Autonomic innervation of the intestine from a baby with megacystis microcolon intestinal hypoperistalsis syndrome: I. Immunohistochemical study.

A typical case of megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is reported. The patient, an infant girl, was fed only by total intravenous nutrition and is now 3 years old. The distribution of several gut peptides was examined in the resected small intestine using an immunohistochemical method. Vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM)-containing nerve fibers were decreased; however, substance P- and leucine enkephalin (Leu-ENK)-containing fibers were increased. The imbalance between several kinds of gut peptides might be one the causes of aperistalsis in MMIHS. This is the first report about the gut peptides of MMIHS.

[Innervation of the nasal mucosa of the guinea pig].

The autonomic and the peptidergic nerve fibers which control the blood flow and gland secretion in the nasal mucosa of the guinea pig were studied using histochemical and immunohistochemical techniques. There was a dense distribution of adrenergic nerve fibers (Adn) around the blood vessels, especially the vein, but a very sparse distribution in the glandular tissue. There was a dense distribution of cholinergic nerve fibers (Chn) both around the blood vessels and in the glandular tissue. The distribution of fibers containing vasoactive intestinal polypeptide (VIP) was dense in the glandular tissue but sparse around blood vessels and very sparse in the subepithelial layer. Fibers containing calcitonin gene-related peptide (CGRP) were densely distributed around blood vessels, glandular tissue and the subepithelial layer. A moderate number of CGRP containing fibers were observed in the intraepithelial layer. Fibers containing substance P (SP) and neurokinin A (NKA) were sparsely distributed around blood vessels, glandular tissue and the subepithelial layer. Radioimmunoassay for NKA and SP in the nasal mucosa revealed a greater content both of NKA and SP in the nasal concha than in the nasal septum. There was a greater content of NKA than SP in both the nasal septum and the nasal concha, with a higher SP/NKA ratio in the nasal concha than in the nasal septum.

Studies on the structure and intrinsic innervation of the normal human prostate.

Prostates from eight adult males (age range 17-63 years) were employed in this investigation. Six glands were obtained from multiorgan transplantation donors at the time of organ harvesting, and two additional glands, at routine post-mortem. Microscopic examination has shown that the majority of acini in the peripheral parts of the gland were individually relatively small and lined by an epithelium two to three cells in thickness. In contrast, acini in the central part of the gland were usually large and contained numerous prominent epithelial folds within the lumen. On the basis of morphology, a clear distinction between peripheral and central acini was not observed. The distribution of autonomic ganglia both within and adjacent to the prostate was studied, and nerve cell bodies containing both acetylcholinesterase (AChE) activity and neuropeptide immunoreactivity were identified. The distribution and density of AChE-positive nerves associated with smooth muscle in either the peripheral or central parts of the prostate were indistinguishable. In addition, the majority of acini in peripheral and central regions possessed a rich subepithelial plexus of autonomic nerves. VIP-positive nerve fibers were found in relation to the epithelium lining acini in central and peripheral regions of the gland. In contrast, nerves possessing NPY immunoreactivity occurred only in relation to the smooth muscle trabeculae of the prostate.

Immunohistochemical detection of GTP-binding regulatory protein (Go) in the autonomic nervous system including the enteric nervous system, superior cervical ganglion and adrenal medulla.

The localization of a GTP-binding regulatory protein, Go, in the autonomic nervous system including the enteric nervous system, superior cervical ganglion, and adrenal medulla, has been immunohistochemically examined by use of affinity-purified antibody against the alpha-subunit of Go. In the small intestine, dense Go-immunoreactive products were localized on the enteric nervous system, i.e. the myenteric plexus of Auerbach and the submucosal plexus of Meissner. In the superior cervical ganglion, presynaptic terminals were strongly immunoreactive to the Go antibody. The adrenal medulla was stained with this antibody, but the adrenal cortex was not immunoreactive to this antibody. Thus, the present study strongly suggests that Go is localized in the autonomic nervous system and plays its role in transmembrane signal transmission in this system.

Neuropeptides in interscapular and perirenal brown adipose tissue in the rat: a plurality of innervation.

The occurrence of neuropeptides in rat brown adipose tissue has been investigated. Immunohistochemical studies on interscapular and perirenal brown fat have demonstrated unequivocally the presence of substance-P (SP)-like, neuropeptide-Y (NPY)-like and calcitonin gene related-peptide (CGRP)-like immunoreactive elements (putative nerves) in adventitial distribution on inter- and intralobular supply arteries and in accompanying nerve bundles. At a more peripheral level, some NPY-like immunoreactive elements and a greater number of CGRP-like immunoreactive elements were observed in the parenchymal field. Somatostatin, bombesin, neurotensin, enkephalin, and vasoactive-intestinal-polypeptide immunoreactivities were not detected. No differences in neuropeptide distribution were noted between interscapular and perirenal brown fat. There is a degree of coincident distribution of SP, NPY and CGRP with that of noradrenergic nervous elements as visualized by condensation histochemistry. Since after 6-hydroxydopamine treatment not all the nerve terminals in rat brown adipose tissue are stigmatized (earlier report), the present results have been discussed in the light of a possible pluralism in innervation of brown adipose tissue.

Substance P and cholecystokinin-like immunoreactive varicosities in somatosensory and autonomic regions of the rat spinal cord: a quantitative study of coexistence.

Substance P- and cholecystokinin-like immunoreactivities have been shown to coexist in some, but not all, dorsal root ganglion cell bodies of the rat. Quantitative immunofluorescence techniques were used in the present study to describe densities of substance P- and cholecystokinin-like immunoreactive varicosities in several spinal cord nuclei. By combining simultaneous immunofluorescent techniques on one tissue section with computerized image processing, coexistence of substance P- and cholecystokinin-like immunoreactivity in varicosities was also quantified. By comparing spinal cord regions between normal and unilateral dorsal rhizotomy affected animals, densities of substance P- and cholecystokinin-like immunoreactive varicosities associated with primary afferent neurons were established. To determine the densities of immunoreactive varicosities that were related to unmyelinated primary afferent fibers, data were compared between normal animals and those treated neonatally with capsaicin. Four major observations were made: (1) Substance P- and cholecystokinin-like immunoreactivity coexist in populations of varicosities in sensory and autonomic regions of rat spinal segment L6. (2) Within the superficial laminae of the dorsal horn, varicosities containing both substance P- and cholecystokinin-like immunoreactivity and cholecystokinin-like immunoreactivity alone are of primary afferent neuron origin, but those containing only substance P-like immunoreactivity are most likely of spinal or descending neuronal origin. (3) Capsaicin-insensitive cholecystokinin-like immunoreactive varicosities were present predominantly in lamina I. These data suggest some cholecystokinin-like immunoreactive varicosities are associated with myelinated primary afferent neurons. (4) Primary afferent fibers containing substance P- and cholecystokinin-like immunoreactivity project to intermediate gray regions of the rat spinal cord. A large proportion of these fibers are capsaicin sensitive, suggesting that they are unmyelinated.

Autonomic neurons supplying the rat eye and the intraorbital distribution of vasoactive intestinal polypeptide (VIP)-like immunoreactivity.

We traced the origin and path of autonomic nerves to the rat eye using, as an aid to dissection, a modified thiocholine method for the histochemical demonstration of cholinesterase. When applied to whole nerves and ganglia supplying the rat eye, this procedure is not specific for cholinergic neurons; instead it stains both sympathetic and parasympathetic nerves, many of which are otherwise too fine to identify in dissection. We found that nerves from the superior cervical and pterygopalatine ganglia form a plexus at the orbital apex corresponding to the retro-orbital plexus found in rabbit, monkey and man. In the rat, nerves from the retro-orbital plexus travel peripherally to the superior surface of the optic-nerve sheath. Here, they fuse with long ciliary nerves and the post-ganglionic nerves from the ciliary ganglion to form another dense nerve-fiber plexus that ultimately supplies the eye. Importantly, the plexus on the optic nerve contains many isolated or aggregated ganglion cells. These are comparable in number to those in the ciliary ganglion itself and are assumed to be accessory ciliary neurons. Using immunohistochemistry, we also sought evidence for vasoactive intestinal polypeptide in these ganglia and nerves. As previously known, many pterygopalatine ganglion cells are immunoreactive for this peptide. Vasoactive intestinal polypeptide (VIP)-like immunoreactive nerve fibers are present in nerves from the retro-orbital plexus to the optic-nerve sheath plexus, in most nerves of the latter plexus, and in most nerves entering the eye. Furthermore, a small proportion of nerve cells in the main and accessory ciliary ganglia also are immunoreactive for VIP. We conclude that in addition to the pterygopalatine ganglion, the ciliary ganglion and its accessory ganglia are sources of VIP-like immunoreactive nerves in the rat eye.

[Gamma-aminobutyric acid receptors in the central nervous system of mammals]. / Retseptory gamma-aminomaslianoi kisloty v tsentral'noi nervnoi sisteme mlekopitaiushchikh.

The review describes properties of gamma-aminobutyric acid (GABA) receptors in the central nervous system of mammalians. Two pharmacologically different receptors--GABAA and GABAB--are involved into GABA-ergic inhibition. GABA receptors (bicuculline-sensitive) regulate chloride permeability of membranes and can be functionally enhanced by benzodiazepine and barbiturate. GABA receptors (bicuculline-insensitive) are suggested to regulate calcium permeability.

Autoradiographic localization and characterization of spinal cord substance P binding sites: high densities in sensory, autonomic, phrenic, and Onuf's motor nuclei.

The presence of the neurotransmitter or neuromodulator, substance P (SP), in the spinal cord implies that a discrete localization of SP receptors also occurs. To map the distribution of and to characterize SP binding sites in the spinal cord, light microscopic autoradiography was used. SP binding sites occurred in the dorsal horn, intermediolateral cell column (IML) and lamina X-region. In the ventral horn, the phrenic, Onuf's and sacral ventromedial motor nuclei were densely labeled. Other regions of the ventral horn were moderately labeled for SP binding sites. The localization of binding sites parallels the regional distribution of SP-containing nerve fibers in the spinal cord. A close correlation between the binding sites for SP and the presence of cholinesterase-stained neurons occurred, and suggest that the SP receptors are located on or proximal to cholinergic neurons. The density of the binding sites in the dorsal horn was highest in the sacral section, followed by the lumbar, thoracic and cervical section. In the lamina X region, however, the density was highest in the thoracic followed by the sacral, lumbar and cervical sections. The high density binding of 125I-Bolton-Hunter reagent labeled SP was inhibited, in a dose-dependent manner, by unlabeled SP. Quantification of the dose-dependent inhibition binding, using computer densitometry, showed differences in the inhibition curves for the cervical lamina X-region and the IML as compared with the other loci containing high density binding sites. The differential sensitivity of the SP receptors to unlabeled SP suggests that there are heterogeneous receptors for SP in the spinal cord, which may be relevant to the role of SP in different spinal cord functions. The binding to specific motor nuclei in the ventral horn also suggest that SP may play a role in the function of specialized striated muscles.

Localization of some neuropeptide- and serotonin-like immunoreactivities in the vegetative network of guinea pig spinal cord.

The localization of Substance P(SP)-, Methionine-Enkephalin(met-Enk)-, Somatostatin(SOM)- Serotonin(SER)-, Cholecystokinin(CCK)-, and Vasoactive intestinal polypeptide (VIP)-like immunoreactivity (-LIR) has been determined immunocytochemically in the thoracic spinal cord intermediate zone of male and female guinea pigs. All neuroactive substances studied are exclusively localized in nerve fibre varicosities and terminals building up the vegetative network of the thoracic spinal cord intermediate zone. This network is situated dorsally to the central canal as a longitudinal plate of approximate thickness of 90-100 microns. Immunoreactive fibres are observed in the two Fasciculi longitudinales laterales and the two Fasciculi longitudinales mediales which are interconnected by transverse and oblique peptide-containing bundles (the terminology used by Petras and Cummings 1972; Galabov and Davidoff 1976). All these bundles interconnect the nuclei intermediolaterales principales and funiculares, the nuclei intercalates spinales and the nuclei intercalates paraependimales in ipsi- and contralateral as well as in rostral and caudal direction. The neurones of these nuclei are surrounded by immunoreactive varicosities and terminals. The quantity of the immunoreactive structures and intensity of the staining varied for the different neuroactive substances. As to the origin of the vegetative network immunoreactive fibres three main possibilities exists: a). From primary afferent neurones situated in the dorsal root ganglia, which send their axons via the dorsal roots (mainly for SP and perhaps for CCK); b). From supraspinal neurones which send their axons descending in the white matter funiculi and in the fasciculi longitudinales laterales and mediales and c). From intrinsic spinal cord neurones, which send their neurites in ascending and descending directions, ipsi- and contralaterally and interconnect the spinal cord segments. The different origin of the vegetative network immunoreactive fibres as well as the complex innervation of the preganglionic sympathetic nerve cells in the intermediate zone of the spinal cord suggests that this network may play an important role in the integration of the central and peripheral vegetative nervous system as well as probably in the integration of the somatic and the vegetative nervous system.

Norepinephrine and serotonin in central autonomic nuclei in the spontaneously hypertensive rat and two normotensive control rats.

Neurochemical analysis of norepinephrine and serotonin with high performance liquid chromatography with electrochemical detection (LCEC) in several microdissected central autonomic nuclei of 4 week old and 16 week old spontaneously hypertensive rats (SHR), normotensive Wistar-Kyoto rats (WKY), and control normotensive Wistar rats (WIS), revealed some differences among these strains, but only one change that correlated with the hypertensive state. Norepinephrine levels in the 4 week old SHR were greater in the parabrachial nuclei and the dorsal motor nucleus of X than in the WKY, but levels in the WIS normotensive also were greater than in the WKY, and equalled levels in the SHR. In the 16 week old rats, no difference was noted between NE levels in the SHR and WKY strains, but levels in the locus coeruleus and parabrachial nuclei of the WIS were greater than in both the SHR and WKY. Serotonin levels in the 4 week old SHR and WKY rats did not differ, while levels in the paraventricular nucleus, locus coeruleus, parabrachial nuclei, and medullary raphe nuclei of the WIS rat were greater than one or both of these inbred strains. In the dorsal motor nucleus of X in 16 week old rats, serotonin levels were greater in the SHR than in either the WKY or WIS controls, suggesting one possible transmitter action that should be explored further for its potential relationship to the hypertensive state.(ABSTRACT TRUNCATED AT 250 WORDS)

Noradrenergic nervous activity in migraine.

The autonomic nervous function in patients with migraine was studied during headache-free intervals. The following observations were made: (1) a decrease in overshoot in Valsalva's maneuver; (2) orthostatic hypotension; (3) low levels of plasma norepinephrine in the steady state; (4) failure in elevation of the plasma norepinephrine level after head-up tilting; (5) dilatation of the pupils after instillation in the eye of 1.25% epinephrine; and (6) a long recovery time in tests by bolus injection of 0.1 microgram of norepinephrine bitartrate per kilogram. The above findings suggest that patients with migraine show sympathetic hypofunction together with denervation hypersensitivity of the iris and the arteries, and that a defective noradrenergic nervous system may play a role in the pathogenesis of migraine.

The regional distribution of neurotensin-like immunoreactivity in central and peripheral tissues of the cat.

The regional distribution of neurotensin-like immunoreactivity (NTLI) was studied by radioimmunoassay in central and peripheral tissues of the cat. In the brain, NTLI showed a wide distribution with highest concentrations in the hypothalamus, the caudate/putamen and the nucleus accumbens. Only low levels of NTLI were measured in the spinal cord and there was no difference between dorsal and ventral horn. In the periphery, NTLI was present in high concentrations in the adrenal medulla and in lower amounts in the superior cervical and the ciliary ganglion. NTLI was present in both lobes of the pituitary gland and throughout the gastrointestinal tract with high concentrations in the ileum. All other peripheral tissues tested contained low but detectable amounts of NTLI. Gel chromatography on Sephadex G-25 was used in order to characterize the immunoreactive material; NTLI in tissue extracts from 3 central and 3 peripheral tissues co-eluted in a single peak at the position of synthetic neurotensin. NTLI is widely distributed throughout cat tissues and there are important differences from the distribution pattern in the rat, the only other species examined in detail to date.

Autonomic innervation of the skin in primary erythermalgia.

We observed the autonomic nerve plexuses in the skin of an erythermalgic patient and a normal individual using the methods of acetylcholinesterase (AChE) histochemistry, catecholamine histofluorescence, and electron microscopy. The density of both AChE-positive and catecholamine-containing nerve terminals in the periarterial and sweat glandular plexuses was greatly reduced in the erythermalgic foot skin compared with those in unaffected skin from the same patient and in the foot skin of a normal individual. Ultrastructurally, the terminal axons containing either agranular (cholinergic) or small dense-cored (adrenergic) vesicles were present in the periarterial and periglandular regions of the erythermalgic skin, but the occurrence of these nerve terminals in the involved skin appeared to be much reduced in frequency compared with uninvolved skin and the skin of a normal individual.

The autonomic innervation of the human greater saphenous vein.

The existence of a double catecholaminergic and cholinergic innervation was demonstrated in the human greater saphenous vein. Catecholamine-containing nerve fibres are organized in a network-like plexus localized at the adventitial-medial border. Acetylcholinesterase-containing nerve fibres are arranged in a plexus found at the adventitial-medial border as well. Catecholamine and acetylcholinesterase-containing nerve fibres, while localized in close apposition since they occupy the same portion of the vein, represent two distinct and independent populations of nerve fibres coming likely from the sympathetic and parasympathetic sections of the autonomic nervous system respectively. Our findings demonstrating a close relationship between catecholaminergic and cholinergic nerve fibres within the wall of the human greater saphenous vein offer morphological support to physiological and pharmacological results reported in the literature of a presynaptic control exerted by cholinergic nerves on norepinephrine release at the level of the saphenous vein.

Peptidergic nerves.

In recent years the autonomic nervous system has been shown to consist of nerves containing a number of different neurotransmitter substances; a system far more complex than was originally thought. It has been demonstrated that a large part of the autonomic nervous system contains peptides, and that, in the gut, these nerves form a major complex that infiltrates the entire length and breadth of the tract. The detailed study of this peptidergic system has been facilitated by the development of specialised immunocytochemical methods, which have yielded information on the distribution and morphology of the various types of nerve. Thus, the distribution of these nerves can often be seen to parallel what is known about the actions of individual peptides. The majority of peptidergic nerves are intrinsic to the gut, forming, with the other intrinsic nerves, what appears to be a largely autonomous unit. This enteric system can be visualised as a "minibrain", under the general influence of the central nervous system but able to function, to a certain extent, by itself. The consequences of a breakdown in the normal functioning of this system can be seen in a number of gut diseases.

Vipergic nerves in the penis.

High concentrations of vasoactive intestinal polypeptide (VIP) were detected by immunocytochemistry and radioimmunoassay in thirty surgical specimens of male external genitalia. VIP was found exclusively in fine autonomic nerves. VIPergic nerves were most densely concentrated in the penis around the pudendal arteries and in the erectile tissue of the corpus cavernosum. Considerable numbers of VIP nerve fibres were also seen in the vas deferens and epididymis. VIP is known to exert regulatory actions on blood-flow, secretion, and muscle tone. Its presence in considerable amounts in the male genital tract suggests that this newly discovered peptide neurotransmitter may be important in the nervous control of male external genitalia.