Pudendal nerve stimulation induces urethral contraction and relaxation

In this study we measured urethral pressure changes in response to efferent pudendal nerve stimulation in rats. All other neural pathways to the urethra were transected, and the urethra was continuously perfused. We found fast twitch-like contractions, superimposed on a slow relaxation. The amplitude of the twitches was independent of the stimulation frequency below 26 Hz, whereas the relaxation depended highly on this frequency. The twitches were caused by striated urethral muscles, and the relaxation was caused by smooth muscles. Both were mediated by acetylcholine. We calculated the effective urethral relaxation as the absolute relaxation multiplied by the time fraction between the twitches. Maximum effective relaxation occurred at 8-10 Hz, exactly the frequency of spontaneous oscillations during bladder voiding in rats. Although the oscillatory sphincter contractions in rats during voiding may be needed in other mechanisms for efficient voiding, our data suggest that they may be a side effect of the actual purpose: urethral relaxation

Cell length measurements in longitudinal smooth muscle strips of the pig urinary bladder

In this study the length of smooth muscle cells in muscle bundles of pig urinary bladder wall was determined after dissection in Tyrode buffers with different calcium concentrations ([Ca2+]). Previous studies have shown that the length of isolated smooth muscle cells decreases with an increase in [Ca2+] in the buffer. Unlike the results in isolated cells, no significant differences in length were found between cells in strips subjected to different [Ca2+]. Cells in bundles dissected from filled bladders were significantly larger than those dissected from emptied bladders. Cells in strips from emptied bladders dissected in 1.8 mM Ca(2+)-Tyrode buffer were shorter than those obtained in Ca(2+)-free buffer. From the measurements it was concluded that: (1) Cell length in intact tissue is directly related to tissue length; series elastic structures external to the cells do not allow significant shortening of the cells. (2) Passive parallel elasticity outside the cells accounts for passive shortening when bladders are emptied manually. (3) Cell length is not related to empty bladder weight. (4) A positive relation exists between empty bladder weight and bladder capacity

A method for isolating smooth muscle cells from pig urinary bladder with low concentrations of collagenase and papain: the relation between calcium concentration and isolated cell length.

The present study describes a method for isolating single smooth muscle cells from pig urinary bladder using a continuous resuspension device. Low concentrations of collagenase and papain were sufficient to obtain a high yield of viable smooth muscle cells, which remained viable for about 3-4 h as tested with fluorescein diacetate. Addition of fetal calf serum increased the lifespan of the isolated cells and the percentage of contractile smooth muscle cells, but caused spontaneous shortening of the cells. The length and volume of the isolated smooth muscle cells depended on the calcium concentration used in the isolation buffer solution. The isolated muscle cells were apparently relaxed if a calcium concentration less than 1.0 mmol/l was used in the isolation medium. In higher calcium concentrations the isolated cells were significantly shorter, probably as a result of a contraction caused by mechanical stimulation of the cells during the isolation procedure

A comparative study of voiding in rat and guinea pig: simultaneous measurement of flow rate and pressure

In this study, the voiding phase of the micturition cycle in the anesthetized rat and guinea pig is analyzed. In both animals, voiding is characterized by an increase in intravesical pressure and then a decrease, which is accompanied by flow through the urethra and emission of urine. An ultrasonic flow probe was used in both species to measure the flow rate in relation to the intravesical pressure. In the (male) rat, so-called high-frequency oscillations are superimposed on the decreasing bladder pressure. These oscillations do not occur in the guinea pig. It is concluded that the high-frequency oscillations are caused by intermittent flow and not by variations in the bladder contraction. The intermittent flow most likely is caused by the relaxation and contraction of the external urethral sphincter and may have a function in territory marking. In our view, it is not likely that the oscillations enhance bladder emptying, as has been suggested in the literature

Inhibitory effects of tibial nerve stimulation on bladder neurophysiology in rats

Tibial nerve stimulation (TNS) is a form of peripheral neuromodulation which has been found effective in treating overactive bladder symptoms, with lesser side effects than first line pharmacotherapy. Despite its widespread clinical use, the underlying mechanism of action is not fully understood. Our aim was to study its effect on the bladder neurophysiology and the trigger mechanism of voiding in the overactive detrusor, simulated by acetic acid (AA) instillation. In urethane anaesthetized male Wistar rats, the tibial nerve was stimulated for 30 min at 5 Hz, pulse width 200 µs and amplitude approximately three times the threshold to induce a slight toe movement. The pressure at which a voiding contraction was triggered (pthres) did not change significantly between the pre- and post-TNS measurements in AA induced detrusor overactivity. It was found that TNS significantly reversed the effects of AA irritation by increasing the bladder compliance and the bladder volume at pthres, as well as suppressed the threshold afferent nerve activity. The slope of the linear relationship between pressure and the afferent activity increased after AA instillation and decreased significantly after stimulation. In addition to its well-known central inhibitory mechanisms, this study has demonstrated that TNS improves bladder storage capacity by delaying the onset of voiding, via an inhibitory effect on the bladder afferent signaling at the peripheral level

Cell length measurements in longitudinal smooth muscle strips of the pig urinary bladder

In this study the length of smooth muscle cells in muscle bundles of pig urinary bladder wall was determined after dissection in Tyrode buffers with different calcium concentrations ([Ca2+]). Previous studies have shown that the length of isolated smooth muscle cells decreases with an increase in [Ca2+] in the buffer. Unlike the results in isolated cells, no significant differences in length were found between cells in strips subjected to different [Ca2+]. Cells in bundles dissected from filled bladders were significantly larger than those dissected from emptied bladders. Cells in strips from emptied bladders dissected in 1.8 mM Ca2+-Tyrode buffer were shorter than those obtained in Ca2+-free buffer. From the measurements it was concluded that: (1) Cell length in intact tissue is directly related to tissue length; series elastic structures external to the cells do not allow significant shortening of the cells. (2) Passive parallel elasticity outside the cells accounts for passive shortening when bladders are emptied manually. (3) Cell length is not related to empty bladder weight. (4) A positive relation exists between empty bladder weight and bladder capacity

Neurophysiological modeling of voiding in rats: urethral nerve response to urethral pressure and flow

In male urethan-anesthetized rats, activity was measured in nerves that run over the proximal urethra. The urethral nerve response to stepwise urethral perfusion could be described by a four-parameter model (fit error < 6%). At the onset of perfusion, the urethra was closed and the pressure increased with the infused volume. The nerve activity (NA) increased linearly with this inserted volume to a maximum (NAmax), which was proportional to the instantaneous pressure. The duration of this first episode (delta t) was inversely proportional to the perfusion rate. After infusion of a fixed volume, the urethra opened and the NA decreased with a time constant phi -1 (approximately 1.8 s) to an elevated level (NAlevel). NAlevel was linearly related to the steady-state pressure. Accordingly, sensors in the urethra are sensitive to pressure rather than to the perfusion rate. The parameters NAmax, NAlevel, and delta t showed very good reproducibility (SD approximately 19% of mean). The measured activity was most likely afferent and conducted to the major pelvic ganglion