (median 82%, p < 0.01). Menthol induced a further decrease (median 50%), signifying a bladder cooling reflex. Detrusor-sphincter activities were dyssynergic during voidings triggered by cold or menthol infusions but were coordinated during control infusions. The bladder cooling reflex was suppressed and the sphincter activity synergic following cold infusions in the awake state. Thus, the bladder cooling reflex is under the active descending inhibitory control in intact, awake animals.

Augmentation of corticogeniculate EPSCs in principal cells of the dorsal lateral geniculate nucleus of the rat investigated in vitro.

Augmentation is a component of short-term synaptic plasticity with a gradual onset and duration in seconds. To investigate this component at the corticogeniculate synapse, whole cell patch-clamp recordings were obtained from principal cells in a slice preparation of the rat dorsal lateral geniculate nucleus. Trains with 10 stimuli at 25 Hz evoked excitatory postsynaptic currents (EPSCs) that grew in amplitude, primarily from facilitation. Such trains also induced augmentation that decayed exponentially with a time constant tau= 4.6 +/- 2.6 s (mean +/- standard deviation). When the trains were repeated at 1-10 s intervals, augmentation markedly increased the size of the first EPSCs, leaving late EPSCs unaffected. The magnitude of augmentation was dependent on the number of pulses, pulse rate and intervals between trains. Augmented EPSCs changed proportionally to basal EPSC amplitudes following alterations in extracellular calcium ion concentration. The results indicate that augmentation is determined by residual calcium remaining in the presynaptic terminal after repetitive spikes, competing with fast facilitation. We propose that augmentation serves to maintain a high synaptic strength in the corticogeniculate positive feedback system during attentive visual exploration.

Unitary EPSCs of corticogeniculate fibers in the rat dorsal lateral geniculate nucleus in vitro.

To investigate unitary corticogeniculate excitatory postsynaptic currents (EPSCs), whole cell patch-clamp recordings were obtained from 20 principal cells in slices of the dorsal lateral geniculate nucleus (dLGN) of DA-HAN rats. EPSCs, evoked by electrical stimulation of corticogeniculate axons, had size distributions with one or more quantal peaks. Gaussian curves fitted to such distributions gave a mean quantal size (q) of -5.0 +/- 0.7 (SD) pA for the EPSCs. Paired-pulse ratio (EPSC2/EPSC1) was 3.3 +/- 0.9 for stimuli separated by 40 ms. The mean quantal size was similar for facilitated EPSCs (-5.2 +/- 0.8 pA), implying an increase in mean quantal content (m). Most corticogeniculate axons were capable of releasing only one or two quanta onto individual principal cells. Mean resting release probability (p) was low, 0.09 +/- 0.04. Binomial models, with the same n but increased p, could account for both the basal and facilitated EPSC size distributions in 6/8 cells. It is suggested that the low resting efficacy of corticogeniculate synapses serves to stabilize this excitatory feedback system. The pronounced facilitation in conjunction with large convergence from many corticogeniculate cells would provide a transient, potent excitation of dLGN cells, compliant with the idea of a visually driven neuronal amplifier.

Outcome of the bladder cooling test in children with nonneurogenic bladder problems.

PURPOSE: The bladder cooling test (BCT) engages a primitive neonatal spinal reflex that becomes suppressed by descending signals in older children and may reappear with suprasacral lesions. We assessed the outcome of the BCT in a large group of children with nonneurogenic bladder problems. MATERIALS AND METHODS: The BCT was evaluated in a consecutive series of 178 girls and 106 boys, 1 month to 18 years old with bladder problems without overt neurology. The test was performed at the end of routine cystometry by a rapid control infusion of body warm saline followed, after fluid evacuation, by the same volume of cold saline (3 to 10C). The test was considered positive if a detrusor contraction greater than 30 cm H2O was evoked by the cold but not the warm fluid. RESULTS: Most children younger than 4 years had a history of pyelonephritis (29 of 34) and/or had vesicoureteral reflux (grade IV to V in 26 of 34). For those younger than 2 years 87% of the BCTs were positive while only 21% of the tests were positive in 2 to 3-year-old children. Most children older than 4 years had idiopathic urge incontinence, and greater than 50% of the BCTs were positive in the youngest (less than 6 years) with a gradual decline to 0% at age 13 years. CONCLUSIONS: Conversion of positive to negative BCTs at about age 2 years presumably represents normal maturation while positive tests in older incontinent children suggest delayed maturation of the central neuronal control of the bladder.

Paired pulse facilitation of corticogeniculate EPSCs in the dorsal lateral geniculate nucleus of the rat investigated in vitro.

To investigate paired pulse facilitation of corticogeniculate EPSCs, whole-cell patch-clamp recordings were made from principal cells in the rat dorsal lateral geniculate nucleus (dLGN) in vitro. Thalamic slices, oriented so that both corticogeniculate and retinogeniculate axons could be stimulated, were cut from young (16- to 37-day-old) DA-HAN rats. Corticogeniculate EPSCs displayed pronounced paired pulse facilitation at stimulus intervals up to 400 ms. The facilitation had a fast and a slow component of decay with time constants of 12 +/- 7 and 164 +/- 47 ms (means +/- S.D.), respectively. Maximum paired pulse ratio (EPSC(2) x EPSC(1)(-1)) was 3.7 +/- 1.1 at the 20-30 ms interval. Similar to other systems, the facilitation was presynaptic. Retinogeniculate EPSCs recorded in the same dLGN cells displayed paired pulse depression at intervals up to at least 700 ms. The two types of EPSCs differed in their calcium response curves. At normal [Ca(2+)](o), the corticogeniculate synapse functioned over the early rising part of a Hill function, while the retinogeniculate synapse operated over the middle and upper parts of the curve. The paired pulse ratio of corticogeniculate EPSCs was maximal at physiological [Ca(2+)](o). The facilitation is proposed to have an important role in the function of the corticogeniculate circuit as a neuronal amplifier.

Intravesical electrical stimulation in the treatment of micturition dysfunction in children.

AIMS: To evaluate the results of intravesical electrical stimulation (IVES) in an open prospective study to treat underactive detrusor in children. The treatment was offered as an alternative to clean intermittent catheterization (CIC). METHODS: Forty-four children were included, 21 girls and 3 boys (6-16 years, md 10) with idiopathic, 9 girls and 11 boys (4-18 years; md 13) with neurogenic underactive detrusor. IVES was given by a catheter electrode in the bladder (cathode) with the anode attached to the suprapubic abdominal skin. Continuous stimulation at 20 or 25 Hz was delivered by battery powered stimulators giving unipolar square-wave pulses (0,2 or 0,7 ms). Stimulation intensity was adjusted individually according to the acceptance of the child (12-64 mA). IVES was initially given at the clinic but 18 children had additional treatment at home. Effect of treatment was monitored by micturiton/incontinence diary, reports of bladder sensation, recordings of urinary flow, residual volume and frequency of urinary tract infections. RESULTS: The IVES-treatment was completed by 39/44 children. Long term normalization of the voiding (md 2,5 years follow up) was obtained for 20/24 children with idiopathic problems (83%) and 8/20 with neurogenic problems (40%). Another four had much improved bladder function. The neurogenic group required more stimulation sessions than the idiopathic group. Of those on CIC, 11/15 who completed IVES could discontinue the catheterization. The frequency of urinary tract infections and incontinence decreased significantly (P < 0.01). CONCLUSIONS: It is concluded that IVES is a promising method to treat the underactive detrusor in children.

Inhibition of the bladder cooling reflex in the awake state: an experimental study in the cat.

PURPOSE: We assessed the bladder cooling reflex in the awake cat. The bladder cooling reflex is consistently observed in anesthetized adult cats but not in awake, neurologically normal humans. This discrepancy could indicate a state dependant control of the reflex or a species difference. This study was designed to differentiate between these alternatives, MATERIALS AND METHODS: Under ketamine-xylazine 5 animals had an indwelling catheter inserted into the bladder. The cooling reflex was tested by injections of cold saline into the bladder (4C to 8C), lowering its wall temperature to about 30C to 32C. The volume used (5 ml) was subthreshold for the Adelta micturition reflex, as confirmed by control injections of body warm saline. The procedure was repeated with the animals fully awake and it was well tolerated by all of them. Reflex responses were assessed by induced bladder pressures. RESULTS: Typical bladder cooling reflexes with peak pressures greater than 3 kPa were evoked in all cats when in narcotic sleep (group mean +/- CI 7.4 +/- 3.1 kPa). No such reflexes were elicited when the animals were awake (2.0 +/- 1.0 kPa). The difference was significant at the level of individual animals. CONCLUSIONS: The bladder cooling reflex is suppressed in adult cats during wakefulness, as in humans. This state dependent control of the bladder cooling reflex adds to its resemblance to the extensor plantar response (Babinski's sign).