Splanchnic Nerve Stimulation Research Articles

Intraoperative electrical stimulation of the pelvic splanchnic nerves during nerve-sparing radical hysterectomy

Objectives. This study sought to determine whether intraoperative electrical stimulation (IES) of pelvic splanchnic nerves (PSNs) while monitoring bladder contraction was useful to predict postoperative bladder function during conventional nerve-sparing radical hysterectomy. Methods. Seventeen patients with stage Ib or IIa cervical cancer underwent conventional radical hysterectomy. IES was performed in all cases, stimulating the roots of PSN, the posterior sheath of the vesicouterine ligament (PVL) and the dorsal area of the ligament. After resection of the uterus, the PSN roots were stimulated again. Bladder function was evaluated by urodynamic study (UDS) preoperatively and 3 months after surgery. Results. The results of IES were consistent with bladder function evaluated by postoperative UDS. In 13 of 17 cases, an increased intravesical pressure was observed with IES of the PSN roots after uterus resection. Nine of 13 cases showed marked detrusor contraction with UDS 3 months after surgery and were able to void without using abdominal pressure except in one case. In the remaining 4 of 17 cases, no response could be detected to IES on either side. Three cases voided using abdominal pressure and one used clean intermittent self-catheterization without spontaneous voiding. Conclusions. IES while monitoring intravesical pressure during radical hysterectomy represents a technically simple and useful procedure for the prediction of postoperative bladder function.

Brain stem mechanisms underlying acupuncture modality-related modulation of cardiovascular responses in rats

The present study was designed to investigate brain stem responses to manual acupuncture (MA) and electroacupuncture (EA) at different frequencies at pericardial P (5-6) acupoints located over the median nerve. Activity of premotor sympathetic cardiovascular neurons in the rostral ventral lateral medulla (rVLM) was recorded during stimulation of visceral and somatic afferents in ventilated anesthetized rats. We stimulated either the splanchnic nerve at 2 Hz (0.1-0.4 mA, 0.5 ms) or the median nerve for 30 s at 2, 10, 20, 40, or 100 Hz using EA (0.3-0.5 mA, 0.5 ms) or at approximately 2 Hz with MA. Twelve of 18 cells responsive to splanchnic and median nerve stimulation could be antidromically driven from the intermediolateral columns of the thoracic spinal cord, T2-T4, indicating that they were premotor sympathetic neurons. All 18 neurons received baroreceptor input, providing evidence of their cardiovascular sympathoexcitatory function. Evoked responses during stimulation of the splanchnic nerve were inhibited by 49 +/- 6% (n = 7) with EA and by 46 +/- 4% (n = 6) with MA, indicating that the extent of inhibitory effects of the two modalities were similar. Inhibition lasted for 20 min after termination of EA or MA. Cardiovascular premotor rVLM neurons responded to 2-Hz electrical stimulation at P 5-6 and to a lesser extent to 10-, 20-, 40-, and 100-Hz stimulation (53 +/- 10, 16 +/- 2, 8 +/- 2, 2 +/- 1, and 0 +/- 0 impulses/30 stimulations, n = 7). These results indicate that rVLM premotor sympathetic cardiovascular neurons that receive convergent input from the splanchnic and median nerves during low-frequency EA and MA are inhibited similarly for prolonged periods by low-frequency MA and EA.

IPSILATERAL SEMINAL EMISSION GENERATED BY ELECTROSTIMULATION OF THE LUMBAR SYMPATHETIC NERVE DURING NERVE SPARING LAPAROSCOPIC RETROPERITONEAL LYMPH NODE DISSECTION FOR TESTICULAR CANCER

To confirm nerve preservation during laparoscopic retroperitoneal lymph node dissection we perform electrostimulation of the lumbar sympathetic nerves relevant to ejaculation. In recent cases we monitored seminal emission using endoscopy of the posterior urethra to observe the response to stimulation. Six patients with testicular tumors underwent intraoperative stimulation of the lumbar sympathetic nerves during laparoscopic retroperitoneal lymph node dissection with a unilateral modified template. A long handled pair of bipolar electrodes was inserted through a laparoscopic port, and ipsilateral and contralateral lumbar sympathetic nerves were individually electrostimulated. Ipsilateral stimulation was performed at the preserved lumbar splanchnic nerves and contralateral stimulation was performed at the union of nerve fibers derived from the lumbar splanchnic nerve just above the superior hypogastric plexus. The side of generated seminal emission was monitored simultaneously by endoscopy of the posterior urethra. In all patients each electrostimulation generated unilateral seminal emission. Each stimulation of a lumbar splanchnic nerve, whether ipsilateral or contralateral, caused seminal emission only from the ejaculatory orifice of the stimulated side. Intraoperative electrostimulation of the lumbar sympathetic nerves generated only ipsilateral seminal emission. This suggests that some efferent sympathetic signals for emission might run ipsilaterally in humans.

Medullary substrate and differential cardiovascular responses during stimulation of specific acupoints.

Electroacupuncture (EA) at P5-P6 acupoints overlying the median nerve reduces premotor sympathetic cardiovascular neuronal activity in the rostral ventral lateral medulla (rVLM) and visceral reflex pressor responses. In previous studies, we have noted different durations of influence of EA comparing P5-P6 and S36-S37 acupoints, suggesting that point specificity may exist. The purpose of this study was to evaluate the influence of stimulating P5-P6 (overlying the median nerve), LI4-L7 (overlying branches of the median nerve and the superficial radial nerve), LI6-LI7 (overlying the superficial radial nerve), LI10-LI11 (overlying the deep radial nerves), S36-S37 (overlying the deep peroneal nerves), or K1-B67 (overlying terminal branches of the tibial nerves) specific acupoints, overlying deep and superficial somatic nerves, on the excitatory cardiovascular reflex and rVLM responses evoked by stimulation of chemosensitive receptors in the cat's gallbladder with bradykinin (BK) or direct splanchnic nerve (SN) stimulation. We observed point-specific differences in magnitude and duration of EA inhibition between P5-P6 or LI10-LI11 and LI4-L7 or S36-S37 in responses to 30-min stimulation with low-frequency, low-current EA. EA at LI6-LI7 and K1-B67 acupoints as well as direct stimulation of the superficial radial nerve did not cause any cardiovascular or rVLM neuronal effects. Cardiovascular neurons in the rVLM, a subset of which were classified as premotor sympathetic cells, responded to brief (30 s) stimulation of the SN as well as acupoints P5-P6, LI10-LI11, LI4-L7, S36-S37, LI6-LI7, or K1-B67, or underlying somatic pathways in a fashion similar to the reflex responses. In fact, we observed a significant linear relationship (r(2) = 0.71) between the evoked rVLM response and reflex change in mean arterial blood pressure. In addition, EA stimulation at P5-P6 and LI4-L7 decreased rVLM neuronal activity by 41 and 12%, respectively, for >1 h, demonstrating that prolonged input into the medulla during stimulation of somatic nerves, depending on the degree of convergence, leads to more or less inhibition of activity of these cardiovascular neurons. Thus EA at acupoints overlying deep and superficial somatic nerves leads to point-specific effects on cardiovascular reflex responses. In a similar manner, sympathetic cardiovascular rVLM neurons that respond to both visceral (reflex) and somatic (EA) nerve stimulation manifest graded responses during stimulation of specific acupoints, suggesting that this medullary region plays a role in site-specific inhibition of cardiovascular reflex responses by acupuncture.

Modulation of hormone secretion by functional electrical stimulation of the intact and incompletely dysfunctional dog pancreas.

The purpose of the present study was to modulate the secretion of insulin and glucagon in Beagle dogs by stimulation of nerves innervating the intact and partly dysfunctional pancreas. Three 33-electrode spiral cuffs were implanted on the vagus, splanchnic and pancreatic nerves in each of two animals. Partial dysfunction of the pancreas was induced with alloxan. The nerves were stimulated using rectangular, charge-balanced, biphasic, and constant current pulses (200 micros, 1 mA, 20 Hz, with a 100-micros delay between biphasic phases). Blood samples from the femoral artery were drawn before the experiment, at the beginning of stimulation, after 5 min of stimulation, and 5 min after the end of stimulation. Radioimmunoassay data showed that in the intact pancreas stimulation of the vagal nerve increased insulin (+99.2 microU/ml) and glucagon (+18.7 pg/ml) secretion and decreased C-peptide secretion (-0.15 ng/ml). Splanchnic nerve stimulation increased insulin (+1.7 microU/ml), C-peptide (+0.01 ng/ml), and glucagon (+50 pg/ml) secretion, whereas pancreatic nerve stimulation did not cause a marked change in any of the three hormones. In the partly dysfunctional pancreas, vagus nerve stimulation increased insulin (+15.5 microU/ml), glucagon (+11 pg/ml), and C-peptide (+0.03 ng/ml) secretion. Splanchnic nerve stimulation reduced insulin secretion (-2.5 microU/ml) and increased glucagon (+58.7 pg/ml) and C-peptide (+0.39 ng/ml) secretion, and pancreatic nerve stimulation increased insulin (+0.2 microU/ml), glucagon (+5.2 pg/ml), and C-peptide (+0.08 ng/ml) secretion. It was concluded that vagal nerve stimulation can significantly increase insulin secretion for a prolonged period of time in intact and in partly dysfunctional pancreas.

The Hippocampal Cholinergic Neurostimulating Peptide, the N-terminal Fragment of the Secreted Phosphatidylethanolamine-binding Protein, Possesses a New Biological Activity on Cardiac Physiology

Phosphatidylethanolamine-binding protein (PEBP), alternatively named Raf-1 kinase inhibitor protein, is the precursor of the hippocampal cholinergic neurostimulating peptide (HCNP) corresponding to its natural N-terminal fragment, previously described to be released by hippocampal neurons. PEBP is a soluble cytoplasmic protein, also associated with plasma and reticulum membranes of numerous cell types. In the present report, using biochemistry and cell biology techniques, we report for the first time the presence of PEBP in bovine chromaffin cell, a well described secretion model. We have examined its presence at the subcellular level and characterized this protein on both secretory granule membranes and intragranular matrix. In addition, its presence in bovine chromaffin cell and platelet exocytotic medium, as well as in serum, was reported showing that it is secreted. Like many other proteins that lack signal sequence, PEBP may be secreted through non-classic signal secretory mechanisms, which could be due to interactions with granule membrane lipids and lipid rafts. By two-dimensional liquid chromatography-tandem mass spectrometry, HCNP was detected among the intragranular matrix components. The observation that PEBP and HCNP were secreted with catecholamines into the circulation prompted us to investigate endocrine effects of this peptide on cardiovascular system. By using as bioassay an isolated and perfused frog (Rana esculenta) heart preparation, we show here that HCNP acts on the cardiac mechanical performance exerting a negative inotropism and counteracting the adrenergic stimulation of isoproterenol. All together, these data suggest that PEBP and HCNP might be considered as new endocrine factors involved in cardiac physiology.

Prolonged inhibition of rostral ventral lateral medullary premotor sympathetic neurons by electroacupuncture in cats

We have shown that electroacupuncture (EA) at the Neiguan–Jianshi (N–J) acupoints over the median nerve reduces myocardial ischemia by modulating the pressor response induced by application of bradykinin on the gallbladder. The present study was designed to investigate the neural substrate underlying the prolonged modulatory effect of EA on visceral afferent input into the rostral ventral lateral medulla (rVLM). Experiments were performed on ventilated anesthetized cats. Neuronal activity was recorded while either stimulating the splanchnic nerve or applying EA at the N–J acupoints. Thirty-three cells responsive to splanchnic nerve and median nerve stimulation were antidromically driven from the intermediolateral columns, T 2–T 4, indicating their function as premotor sympathetic neurons. These neurons also received baroreceptor input demonstrating that they were cardiovascular sympathoexcitatory cells. Arterial pulse-triggered averaging and coherence analysis demonstrated a correlation between cardiac-related discharge activity with 2.8±0.3 Hz rhythms and arterial blood pressure. Stimulation (2 Hz, 1–4 mA, 0.5 ms) of the splanchnic nerve for 30 s evoked excitatory responses. These neuronal responses were reduced during and after 30-min stimulation of EA at the Neiguan–Jianshi acupoints. These splanchnic nerve-induced excitatory responses in neurons subjected to 30 min of EA were reduced by 68%. Iontophoresis of naloxone promptly reversed the EA-induced inhibitory effect by 52%. Neuronal activity in the rVLM induced by splanchnic nerve stimulation was reduced for 50 (or more) min after termination of EA in 7 of 12 rVLM neurons. Our results indicate that rVLM premotor sympathetic cardiovascular neurons receive convergent input from the gallbladder through the splanchnic nerve and N–J acupoints through the median nerves. Through an opioid mechanism, EA inhibits splanchnic nerve-induced excitatory responses of these rVLM neurons. Many of these neurons receiving convergent visceral and somatic input exhibit long-lasting inhibition by EA.

Spontaneous regeneration of the seriously injured sympathetic pathway projecting to the prostate over a long period in the dog.

To explore the spontaneous regeneration, over a long period, of the seriously injured sympathetic pathway controlling the prostate. The hypogastric nerve (HGN), which is part of the sympathetic pathway from the spinal cord to the prostate, was partly removed over half of its length on both sides in six dogs. Four years after surgery the responses of the prostate to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN proximal to the site removed, were assessed. In six dogs, 10 of the 17 LSNs (second to fourth) and four of the 10 HGNs stimulated elicited prostatic contraction. The pathways via the ipsilateral HGN and/or the contralateral HGN from the LSNs to the prostate were identified as having regenerated in four of six dogs. These results indicate that the sympathetic pathways via the HGN to the canine prostate can regenerate spontaneously over a long period after serious injury, and that their cross-innervation system can also be repaired.

Control of canine membranous urethra, bulbocavernosus and ischiocavernosus muscles by lumbosacral sympathetic pathways

The membranous urethral muscles, bulbocavernosus muscle (BCM) and ischiocavernosus muscle (ICM) play a significant role on expulsion at ejaculation. Projection of the sympathetic pathways in the retroperitoneum to these muscles was investigated in the dog. Intraluminar pressure of the membranous urethra and contractions of the BCM and ICM to electrical stimulation of the lumbar splanchnic nerve (LSN) or the lumbosacral sympathetic chain (LSC) were examined. Stimulation of the LSN was performed after transection of the unilateral hypogastric nerve (HGN) to clarify its cross-innervation. Stimulation of each of the 2nd to 4th LSNs elicited elevation of the intraluminal pressure of the membranous urethra. In eight dogs whose right HGNs were transected, 15 of the 19 right LSNs, and 12 of the 16 left LSNs examined elicited elevation of the membranous urethral pressure, respectively. In five dogs whose left HGNs were transected, 6 of the 9 right LSNs and 10 of the 12 left LSNs stimulated elicited elevation of the pressure. Stimulation of the HGN caused no detectable contractions of the BCM and ICM in all six dogs examined. Stimulation of the LSC elicited responses of the BCM and ICM in all six dogs examined and the membranous urethra in three of six dogs. The maximum value of the response of the BCM was greater than that of the ICM. After consecutive transection of the ipsilateral pelvic nerve, stimulation of the LSC showed no significant changes in the responses of the BCM and ICM. The above results indicate the following: (1) each of the L2–L4 LSNs sends signals to the membranous urethra via two routes, passing through the ipsilateral or contralateral HGN; (2) the above nerve does not send effective signals to the BCM and ICM; (3) the LSC sends signals to the membranous urethra, BCM and ICM; (4) the signals from the LSC mainly pass through the pudendal nerve to the BCM and ICM; and (5) the signals were dominant to the BCM compared with the ICM.

Modulation by C 2 ceramide of the nicotinic transmission within the coeliac ganglion in the rabbit

We have investigated the modulation by ceramide of the nicotinic activation of the prevertebral sympathetic neurons. Our study was performed in vitro in rabbit isolated coeliac ganglion, using intracellular recording techniques. We have used C 2 ceramide, a permeant analog of ceramide. The effects of C 2 ceramide were first assessed when nicotinic activation was elicited without modulatory mechanisms (fast excitatory postsynaptic potentials triggered by stimulation of the thoracic splanchnic nerves with a single pulse). In all the neurons tested, C 2 ceramide triggered an increase in the amplitude of the fast excitatory postsynaptic potentials demonstrating a direct facilitatory effect on the nicotinic activation. We then investigated the effects of C 2 ceramide on modulatory mechanisms of this activation. These mechanisms occur when a train of pulses of supramaximum intensity is applied on the splanchnic nerves. During the train, a gradual depression of fast nicotinic activation occurred: the pulses failed to systematically elicit action potentials. We have previously demonstrated that this regulatory phenomenon is partly modulated by nitric oxide which exerts a dual effect: facilitation or inhibition of the nicotinic activation. In all the neurons tested, C 2 ceramide decreased the number of action potentials fired during a train of pulses, demonstrating an indirect inhibitory effect on the nicotinic activation. The use of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (nitric oxide scavenger) suppressed the inhibitory effect of C 2 ceramide, demonstrating that this effect is mediated through the nitric oxide pathway. C 2 dihydro-ceramide, an inactive analog of ceramide, was without effect on the nicotinic activation of the ganglionic neurons. These results demonstrate that ceramide exerts a complex modulation of the nicotinic activation of the prevertebral neurons: direct facilitation and indirect inhibition involving the nitric oxide pathway. In fact, C 2 ceramide plays a key gating role in the dual effect of the nitric oxide pathway by activating the inhibitory effect. The existence of this gating mechanism involving ceramide and nitric oxide opens new perspectives in terms of our understanding of the modulation of synaptic transmission within the prevertebral ganglia. Our study demonstrates that sphingolipids are involved in complex modulations of the synaptic activation within the prevertebral ganglia, and thus contribute to their integrative properties.

PACAP release from the canine adrenal gland in vivo: its functional role in severe hypotension.

This study was to investigate if endogenous pituitary adenylate cyclase-activating polypeptide (PACAP) can be released during direct splanchnic nerve stimulation in vivo and to determine whether PACAP in the adrenal gland can modulate the medullary response to sympathoadrenal reflex. The output of adrenal catecholamine and PACAP-38-like immunoreactivity (PACAP-38-ir) increased in a frequency-dependent manner after direct splanchnic nerve stimulation (0.2-20 Hz). Both responses were highly reproducible, and PACAP-38-ir output closely correlated with catecholamine output. Sodium nitroprusside (SNP; 0.1 mg/kg iv bolus) caused a severe hypotension resulting in marked increases in catecholamine secretion. In the presence of local PACAP-27 (125 ng), the maximum catecholamine response to SNP was significantly potentiated in a synergistic manner compared with that obtained in the group receiving SNP or PACAP-27 alone. The study indicates that endogenous PACAP-38 can be released particularly when the sympathoadrenal system is highly activated and that the local exogenous PACAP-27 enhanced the reflex-induced catecholamine release, suggesting collectively a facilitating role of PACAP as neuromodulator in the sympathoadrenal function in vivo.

Stimulation of nerves innervating the dog's pancreas.

Our aim was to modulate secretion of insulin and glucagon into the blood of a diabetic but otherwise healthy dog with stimulation of nerves innervating the pancreas. The 33 electrode spiral cuffs were implanted in an adult beagle canine. The first cuff was installed on the vagus nerve, the second one on the splanchnic nerve, and the last one on the pancreatic nerve. To cause insulin-dependent Type I diabetes, partial dysfunction of the pancreas was induced. Nerves were stimulated using biphasic, rectangular current pulses (10 mA, 200 micros, 20 Hz). Samples from the femoral artery were drawn before the experiment, after 5 min, and 5 min after the stimulation stopped. The results of radioimmunological assay of blood samples showed that in the intact pancreas, stimulation of the vagus nerve caused a considerable increase in insulin secretion, no significant change in glucagon secretion, and a decrease in C-peptide secretion. Splanchnic nerve stimulation did not cause considerable change in insulin and C-peptide secretion while considerable increase in glucagon secretion was noticed. Pancreatic nerve stimulation did not considerably change the secretion of any of the three hormones. In the dysfunctioned pancreas, vagal nerve stimulation caused an increase in insulin and glucagon secretion and a minor increase in C-peptide secretion. Splanchnic nerve stimulation caused a minor decrease in insulin secretion, a considerable increase in glucagon secretion, and a small increase in C-peptide secretion. Pancreatic nerve stimulation did not cause a considerable change in insulin secretion while a minor increase in glucagon and C-peptide secretion was observed.

Autoinhibitory function of the sympathetic prejunctional neuropeptide Y Y 2 receptor evidenced by BIIE0246

The significance of neuropeptide Y Y 2 receptors in sympathetic nonadrenergic transmission was investigated using the novel selective antagonist BIIE0246 (( S)- N2-[[1-[2-[4-[( R, S)-5,11-dihydro-6(6 h)-oxodibenz[ b, e]azepin-11-yl]-1-piperazinyl]-2-oxoethyl]cyclopentyl]acetyl]- N-[2-[1,2-dihydro-3,5 (4 H)-dioxo-1,2-diphenyl-3 H-1,2,4-triazol-4-yl]ethyl]-argininamide). In anaesthetized pigs pretreated with reserpine, and after transection of sympathetic nerves (depleted of noradrenaline), electrical stimulation of renal and splanchnic sympathetic nerves evoked vasoconstriction in, and overflow of neuropeptide Y-like immunoreactivity from, kidney and spleen, respectively. In the presence of BIIE0246, the nerve-evoked overflows of neuropeptide Y-like immunoreactivity were markedly increased and the splenic vasoconstrictor response prolonged. In addition, BIIE0246 caused splenic vasodilatation per se in this model where basal levels of circulating neuropeptide Y exceed 40 pM. It is concluded that endogenous neurogenical neuropeptide Y regulates its own release via activation of sympathetic prejunctional inhibitory neuropeptide Y Y 2 receptors in both spleen and kidney in the reserpinized pig. Moreover, when circulating levels of neuropeptide Y are moderately increased, activation of neuropeptide Y Y 2 receptors seems to contribute to basal splenic vascular tone.

Nicotinic receptors involved in gastric noradrenaline release evoked by electrical stimulation of the splanchnic nerve in rats

In the present experiment, we tried to compare the functional nicotinic receptors activated by electrical stimulation of the greater splanchnic nerve (containing preganglionic sympathetic nerves) to those activated by (−)-nicotine, using the isolated rat stomach. The stomach was perfused with Krebs–Ringer solution and endogenous noradrenaline released into the perfusate was electrochemically measured using high-performance liquid chromatography. The release of noradrenaline evoked by repeated application of 30 mM (−)-nicotine rapidly declined. However, the release of noradrenaline evoked by electrical stimulation of the splanchnic nerve at 2.5 Hz was not disturbed by the appearance of tachyphylaxis for (−)-nicotine. The (−)-nicotine-induced release of noradrenaline was abolished by diltiazem, but this reagent had no effect on the electrically evoked release of noradrenaline. The electrically evoked release of noradrenaline was not influenced by atropine, but was reduced to approximately 50% by hexamethonium. This electrically evoked release of noradrenaline was not influenced by α-bungarotoxin, α-conotoxin ImI (blockers of α7 nicotinic receptors) or dihydro-β-erythroidine (a blocker of α4β2 nicotinic receptors), but was reduced to about 50% by mecamylamine (a blocker of α3β4 nicotinic receptors). The (−)-nicotine-induced release of noradrenaline has already been shown to be partially blocked by dihydro-β-erythroidine and to be abolished by mecamylamine as shown by Yokotani et al. [Eur. J. Pharmacol. 402 (2000) 223.]. These results suggest that the gastric release of noradrenaline in response to electrical stimulation of the greater splanchnic nerve is mediated by cholinergic (probably ganglionic α3β4 nicotinic receptor-mediated) and non-cholinergic mechanisms in rats. However, the functional nicotinic receptor activated by electrical stimulation of the splanchnic nerve seems to be different in character from that activated by (−)-nicotine.

Differential responses of regional sympathetic activity and blood flow to visceral afferent stimulation

The sympathetic nervous system is essential for the cardiovascular responses to stimulation of visceral afferents. It remains unclear how the reflex-evoked sympathetic output is distributed to different vascular beds to initiate the hemodynamic changes. In the present study, we examined changes in regional sympathetic nerve activity and blood flows in anesthetized cats. Cardiovascular reflexes were induced by either electrical stimulation of the right splanchnic nerve or application of 10 microg/ml of bradykinin to the gallbladder. Blood flows were measured using colored microspheres or the Transonic flow meter system. Sympathetic efferent activity was recorded from the left splanchnic, inferior cardiac, and tibial nerves. Stimulation of visceral afferents decreased significantly blood flows in the celiac (from 49 +/- 4 to 25 +/- 3 ml/min) and superior mesenteric (from 35 +/- 4 to 23 +/- 2 ml/min) arteries, and the vascular resistance in the splanchnic bed was profoundly increased. Consistently, stimulation of visceral afferents decreased tissue blood flows in the splanchnic organs. By contrast, activation of visceral afferents increased significantly blood flows in the coronary artery and portal vein but did not alter the vascular resistance of the femoral artery. Furthermore, stimulation of visceral afferents increased significantly sympathetic efferent activity in the splanchnic (182 +/- 44%) but not in the inferior cardiac and tibial nerves. Therefore, this study provides substantial new evidence that stimulation of abdominal visceral afferents differentially induces sympathetic outflow to the splanchnic vascular bed.

Sympathetic efferent pathways projecting to the vagina in the dog

The level of the thoracolumbar sympathetic outflow projecting to the vagina and the laterality of the signals passing through the splanchnic nerves to the vagina have not been well understood. This study was undertaken to explore the sympathetic innervation of the canine vagina by measuring intraluminal pressure of the vagina and contraction of the vaginal wall by electrical stimulation of each of the thoracolumbar splanchnic nerves with or without transection of unilateral hypogastric nerve (HGN). The 2nd–4th lumbar splanchnic nerve (LSN) stimulation elicited elevation of intraluminal pressure of the vagina and contraction of the vaginal wall. In contrast, intermesenteric plexus stimulations caused no response in both the vaginal pressure and the wall itself. On the right side, 12 of 27 LSNs that were stimulated elicited elevation of vaginal pressure and 16 of 32 LSN stimulations caused contraction of the wall, 10 increases in tension and 6 reductions. On the left side, 14 of 26 LSN stimulations elicited elevation of vaginal pressure and 12 of 27 LSN stimulations caused contraction of the wall, 5 increases in tension and 7 reductions. After transection of the right HGN, 6 of 14 right LSN stimulations elicited elevation of vaginal pressure and 10 of 15 right LSN stimulations caused contraction of the wall, 9 increases in tension and 1 reduction. After transection of the left HGN, 13 of 19 left LSN stimulations elicited elevation of vaginal pressure and 10 of 18 left LSN stimulations caused contraction of the wall, 4 increases in tension and 6 reductions. The results indicate that each of the L2–L4 LSNs on either side sends signals to the canine vagina via multiple routes with a crossing site at the level of the caudal mesenteric plexus (CMP).

Spontaneous reconstruction of the canine hypogastric nerve over a long period after removing half of its length

Spontaneous reconstruction of the sympathetic pathway projecting to the seminal tract after serious injury has not been well understood. Multiple cross-innervation mechanisms from the spinal cord via the hypogastric nerve to the seminal tract have been demonstrated currently. This study was undertaken to explore long-term spontaneous reconstruction of the canine hypogastric nerve (HGN), which controls ejaculation, after removing half of its length. Further, the study tried to identify the crossed-pathway(s) reconstructed. Responses of the vas deferens/epididymis and bladder neck to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN were examined. In six dogs whose hypogastric nerve was injured bilaterally as described above 4 years before, corresponding to more than 20 years in human, nine (43%) and 13 (57%) of the 21 LSNs stimulated elicited elevation of vasal and bladder neck pressure, respectively. By combining re-transection of a HGN, the following pathways to the vas deferens/epididymis were identified to have been reconstructed: (1) to the ipsilateral vas deferens/epididymis without crossing to the other side; (2) to the contralateral vas deferens/epididymis by crossing to the other side at the caudal mesenteric plexus (CMP); (3) to the contralateral vas deferens/epididymis by crossing to the other side from the ipsilateral HGN at the commissural branches between the right and left pelvic plexuses (CBPP); and (4) to the ipsilateral vas deferens/epididymis by crossing twice at the CMP to the other side and at the CBPP again from the contralateral HGN to the ipsilateral side. The similar patterns of reconstruction were also observed in the bladder neck. The above results indicate that the sympathetic pathways via the HGN to the canine seminal tract can be reconstructed spontaneously in a high rate over a long period after serious injuries and that their cross-innervation system can be repaired.

PACAP-(1–38) as neurotransmitter in the porcine adrenal glands

The concentration of pituitary adenylyl cyclase-activating polypeptide [PACAP-(1-38)] in porcine adrenal glands amounted to 14 +/- 3 pmol/g tissue. PACAP immunoreactive (PACAP-IR) fibers innervated adrenal chromaffin cells (often co-localized with choline acetyltransferase). Subcapsular fibers traversed the cortex-innervating endocrine cells and blood vessels [some co-storing mainly calcitonin gene-related peptide but also vasoactive intestinal polypeptide (VIP)]. PACAP-IR fibers were demonstrated in the splanchnic nerves, whereas IR adrenal nerve cell bodies were absent. In isolated, vascularly perfused adrenal gland, splanchnic nerve stimulation (16 Hz) and capsaicin (10(-5) M) increased PACAP-(1-38) release (1.6-fold and 6-fold respectively, P = 0.02). PACAP-(1-38) dose-dependently stimulated cortisol (2 x 10(-10) M; 24-fold increase, P = 0.02) and chromogranin A fragment (2 x 10(-9) M; 15-fold increase, P = 0.05) secretion. Both were strongly inhibited by the PAC(1)/VPAC(2) receptor antagonist PACAP-(6-38) (10(-7) M). PACAP-(6-38) also inhibited splanchnic nerve (10 Hz)-induced cortisol secretion but lacked any effect on splanchnic nerve-induced pancreastatin secretion. PACAP-(1-38) (2 x 10(-10) M) decreased vascular resistance from 5.5 +/- 0.6 to 4.6 +/- 0.4 mmHg. min. ml(-1). PACAP-(6-38) had no effect on this response. We conclude that PACAP-(1-38) may play a role in splanchnic nerve-induced adrenal secretion and in afferent reflex pathways.

Facilitation and inhibition by endothelin-1 of adrenal catecholamine secretion in anesthetized dogs

We examined the participation of endothelin ET A and ET B receptors in modulation by endothelin-1 of adrenal catecholamine secretion during cholinergic activation in pentobarbital-anesthetized dogs. Drugs were infused intra-arterially into the adrenal gland. Splanchnic nerve stimulation (1 and 3 Hz) increased adrenal catecholamine output in a frequency-dependent manner. Endothelin-1 (0.2, 0.6, and 2 ng/kg/min) enhanced the catecholamine response induced by the 3-Hz nerve stimulation. Under pretreatment with an endothelin ET A receptor antagonist ( R)-2-[( R)-2-[( S)-2-[[1-(hexahydro-1 H-azepinyl)]carbonyl]amino-4-methylpentanoyl]amino-3-(2-pyridyl) propionic acid (FR139317) (1 μg/kg/min), endothelin-1 suppressed the 1- and 3- Hz nerve stimulation-induced catecholamine response in a dose-dependent manner. No inhibitory or facilitatory effect of endothelin-1 was observed under simultaneous pretreatment with FR139317 and an endothelin ET B receptor antagonist N- cis 2,6-dimethylpiperidinocarbonyl- l-γ-methylleucyl- d-1-methoxycarbonyltryptophanyl- d-norleucine (BQ-788) (1 μg/kg/min) or under pretreatment with BQ-788 alone. These results suggest that in the dog adrenal gland, endothelin-1 facilitates and inhibits adrenal catecholamine secretion during cholinergic activation by stimulating endothelin ET A and ET B receptors, respectively.

Alpha-adrenoceptor-mediated penile erection in dogs: in vivo and in vitro observations.

We have evaluated the role of adrenergic components in the pelvic splanchnic nerve on the erectile function in the dog. Electrical stimulation of pelvic splanchnic nerves increased blood flow in the internal pudendal artery and also elevated the cavernous pressure. These increases were blocked in part by phentolamine or methylene blue, but not by propranolol or atropine. The effects of cholinergic and adrenergic agonists and antagonists on mechanical responses were also examined in muscle strips obtained from various arteries in the intrapelvic region including the internal pudendal artery. Norepinephrine induced contraction in the iliac artery and relaxation in the internal pudendal artery, and both the contraction and relaxation responses were blocked by phentolamine but not by propranolol. These findings suggest that in the dog, alpha-adrenergic components projected through the pelvic splanchnic nerve may contribute to penile erection, together with cyclic GMP-mediated mechanisms.