Neuronal nitric oxide synthase in the neural pathways of the urinary bladder

Nitric oxide (NO) is a unique biological messenger molecule. It serves, in part, as a neurotransmitter in the central and peripheral nervous systems. Neurons containing NO have been identified histochemically by the presence of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) reactivity or immunohistochemically by the antibody for neuronal NO synthase (n-NOS). Previous histochemical or pharmacological studies have raised the possibility that NO may play an important role in the neural pathways of the lower urinary tract. There is also considerable evidence to suggest that n-NOS is plastic and could be upregulated following certain lesions in the lower urinary tract. The present review summarises the distribution of n-NOS containing neurons innervating the urinary bladder and the changes of the enzyme expression in some experimentally induced pathological conditions.     

Immunohistochemical localisation of neuronal nitric oxide synthase in the rainbow trout kidney

The distribution of nitrergic nervous structures in the trout kidney was studied by peroxidase-linked ABC immunostaining procedures using a polyclonal antibody raised against the neuronal isoform of nitric oxide synthase. The nitrergic plexus reaches the kidney along the vasculature, mainly running with the postcardinal vein where nitrergic fibres, microganglia like cellular clusters and isolated neurones were detected. The atubular head-kidney only showed isolated nitrergic fibres close to the larger arteries. On the other hand, the collecting tubules, collecting ducts, large arteries and glomerular arterioles of the tubular middle and posterior trunks were innervated by nitrergic fibres even though immunoreactive neurones were also observed in close apposition to some tubular elements and large arteries. These results suggest that, according to morphofunctional differences between the fish and mammalian kidneys, nitrergic neural structures may be involved in the control of particular renal functions in the rainbow trout.     

Modulation of neuroeffector transmission in guinea-pig pulmonary artery and vas deferens by exogenous nitric oxide

Cederqvist , B. & Gustafsson , L. E. 1994. Modulation of neuroeffector transmission in guinea-pig pulmonary artery and vas deferens by exogenous nitric oxide. Acta Physiol Scand 150, 75–81. Received 11 March 1993, accepted 30 July 1993. ISSN 0001–6772. Department of Physiology and Institute of Environmental Medicine, Karolinska Institute, Stockholm, Sweden. Blockade of nitric oxide (NO) synthesis enhances contractile responses to transmural nerve stimulation in guinea-pig pulmonary artery, indicating neuromodulation by endogenous nitric oxide. In the present study, neuromodulatory effects of exogenous NO were examined in guinea-pig pulmonary artery and vas deferens. Application of NO as acid nitrite, to the guinea-pig pulmonary artery, inhibited the contractile response to transmural nerve stimulation as well as contractions to exogenous noradrenaline. The inhibition occurred in a dose-dependent manner. Acid nitrite did not affect stimulation-induced release of [³H]noradrenaline in guinea-pig pulmonary artery. In the guinea-pig vas deferens, contractile ‘twitch’ responses to brief (25 pulses) transmural nerve stimulation and stimulation-induced release of [³H]noradrenaline was unaffected by acid nitrite. However, ‘twitch’ responses to prolonged stimulation were inhibited by acid nitrite, and tonic ‘hump’ responses were either enhanced or unaffected by acid nitrite. In conclusion, exogenous nitric oxide exerted an inhibition of adrenergic neurotransmission post-junctionally in guinea-pig pulmonary artery. In guinea-pig vas deferens, exogenous nitric oxide affected adrenergic and/or non-adrenergic non-cholinergic neurotransmission in a complex fashion, however without alteration in noradrenaline release. The data support a role for NO in modulation of neuroeffector transmission, especially by modulation of effector sensitivity, whereas modulation of noradrenaline release seems an unlikely role for NO in these and similar peripheral tissues.     

The innervation of rainbow trout (Oncorhynchus mykiss) liver: protein gene product 9.5 and neuronal nitric oxide synthase immunoreactivities

We have explored the innervation of the rainbow trout ( O. mykiss ) liver using immunohistochemical procedures and light microscopy to detect in situ protein gene product 9.5 and neuronal nitric oxide synthase immunoreactivities (PGP-IR and NOS-IR). The results showed PGP-IR nerve fibres running with the extralobular biliary duct (EBD), hepatic artery (EHA) and portal vein (EPV) that form the hepatic hilum, as well as following the spatial distribution of the intrahepatic blood vessel and biliary channels. These nerve fibres appear as single varicose processes, thin bundles, or thick bundles depending on their diameter and location in the wall of the blood vessel or biliary duct. No PGP-IR fibres were detected in the liver parenchyma. NOS-IR nerve fibres were located only in the vessels and ducts that form the hepatic hilum (EBD, EHA, EPV); in addition, NOS-IR nerve cell bodies were found isolated or forming ganglionated plexuses in the peribiliary fibromuscular tissue of the EBD. No PGP-IR ganglionated plexuses were detected in the EBD. The location of the general (PGP-IR) and nitrergic (nNOS-IR) intrinsic nerves of the trout liver suggest a conserved evolutionary role of the nervous control of hepatic blood flow and hepatobiliary activity.     

Electron microscopic study of intrinsic cardiac ganglia in the adult human

The aim of the present study was to describe in detail the ultrastructure of intrinsic cardiac ganglionic cells in the healthy human as these cells appear to be directly involved in the development of tachycardia, atrioventricular block, ventricular fibrillation, and sudden cardiac death. Tissues examined in this study were obtained from hearts of 10 adult humans of either sex aged 22-80 years at autopsy performed no more than 8 h after death. The examined human intrinsic cardiac nerve cells were in most respects typical autonomic neurons surrounded by a sheath of satellite cells that was either uni- or multilayered. In addition to regular unmyelinated axons, prominent large axon terminals containing lamellated dense bodies, mitochondria and vesicles in the cytoplasm were observed in the ganglion neuropil. Synaptic profiles were more common in the ganglion neuropil than on neuronal somata. According to axon terminal contents, synaptic profiles were of three types. The most common Type 1 synaptic profiles contained a predominance of small clear, with a few larger dense-cored vesicles and mitochondria. Type 2 synaptic profiles, in addition to the same components as in Type 1, had glycogen-like particles. Type 3 vesicle-containing profiles clearly differed from both the previous ones as they were the largest in diameter and included plentifiul large clear pleomorphic or dense-cored vesicles together with small clear and larger dense-cored vesicles, mitochondria, dense and multivesicular bodies. Independently of age of the human, the most frequent neuronal abnormality was an abundant accumulation of inclusions inside of somata and dendrites that, in profile, appeared like circular membranous or fine granular bodies variable in electron density. In addition to inclusions, some neuronal somata and dendrites had strongly swollen mitochondria filled up with granular material in spite of their close association with normal looking ganglionic neurons. Structures resembling an axon growth cone in profile were revealed inside of cardiac ganglia derived from an 80 year old man. In conclusion, the present results provide baseline information on the normal ultrastructure of intracardiac ganglia in healthy humans which may be useful for assessing and interpreting the degree of damage of ganglionic cells both in autonomic and sensory neuropathies of the human heart.     

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 β-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.     

Immunohistochemical mapping of nitric oxide synthase in the rat hypothalamus and colocalization with neuropeptides

The localization and distribution of nitric oxide synthase in the hypothalamus have been studied with an immunohistochemical technique using antibodies to neuronal rat nitric oxide synthase. Subsequent double-labeling experiments examined the colocalization patterns of nitric oxide synthase and several peptides. Our results demonstrate a widespread occurrence of nitric oxide synthase-immunoreactive nerve cell bodies and processes throughout the hypothalamus, especially in various parts of the preoptic region, in the supraoptic and paraventricular nuclei, the lateral hypothalamic area, the ventromedial and dorsomedial nuclei, the arcuate nucleus and various parts of the mammillary region. Double labeling experiments showed that nitric oxide synthase-like immunoreactivity coexists with substance P-like immunoreactivity in the medial preoptic area, with oxytocin-, cholecystokinin- and galanin message-associated peptide-like immunoreactivity in the supraoptic nucleus, with enkephalin-, oxytocin- and corticotropin releasing factor-like immunoreactivity in the paraventricular nucleus and with enkephalin-like immunoreactivity in the arcuate nucleus. Furthermore, in the ventromedial nucleus, nitric oxide synthase-like immunoreactivity coexisted with enkephalin-, substance P-, and somatostatin-like immunoreactivity, and in the dorsomedial nucleus with enkephalin-, galanin message-associated peptide- and substance P-like immunoreactivity. In the mammillary region nitric oxide synthase-like immunoreactivity coexisted with enkephalin-, cholecystokinin-, and substance P-like immunoreactivity. Among these neuropeptides, enkephalin and substance P were most frequently found in nitric oxide synthase-immunoreactive neurons. We conclude that nitric oxide synthase-immunoreactive neurons contain neuropeptides in various parts of the hypothalamus, and that nitric oxide in the hypothalamus may be involved in a variety of neuroendocrine and autonomic functions.     

Fine-structural and degenerative features in adult and aged human synpathetic ganglion cells

Sympathetic ganglia, either cervical and upper thoracic or lumbar, were removed from adult and aged patients suffering from circulatory deficiencies in their upper on lower extremities, respectively. The ganglion cells had several features that can be associated with age. Lipofuscin was ample in all ganglion cells and was usually polarly concentrated. Its amount tended to increase with age and it was present also in the glia cells. Lipofuscin autofluorescence often prevented the visualization of the formaldehyde induced fluorescence for catecholamines. At the electron microscopic level, pigment bodies were seen to be composed of three different kinds of osmiophilic properties: (1) gray component that had (2) dark patches dispersed into it and (3) pale, oval, incorporated droplets. In principal ganglion cells, the frist two formed the major part, the pale one taking over in the small granule-containing cells. Various inclusion bodies included a cylinder-shaped type that had a varying pattern of rod-like structures inside it. Myelin figures (laminar bodies) were sometimes found to fill neurite profiles, occasionally with random mitochondrial accumulations. These bore a distant resemblance to the primitive type of neuritic (senile) plaques, although none of the patients was diagnosed to have, for example, Alzheimer's disease or senile dementia of the Alzheimer type. Nevertheless, it appears to us that it might be possible to find coexistent neuropathological changes in peripheral sympathetic ganglia in diseases affecting primarily the central nervous system.     

The origins of the adrenergic fibres which innervate the internal anal sphincter, the rectum, and other tissues of the pelvic region in the guinea-pig

Summary The adrenergic innervation of the pelvic viscera was examined by the fluorescence histochemical technique, applied to tissue from untreated guinea-pigs and from guinea-pigs in which nerve pathways had been interrupted at operation. It was found that adrenergic neurons in the inferior mesenteric ganglia give rise to axons which run in the colonic nerves and end in the myenteric and submucous plexuses and around the arteries of the distal colon. In the rectum, part of the innervation of the myenteric plexus and all of the innervation of the submucous plexus comes from the inferior mesenteric ganglia. The rest of the adrenergic innervation of the myenteric plexus comes from the posterior pelvic ganglia or the sacral sympathetic chains. The innervation of the blood vessels of the rectum is from the posterior pelvic ganglia. Adrenergic nerves run from the sacral sympathetic chains and pass via nerves accompanying the rectal arteries to the internal anal sphincter. Other adrenergic fibres to the internal anal sphincter either arise in, or pass through, the posterior pelvic plexuses. The anal accessory muscle is innervated by adrenergic axons arising in the posterior pelvic plexuses. Adrenergic nerves which run in the pudendal nerves, probably from the sacral sympathetic chains, innervate the erectile tissue of the penis.This work was supported by grants from the Australian Research Grants Committee and the National Health and Medical Research Council. We thank Professor G. Burnstock for his generous support.     

The relation between in vitro. efflux of noradrenaline from platelets and vas deferens in man

The efflux of isotope-labelled noradrenaline from platelets and vas deferens was compared in 29 healthy males. Platelets and a preparation of tissue from vas deferens were incubated with isotope-labelled noradrenaline until equilibrium in the uptake was obtained. The spontaneous efflux of noradrenaline in buffer was measured for 20 min. There was a significant positive correlation between the efflux of noradrenaline from platelets and vas deferens (r 0.56, P < 0.001).     

The ramifications of adrenergic nerve terminals in the rectum, anal sphincter and anal accessory muscles of the guinea-pig

Summary The anatomy and the adrenergic innervation of the rectum, internal anal sphincter and of accessory structures are described for the guinea-pig. The distribution of adrenergic nerves was examined using the fluorescence histochemical technique applied to both sections and whole mount preparations. The longitudinal and circular muscle of the rectum and the muscularis mucosae are all supplied by adrenergic nerve terminals. The density of the adrenergic innervation of the muscularis externa increases towards the anal sphincter. There is a very dense innervation of the internal anal sphincter, of the anal accessory muscles and of the corrugator ani. Non-fluorescent neurons in the ganglia of the myenteric plexus are supplied by adrenergic terminals. The ganglia become smaller and sparser towards the internal anal sphincter and non-ganglionated nerve strands containing adrenergic axons run from the plexus to the sphincter muscle. Adrenergic fibers innervate two interconnected ganglionated plexuses in the submucosa. Very few adrenergic nerve cells were found in the myenteric plexus and they were not found at all in the submucosa. The extrinsic arteries and veins of the pelvic region are heavily innervated by adrenergic nerves. Within the gut wall the arteries are densely innervated but there is little or no innervation of the veins.This work was supported by grants from the Australian Research Grants Committee and the National Health and Medical Research Council. We thank Professor G. Burnstock for his generous support.     

Prostaglandins and nitric oxide in regional kidney blood flow responses to renal nerve stimulation

We examined the roles of cyclooxygenase products and of interactions between the cyclooxygenase and nitric oxide systems in the mechanisms underlying the relative insensitivity of medullary perfusion to renal nerve stimulation (RNS) in anaesthetized rabbits. To this end we examined the effects of ibuprofen and N^G-nitro-l-arginine (L-NNA), both alone and in combination, on the responses of regional kidney perfusion to RNS. Under control conditions, RNS produced frequency-dependent reductions in total renal blood flow (RBF; –82±3% at 6 Hz), cortical laser-Doppler flux (CLDF; –84±4% at 6 Hz) and, to a lesser extent, medullary laser-Doppler flux (MLDF; –46±7% at 6 Hz). Ibuprofen did not affect these responses significantly, suggesting that cyclooxygenase products have little net role in modulating renal vascular responses to RNS. L-NNA enhanced RBF (P=0.002), CLDF (P=0.03) and MLDF (P=0.03) responses to RNS. As we have shown previously, this effect of L-NNA was particularly prominent for MLDF at RNS frequencies 1.5 Hz. Subsequent administration of ibuprofen, in L-NNA-pretreated rabbits, did not affect responses to RNS significantly. We conclude that counter-regulatory actions of NO, but not of prostaglandins, partly underlie the relative insensitivity of medullary perfusion to renal nerve activation.     

Morphometric and ultrastructural changes with ageing in mouse peripheral nerve

Qualitative and quantitative information is reported on the morphological changes that occur in nerve fibres and nonneuronal cells of peripheral nerve during the lifetime of the mouse. Tibial nerves of mice aged 6–33 mo were studied. With ageing, collagen accumulates in the perineurium and lipid droplets in the perineurial cells. Macrophages and mast cells increase in number, and onion bulbs and collagen pockets are frequently present. Schwann cells associated with myelinated fibres (MF) slightly decrease in number in parallel with an increase of the internodal length from 6 to 12 mo, but increase in older nerves when demyelination and remyelination are common. The unmyelinated axon to myelinated fibre (UA/MF) ratio was about 2 until 12 mo, decreasing to 1.6 by 27 mo. In older mice, the loss of nerve fibres involves UA (50% loss of 27–33 mo cf. 6 mo) more markedly than MF (35%). In aged nerves wide incisures and infolded or outfolded myelin loops are frequent, resulting in an increased irregularity in the morphology of fibres along the internodes. In the mouse there is an adult time period, 12–20 mo, during which several features of degeneration progressively appear, and an ageing period from 20 mo upwards when the nerve suffers a general disorganisation and marked fibre loss.     

QT离散度与心自主神经的关系

ＱＴ离散度 (ＱＴｄｉｓｐｅｒｓｉｏｎ ,ＱＴｄ)定义为同步记录 1 2导联心电图中测量的最大ＱＴ间期 (ＱＴｍａｘ)与最小ＱＴ间期 (ＱＴｍｉｎ)之差[1 ] ,反映了心室肌复极不同步性和电不稳定性[2 - 4 ] 。这一概念已得到了心内膜和心外膜单相动作电位与体表心电图的对照研究[4,5] 及动物实验[6] 证实 ,大量临床研究也证明了ＱＴｄ与心室肌复极不同步性的高度相关[7- 9] 。近年来 ,对于ＱＴｄ的电生理基础及其本质引起广泛关注 ,人们认识到自主神经系统与心性猝死密切相关 ,交感过度激活可导致致命性心律失常发作 ,而迷走激活却有保护和抗…     

Evidence for a detrimental role of nitric oxide synthesized by endothelial nitric oxide synthase after peripheral nerve injury

Physical injury to a nerve is the most common cause of acquired peripheral neuropathy. Identification of molecules involved in degenerative and regenerative processes is a key step toward development of therapeutic tools in order to accelerate motor, sensory and/or autonomic function recovery. We have studied the role of nitric oxide (NO) using as a model the severe crushing of a motor nerve in adult rats. This type of injury up-regulates the three isoforms of nitric oxide synthase (NOS) in the affected nerve. Chronic systemic inhibition of NOS accelerated the onset of functional muscle reinnervation evaluated by the recording of compound muscle action potential evoked by electrical stimulation of the injured nerve. Besides, it increased the number of back-labeled motoneurons by application, 2 days after injury, of a retrograde marker 10 mm distal to the crushing site. These effects were mimicked by chronic specific inhibition of the endothelial isoform of nitric oxide synthase (eNOS), but not by specific inhibitors of the neuronal or inducible isoform. Next, we intraneurally injected a replication-deficient adenoviral vector directing the expression of a dominant negative mutant of eNOS (Ad-TeNOS). A single injection of Ad-TeNOS on the day of crushing significantly accelerated functional recovery of neuromuscular junction and increased axonal regeneration. Moreover, Ad-TeNOS did not compromise motoneuron viability or stability of reestablished neuromuscular junctions. Taken together, these results suggest that NO of endothelial origin slows down muscle reinnervation by means of detrimental actions on axonal regeneration after peripheral nerve injury. These experiments identify eNOS as a potential therapeutic target for treatment of traumatic nerve injuries and highlight the potential of gene therapy in treating injuries of this type using viral vectors to suppress the activity of eNOS.     

Cyclooxygenase and nitric oxide synthase in the presympathetic neurons in the paraventricular hypothalamic nucleus are involved in restraint stress-induced sympathetic activation in rats

Stress is one of the important factors to activate the sympathetic nervous system. We recently reported that central administration of corticotropin-releasing factor (CRF), known as a stress-related neuropeptide, increases the expression of both cyclooxygenase (COX) and nitric oxide synthase (NOS) in presympathetic neurons in the paraventricular hypothalamic nucleus (PVN). In the present study, therefore, we investigated whether brain COX and NOS can also mediate restraint stress (RS)-induced sympathetic activation by assessing the plasma catecholamine levels and neuronal activation of presympathetic neurons in the PVN. In addition, we examined effects of RS on the expression of both COX and NOS isozymes in the presympathetic PVN neurons. Intraperitoneal administration of an inhibitor for COX-1, COX-2 or inducible NOS (iNOS), but not for neuronal NOS (nNOS), reduced RS-induced elevation of plasma catecholamine levels and Fos expression in the presympathetic PVN neurons. Moreover, RS increased the expression of COX-1, COX-2 and iNOS in the presympathetic PVN neurons, whereas nNOS expression did not change. These results suggest that COX-1, COX-2 and iNOS in the presympathetic PVN neurons mediate acute RS-induced sympathetic activation.     

Borna disease virus P protein inhibits nitric oxide synthase gene expression in astrocytes

Borna disease virus (BDV) is one of the potential infectious agents involved in the development of central nervous system (CNS) diseases. Neurons and astrocytes are the main targets of BDV infection, but little is known about the roles of BDV infection in the biological effects of astrocytes. Here we reported that BDV inhibits the activation of inducible nitric oxide synthase (iNOS) in murine astrocytes induced by bacterial LPS and PMA. To determine which protein of BDV is responsible for the regulation of iNOS expression, we co-transfected murine astrocytes with reporter plasmid iNOS-luciferase and plasmid expressing individual BDV proteins. Results from analyses of reporter activities revealed that only the phosphoprotein (P) of BDV had an inhibitory effect on the activation of iNOS. In addition, P protein inhibits nitric oxide production through regulating iNOS expression. We also reported that the nuclear factor kappa B (NF-kappaB) binding element, AP-1 recognition site, and interferon-stimulated response element (ISRE) on the iNOS promoter were involved in the repression of iNOS gene expression regulated by the P protein. Functional analysis indicated that sequences from amino acids 134 to 174 of the P protein are necessary for the regulation of iNOS. These data suggested that BDV may suppress signal transduction pathways, which resulted in the inhibition of iNOS activation in astrocytes.     

Identification of nitric oxide synthase in human and bovine oviduct

Nitric oxide synthase (NOS) is responsible for the biologicalproduction of nitric oxide (NO) in several organs. NOS activityhas also been localized in the reproductive tract, althoughdirect evidence for its presence in the human or bovine oviductis still lacking. In the present study, four different techniqueswere used to identify the presence of NOS activity in human(n = 11) and bovine (n = 9) oviduct: (i) conversion of [³H]-L-arginineto [³H]-L-citrulline; (ii) production of nitrite/nitrate (NO₂/NO₃;stable NO metabolites); (iii) identification of NADPH-diaphoraseactivity; and (iv) immunostaining with antiserum to endothelialNOS. Cytosolic extracts from human ampullary segments of theFallopian tube, obtained from post-partum patients (n = 4),converted [³H]-L-arginine to [³H]-L-citrulline (21.0 ±8.8 fmol/mg protein/min). This conversion rate was significantly(P <0.05) reduced in the presence of either EDTA or N-monomethyl-L-argininemonoacetate (L-NMMA), an inhibitor of NOS activity. When bovine(n = 3) ampullary segments were incubated for 36 h in Hanks'balanced salt solution, the concentration of NO₂/NO₃ in themedium was increased (P <0.05) if segments were pretreatedwith lipopolysaccharide (LPS; an inducer of inducible NOS),but not after treatment with LPS + L-NMMA. Additionally, epithelialcells cultured from ampullary segments showed positive stainingboth for NADPH-diaphorase activity and with antiserum to endothelialNOS. The results of the present study provide direct evidencefor the presence of both the Ca²⁺ -dependent constitutive formof NOS, as well as the inducible form of NOS activity in humanand bovine oviduct. Since the oviduct plays a key role in thereproductive process, it is possible that the two forms of NOSmay be involved in the physiological regulation of oviduct function. bovine/human/nitric oxide/nitric oxide synthase/oviduct