Pelvic Afferent Fibers Research Articles

A mechanistic study on urine retention in d-amphetamine addicts.

Chronic amphetamine intake leads to neurogenic bladder and chronic urinary retention. The mechanism underlying persistent urinary retention is unclear. The pelvic-urethral reflex (PUR) is essential for the urethra to develop sufficient resistance to maintain urine continence, an important function of the urinary system. Recent studies on PUR activities have indicated that repetitive/tetanic stimulation of the pelvic afferent fibers induces spinal reflex potentiation (SRP) in PUR activities, which further increases urinary retention. In this study, results showed that test stimulation (TS, 1/30 Hz) evoked a baseline reflex activity, while repetitive stimulation (RS, 1 Hz) induced reflex potentiation in the external urethral sphincter. Intrathecal d-amphetamine (AMPH, 30 μM) did not but higher AMPH concentration (100 μM) induced SRP in TS-induced reflex activity. H89 (10 μM, a protein kinase A inhibitor), but not chelerythrine chloride (CTC, 10 μM, a protein kinase C inhibitor), prevented the 100 μM AMPH-elicited SRP. At 30 μM, forskolin, an activator of adenylyl cyclase, elicited SRP. The co-administration of 10 μM forskolin and 30 μM AMPH induced SRP in TS-induced reflex activity. These results implied that the repetitive/tetanic stimulation of the pelvic afferent fibers could induce SRP in PUR activities, so that the urethra can produce sufficient resistance and played a significant role in urinary retention. Findings in this study demonstrated that amphetamine could induce bladder dysfunction by triggering protein kinase A activation, and provide a practical basis for the development of treatment for amphetamine-associated urinary retention.

Intraspinal sprouting of unmyelinated pelvic afferents after complete spinal cord injury is correlated with autonomic dysreflexia induced by visceral pain

Autonomic dysreflexia is a potentially life-threatening hypertensive syndrome following high thoracic (T) spinal cord injury (SCI). It is commonly triggered by noxious pelvic stimuli below the injury site that correlates with increased sprouting of primary afferent C-fibers into the lumbosacral (L/S) spinal cord. We have recently demonstrated that injury-induced plasticity of (L/S) propriospinal neurons, which relay pelvic visceral sensations to thoracolumbar sympathetic preganglionic neurons, is also correlated with the development of this syndrome. To determine the phenotype of pelvic afferent fiber sprouts after SCI, cholera toxin subunit beta (CTb) was injected into the distal colon 2 weeks post–T4 transection/sham to label colonic visceral afferents. After 1 week of transport, the (L/S) spinal cords were cryosectioned and immunohistochemically stained for CTb, the nociceptive-specific marker calcitonin gene-related peptide (CGRP), and the myelinated fiber marker RT97. Quantitative analysis showed that the density of CGRP+ afferent fibers was significantly increased in the L/S dorsal horns of T4-transected versus sham rats, whereas RT97+ afferent fiber density showed no change. Importantly, CTb-labeled pelvic afferent fibers were co-localized with CGRP+ fibers, but not with RT97+ fibers. These results suggest that the sprouting of unmyelinated nociceptive pelvic afferents following high thoracic SCI, but not myelinated fibers, contributes to hypertensive autonomic dysreflexia induced by pelvic visceral pain.

Expression and function of rat urothelial P2Y receptors

The control and regulation of the lower urinary tract are partly mediated by purinergic signaling. This study investigated the distribution and function of P2Y receptors in the rat urinary bladder. Application of P2Y agonists to rat urothelial cells evoked increases in intracellular calcium; the rank order of agonist potency (pEC(50) +/- SE) was ATP (5.10 +/- 0.07) > UTP (4.91 +/- 0.14) > UTPgammaS (4.61 +/- 0.16) = ATPgammaS (4.70 +/- 0.05) > 2-methylthio adenosine 5'-diphosphate = 5'-(N-ethylcarboxamido)adenosine = ADP (<3.5). The rank order potency for these agonists indicates that urothelial cells functionally express P2Y(2)/P2Y(4) receptors, with a relative lack of contribution from other P2Y or adenosine receptors. Real-time PCR, Western blotting, and immunocytochemistry confirmed the expression of P2Y(2) and to a lesser extent P2Y(4) in the urothelium. Immunocytochemical studies revealed expression of P2Y(2) staining in all layers of the urothelium, with relative absence of P2Y(4). P2Y(2) staining was also present in suburothelial nerve bundles and underlying detrusor smooth muscle. Addition of UTP and UTPgammaS was found to evoke ATP release from cultured rat urothelial cells. These findings indicate that cultured rat urothelial cells functionally express P2Y(2)/P2Y(4) receptors. Activation of these receptors could have a role in autocrine and paracrine signaling throughout the urothelium. This could lead to the release of bioactive mediators such as additional ATP, nitric oxide, and acetylcholine, which can modulate the micturition reflex by acting on suburothelial myofibroblasts and/or pelvic afferent fibers.

Expression of receptors for glial cell line‐derived neurotrophic factor family ligands in sacral spinal cord reveals separate targets of pelvic afferent fibers

Nerve growth factor has been proposed to mediate many structural and chemical changes in bladder sensory neurons after injury or inflammation. We have examined the expression of receptors for the glial cell line-derived neurotrophic factor (GDNF) family within sensory terminals located in the sacral spinal cord and in bladder-projecting sacral dorsal root ganglion neurons of adult female Sprague-Dawley rats. Nerve fibers immunolabelled for GFRalpha1 (GDNF receptor), GFRalpha2 (neurturin receptor), or GFRalpha3 (artemin receptor) showed distinct distribution patterns in the spinal cord, suggesting separate populations of sensory fibers with different functions: GFRalpha1-labeled fibers were in outer lamina II and the lateral-collateral pathway and associated with autonomic interneurons and preganglionic neurons; GFRalpha2-labeled fibers were only in inner lamina II; GFRalpha3-labeled fibers were in lamina I, the lateral-collateral pathway, and areas surrounding dorsal groups of preganglionic neurons and associated interneurons. Immunofluorescence studies of retrogradely labelled bladder-projecting neurons in sacral dorsal root ganglia showed that approximately 25% expressed GFRalpha1 or GFRalpha3 immunoreactivity, the preferred receptors for GDNF and artemin, respectively. After cyclophosphamide-induced bladder inflammation, fluorescence intensity of GFRalpha1-positive fibers increased within the dorsal horn, but there was no change in the GFRalpha2- or GFRalpha3-positive fibers. These studies have shown that GDNF and artemin may target bladder sensory neurons and potentially mediate plasticity of sacral visceral afferent neurons following inflammation. Our results have also revealed three distinct subpopulations of sensory fibers within the sacral spinal cord, which have not been identified previously using other markers.

&lt;b&gt;Urothelium could be important for vesical pain &lt;/b&gt;

Urothelium has been a new key for vesical pain and/or the unusual sensation form the urinary bladder. Afferent fibers in the smooth muscle are mostly A delta fibers and sensitive to distention of the urinary bladder, In contrast, C-fibers in the pelvic afferent fibers were called as “silent fibers”, since the functional significance during the normal micturition was not clear. Recent histological study demonstrated unmyelinated fibers were located submucosal area and urothelium. These contained CGRP and/or substance P. TRPV1 channel in the urothelium released ATP, and P2X3 receptors in the afferent nerve terminals elicited the neural activities. Alpha-1D receptors were also in the urothelium and released ATP. Acetylcholine receptors (M2 and M3 and nicotinic receptors) presented in the urothelium. Acetylcholine was also released from the urothelium. During the inflammation of the urinary bladder, prostaglandin E2 in the urine was increased. EP1 receptors, prostaglandin receptors, presented in the urothelium and primary afferent nerve terminals. These also increased the afferent activity. Variety of neurotransmitter receptors and channels in the urothelium releasing ATP and acetylcholine should be important for controlling the micturition reflex in the pathological condition including the visceral pain.

P2X3Knock-Out Mice Reveal a Major Sensory Role for Urothelially Released ATP

The present study explores the possible involvement of a purinergic mechanism in mechanosensory transduction in the bladder using P2X(3) receptor knock-out (P2X(3)-/-) and wild-type control (P2X(3)+/+) mice. Immunohistochemistry revealed abundant nerve fibers in a suburothelial plexus in the mouse bladder that are immunoreactive to anti-P2X(3). P2X(3)-positive staining was completely absent in the subepithelial plexus of the P2X(3)-/- mice, whereas staining for calcitonin gene-related peptide and vanilloid receptor 1 receptors remained. Using a novel superfused mouse bladder-pelvic nerve preparation, we detected a release of ATP proportional to the extent of bladder distension in both P2X(3)+/+ and P2X(3)-/- mice, although P2X(3)-/- bladder had an increased capacity compared with that of the P2X(3)+/+ bladder. The activity of multifiber pelvic nerve afferents increased progressively during gradual bladder distension (at a rate of 0.1 ml/min). However, the bladder afferents from P2X(3)-/- mice showed an attenuated response to bladder distension. Mouse bladder afferents of P2X(3)+/+, but not P2X(3)-/-, were rapidly activated by intravesical injections of P2X agonists (ATP or alpha,beta-methylene ATP) and subsequently showed an augmented response to bladder distension. By contrast, P2X antagonists [2',3'-O-(2,4,6-trinitrophenyl)-ATP and pyridoxal 5-phosphate 6-azophenyl-2',4'-disulfonic acid] and capsaicin attenuated distension-induced discharges in bladder afferents. These data strongly suggest a major sensory role for urothelially released ATP acting via P2X(3) receptors on a subpopulation of pelvic afferent fibers.

Discharges of bulbar respiratory neurons during rhythmic straining evoked by activation of pelvic afferent fibers in dogs

Each cycle of rhythmic straining evoked through the reflex center in th Kölliker-Fuse nucleus by stimulation of pelvic afferents in decerebrate dogs usually began in early expiration. During the rhythmic straining cycle, postinspiratory discharges of the phrenic nerve increased simultaneously with a burst of discharges of the nerves innervating the rectus abdominis and adductors of the glottis. While about half of the bulbar expiratory units discharged concurrently with the rhythmic straining, almost none of the inspiratory units examined did so. Nearly all expiratory bulbospinal units discharged concurrently, but none of the inspiratory bulbospinal units did so. These results show that expiratory neurons in the caudal bulb relay commands for rhythmic straining from the pontine reflex center to motor neurons of expiratory muscles, but that bulbar inspiratory neurons do not relay the commands to inspiratory motor neurons. Discharges concurrent with rhythmic straining were also evoked in all 4 postinspiratory units of the ventral group. 3 every early onset expiratory units and all 9 inspiratory-expiratory units of the dorsal group. Possible roles played by these respiratory neurons in the organization of rhythmic straining were discussed.

Ascending and descending pathways of reflex straining in the dog.

The location of ascending and descending pathways of the straining reflex was studied by making bulbar cuts in dogs. Rhythmic increases, which were elicited simultaneously in nervous outflows to the diaphragm and rectus abdominis by activation of pelvic afferent fibers, were used to indicate rhythmic straining. The increases were not affected by cutting the dorsal funiculus at the bulbospinal junction, but stopped after cutting both lateral funiculi and by longitudinally cutting the roof of the central canal just caudal to the obex. The increases were impaired by unilateral section of the bulbospinal junction, but were abolished completely after successive ipsilateral hemisection of the rostral bulb. Moreover, rhythmic increases, which appeared synchronously with rhythmic straining in unit discharges recorded from the Kölliker-Fuse nucleus and neighboring structures, disappeared after the longitudinal bulbar cut. However, reflex discharges elicited in pelvic vesical and rectal branches by stimulation of corresponding pelvic afferent fibers were not reduced. These results show that the indispensable part of the ascending pathway of rhythmic straining passes through the lateral funiculus and decussates at the lower bulb, but the pathway of micturition and defecation reflexes do not cross at that level.