Differences in sympathetic nervous system activity and NMDA receptor levels within the hypothalamic paraventricular nucleus in rats with differential ejaculatory behavior.

Differences in intravaginal ejaculation latency reflect normal biological variation, but the causes are poorly understood. Here, we investigated whether variation in ejaculation latency in an experimental rat model is related to altered sympathetic nervous system (SNS) activity and expression of N-methyl-D-aspartic acid (NMDA) receptors in the paraventricular nucleus of the hypothalamus (PVN). Male rats were classified as "sluggish," "normal," and "rapid" ejaculators on the basis of ejaculation frequency during copulatory behavioral testing. The lumbar splanchnic nerve activity baselines in these groups were not significantly different at 1460 ± 480 mV, 1660 ± 600 mV, and 1680 ± 490 mV, respectively (P = 0.71). However, SNS sensitivity was remarkably different between the groups (P < 0.01), being 28.9% ± 8.1% in "sluggish," 48.4% ± 7.5% in "normal," and 88.7% ± 7.4% in "rapid" groups. Compared with "normal" ejaculators, the percentage of neurons expressing NMDA receptors in the PVN of "rapid" ejaculators was significantly higher, whereas it was significantly lower in "sluggish" ejaculators (P = 0.01). In addition, there was a positive correlation between the expression of NMDA receptors in the PVN and SNS sensitivity (r = 0.876, P = 0.02). This study shows that intravaginal ejaculatory latency is associated with SNS activity and is mediated by NMDA receptors in the PVN.

Differences in the expression of transient receptor potential channel V1, transient receptor potential channel A1 and mechanosensitive two pore-domain K+ channels between the lumbar splanchnic and pelvic nerve innervations of mouse urinary bladder and colon.

The bladder and distal colon are innervated by lumbar splanchnic (LSN) and pelvic nerves (PN) whose axons arise from dorsal root ganglia (DRG) neurons at thoracolumbar (TL) and lumbosacral (LS) spinal levels, respectively. In an attempt to understand the molecular basis of differences between LSN and PN mechanosensitive afferents, we analyzed the gene expression of two potentially counteracting ion channel groups involved in mechanosensation, transient receptor potential channels (TRPV1 and TRPA1) and mechanosensitive two pore-domain K(+) (K(2P)) channels (TREK-1, TREK-2 and TRAAK), in TL and LS DRG neurons innervating mouse bladder or distal colon. The proportion of TRPV1-expressing cells (41∼61%) did not differ between TL and LS neurons innervating bladder or colon. TRPA1 was seldom detected in bladder LS neurons whereas it was expressed in 64∼66% of bladder TL, colon TL and colon LS neurons. Coexpression of TRPV1 and TRPA1 was frequent. TREK-1-expressing cells were more prevalent in LS than TL ganglia in both bladder- and colon-DRG neurons. All three K(2P) channels were detected more frequently in TRPV1-positive neurons in TL ganglia. More than half of TL neurons expressing only TRPA1 were devoid of any of the three K(2P) channels, whereas all TL neurons expressing both TRPA1 and TRPV1 expressed at least one of the K(2P) channels. These results reveal clear differences between LSN and PN sensory pathways in TRPA1 and TREK-1 gene expression and in the gene expression of K(2P) channels in TRPV1-expressing neurons. This study further documents heterogeneity of visceral afferents based on combinations of the five channels examined.

Biophysical and histological determinants underlying natural firing behaviors of splanchnic sympathetic preganglionic neurons in neonatal rats.

Isolated thoracic spinal cords of neonatal rats spontaneously generate splanchnic sympathetic nerve discharge (SND) with a quasiperiodic rhythm approximately 1-Hz. Using in vitro nerve-cord preparations that retained T6-T12 spinal segments, we investigated whether the natural firing behavior of sympathetic preganglionic neurons (SPNs) encoded the SND rhythm and what were the main biophysical and histological determinants of SPN firing. Under extracellular recording conditions, electrical stimulation of splanchnic nerves elicited antidromic responses in 212 SPNs. Among them, 92 SPNs were quiescent; 120 active SPNs had an average firing rate of 0.72+/-0.04 Hz, which was close to the quasiperiodic rhythm of SND. SPNs with rhythmic burst firing were rare. Probability plots of interspike intervals were constructed to extract mathematical features underlying SPN firing. Most active SPNs (88%) had a firing well described by unimodal Gaussian, suggesting a predominantly tonic pattern with normal variations. Biophysical properties of 112 SPNs were measured under whole-cell recording conditions. The charging time constant, tau, is positively correlated with the average firing rate. Histological properties were examined in 45 SPNs with intracellular diffusion of Lucifer Yellow or biocytin. SPNs with pyramidal somata and multipolar dendrites tend to be spontaneously active. In contrast, those with bipolar somata and fewer dendritic branches were quiescent in firing. These observations suggest that activity levels of SPNs are correlated with their capacity for temporal and spatial summation of synaptic inputs. How the seemingly tonic firing of individual SPNs is integrated into whole-nerve SND with quasiperiodic rhythms is discussed.

Characterization of vitamin A-storing cells in mouse fibrous kidneys using Cygb/STAP as a marker of activated stellate cells.

The expression of the cytoglobin/stellate cell activation-associated protein (Cygb/STAP) was recently confirmed in all splanchnic vitamin A-storing cells--including hepatic stellate cells (HSCs)--in normal conditions. In the hepatic fibrous lesion, the expression of Cygb/STAP has been shown to be upregulated in activated HSCs and myofibroblasts (MFs), which have synthesized extracellular matrices. Furthermore, splanchnic vitamin A-storing cells have been reported to be distributed in the kidney. In this study, we clarify the contribution of vitamin A-storing cells to renal fibrosis by focusing on Cygb/ STAP. Adult mice were subjected to unilateral ureteral obstruction (UUO) and kidneys were harvested 1, 3, 7, and 10 days after UUO. Numbers of Cygb/STAP-immunopositive cells as well as Cygb/STAP mRNA 3 days after UUO (UUO day 3 kidney) increased. Vitamin A-autofluorescence was observed in intertubular spaces of controls but gradually declined in a time-dependent manner after UUO. Cygb/STAP+ cells were not completely identical with alpha-smooth muscle actin (alphaSMA)-positive cells in the control or UUO day 7 kidneys. Immunohistochemical analysis for Cygb/STAP and fibulin-2 (Fib), a specific marker for distinguishing MFs from activated HSCs, revealed that the number of Fib+STAP+ cells (MFs) and Fib-STAP+ cells (splanchnic vitamin A-storing cells) significantly increased in UUO day 3 and UUO day 7 kidneys compared with the controls. Our present findings support the concept that Cygb/STAP can be a unique marker for splanchnic fibroblast-like cells, namely the vitamin A-storing cell lineage, and suggest that splanchnic vitamin A-storing cells contribute to renal fibrogenesis in the obstructed kidney.

Human pelvic extramural ganglion cells: a semiquantitative and immunohistochemical study.

In pelvic surgery, much attention is paid to nerve bundles but not to ganglion cells. Using serial section histology of 14 postmortem-treated hemipelvis (eight males, six females; mean, 79 years old), we examined the population number, distribution, and tyrosine hydroxylase-immunoreactivity (TH-IR; suggesting sympathetic neurons) of extramural pelvic ganglion cells. All pelvic ganglion cells were uniformly sized (25-30 microm) contrasting with small intramural rectal neurons. Abundant ganglion cells (30,000-140,000 unilaterally) existed not only along the pelvic viscera except for the rectum, but also along the hypogastric nerve, pelvic splanchnic nerve, pelvic plexus, and associated branches excluding those within the mesorectum. The intrapelvic ganglion cells outside the sympathetic trunk did not form macroscopically identifiable ganglia, but made small clusters (0.1-2.0 mm of maximum diameter) or were diffusely scattered within nerve bundles. More than half of these cells appeared TH-IR positive, although the positive/negative proportion differed between nerves and specimens. Greater numbers of ganglion cells were found in dorsosuperior sites (e.g., around the seminal vesicle) rather than in ventroinferior sites (e.g., along the urethra) in males, and vice versa in females. However, in total cell numbers, interindividual variations were evident rather than intergender difference. Due to significant interindividual variations in cell number, differences are likely to exist between patients in "resistance" to surgical stresses. We hypothesized that pelvic ganglion cells are liable to be damaged due to drying along the surgical margin, hypoxia in venous bleeding, pressure from surgical retractors, extension stress with taping and excess traction and/or direct injury with electrical scalpels.

Differential chemosensory function and receptor expression of splanchnic and pelvic colonic afferents in mice.

Lumbar splanchnic (LSN) and sacral pelvic (PN) nerves convey different mechanosensory information from the colon to the spinal cord. Here we determined whether these pathways also differ in their chemosensitivity and receptor expression. Using an in vitro mouse colon preparation, individual primary afferents were tested with selective P2X and transient receptor potential vanilloid receptor 1 (TRPV1) receptor ligands. Afferent cell bodies in thoracolumbar and lumbosacral dorsal root ganglia (DRG) were retrogradely labelled from the colon and analysed for P2X3- and TRPV1-like immunoreactivity (LI). Forty per cent of LSN afferents responded to alpha,beta-methylene adenosine 5'-triphosphate (alpha,beta-meATP; 1 mm), an effect that was concentration dependent and reversed by the P2X antagonist pyridoxyl5-phosphate 6-azophenyl-2',4'-disulphonic acid (PPADS) (100 microm). Significantly fewer PN afferents (7%) responded to alpha,beta-meATP. Correspondingly, 36% of colonic thoracolumbar DRG neurones exhibited P2X3-LI compared with only 19% of colonic lumbosacral neurones. Capsaicin (3 microm) excited 61% of LSN afferents and 47% of PN afferents; 82% of thoracolumbar and 50% of lumbosacral colonic DRG neurones displayed TRPV1-LI. Mechanically insensitive afferents were recruited by alpha,beta-meATP or capsaicin, and were almost exclusive to the LSN. Capsaicin-responsive LSN afferents displayed marked mechanical desensitization after responding to capsaicin, which did not occur in capsaicin-responsive PN afferents. Therefore, colonic LSN and PN pathways differ in their chemosensitivity to known noxious stimuli and their corresponding receptor expression. As these pathways relay information that may relate to symptoms in functional gastrointestinal disease, these results may have implications for the efficacy of therapies targeting receptor modulation.

The pattern of c-Fos immunoreactivity in the hindbrain of the rat following stomach distension.

It has been previously shown that the walls of the stomach contain vagal and splanchnic afferents, connected to low and high threshold (LT and HT) gastric receptors, that convey physiological and noxious information to areas of the hindbrain involved mainly in the control of gastrointestinal function. Because distension of the stomach also reflexly increases the sympathetic drive to the cardiovascular system, the present study was planned to examine the pattern of activation of all nuclei encountered throughout the hindbrain in response to gastric distension. In anaesthetized rats, the stimulus was controlled by employing different transmural pressures and frequencies of distension, and c-Fos immunohistochemistry was used to characterize neuronal activation. Low intensity stimulation induced c-Fos expression in the cranial part of nucleus of solitary tract (NTS), the nucleus ambiguus (NA), the lateral reticular area (LRt) and the ventrolateral medulla (RVL/CVL). At low frequency of stimulation c-Fos positive nuclei (p.n.) were found in NTS only. At high frequency of stimulation an increase in c-Fos immunoreactivity was found. High intensity stimulation induced c-Fos expression in area postrema (AP), the lateral vestibular nucleus (LVe) and the caudal part of the NTS. At low frequency, only the number of c-Fos p.n. was increased. Increasing the frequency of stimulation induced c-Fos expression in further nuclei such as the parabrachial nucleus (PBN), the inferior olive subnuclei (IOn), the oral part of spinal trigeminal nucleus (Sp5O) and locus coeruleus (LC). At higher frequencies c-Fos immunoreactivity decreased in NTS and LRt, disappeared in VLM and increased in NA. Thus stomach distension activated several neuronal excitatory and inhibitory circuits that are involved in the control of gastrointestinal function as well as in cardiovascular, respiratory and pain regulation. The differences in c-Fos immunoreactivity induced by changing the distension patterns suggested interactions between groups of vagal and splanchnic afferents.

Anatomical study of lumbar spine innervation.

To precisely evaluate low back pain, identification of the detailed innervation of the lumbar spine is necessary. On twenty-five sides of adult cadavers we investigated various patterns of rami communicantes (RC) and their relationship to the psoas major muscle (PM). In ten sides, we focused our dissection on the minute nerve supply of the anterior (ALL) and posterior longitudinal ligaments (PLL), vertebral bodies and the intervertebral discs (IVD). According to the mode of piercing PM, two types of RC were observed: superficial oblique rami (SOR) and deep transverse rami (DTR). SOR ran obliquely between superficial heads of PM, connecting sympathetic trunk (ST) and T12-L2 (3) spinal nerves non-segmentally. DTR ran segmentally close to the vertebral bodies and were situated deep to the PM slips. On the lateral side of the lumbar spine, the vertebral bodies and IVD received branches from DTR and ventral rami segmentally, as well as branches from the sympathetic trunk (ST) and, in the upper lumbar region, SOR non-segmentally. On the anterior aspect of the lumbar spine, ALL received branches from ST and splanchnic nerves non-segmentally. Within the vertebral canal, the posterior aspect of IVD and PLL received the sinu-vertebral nerves originating from DTR. These findings suggest the coexistence of two different types of innervation: one originating directly from the spinal nerve segmentally, and one reaching vertebral structures via the sympathetic nerves non-segmentally. Therefore, sympathetic nerves are likely involved in the proprioception of the spinal column.

Cholecystokinin selectively affects presympathetic vasomotor neurons and sympathetic vasomotor outflow.

Cholecystokinin (CCK) is a potential mediator of gastrointestinal vasodilatation during digestion. To determine whether CCK influences sympathetic vasomotor function, we examined the effect of systemic CCK administration on mean arterial blood pressure (MAP), heart rate (HR), lumbar sympathetic nerve discharge (LSND), splanchnic sympathetic nerve discharge (SSND), and the discharge of presympathetic neurons of the rostral ventrolateral medulla (RVLM) in alpha-chloralose-anesthetized rats. CCK (1-8 microg/kg iv) reduced MAP, HR, and SSND and transiently increased LSND. Vagotomy abolished the effects of CCK on MAP and SSND as did the CCK-A receptor antagonist devazepide (0.5 mg/kg iv). The bradycardic effect of CCK was unaltered by vagotomy but abolished by devazepide. CCK increased superior mesenteric arterial conductance but did not alter iliac conductance. CCK inhibited a subpopulation (approximately 49%) of RVLM presympathetic neurons whereas approximately 28% of neurons tested were activated by CCK. The effects of CCK on RVLM neuronal discharge were blocked by devazepide. RVLM neurons inhibited by exogenous CCK acting via CCK-A receptors on vagal afferents may control sympathetic vasomotor outflow to the gastrointestinal tract vasculature.

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.

Effect of splanchnectomy on jejunal motility and fos expression in brain stem after intestinal anaphylaxis in rat.

This study was to determine whether alterations in jejunal motility observed after antigen challenge of sensitized rats occurred after extirpation of the celiac-superior mesenteric ganglia. Hooded-Lister rats were prepared with an intact or extirpated celiac-superior mesenteric ganglion, an isolated Thiry-Vella loop of ileum for instillation of antigen, and jejunal electrodes for myoelectric recording. Animals were sensitized by injection of 10 microg egg albumin (EA, ip), and specific anti-EA IgE titers were determined to be >1:64. In both control and splanchnectomized rats, normal fasting migrating myoelectric complexes (MMC) were observed before challenge with EA. MMCs were disrupted, and diarrhea was observed immediately after EA challenge of control but not splanchnectomized animals. Brain stems were removed and processed for Fos immunoreactivity. The absence of perivascular neuropeptide Y immunoreactivity in the submucosa was used to confirm the success of splanchnectomy. The number of Fos-immunoreactive neuronal nuclei was significantly reduced in the brain stem after splanchnectomy. Thus the mesenteric sympathetic ganglia are an integral part of the extramural neuronal pathways required for altered motility in this model of intestinal anaphylaxis.

Unmyelinated nerve fibre analysis of the human lesser splanchnic nerve.

The aim of the present study is to analyse unmyelinated nerve fibres of the human lesser splanchnic nerve in relation to the ageing process. With the help of an image-analyser, we examined 30 human lesser splanchnic nerves. The analysis was conducted with the use of a new staining method that makes it possible to discriminate various structures of the nervous tissue. Our report provides for the first time information on the ageing process of the human lesser splanchnic nerve fibres. The results indicate that a decrease in transverse area and perimeter of unmyelinated axons is one of the important changes occurring in the human lesser splanchnic nerve during the ageing process.

Myelinated nerve fiber analysis of the human greater splanchnic nerve.

The purpose of the present study is to analyze the human greater splanchnic nerve in relation to aging. We adopted a new staining method which makes it possible to discriminate various structures of the nervous tissue. We examined 25 human greater splanchnic nerves from cadavers for anatomy dissection. We measured the number, area and perimeter of axons. The results reveal that: (1) there is no correlation between age and the number of axons; (2) the mean area and perimeter of axons increase with age, but not the total area and perimeter. We compared these results with those for the lesser splanchnic nerve. These morphological changes in the greater splanchnic nerve may indicate a kind of compensation through axon hypertrophy for hypofunctions in abdominal organ control.

Antagonist precipitated clonidine withdrawal in rat: effects on locus coeruleus neurons, sympathetic nerves and cardiovascular parameters.

The goal of the present study was to examine the effect of clonidine withdrawal on the neural control of blood pressure. Rats were treated for 7-13 days with clonidine via osmotic minipumps (200 microg kg(-1) day(-1), s.c.). Controls received saline or were sham operated. Withdrawal was precipitated by the alpha2-adrenergic receptor (alpha2-AR) antagonist atipamezole. Most experiments were done under halothane anesthesia. Chronic treatment with clonidine did not change mean arterial pressure (MAP) or heart rate (HR) but raised femoral artery resistance and the activity of locus coeruleus neurons slightly. Atipamezole given to rats treated chronically with clonidine produced the following effects: no change in MAP, severe tachycardia, sustained increase in splanchnic sympathetic nerve discharge (SND; +75 +/- 13%), transient increase in lumbar SND (+23 +/- 7%), ON-OFF activity pattern in the locus coeruleus (LC). The ON phase of LC activity was synchronized with upswings of SND and with small changes in MAP. A second alpha2-AR antagonist, methoxyidazoxan, produced effects identical to those of atipamezole. Atipamezole given to control rats produced no effect on MAP, HR, SND or LC activity. Atipamezole reversed the hypotension, sympathoinhibition and bradycardia produced by acute administration of clonidine. In awake rats treated chronically with clonidine, atipamezole did not change MAP but produced arterial pressure lability and tachycardia. In conclusion, under anesthesia, selective alpha2-AR antagonists elicit a clonidine withdrawal syndrome that displays autonomic characteristics reminiscent of the spontaneous withdrawal syndrome found in awake rats. The most prominent features of this syndrome are tachycardia, sympathoactivation, lack of hypertension and an oscillating activity pattern of brainstem neurons leading to abrupt changes in SND and in MAP.

An electrophysiological study on the autonomic innervation of the mesenteric lymph node in the rat.

Splanchnic innervation of the mesenteric lymph node was studied by means of electrophysiological technique in the rat. The effect of intravenous (i.v.) injection of recombinant human interleukin-1beta (rhIL-1beta) on the activity of efferent nerve fibers innervating the mesenteric lymph node was observed in the urethane anesthetized rat. An i.v. injection of 10 ng rhIL-1beta caused a gradual increase in efferent activity which lasted longer than 90 min. Dose related responses were observed at doses of 1, 10 and 100 ng. The least effective dose was about 10 ng. The conduction velocities estimated in mesenteric nerve fibers to the lymph node distributed in the range of 1.9-0.9 m/s, and the mean velocity was 1.39+/-0.34 m/s (n = 5). These observations implicate the involvement in the neural modulation of immune function of the mesenteric lymph node.

Myelinated nerve fibre analysis of the human small splanchnic nerve.

The aim of the present study is to analyse nerve fibres of the human small splanchnic nerve in relation to the ageing process. The analysis was conducted with the use of a new staining method that makes it possible to discriminate various structures of the nervous tissue. An image-analysing digitiser, a microscope with a drawing tube and a personal computer for storing data and performing statistical analyses were employed in this study. We examined 30 human small splanchnic nerves which were taken from cadaver specimens (20 males and 10 females) aged from 44 to 96 years. Our report may provide a first information concerning the ageing process of the human small splanchnic nerve. The results indicate that a decrease in transverse area and perimeter of myelinated axons in one of the important changes occurring in the human small splanchnic nerve with the ageing process.

Changes in proenkephalin gene expression in the developing hamster.

Proenkephalin (Penk) gene expression is high in the adult hamster adrenal medulla and it is comparable to that found in both the hamster and rat striatum. In addition, Penk gene expression in the hamster adrenal medulla is more typical of adult mammalian adrenals than the rat. Since the nature of Penk gene expression in the developing hamster adrenal is not known, it was examined and compared to that found in the striatum were adult levels in the adrenal and striatum are similar. The results show that Penk gene expression progressively increases in the developing hamster adrenal to peak on postnatal day 4. There is then a small decline to adult levels by postnatal day 12 when the morphology of the developing adrenal resembles the adult. Functional splanchnic nerve activity, as assessed by the ability of reserpine to induce increases in adrenal tyrosine hydroxylase mRNA, is not present until after postnatal day 4. Therefore, early increases in Penk gene expression are independent of splanchnic nerve activity. Adrenal EC peptides resulting from the developmental increases in Penk gene expression appear to be unprocessed and proenkephalin-like. This is based on the very low levels of free enkephalin (met-enkephalin) detected in the adrenals from both newborn and adult hamsters (1-5% of total EC peptide levels). In the developing hamster striatum, Penk gene expression remains low and unchanged until postnatal day 4 and increases six-fold by adulthood. Free enkephalin (met-enkephalin) levels remain high (between 36 and 88% of total EC peptide levels) in the developing and adult hamster striatum. Therefore the results show early increases in adrenal Penk gene expression in the developing hamster that are independent of splanchnic nerve activity and adult Penk gene expression which is high and dependent on splanchnic nerve activity. This differs from what is observed in the frequently studied rat. However, developmental changes in the hamster striatum are similar to those in the rat.

H2O2 sensitivity of afferent splanchnic C fiber units in vitro.

1. Single-unit impulse activity evoked by transient, focal application of hydrogen peroxide (H2O2) to identified visceral receptive fields has been characterized in an in vitro rat splanchnic nerve-mesentery preparation. In addition to H2O2 responsiveness, units were characterized in terms of sensitivity to mechanical stimuli, warming, and bradykinin. 2. Mesenteric receptive fields of single splanchnic afferent C fibers in vitro were located with the use of warm (approximately 45 degrees C saline) or mechanical search stimuli. After delimitation of the warm-sensitive and/or mechanosensitive receptive field, units were tested for responsiveness to transient, focal application of H2O2. Microliter volumes (usually 1 microliter) of H2O2 (88-880 mM) evoked responses in 25 of 42 (60%) units with identified warm-sensitive and/or mechanosensitive receptive fields, and in an additional 10 units for which H2O2 was the only effective stimulus. 3. Tachyphylaxis to repeated H2O2 stimulation was observed with interstimulus intervals <30 min, but did not indicate irreversible inactivation of the terminal, because 1) during this period warm and mechanical stimuli elicited responses equal to or greater than those before H2O2 treatment, and 2) H2O2 sensitivity was restored after units were allowed to recover. 4. Eight units unresponsive to an initial dose of H2O2 responded vigorously to a repeated application at the same site, suggesting a potentiating effect of prior H2O2 exposure. 5. Sixty-two percent (8 of 13) of H2O2-responsive units, but no (0 of 6) H2O2-unresponsive units responded to transient, focal bradykinin (9-90 nM) application. 6. An indirect mode of H2O2-evoked afferent excitation in some units was suggested by several observations, including the prolonged (up to 8 min) duration of the response of some units to transient H2O2 application, and the occasionally long (>2 min) response latencies to focal application of H2O2 to defined receptive fields. 7. Excitation of splanchnic neurons by H2O2 may be relevant to the modulation of reactive oxygen species production by immunocompetent cells, because sensory neuropeptides contained in these afferent fibers are known to influence the respiratory burst of macrophages and neutrophils.

Role of vagal and splanchnic capsaicin-sensitive afferents in enterogastric inhibition of acid secretion in rats.

The role of capsaicin-sensitive afferent innervation and neural pathways involved in the enterogastric inhibition of gastric acid secretion by luminal acid was investigated in urethan-anesthetized rats. Intestinal perfusion with graded concentrations of HCl (50, 75, and 100 mM) for 1 h dose dependently inhibited both thyrotropin-releasing hormone (TRH) analogue- and pentagastrin-stimulated acid output (P < 0.01). The inhibitory effect of intestinal perfusion with HCl (100 mM) on pentagastrin-stimulated acid secretion was blocked by bilateral vagotomy, whereas celiac ganglionectomy had no effect. Systemic capsaicin pretreatment (125 mg/kg sc) reduced the antisecretory effects of luminal acid on both TRH analogue- and pentagastrin-stimulated acid secretion. Neither selective perivagal nor selective periceliac capsaicin treatments (1% solution) modified the antisecretory effect of intestinal perfusion with HCl (75 mM) on TRH analogue-stimulated acid secretion. However, combined selective perivagal plus periceliac capsaicin treatment reduced it to the same extent as systemic capsaicin treatment. We conclude that enterogastric inhibition of acid secretion by luminal acid in urethan-anesthetized rats is mediated by extrinsic reflexes involving both vagal and splanchnic capsaicin-sensitive afferent fibers.

Raphe pallidus excites a unique class of sympathetic preganglionic neurons.

The responses of splanchnic sympathetic preganglionic neurons (SPNs) to stimulation in raphe pallidus and in rostral ventrolateral medulla (RVLM) were compared to determine the basis for the excitatory responses evoked in the whole splanchnic preganglionic nerve bundle. Most (88%) of the SPNs with a short-latency (32 ms) excitatory response to RVLM stimulation were unaffected by raphe pallidus stimulation, although 12% were excited at a long latency (123 ms). Each of the SPNs with long-latency (114 ms) excitatory responses to RVLM stimulation was also excited by raphe pallidus stimulation at latencies (106 ms) that were 7 ms (P < 0.01) shorter than those evoked from the RVLM. Antidromic activation of raphe pallidus neurons from both the T8 intermediolateral nucleus (98 ms) and from the RVLM (18 ms) indicated that their spinally projecting axons emit collaterals (mean conduction time: 12 ms) into the ventrolateral medulla. In conclusion, the short-latency (70 ms) splanchnic nerve excitation evoked by RVLM stimulation is mediated primarily by SPNs that do not respond to raphe pallidus stimulation. Similarly, the long-latency (162 ms) splanchnic excitation evoked from the raphe pallidus is mediated primarily by SPNs that do not respond to the rapidly conducting sympathoexcitatory pathway from the RVLM. The long-latency (169 ms), RVLM stimulus-evoked excitation of the splanchnic nerve may arise from action potentials conducted on the axonal branches of raphe spinal neurons.