Cholinergic and adrenergic innervation of the bladder obstructed by urethral stenosis in the rat

In the obstructed bladder by urethral stenosis the morphometry studies show a decrease of the cholinergic and adrenergic intrinsic innervation in a different way according the tissue layer and the experimental delay. Adrenergic nerves are more affected than cholinergic ones but they recover more quickly. In the adventitia and the sub-mucosa first affected by the high pressures inside the bladder the density of the autonomous nerves decreases from the first week. The perivascular and muscular nervous fibers are less sensitive and the similar shape of their innervation histogram gives evidence of a tight bond between their adrenergic network. Beside their motor participation the adrenergic terminal fibers may play a role in the cellular trophicity of the muscle.     

Comparative study of the autonomic innervation of the mammalian ovary,with particular regard to the follicular system

Summary The autonomic innervation of the ovary was studied in 12 mammalian species utilizing the cholinesterase method in combination with pseudocholinesterase inhibition for the cholinergic component, and glyoxylic acid histochemistry together with fluorometric determination of noradrenaline for the adrenergic component. Ovaries from cow, sheep, cat, and guinea pig were very richly supplied with adrenergic nerves in the cortical stroma, particularly enclosing follicles in various stages of development. In the follicular wall the nerve terminals were located in the theca externa, where they ran parallel to the follicular surface. Numerous adrenergic terminals also surrounded ovarian blood vessels. The adrenergic innervation was of intermediary density in the human ovary and in the pig, dog, cat, and opossum. Ovaries from rabbit, mouse and hamster had a sparse adrenergic nerve supply. The amount of intraovarian adrenergic nerves agreed well with the tissue concentration of noradrenaline in the various species. The cholinergic innervation was generally less well developed, but had the same distribution as the adrenergic system around blood vessels and in the ovarian stroma, including follicular walls.     

A comparative study of the adrenergic innervation of the iris and ciliary structures in 18 species in phylogenesis

A detailed topography of adrenergic innervation in invertebrates (lobster), low vertebrates (fish, amphibians, reptiles, birds), and nine species of mammals is presented. Flack and Hillarp's specific fluorescent histochemical method using freeze-dried material was used. Phylogenetically, adrenergic innervation appeared earlier under the ciliary epithelium and in the muscle than surrounding the vessels, and in all species many fibers were without any connection to the vessel walls. Adrenergic innervation was very rich in the dilator muscle extending toward the epithelium of the posterior chamber; a surprisingly rich network was found in the sphincter muscle and also in ciliary spaces of some species. Numerous fluorescent mast cells were visualized in the pecten of the bird eye and in the ciliary tissue of the sheep and cow.     

Adrenergic innervation of the male reproductive ducts in some mammals

Summary The distribution of adrenergic nerves to the male reproductive ducts were studied by applying the highly specific method of Falck and Hillarp for the cellular demonstration of the adrenergic transmitter. In rats, guinea-pigs, and rabbits, the ducts of the testis and the caput epididymidis were found to lack adrenergic varicose terminals, as they also lack cholinesterase positive structures. Adrenergic terminals extend in the vas deferens and epididymis from the prostatic end as far as the upper mid-corpus level of the ductus epididymidis, occurring in the muscle layers and, except in the rat, in the lamina propria. In the cat, ductuli efferentes and caput epididymidis also receive an adrenergic innervation.For skilful technical assistance we are grateful to Mrs. Ulla Flyger and Miss Berith Hansson. The investigation has been supported by a research grant (B 66-257) from the Swedish Medical Research Council.On sabbatical leave from the Biology Department, University of Oregon, Eugene, Oregon, with the aid of a Senior Research Fellowship from the National Council of Child Health and Human Development, National Institutes of Health, U.S. Department of Health, Welfare and Education. The use of facilities of the Department of Physiology I, Karolinska Institutet, with the kind permission of Professor U. S. von Euler and Research Docent R. Eliasson during the course of this study also is greatly appreciated.     

The development of innervation in the rat atrioventricular node

Summary The problem of development of the innervation of the rat atrioventricular node has been investigated by electron microscopy. Nerve bundles appear in relation to the node as early as the second postnatal day and vesiculated axons are seen throughout the entire node by the fourth day. Intimate contacts between nodal cells, axons and terminal varicosities are frequently observed.Use of the 5-hydroxydopamine tracer technique has enabled the identification of both cholinergic and adrenergic axons. It is concluded that the node has a dual innervation although cholinergic endings far outnumber those classified as adrenergic on the sixth postnatal day.These results are quite different to earlier findings made at the light microscope level and the discrepancies are discussed with respect to the histochemical techniques used. The suggestion that nodal differentiation is induced by nerves is considered in relation to the differences in cholinesterase activity exhibited by nodal cells during normal development and following neonatal sympathectomy.     

Adrenergic innervation of the ureters, urinary bladder, and urethra in pigs

Studies were conducted on 4 sexually mature and 4 immature pigs. Scraps of the ureters, urinary bladder, and urethra were cut with a freezing microtome. Fluorescence method of Torre and Surgeon (1976) was used to reveal the adrenergic innervation. It was found that the ureters were weakly supplied with the adrenergic nerves; most of the nerves were located in the muscular and submucosal membranes. Apex of the urinary bladder possessed the weakest innervation. More nerves were found in particular layers of the bladder corpus whereas bladder trigonum and cervix possessed numerous nerves. Adrenergic innervation of the urethra was similar to that of the urinary bladder's cervix. Adrenergic nerves were present in the serous and muscular membranes of both the urinary bladder and the urethra. Part of the nerve fibres was connected with blood vessels of the organs under study.     

Fluorescence microscopical and chemical characterization of the adrenergic innervation in mammalian liver tissue

Liver tissue from 12 different mammalian species was studied with a fluorescence histochemical technique for the cellular localization of amines (Falck-Hillarp technique) and with a chemical method for the determination of norepinephrine (HPLC-technique). Adrenergic nerve plexus were found in interlobular blood vessels derived from the portal vein and hepatic artery. Varicose adrenergic nerve fibres were, generally, seen to branch from the fibres around the blood vessels and to enter the liver parenchyma, where they formed a randomly distributed intralobular network. The density of these intralobular fibres showed marked species variation. Human liver and liver from the rhesus monkey, baboon, cynomolgus monkey and guinea pig showed a high density of parenchymal adrenergic nerves. Rabbit, cat, pig, cow and horse liver formed an intermediate group, having fewer varicose adrenergic nerve fibres but an unequivocal distribution of these nerves to the liver parenchyma. In rat and mouse liver no parenchymal innervation could be demonstrated. The density of the parenchymal innervation generally correlated with the concentration of norepinephrine in the liver tissue.     

Histochemical and ultrastructural data on the adrenergic and cholinergic innervation of the epididymis in the mouse

The adrenergic and cholinergic innervation of the epididymal duct in the mouse has been investigated using histochemical methods and electron microscopy. This study demonstrates that the epididymal innervation of the mouse does not really differ from that of other mammals. Cholinergic nerves are mainly vascular, even in the cauda where cholinesterase activity is increased within the tubular musculature. Catecholamine fibres ensure the motricity of the wall of the canalicular system, particularly the terminal segment where the smooth muscle fibres are specifically differentiated.     

Adrenergic Innervation of Bowel in Hirschsprung's Disease

The adrenergic innervation of normal and aganglionic regions of bowel from patients with Hirschsprung''s disease was investigated by a fluorescent histochemical technique. In normal bowel the adrenergic nerves end about intramural ganglion cells. In aganglionic bowel the adrenergic nerves form a dense varicose plexus in both muscularis externa and muscularis mucosae. It is suggested that the cause of megacolon in Hirschsprung''s disease is due to a lack of nervous pathways controlling the intrinsic reflexes, which is probably congenital in origin.     

Sympathetic innervation of the pulmonary artery,ascending aorta,and coronar glomera of the rabbit

Summary Adrenergic nerve fibres were demonstrated in the connective tissue of the rabbit coronar glomera by means of the formaldehyde-induced fluorescence technique for catecholamines. This type of innervation is similar to the adrenergic nerve supply to the rabbit and cat carotid body. Adrenergic fibres terminate subendothelially and only a few can be traced to type I cells in the glomera coronaria. The sympathetic innervation of the ascending aorta is exceedingly sparse in contrast to the pulmonary trunk, while vasa vasorum of the ascending aorta exhibit a dense sympathetic innervation.     

Adrenergic innervation of the kidneys in man and various laboratory animals

By means of the neurohistochemical method for slice incubation in 2% solution of glyoxylic acid, innervation of the kidneys of a 57-year-old man after a sudden cardiac death has been investigated, as well as innervation of the kidneys in white rat, rabbit, guinea pig and cat. A rich adrenergic innervation in the organ's blood vessels has been revealed. In particular, adrenergic nervous fibers have been found along the course of afferent glomerular arterioles. Together with innervation of the proximal and distal convoluted tubules, a high density of the terminal adrenergic nervous plexus is revealed along the course of the nephron loops. Adrenergic nervous plexuses of high density are found in the area of the initial part of the urinary excretory pathways and their connection with nervous plexuses of the kidney itself.     

Vasomotor innervation in the hind limb muscles of the normal and 6-hydroxydopamine treated cat

Summary Vasomotor nervous distribution to arteries in the cat gastrocnemius and soleus muscles was investigated in the normal and 6-hydroxydopamine treated animal. Muscle samples were processed for the demonstration of acetylcholinesterase by a thiocholine method and for the presence of catecholamines by a formol fluorescence technique. In isolated perfused muscle preparations noradrenaline or adrenaline increased perfusion pressure while acetylcholine or isoprenaline decreased this pressure. A beaded fluorescent periarterial plexus (not visualised after reserpine treatment) was observed. Vasomotor axons, all of which were ACHE-negative, exhibited degenerative changes following 6-hydroxydopamine injection, while ACHE-positive axons outside the adventitial sheath were unaffected. No indication of cholinergic periarterial nerves was found and our evidence suggests that the vasomotor innervation is adrenergic sympathetic in nature.This work was supported by a grant from the Medical Research Council. During the tenure of a Wellcome Trust Fellowship, Dr. F. Joó is on leave of absence from Jozsef Attila University, Szeged, Hungary. The electron microscope is on permanent loan to one of us (J. D. L.) from the Wellcome Trust. We are most grateful to Mrs. G. Howells and Miss D. Morgan for technical assistance and to Mr. P. Hire for photographic help.     

Innervation of the renal cortex.

Morphologic studies of renal innervation have utilized the methods of histochemistry and electron microscopy. Much information has been derived from examination of the renal cortex in monkey and rat. Fluorescence histochemistry shows a rich adrenergic innervation. Acetylcholinesterase can be demonstrated histochemically in the renal nerves by light and electron microscopy. Studies in the rat using 6-hydroxydopamine, a drug that selectively destroys adrenergic nerves, indicate that the glomerular arterioles and surrounding tubules are innervated by adrenergic nerves containing acetylcholinesterase. Distinct neurovascular and neurotubular junctions are observed the electron microscope. They are anatomically consistent with being the sites of synaptic transmission. Ultrastructural analysis of serial sections reveals that single individual axons contact multiple vascular cells and renal tubules. We now have a considerable body of information concerning the morphology of renal innervation are are beginning to appreciate the role of the renal nerves in kidney function.     

Function of the anal sphincters following anterior resections of rectum

The purpose of the present work is to evaluate the function of the anal sphincter following anterior resections of the rectum. Our data have shown that the anorectal reflex did not depend on the presence of the rectal mucosa. The external anal sphincter contraction involves a reflex which is initiated by stretch receptors in the levator ani muscle. The internal anal sphincter relaxation is likely a local reflex involving nerve pathways not yet completely defined.     

Integrative control of rectoanal reflex in guinea pigs through lumbar colonic nerves

The aim of the present study was to analyze the neuromodulation of rectoanal reflex activity by lumbar sympathetic nerves in guinea pigs. The mechanical activities of the rectum were recorded with a balloon connected to a pressure transducer, and those of the internal anal sphincter (IAS) were recorded with a custom-made strain gauge force transducer. Gradual and sustained rectal distension evoked the rectoanal reflex, causing cholinergic contractions of the rectum and synchronous nitrergic relaxations of the IAS. Section of the lumbar colonic nerves enhanced both rectal contractions and IAS relaxations. Section of the 13th thoracic cord abolished both rectal contractions and IAS relaxations, but section of the lumbar colonic nerves restored them. Lumbar sympathectomy and pithing sacral cords greatly diminished these rectal contractions and IAS relaxations, but the intrinsic reflex component remained. NG-nitro-L-arginine methyl ester enhanced the intrinsic reflex-mediated contraction of the rectum and abolished reflex-mediated relaxation of the IAS and converted into cholinergic contractions. The present results indicate that the extrinsic lumbar inhibitory outflow causes marked inhibition of the rectoanal reflex via the lumbar colonic nerves.     

Autonomic innervation of the ocular choroid membrane in the chicken

Summary The distribution pattern of adrenergic fibres innervating the ocular choroid membrane of the chicken was studied by means of fluorescence and electron microscopy. In addition, the origin of these fibres was investigated after superior cervical ganglionectomy. Adrenergic axons reach the choroid, partly forming the perivascular plexuses and partly running in the choroid nerves and the choroidal branches of the ciliary nerves. The axon terminals distribute to the smooth muscle cells of the arterial wall and to the extensive system of smooth muscle cells of the intervascular stroma. After unilateral ganglionectomy, fluorescent fibres almost completely disappeared, and degenerative changes could be observed in the terminal varicosities on both smooth muscle cell populations. These findings suggest that the adrenergic axons either originate from neurones within the ipsilateral superior cervical ganglion, or pass through this ganglion. The persistence of normal terminals in short- and long-term ganglionectomised animals shows that the vasal and intervascular muscle cells of the choroid membrane are provided with both an adrenergic and a cholinergic innervation.This work was supported by grant No 80.00442.04 from the Italian National Research Council (CNR)     

Studies on the acetylcholinesterase (ache)-positive and -negative autonomic axons supplying smooth muscle in the normal and 6-hydroxydopamine (6-OHDA)treated rat iris

Summary The adrenergic and cholinergic innervation to the rat iris has been studied at a light and electron microscopic level. Catecholamine fluorescence histochemistry showed adrenergic nerves to be present in both the dilatator and the constrictor pupillae regions. At a fine structural level the terminal innervation of the iris was studied and criteria for the differentiation between presumptive adrenergic and presumptive cholinergic axon terminals were examined. To aid this examination presumptive adrenergic axons were either labelled with the false adrenergic transmitter, 5-hydroxydopamine, or chemical sympathectomy performed using 6-hydroxydopamine. The value of using acetylcholinesterase staining as a marker for cholinergic nerve terminals was also studied.Results showed a mixed adrenergic/cholinergic innervation to the dilatator pupillae. In the constrictor pupillae an exclusively cholinergic innervation was found although adrenergic and cholinergic nerves were found supplying the blood vessels and at the dilatator-constrictor interface. These findings are discussed with regard to innervation-function relationships in the iris.     

Adrenergic innervation of fetal pig adipose tissue. Histochemical and ultrastructural studies

Histochemistry and electron microscopy were used to study the adrenergic innervation of subcutaneous adipose tissue in fetal pigs. Adrenergic innervation was present around arteries, arterioles, and adipocyte-associated capillaries. Nerve fibers were infrequently observed around veins, venules, and adipocytes. Ultrastructural features of nerves included mitochondria, clear synaptic vesicles, and a small number of vesicles with a core of electron-dense material. Innervation of adipose tissue was similar in 70-, 90-, and 110-day-old fetuses. Examination of fetuses decapitated at 45 days of gestation and removed at 110 days showed that adrenergic innervation was absent in adipose tissue of decapitated fetuses. Adrenergic innervation was also absent in adipose tissue from fetuses hypophysectomized at 72-73 days of gestation and examined at 110 days. These data indicate that fetal porcine adipose tissue may be under neural control by day 70 of gestation. Furthermore, an intact pituitary is necessary for the development of adrenergic innervation in fetal adipose tissue.     

The neuronal and extra-neuronal localisations of biogenic amines in the cervical region of the domestic fowl (Gallus Gallus domesticus L.)

Summary The Falck-Hillarp technique has been used to demonstrate the neuronal and extra-neuronal localisations of biogenic amines in the cervical region of the domestic fowl. Adrenergic cell bodies were found in the superior cervical ganglion and in the ganglia of the cervical paravertebral chain. The axons of the latter ran into the corresponding spinal nerves and thus to the periphery. Very few adrenergic fibres were found in the interganglionic portions of the cervical paravertebral chain. The precarotid branch of the glossopharyngeal nerve, and the vagus nerve, below its junction with the former, contained numbers of adrenergic fibres. The retrocarotid nerve-trunk from the superior cervical ganglion was composed of adrenergic fibres. With the exception of the parathyroid gland, the adrenergic nerves seen in the branchial derivatives (thymus, thyroid and ultimobranchials) appeared to be associated with blood vessels. Under normal conditions the cells of the ultimobranchial body were nonfluorescent, but after injection of 6-hydroxydopamine the cells were brightly fluorescent. The carotid body was devoid of adrenergic nerves other than those with blood vessels, but the cells of the carotid body were brightly fluorescent. Various fluorescent cell types were found throughout the cervical region, particularly in association with the vasculature. I should like to thank Prof. G. Burnstock (Department of Zoology, Melbourne University) in whose department this work was carried out and Dr. R. D. Hodges (Wye College, London University) for his indispensable advice on the disposition of the avian ultimobranchial body. The author held a Postdoctoral Research Fellowship of the National Heart Foundation of Australia during part of this study.     

Seasonal changes in the adrenergic fibers in the wall of the microvessels of the cheek pouch of the hamster, Mesocricetus auratus

Adrenergic innervation has been investigated by fluorescent histochemical method with glyoxylic acid. Adrenergic fibers were found in summer period on the arterioles with a diameter varying from 100 to 30 micron, their fluorescence being significantly lower than that in the vessels of microcirculatory bed of the mesentery, tongue, femoral muscles of the same animal. No adrenergic fibers were revealed on the arterioles with a smaller diameter, on veins and venules. In winter period, fluorescence is completely absent from all the vessels of the cheek pouch, which may be due to the decrease in the level of catecholamines in the nerves and other tissues during hibernation.