Neuropeptide Y-like immunoreactivity in intramural ganglia of the newborn guinea pig urinary bladder

Immunoreactive neuropeptide Y (NPY) was demonstrated in neuronal elements in the urinary bladder wall of the newborn guinea pig. Numerous intramural ganglia were found lying among the smooth muscle bundles and in the submucosa, and NPY-like immunoreactive nerve cell bodies were demonstrated within all of these ganglia. Nerve fibres containing NPY were also richly distributed in the detrusor muscle, submucosa and around blood vessels. In dissociated cell cultures from newborn guinea pig detrusor muscle, a subpopulation (70-85%) of both mononucleate and binucleate intramural neurones was shown to contain NPY-like immunoreactivity. A low percentage (1-6%) of the intramural bladder neurones contained dopamine-beta-hydroxylase. In conclusion, while some NPY-containing nerve fibres in the wall of the bladder are of sympathetic origin, especially those supplying blood vessels, the results of this present study establish that many of these NPY-containing nerve fibres originate from non-adrenergic cell bodies within the intramural bladder ganglia.     

Study of the innervation of blood vessels of the digestive organs by fluorescence microscopy]

The blood vessels of the samll intestine and the gallbladder were shown to possess a great amount of adrenergic nerve fibres which, when penetrating the thickness of the wall of the above organs, become thinner and the distributed between the tissue structures of the organs as the thinnest monoaxonal network. The method of Falck--Hillarp--Krokhina was used. Among the vessel nerves there are perivascular nerves accompanying the vessels along their total legnth, juxtavascular and intramural nervous bundles of the sumpathetic nature detected by the fluorescent-microscopy method. Large arteries are disposed in a considerably thicker network of specifically fluorescing fibres than veins and small arteries.     

Noradrenergic and peptidergic innervation of the mammary gland in the immature pig

The presence and pattern of coexistence of some biologically active substances in nerve fibres supplying the mammary gland in the immature pig were studied using immunohistochemical methods. The substances studied included: protein gene product 9.5 (PGP), tyrosine hydroxylase (TH), somatostatin (SOM), neuropeptide Y (NPY), galanin (GAL), calcitonin gene-related peptide (CGRP) and substance P (SP). The mammary gland was found to be richly supplied by PGP-immunoreactive (PGP-IR) nerve fibres that surrounded blood vessels, bundles of smooth muscle cells and lactiferous ducts. The vast majority of these nerves also displayed immunoreactivity to TH. Immunoreactivity to SOM was observed in a moderate number of nerve fibres which were associated with smooth muscles of the nipple and blood vessels. Immunoreactivity to NPY occurred in many nerve fibres associated with blood vessels and in single nerves supplying smooth muscle cells. Solitary GAL-IR axons supplied mostly blood vessels. Many CGRP-IR nerve fibres were associated with both blood vessels and smooth muscles. SP-IR nerve fibres richly supplied blood vessels only. The colocalization study revealed that SOM, NPY and GAL partly colocalized with TH in nerve fibres supplying the porcine mammary gland.     

So-called muscular "bridges" and their interrelationship with blood vessels]

Fifty hearts of corpses of humans of different age groups were studied by means of macro-microscopical anatomical dissection of muscle fibres and cardiac blood vessels. The muscle bridges were found over the coronary arteries in all preparations of the heart. They were most frequently met on the anterior interventricular branch of the left coronary artery. The topography of muscle bridges is described and the miocardium muscle bundles participating in their formation are revealed. The character of the course of fibres in the muscle bridge is related to the course of the fibres of the muscle bundle forming it.     

Substance P-immunoreactive nerve fibres in the anterior segment of the rabbit eye

Summary Substance P-immunoreactive nerve terminals were found in several locations in the anterior segment of the rabbit eye. In the iris they occurred in the sphincter muscle and were randomly distributed in the iris stroma with some fibres running close to the dilator muscle. In the ciliary body these immunoreactive elements were few and occurred within bundles of nerve fibres, while in the ciliary processes they were more numerous with a predominantly subepithelial location. Blood vessels in the anterior uvea were often surrounded by substance P-immunoreactive fibres. No substance P-fibres were found in the cornea, while the sclera contained very few such elements.Using conventional in vitro techniques it was found that the sphincter pupillae muscle of the iris responded to electrical stimulation with a contraction that was resistant to cholinergic and adrenergic blockade, but was inhibited by the neuronal blocker tetrodotoxin. This indicates the existence of a non-cholinergic, non-adrenergic neuronal mediator of the contractile response. Exogenously applied substance P produced a long-lasting contraction of the spincter muscle, an observation compatible with the view that substance P is the noncholinergic, non-adrenergic neurotransmitter involved.     

The origin and distribution of adrenergic nerve fibres in the guinea-pig colon

Summary A detailed study of the origin and distribution of sympathetic fibres in the distal colon of the guinea-pig has been made using the fluorescent histochemical method for localizing catecholamines. The extrinsic adrenergic fibres of the colonie sympathetic nerves follow the inferior mesenteric artery and its branches to the colon. Some of the extrinsic adrenergic fibres are associated with the parasympathetic fibres of the pelvic nerves near the colon. Complete adrenergic denervation follows the removal of the inferior mesenteric ganglion or the destruction of the nerves running with the inferior mesenteric artery.No fluorescent fibres, other than those associated with blood vessels, were observed in air-dried stretch preparations of the isolated longitudinal muscle. However, a substantial number of varicose, terminal fibres, not associated with blood vessels, were observed in the circular muscle. Some varicose fibres, apart from those associated with ganglion cells, were observed in the myenteric plexus. These fibres were seen in the bundles of nerves running between the nodes of the plexus and also as single fibres which branched from the plexus to end in areas free of ganglion cells.Three plexuses of adrenergic nerve fibres have been distinguished in the submucosa: a dense plexus of terminal fibres innervating both the veins and arteries; a plexus consisting of innervated nodes of ganglion cells, connected by bundles of fluorescent and non-fluorescent nerves; and a plexus of varicose and non-varicose fibres, which is not associated with ganglion cells. Some groups of ganglion cells in the submucosa were without adrenergic innervation.A plexus of varicose fibres forms a meshwork in the lamina propria of the mucosa. The muscularis mucosae is sparsely innervated. Most of the blood vessels in the mucosa are not associated with adrenergic fibres.     

Pituitary adenylate cyclase activating polypeptide (PACAP) in the rat mammary gland

Pituitary adenylate cyclase activating polypeptide (PACAP), a member of the vasoactive intestinal polypeptide (VIP) family of peptides, is present in the brain and in neuronal elements of a number of peripheral organs. Since no information on PACAP in the mammary gland exists, we have investigated, by radioimmunoassay and immunohistochemistry, the occurrence and distribution of PACAP immunoreactivity in the mammary gland of lactating and non-lactating rats. A specific monoclonal mouse anti-PACAP antibody'has been used to show that the peptide is located in nerve fibres associated with bundles of circular and longitudinal smooth muscle surrounding the lactiferous duct of the nipple. PACAP-immunoreactive nerve fibres and nerve bundles are present in the subepidermal connective tissue of the nipple and in the mammary parenchyma, some of the fibres being in close contact with blood vessels. Occasionally, a few delicate varicose fibres are associated with secretory alveoli and lactiferous ducts. The majority of PACAP-positive nerve fibres are, however, located in the glabrous skin of the nipple and the hairy skin adjacent to the nipple forming a subepithelial plexus from which delicate varicose nerve fibres enter the overlying epithelium. Double immunostaining for PACAP and a marker for sensory neurons, calcitonin gene-related peptide, has disclosed that the two peptides are almost completely co-localized. A minor population of the PACAP-immunoreactive nerve fibres shows co-existence with VIP. Although no obvious changes at the immunohistochemical level could be observed during pregnancy or lactation, elevated concentrations of immunoreactive PACAP-38 in mammary extracts have been found during lactation. Our data suggest that PACAP is involved in the nervous control of mammary gland function, probably in the transmission of suckling stimuli.     

Peptide-containing nerves in the urinary bladder of the toad,Bufo marinus

The distributions of peptide-containing nerves in the urinary bladder of the toad, Bufo marinus, were studied by means of fluorescence immunohistochemistry of whole-mount preparations. The bundles of smooth muscle in the bladder are well supplied by varicose nerve fibres displaying somatostatin-like immunoreactivity; these fibres probably arise from intrinsic perikarya. The urinary bladder also has a well-developed plexus of nerves containing substance P-like immunoreactive material; these elements probably represent sensory nerves of extrinsic origin. Nerve fibres showing immunoreactivity to vasoactive intestinal polypeptide (VIP) or enkephalin are rare within the urinary bladder of the toad. It is considered unlikely that any of these peptides directly mediates the hyoscine-resistant excitatory response of the smooth muscle to nerve stimulation in the toad bladder.     

The distribution and origin of calcitonin gene-related peptide-containing nerve fibres in feline dental pulp

Summary The origin and distribution of calcitonin gene-related peptide (CGRP)-like immunoreactivity in feline dental pulp were studied using indirect immunofluorescence. Nerve fibres with varicosities exhibiting CGRP-like immunoreactivity were observed to enter the pulp with blood vessels. Many CGRP-containing nerve fibres were found to extend along blood vessels in the central pulp, and some of these fibres exhibited a network arrangement in the walls of dental pulp blood vessels. However, some of fibres were apparently not associated with blood vessels. Some thin, CGRP-containing nerve fibres formed a part of the nerve plexus in the subodontoblastic area and penetrated into the odontoblastic layer. In animals that had undergone transection of the inferior alveolar nerve, no CGRP-containing nerve fibres were observed. Application of a double-immunofluorescence staining technique also revealed that the distribution of CGRP-containing nerve fibres is very similar to that of substance P-containing nerve fibres.     

The distribution and origin of calcitonin gene-related peptide-containing nerve fibres in feline dental pulp. Relationship with substance P-containing nerve fibres

The origin and distribution of calcitonin gene-related peptide (CGRP)-like immunoreactivity in feline dental pulp were studied using indirect immunofluorescence. Nerve fibres with varicosities exhibiting CGRP-like immunoreactivity were observed to enter the pulp with blood vessels. Many CGRP-containing nerve fibres were found to extend along blood vessels in the central pulp, and some of these fibres exhibited a network arrangement in the walls of dental pulp blood vessels. However, some of fibres were apparently not associated with blood vessels. Some thin, CGRP-containing nerve fibres formed a part of the nerve plexus in the subodontoblastic area and penetrated into the odontoblastic layer. In animals that had undergone transection of the inferior alveolar nerve, no CGRP-containing nerve fibres were observed. Application of a double-immunofluorescence staining technique also revealed that the distribution of CGRP-containing nerve fibres is very similar to that of substance P-containing nerve fibres.     

An immunohistochemical study of serotonin-containing nerves in the colon of rats

The localization of the serotonin-like immunoreactive nerves of the rat colon was investigated by means of immunohistochemistry, utilizing an antibody against serotonin. In non-treated colons, serotonin-positive neuropils were consistently detected around the myenteric plexus. In pargyline-treated colons, serotonin-like fibres were demonstrated in association with either the small intramural blood vessels of the submucosa or the extramural nerve bundles. Treatment with 5-hydroxytryptophan (5-HTP) permitted the visualization of additional serotonin-immunoreactive fibres around the large extramural blood vessels. Immunoreactive nerve cell bodies were demonstrated in the myenteric plexus of colons treated with 5-HTP or colchicine. From these observations, it is suggested that the serotoninergic nerves of the rat colon comprise both intrinsic and extrinsic elements.     

An immunohistochemical study of serotonin-containing nerves in the colon of rats

Summary The localization of the serotonin-like immunoreactive nerves of the rat colon was investigated by means of immunohistochemistry, utilizing an antibody against serotonin. In non-treated colons, serotonin-positive neuropils were consistently detected around the myenteric plexus. In pargyline-treated colons, serotonin-like fibres were demonstrated in association with either the small intramural blood vessels of the submucosa or the extramural nerve bundles. Treatment with 5-hydroxytryptophan (5-HTP) permitted the visualization of additional serotonin-immunoreactive fibres around the large extramural blood vessels. Immunoreactive nerve cell bodies were demonstrated in the myenteric plexus of colons treated with 5-HTP or colchicine. From these observations, it is suggested that the serotoninergic nerves of the rat colon comprise both intrinsic and extrinsic elements.     

The differential distribution of vasoactive intestinal polypeptide in the normal human female genital tract

Summary VIP-like immunoreactive material is present in the female reproductive tract, with a distinct pattern of distribution. The highest concentrations of extractable material and immunoreactive nerve fibres were found in the cervix and vagina. In the cervix these fibres were seen below the surface epithelium and around cervical glands as well as in association with blood vessels and smooth muscle bundles. In the vagina the nerve fibres were most abundant in the superficial regions of the lamina propria. Scattered fibres were also present in the rest of the uterus and in the fallopian tubes. Chromatographic evidence indicates that this VIP-like material is of a similar molecular size to that extracted from other organs. Possible roles for VIP in the regulation of myometrial activity and of cervical and vaginal dilation and secretion are proposed.     

The palisade endings of cat extraocular muscles: a light and electron microscope study.

The palisade endings (PEs), a particular type of nerve ending found only in extraocular muscles of mammals, have been studied using both silver-stained teased preparations and electron microscope techniques. They have been found, in act, in both the proximal and distal muscle insertions of the four recti and the two oblique mucles. PEs are exclusively associated with some of the mitochondria-poor, multiply-innervated muscle fibres present in the globar layer os these muscles, and consist of a multitude of terminal branches embracing the extremity of the muscle fibre and penetrating the infoldings formed by the muscle fibre at its tendinous attachment. The whole formation is surrounded by a thin capsule. These nerve endings present striking similarities to the developing Golgi tendon organ; the terminal branches lying among the collagen fibrils and occasionally making 'sensory-like' close contacts with the muscle fibre are disposed in such a way that they could easily have a sensory role. It was concluded that PEs present sufficient morphological evidence to be considered as sensory, encapsulated, myotendinous receptors, each related to a single multiply-innervated muscle fibre.     

Structure of the perineurium of the peripheral nervous system]

The structure of the perineurium in different parts of the peripheral nervous system of rats, rabbits and cats was studied by light-optical and electron microscopic methods. The structure of the perineurium in all the animals studied is sim8lar and consists of different number of the epithelial type layers of the perineural cells, with bundles of cooagnous fibres between them. The greatest anount of layers is found in the perineurium of the sensory and vegetative ganglia, their amount being less between the nerve trunks and bundles. Solitary sensory mielinated nerve fibres are surrounded with a perineural etui consisting of one or two cellular layers. The thickness of the perineural cells varies from 300 to 1500 A and only in the nucleus field it is equal to 1-2 mu. Every layer of the perineural cells is surrounded by a basal membrane. In their cytoplasm there are many pinocytic vesicles in addition to main organells. Between the perineural cells there exist close contacts. The internal layer of the perineurium is the place of origin of intraganglionic septa and in certain distance surrounds the vessels entering the ganglion. Ultrastructurally the perineural cells are similar to the endothelium of the vessels.     

Distribution of cholinergic and catecholaminergic nerves in the colon of the rat with aganglionosis.

We compared localization and distribution of putative cholinergic fibers by acetylcholinesterase and of adrenergic fibers visualized by the glyoxylic acid technique in the aganglionic segment using whole mount preparations of aganglionosis rat (AGR) and compared them with those of normal littermates. We also attempted simultaneous staining of acetylcholinesterase (AChE) and catecholamine fluorescence (C-F) on the same whole mount preparations to compare the differences in distribution pattern. All AGR used in this study had narrowed segments of the bowel extending from the distal ileum to the anus, and had no ganglion cells in these narrowed segments. In the intermuscular space, normally occupied with myenteric ganglion, of the narrowed distal colon and rectum, various sizes of nerve bundles and fibers reactive for AChE and C-F appeared to make coarse and irregular networks. These thick nerve bundles appeared to ascend to the proximal colon and disappeared in the cecum. In the distal ileum, almost totally absence of AChE positive nerve fibers, but a few fine C-F fibers, probably associated with blood vessels, were observed. By the method of simultaneous staining of AChE and C-F method in the whole mount preparations, the thick nerve bundles in the narrowed segments showed both of AChE positive and C-F positive. However, there were differences in peripheral fine nerve fibers in the segment; especially numerous perivascular C-F positive nerve fibers, but a few AChE positive ones were found. In the upper aganglionic narrowed segments, greatly diminished numbers of AChE positive and C-F positive nerve fibers were found in the circular muscle layer and in the submucosal layer. In the lower aganglionic narrowed segments, there were thick nerve bundles, forming irregular interlaced network. The role of these extrinsic nerve fibers in aganglionic segments is unclear.     

A histochemical study of the innervation of cerebral blood vessels in the snake

Summary Dual innervation of snake cerebral blood vessels by adrenergic and cholinergic fibres was demonstrated with the use of histochemical methods. Although the nerve plexuses are somewhat less dense, the essential features of innervation of the blood vessels are similar to those of mammals with the exception that the adrenergic plexuses are more prominent than the cholinergic plexuses. The major arteries of the cerebral carotid system have a rich nerve supply. However, the innervation is less rich in the basilar and poor in the spinal (vertebral) arteries. Although the arteries supplying the right side of head are poorly developed, three pairs of arteries, cerebral carotids, ophthalmics and spinals, supply the snake brain. The carotids and ophthalmics are densely innervated and are accompanied by thick nerve bundles, suggesting that the nerves preferentially enter the skull along those arteries. Some parenchymal arterioles are also dually innervated. Connection between the brain parenchyma and intracerebral capillaries via both cholinergic and adrenergic fibres was observed. In addition cholinergic nerve fibres, connecting capillaries and the intramedullary nerve fibre bundles, were noticed. Capillary blood flow may be influenced by both adrenergic and cholinergic central neurons. The walls of capillaries also exhibit heavy acetylcholinesterase activity. This may indicate an important role for the capillary in the regulation of intracerebral blood flow.     

Variations in atrioventricular valve innervation in four species of mammals

In this series of studies, the innervation patterns of whole-mount preparations of bicuspid and tricuspid valves were studied by light microscopy in the mouse, rat, guinea pig, and opossum. The acetylcholinesterase-positive networks of nerve fibers showed many similarities in the basic patterns of valve innervation in all of the species studied, but several interspecies variations were observed. The basal zone of the valve adjacent to the fibromuscular atrioventricular ring displayed the most dense plexus of nerves, with acetylcholinesterase-positive fibers being seen across the width of the valve. In the intermediate zone of the valve, less dense plexuses of nerve fibers were found; and these were more numerous in the cuspal areas and less numerous in the intervening commissural areas. In the distal portions of the valve, nerve networks arborized extensively, with some of their nerve fibers extending toward the chordae tendineae and the free edges of the valve cusps. Only in the guinea pig and opossum did these fibers reach the free margin of the valve cusp, where they either ended directly as free nerve endings or lay parallel to the free edge of the cusp, often running between adjacent chordae tendineae. Although the patterns of innervation were similar in both bicuspid and tricuspid valves, the innervation density of the bicuspid valve was greater than that of the tricuspid valve for each species examined. A distinguishing feature of guinea pig and opossum tricuspid valves was that their chordae tendineae were relatively more prominent and more densely innervated than the bicuspid chordae tendineae. Free nerve endings with no light microscopic evidence of specialization were present throughout the bicuspid and tricuspid valves of all species studied. Some nerve endings in the opossum showed evidence of specialization, with brush-like arborizations leading to presumed free terminals seen chiefly in the distal zone of the valve cusps. Although some general tendencies were apparent, we have demonstrated that interspecies heterogeneity exists in the terminal networks of the atrioventricular valves of mouse, rat, guinea pig, and opossum.     

Substance P immunoreactivity in rat von Ebner's gland

Summary The present immunohistochemical study revealed substance P-immunoreactive neuronal elements in the von Ebner's gland of rats. Immunoreactive ganglion cells were observed as single cells or groups of several immunoreactive ganglion cells among intra-lingual muscles, at the base of the vallate papillae and near the von Ebner's gland. Very numerous substance P-immunoreactive varicose nerve fibres ran closely associated with the serous cells and excretory duct cells, and were seen to run along blood vessels in the gland. Since substance P-immunoreactive ganglion cells were present near the glands, the immunoreactive varicose nerve fibres in the von Ebner's gland may be partly derived from the intra-lingual ganglion cells. These substance P-immunoreactive varicose nerve fibres may have an effect on the secretory activity of the serous cells and duct cells, and on the vasodilation of blood vessels of the von Ebner's gland. Actin immunoreactivity was seen in numerous myoepithelial cells embracing serous cells and duct cells, and in the smooth muscle cells of blood vessels of the gland. By using a double immunolabelling technique with anti-substance P and anti-actin sera, substance P-immunoreactive varicose nerve fibres were found to be in close contact with myoepithelial cells.     

Venous valves in the large branches of superior vena cava

In 97 human cadavers the valves of the following blood vessels were investigated with regard to their cusps and their sizes and positions: the internal jugular veins, the subclavian veins, the brachiocephalic veins, and the superior vena cava. The cusps of each of the valves, which consist of two or three parts, are neither always of equal size nor obligatorily sufficient. Unipartite valves may be sufficient as well as insufficient. Internal jugular veins: The inferior bulb of the internal jugular vein is provided with valves which in 6% of the cases consist of three parts, in 66% of two parts, and in 15% of only one cusp. The concave margins of most of them go down as far as the venous angle. The convex edges attached to the wall of the vein extend to a higher level on the right side than on the left. In 13% there do not exist any valves. Varieties are described separately in this paper. Subclavian veins: Valves are found along the length of the vessel. Only few of them reach the venous angle. In rare cases there exist two valves: one at the beginning, the other at the end of the subclavian vein. In 4% of the cases the valves consist of three, in 75% of two cusps. In 12% they are unipartite. In 9% there are no valves to be found. The right side is more often without valves than the left. Brachiocephalic veins: Only a minority of these vessels is provided with valves. Most of these consist of one cusp, are insufficient, and are situated in the left innominate vein.(ABSTRACT TRUNCATED AT 250 WORDS)