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A spinal cord slice preparation for analyzing synaptic responses to stimulation of pelvic and pudendal nerves in mature rats.

The dorsal commissural nucleus (DCN) in the lumbosacral spinal cord (L6-S1) receives primary afferent fibers from both pelvic and pudendal nerves in rats. However, the physiological and pharmacological properties of synaptic responses of the DCN neurons to stimulation of those nerves remain unclear. We have developed a longitudinal spinal cord (L6-S1) slice preparation from mature rats that retained both nerves attached. Blind whole-cell recordings were made from the DCN neurons in this preparation. In most neurons, mono- and/or poly-synaptic fast excitatory postsynaptic potentials (EPSPs) were evoked by electrical stimulation of either the pelvic or pudendal nerve. These EPSPs were mediated by activation of Abeta/Adelta and/or C fibers (conduction velocities, 0.5-17.3 m/s), and were abolished by CNQX. Fast EPSPs elicited by either pelvic or pudendal nerve stimulation were occasionally accompanied by bicuculline- and strychnine-sensitive IPSPs. In one-third of the neurons tested, mono- and/or poly-synaptic EPSPs were elicited by the stimulation of both the pelvic and pudendal nerves, indicating convergence of the visceral and somatic primary afferent inputs from the pelvic region onto the DCN neurons. The preparation is applicable to study the mechanism of the integration of the visceral and somatic inputs in the spinal cord.

Age Factors↗

The three-dimensional structure of neurons in the guinea pig inferior mesenteric and pelvic hypogastric ganglia.

The three-dimensional (3-D) morphology of sympathetic inferior mesenteric ganglion (IMG) neurons and sympathetic-parasympathetic pelvic hypogastric ganglion (PHG) neurons was studied using confocal laser scanning microscopy. Cell bodies of IMG neurons were disc-shaped and were arranged orderly in layers. The dendritic arbor of individual neurons was confined to a plane with a thickness that did not exceed the thickness of the parent cell body. The actual dendritic surface area (71,400 micron 2) and volume (81,500 micron 3) of the IMG neurons were up to 100-fold larger than previously reported for similar sympathetic neurons using data of 2-D measurements and estimations of the third dimension. PHG neurons had a much smaller dendritic surface area (4100 micron 2) and volume (2400 micron 3) compared to IMG neurons. The ratio dendritic/somal surface area for individual IMG and PHG neurons ranged from 5:1 to 14:1 and from 0.1:1 to 6:1, respectively. The total dendritic path-length was 8-42 times greater for IMG than for PHG neurons. Neurons in the IMG were either stellate with radiating dendrites or bipolar-shaped with dendrites emerging from the two poles of the cell body. Neurons in the PHG were of two morphological types. One type (nearly 2/3 of all the imaged PHG neurons) had two to seven relatively long dendrites and an axon; the other type had only one to three short unbranched dendrites and an axon. The spatial organization of neurons within the ganglia and the structural features of individual neurons are likely to have important implications regarding connectivity patterns between neurons within the ganglion as well as on how information is processed by the ganglion.

Animals↗

Reflex haemodynamic responses caused by distension of the uterus in anaesthetized pigs.

The present study was undertaken in anaesthetized pigs to determine whether distension of the uterus reflexly affects the aortic blood pressure, heart rate, left ventricular inotropic state and the coronary circulation. Experiments were performed in 17 pigs anaesthetized with alpha-chloralose and artificially ventilated. Coronary blood flow was measured with an electromagnetic flowmeter positioned around the origin of the left circumflex coronary artery. The uterus was distended by injecting 20 ml warm Ringer solution into a balloon positioned within the uterus (mean transmural pressure of about 17 mmHg). Distension of the uterus without controlling any haemodynamic variable caused an increase in aortic blood pressure. When this response was prevented, an increase in heart rate was obtained in each animal. When the heart rate and blood pressure responses were prevented, the distension did not cause significant changes in the maximum rate of change of left ventricular pressure, but always caused a decrease in mean coronary blood flow. In five pigs, the increase in heart rate and the decrease in mean coronary blood flow were graded by step increments of distension. In six pigs, the haemodynamic responses to distension of the uterus were not affected by the administration of atropine. In 12 pigs, which included the six given atropine, the increase in heart rate was abolished by the administration of propranolol and the increase in aortic blood pressure and the decrease in mean coronary blood flow were abolished by the subsequent administration of phentolamine. In the remaining five pigs, the haemodynamic responses caused by uterine distension were abolished by the administration of bretylium tosylate. The present study showed that distension of the uterus in anaesthetized pigs primarily caused reflex increases in heart rate and aortic blood pressure and coronary vasoconstriction. These reflex responses were mediated by efferent sympathetic mechanisms.

Afferent Pathways↗

Uterine contractility and blood flow are reflexively regulated by cutaneous afferent stimulation in anesthetized rats.

The effects of cutaneous mechanical afferent stimulation of various skin areas on uterine contractility and blood flow were examined in anesthetized non-pregnant rats. The contractility of the uterus was measured by the balloon method in the uterus. The uterine blood flow was measured by laser Doppler flowmetry. Noxious pinching stimulation of the perineum for 1 min induced an abrupt contraction of the uterus during stimulation. Pinching of a hindpaw or perineum and innocuous brushing of the perineum for 1 min increased uterine blood flow. Stimulation of other skin areas produced no changes in uterine contractility or blood flow. Most uterine responses were abolished by severance of the pelvic nerves, which innervated the uterus. The activity of pelvic parasympathetic efferent nerves to the uterus increased following perineal pinching. All these cutaneous stimulation-induced responses of uterine contractility, blood flow and pelvic efferent nerve activity still existed, and were even augmented, after acute spinalization. These results indicate that cutaneous mechanical sensory stimulation can regulate uterine contractility and blood flow by a segmental spinal reflex mechanism via uterine parasympathetic efferent nerves.

Anesthesia↗

Transmitter profile and spinal inputs of pelvic ganglion cells projecting with preganglionic axons along the hypogastric and pelvic nerves of the male rat.

Pelvic autonomic ganglion cells receive spinal preganglionic inputs via the hypogastric (lumbar) or pelvic (sacral) nerves. Damage to these nerves stimulates axogenesis (sprouting) from pelvic ganglion cells and two possible triggers are deafferentation (decentralisation) or, if some ganglion cells project centrally in these nerves, axotomy. We have used a combination of retrograde tracing and immunohistochemistry in male rats to identify the number of pelvic ganglion cells that project centrally along these nerves, their transmitter type and the spinal level of their preganglionic inputs. Only a small number (<1%) of pelvic ganglion cells project along these nerves; 29-65 project in each hypogastric nerve and 41-71 in each pelvic nerve. These neurons comprise of both cholinergic and noradrenergic classes and the majority receive preganglionic inputs from the nerve in which they also project. These results suggest that damage of the hypogastric and pelvic nerves close to the pelvic ganglion is unlikely to cause axotomy of many pelvic ganglion cells. Therefore deafferentation rather than axotomy is likely to be the primary trigger of axogenesis occurring in pelvic ganglia after these lesions.

Afferent Pathways↗

Bladder inflammation and hypogastric neurectomy influence uterine motility in the rat.

How pathophysiology of one pelvic organ influences the physiology of another is poorly understood. Here we compared the influence of bladder inflammation and hypogastric neurectomy (HYPX) on uterine contractions and bladder reflexes in urethane-anesthetized rats. Uterine contractions were measured via a latex balloon in one uterine horn. Bladder reflexes were assessed by micturition thresholds (MT) obtained cystometrographically. Whereas bladder inflammation significantly increased bladder reflexes (i.e., reduced MTs), it significantly decreased uterine contraction rate. Whereas HYPX produced small significant decreases in MT, it decreased the rate and significantly increased the amplitude of uterine contractions. These results indicate that bladder pathophysiology can influence uterine contractions and that some of this influence may be via the hypogastric nerve. Such viscero-visceral interactions likely involve spinal cord mechanisms and may have considerable clinical relevance.

Animals↗

Effects of intrathecal injection of tamsulosin and naftopidil, alpha-1A and -1D adrenergic receptor antagonists, on bladder activity in rats.

The effects of intrathecal injection of tamsulosin (an alpha-1A adrenergic receptor antagonist) and naftopidil (an alpha-1D adrenergic receptor antagonist) on isovolumetric bladder contraction were investigated in rats under urethane anesthesia. Intrathecal injection of tamsulosin (10-30 microg) or naftopidil (0.1-30 microg) transiently abolished isovolumetric rhythmic bladder contraction. Following the recovery of bladder contraction, the interval between contractions was the same as the control value before the injection. The amplitude of bladder contraction was decreased by intrathecal injection of naftopidil (3-30 microg), but not by tamsulosin. Therefore, in addition to the antagonistic action of these agents on the alpha-1 adrenergic receptors of prostatic smooth muscle, both agents (especially naftopidil) may also act on the lumbosacral cord, and thus may improve collecting disorders in patients with benign prostatic hyperplasia.

Adrenergic alpha-1 Receptor Antagonists↗

Localization of hindlimb vasomotor neurones in the lumbar spinal cord of the guinea pig.

After retrograde labelling with horseradish peroxidase, sympathetic preganglionic neurones projecting to paravertebral ganglion cells with destinations primarily in the hindlimb were found to lie laterally in the intermediate region of the lumbar spinal cord. The majority of the labelled cell bodies were located near the edge of the grey matter or lateral to it within the white matter. In the most caudal segments (L3-L4) neurons extended right across the lateral funiculi. This distribution of neurones with predominantly vasoconstrictor functions differs markedly from that observed after labelling preganglionic fibres that project in the hypogastric nerve to the pelvic viscera.

Animals↗

Low and high frequency electroacupuncture at Hoku elicits a distinct mechanism to activate sympathetic nervous system in anesthetized rats.

To address the effect of electroacupuncture (Ea) on autonomic nerve activity, the responses of rhythmic micturition contraction (RMC), urine excretion (UE), blood pressure (BP), renal sympathetic nerve activity (RNA) and pelvic parasympathetic nerve activity (PNA) to Ea were investigated in urethane-anesthetized rats. The acupoint Hoku (Li-4) was tested with two different stimulation frequencies (2 Hz and 20 Hz). Elongation of the RMC cycle and an increase in UE associated with the elevation of BP and RNA was elicited during Ea at Hoku. However, the pressor response induced by low frequency Ea (LFEa) was different from that by high frequency Ea (HFEa), i.e. a tonic effect was elicited by LFEa, while a phasic one was induced by HFEa. These results imply that: (1) Ea at Hoku may selectively activate the sympathetic, but not the parasympathetic nervous system, (2) Ea at Hoku with a different stimulation frequency may elicit a distinct mechanism to activate the sympathetic nervous system and (3) Ea at Hoku may ameliorate the hyperactive bladder in clinical therapy.

Acupuncture Points↗

Sacral spinal cord neurons responsive to bladder pelvic and perineal inputs in cats.

In chloralose-anesthetized or decerebrate male cats, 70% of 73 sacral spinal cord neurons activated from the bladder branch of the pelvic nerve also received excitatory inputs from urethra and/or perineal cutaneous nerves (sensory pudendal in 55% and superficial perineal in 84% of neurons). Only 29% of these neurons were excited by the hindlimb skin and muscle nerves tested. The pelvic nerve-responsive neurons received monosynaptic urethral/perineal input in 25% of cases and required temporal summation of this input in 47% of cases. Of 211 neurons responding to superficial perineal nerve stimulation, 101 were not excited by the other nerves tested. Neurons activated by superficial perineal nerve stimulation were found predominantly in S2. It is likely that the superficial perineal nerve represents an important pathway whereby perineal stimulation influences bladder function.

Animals↗

Interleukin-1beta causes a biphasic response in neurons of rat major pelvic ganglia.

The effect of interleukin-1beta (IL-1beta) on peripheral autonomic neurons was examined with intracellular microelectrodes, in vitro. Recombinant human IL-1beta (6-300 pM) produced a depolarization, associated with decrease in input resistance, followed by a hyperpolarization, associated with increase in input resistance, in neurons of rat major pelvic ganglia (MPG). IL-1beta 163-171 (10-100 pM), the active domain of human IL-1beta, also produced a biphasic response. The IL-1beta-induced responses reversed polarity at the equilibrium potential for Cl-. The IL-1beta-induced responses were blocked by picrotoxin (100 microM) but not by bicuculline (20 microM). Imidazole-4-acetic acid (14AA, 100 microM), a GABA(C) receptor antagonist, reduced the IL-1beta-induced responses. The results suggest that the IL-1beta-induced biphasic response is mediated through GABA(C) receptors in rat MPG neurons.

Animals↗

Testosterone and nerve growth factor have distinct but interacting effects on structure and neurotransmitter expression of adult pelvic ganglion cells in vitro.

Circulating testosterone has potent effects on the structure and function of many pelvic ganglion cells in adult rats in vivo. However not all androgen-sensitive pelvic neurones possess androgen receptors and testosterone effects may therefore be indirect, by an action on the target organs. Here we have examined if testosterone influences neuronal structure in vitro in pelvic ganglion cells cultured from adult male rats. We have also used multiple label immunofluorescence to monitor the expression of transmitter-synthesising enzymes and peptides under various culture conditions. Testosterone was a more potent stimulant of noradrenergic soma growth in culture than nerve growth factor. Whereas nerve growth factor increased the number, branching and length of neurites, testosterone stimulated growth of a small number of very short processes, each of which bore numerous short protrusions. Testosterone also impeded the longer neurite growth induced by nerve growth factor. Many pelvic ganglion cells altered their expression of transmitters/neuropeptides under different culture conditions. In particular, under control conditions or during nerve growth factor treatment, vasoactive intestinal peptide was up-regulated in noradrenergic and cholinergic neurones; testosterone impeded this up-regulation in noradrenergic neurones. Choline acetyltransferase immunoreactivity could only be visualised when nerve growth factor was present in the cultures, and cholinergic neurones showed less neurite outgrowth than noradrenergic neurones under all culture conditions. Nerve growth factor did not stimulate levels of this enzyme as strongly if testosterone was present. This study has shown that testosterone has potent effects on the structure of many pelvic ganglion cells in vitro. It is possible that these effects are mediated indirectly, e.g. by stimulating glial-derived substances, however our results suggest that the effects are not mediated by nerve growth factor. The results also show that testosterone influences some of the actions of nerve growth factor, suggesting that there may be complex interactions between steroid signalling and neurotrophic factors in maintaining neuronal structure and function in vivo.

Acetylcholine↗

[Ilio-inguinal Ilio-hypogastic nerve block with a single puncture: an alterantive for anesthesia in emergency inguinal surgery].

The authors describe the anaesthetic procedure for a strangulated hernia repair needing resection and anastomosis of the small bowel in an adult patient. This procedure was performed with an ilio-inguinal/ilio-hypogastric nerve block according to a paediatrical simplified technique with a single puncture. For this patient who had relative contraindications for central blocks, this regional technique allowed to avoid general anaesthesia with its gastric aspiration and predictible difficult intubation risks. This block associated with a very light sedation was sufficient for all the surgical procedure, and postoperative analgesia was efficient over 3 hours. This simplified nerve block, better than the conventional approach for the clinical practice, represents a recommended alternative for hernia repair in emergency for high risk patients who could have a general anaesthesia or a central block.

Aged↗

Endoscopic treatment for endometriosis.

Endometriosis is best regarded as a chronic disease that can vary in symptomatology over time. Endoscopic therapy for relief of pelvic pain as well as infertility is a therapeutic option. The formation of a rational treatment plan before surgery will ensure a minimum number of reproductive surgeries over the patient's lifetime.

Chronic Disease↗

Nitric oxide synthase, choline acetyltransferase, catecholamine enzymes and neuropeptides and their colocalization in the anterior pelvic ganglion, the inferior mesenteric ganglion and the hypogastric nerve of the male guinea pig.

By the indirect immunofluorescence method, the distribution of nitric oxide synthase (NOS)-like immunoreactivity (LI) and its possible colocalization with neuropeptide immunoreactivities, with two enzymes for the catecholamine synthesis pathway, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), as well as the enzyme for the acetylcholine synthesis pathway, choline acetyltransferase (ChAT) were studied in the anterior pelvic ganglion (APG), the inferior mesenteric ganglion (IMG) and the hypogastric nerve in the male guinea pig. The analyses were performed on tissues from intact animals, as well as after compression/ligation or cut of the hypogastric nerve. In some cases the colonic nerves were also cut. Analysis of the APG showed two main neuronal cell populations, one group containing NOS localized in the caudal part of the APG and one TH-positive group lacking NOS in its cranial part. The majority of the NOS-positive neurons contained ChAT-LI. Some NOS-positive cells did not contain detectable ChAT, but all ChAT-positive cells contained NOS. NOS neurons often contained peptides, including vasoactive intestinal peptide (VIP), neuropeptide tyrosine (NPY), somatostatin (SOM) and/or calcitonin gene-related peptide (CGRP). Some NOS cells expressed DBH, but never TH. The second cell group, characterized by absence of NOS, contained TH, mostly DBH and NPY and occasionally SOM and CGRP. Some TH-positive neurons lacked DBH. In the IMG, the NOS-LI was principally in nerve fibers, which were of two types, one consisting of strongly immunoreactive, coarse, varicose fibers with a patchy distribution, the other one forming fine, varicose, weakly immunoreactive fibers with a more general distribution. In the coarse networks, NOS-LI coexisted with VIP- and DYN-LI and the fibers surrounded mainly the SOM-containing noradrenergic principal ganglion cells. A network of ChAT-positive, often NOS-containing nerve fibers, surrounded the principal neurons. Occasional neuronal cell bodies in the IMG contained both NOS- and ChAT-LI. Accumulation of NOS was observed, both caudal and cranial, to a crush of the hypogastric nerve. VIP accumulated mainly on the caudal side and often coexisted with NOS. NPY accumulated on both sides of the crush, but mainly on the cranial side, and ENK was exclusively on the cranial side. Neither peptide coexisted with NOS. Both substance P (SP) and CGRP showed the strongest accumulation on the cranial side, possibly partly colocalized with NOS. It is concluded that the APG in the male guinea-pig consists of two major complementary neuron populations, the cholinergic neurons always containing NOS and the noradrenergic neurons containing TH and DBH. Some NOS neurons lacked ChAT and could represent truly non-adrenergic, non-cholinergic neurons. In addition, there may be a small dopaminergic neuron population, that is containing TH but lacking DBH. The cholinergic NOS neurons contain varying combinations of peptides. The noradrenergic population often contained NPY and occasionally SOM and CGRP. It is suggested that NO may interact with a number of other messenger molecules to play a role both within the APG and IMG and also in the projection areas of the APG.

Animals↗

Surgical management of groin pain of neural origin.

BACKGROUND: An approach to surgical management of the patient with groin pain is described based on our experience with 54 patients, six of whom had bilateral symptoms. History and physical examination are sufficient to relate the pain to one or more of the lateral femoral cutaneous (LFC), ilioinguinal (II), iliohypogastric (IH), or genitofemoral (GF) nerves. STUDY DESIGN: Retrospective analysis of patients with groin pain is reported, with emphasis on cause, involved nerves, and outcomes of operative management. The LFC was decompressed. The II, IH, and GF nerves were resected. Outcomes were graded as excellent, good, and poor in terms of pain relief and functional restoration. RESULTS: For the entire series of patients with painful groins, excellent relief of pain was achieved in 68% and restoration of function achieved in 72%. Ten percent had a poor result. The best results were for II and IH, which were 78% and 83% excellent for both pain relief and restoration of function, with 11% and 17% having a poor result, respectively. The worst results were for the small group of patients with a GF problem, 50% of whom had an excellent and 25% a poor result. Patients who were likely to get an LFC entrapment were those with a nerve located above or within the inguinal ligament. Complications included bruising and cautery injury to the LFC. CONCLUSIONS: Groin pain of neural origin can be relieved with a high degree of patient satisfaction by considering whether one or more of four different nerves are the source of that pain, by realizing that symptoms can be referred to regions other than the groin, such as the pelvic viscera (IH), the knee (LFC), and the testicle (GF), and by treating the appropriate nerve(s) by either neurolysis (LFC) or resection.

Adult↗