Monoaminergic mechanisms in the nervous system of Lumbricus terrestris (L.)

Summary The localization and intraneuronal distribution of the monoaminergic transmitters in the nervous system of the earthworm, Lumbricus terrestris, have been investigated in detail with the aid of the histochemical fluorescence method of Falck and Hillarp.In the ventral nerve cord, many yellow fluorescent, 5-hydroxytryptamine containing neurons are found, but only few green fluorescent noradrenaline containing cell bodies, which, however, are numerous in the peripheral nervous system. There is an abundance of both fibre types in the neuropile.The 5-hydroxytryptaminergic neurons probably have a motor (possibly inhibitor) function; the adrenergic neurons in the body segments are supposed to have a receptor (exteroceptive and possibly proprioceptive) function.In the cerebral ganglion, both 5-hydroxytryptamine and noradrenaline containing neurons are found in large numbers, and there are closely packed numerous fibres of both types in the neuropile. Their function is more obscure, though an associative function can be presumed for some adrenergic neurons; smaller 5-hydroxytryptaminergic neurons might have a motor (perhaps inhibitor) function.Adrenergic sensory cells are found in the body integument, most frequently in the clitellum segments, in the prostomium, and in the roof of the buccal cavity. These cells give off varicose fibres that form a basi-epithelial network which is in communication with the green fluorescent sensory fascicles in the ventral nerve cord via the epidermal nerves, the ring nerves, and the segmental nerves. No direct adrenergic sensory-effector innervation of either circular and/or longitudinal musculature or gland cells seems to exist. No adrenergic free nerve endings in the body integument have been observed. Instead, there must be a synaptic contact with the motoneurons, either directly in the neuropile or via an interjacent neuron.No synaptic contacts have been observed in the ventral nerve cord between adrenergic or 5-hydroxytryptaminergic fibres and either the giant fibres or fluorescent or nonfluorescent perikarya.An adrenergic innervation of the pharynx musculature has been found, and sensory cells of a different type are present in and below the epithelium; here, a direct senso-motoric innervation of the pharyngeal musculature cannot be excluded. It is established that the adrenergic neurons in the stomatogastric nervous system have an exciting function on the pharynx, whereas a direct monoaminergic influence of the muscular movements of the intestine probably does not exist.Abbreviations Used A adrenaline - CA catecholamine - DA dopamine - 5-HT 5-hydroxytryptamine - MA monoamine - NA noradrenaline The research reported in this document has been sponsored by the Air Force Office of Scientific Research under Grant AF EOAR 67-15 through the European Office of Aerospace Research (OAR), United States Air Force, by the Swedish Natural Science Research Council (99-34, 6627), and by the Swedish Medical Research Council (B67-12X-712-02A).     

Cellular localization of monoamines in the median eminence and the infundibular stem of some mammals

Summary A histochemical analysis of the monoamines which are strongly accumulated in the median eminence and the proximal part of infundibular stem of all species examined (mouse, rat, guinea pig, hamster, rabbit, and cat) was performed with the help of a highly specific and sensitive fluorescence method. Strong evidence was obtained for the view that the monoamines are localized in very high concentrations to the terminal parts of non-sympathetic nerve fibres, which — mainly at least — converge to the primary plexus of the hypophyseal portal system. The capillaries are densely and closely surrounded by the nerve fibres.Pharmacological experiments, involving the administration of reserpine, nialamide, m-tyrosine and -methyl-m-tyrosine, furnished good evidence for the view that primary catecholamines, probably mainly DA but also NA, are the predominant monoamines present. The experiments also revealed the existence of catecholamine-containing nerve cells in the arcuate nuclei and the ventral portion of the anterior periventricular nuclei. These nerve cells, situated in the regions where the tubero-infundibular tract arises, may be the cell bodies of adrenergic neurons to which the amine-containing nerve fibres at least partly belong.The findings indicate that primary catecholamines are released to the primary plexus of the hypophyseal portal system and thus transported to the anterior lobe. These amines may consequently act as neuro-humoral transmittors for the regulation of the activity of the anterior pituitary. — No direct adrenergic innervation of the cells in the pars tuberalis and anterior lobe was found. The portal vesstes in the pars tuberalis receive a very sparse adrenergic innervation and the vessels in the anterior lobe receive no or very few adrenergic nerves. Pars intermedia, on the other hand, may have a non-sympathetic adrenergic innervation.The Following Abbreviations are Used DA Dopamine - NA Noradrenaline - A Adrenaline - 5-HT 5-hydroxytryptamine For generous supplies of drugs we are indepted to Swedish Ciba, Stockholm (reserpine), and Swedish Pfizer, Stockholm (nialamide). The investigation was supported by research grants from the United States Public Health Service (NB 02854-03), the Swedish Medical Research Council, and the Therese and Johan Andersson Memorial Foundation. The excellent technical assistance of Miss M. Gustafsson is gratefully acknowledged.     

Remarkable adrenergic nerves in the exocrine pancreas

Summary The adrenergic nerves in the pancreas of mice, rats, guinea-pigs, rabbits, and cats were investigated with the fluorescence method of Falck and Hillarp. The relations between the adrenergic fibres and the vessels were studied by the injection of india ink into the vessels.Besides the normal manifestation of adrenergic fibres at the large vessels, some vessels of capillary size were also accompanied by adrenergic fibres. These fibres had a very weak fluorescence, and showed up regularly only when the animal had been treated with Nialamide and L-DOPA or dopamine to increase the catecholamine content of the adrenergic fibres. The weakness of the fluorescence is perhaps due to low transmitter concentration or to small size of the nerve fibres, or to both. A rough estimate indicated that either the transmitter concentration of the nerve fibre is at least approximately 100 times below that seen in adrenergic nerves in other tissues, or that the radius of the varicosities of the nerve fibres is less than 0.2 . Neither alternative has previously been recognized.The secretory acini of the pancreas seem to lack a direct adrenergic supply. In the intrapancreatic ganglia, non-fluorescent nerve cells were reached by adrenergic terminals. No adrenergic nerve cells were detected in the pancreas of rats and cats. Small intensely fluorescent catecholamine-containing cells were observed in connexion with the intrapancreatic ganglia of rats.The research reported in this document has been sponsored by the Air Force Office of Scientific Research under grant AF EOAR 67-15 through the European Office of Aerospace Research (OAR), United States Air Force, by the United States Public Health Service (grant NB 06701-01) by the Swedish Medical Research Council (project B 67-12X-712-02A), and by the Faculty of Medicine, University of Lund, Sweden.     

Adrenergic and cholinesterase-containing neurons of the heart

Summary The adrenergic and acetylcholinesterase-containing nerves of the hearts of mice, rats, guinea-pigs, rabbits, and cats were studied. The fluorescence technique of Falck and Hillarp was used for the demonstration of adrenergic nerves, whereas a modified Koelle cholinesterase technique was used for the cholinesterase-containing nerves. The inhibitors used were Mipafox, iso-OMPA and Nu 683. Microspectrofluorometry was used to identify the structures containing dopamine.Adrenergic as well as acetylcholinesterase-containing fibres were found in all parts of the heart, most abundantly in the atria. Dense nerve plexa supplied the sinoarial and atrioventricular nodes. There was a plexus of both fibre types in the endocardium and on the atrial side of the valves. In the valves, it could be shown that adrenergic and cholinesterase-containing fibres ran closely parallel to each other. Indirect evidence suggested that this applies also to the myocardium.No nerve fibres containing dopamine were revealed in the microspectrofluorometer. The dopamine previously found in the atria seems, instead, to be situated in so-called small intensely fluorescent cells.No adrenergic ganglion cells were found in the heart despite extensive search. The vagus of rabbits was found to contain only few adrenergic preterminals.     

Histochemical studies on the adrenergic innervation of the nictitating membrane of the cat

Summary The sympathetic adrenergic innervation has been studied with the fluorescence method of Falck and Hillarp. The smooth muscle of the membrane has been found to be richly innervated which is well correlated to the high amounts of noradrenaline (NA) found in that organ with biochemical methods. The distribution of the nerves in a ground plexus and the close contacts between the nerves and the muscle cells have been discussed in relation to the physiology of this sympathetically innervated organ.     

Adrenergic and cholinergic nerves of bovine,guinea pig and hamster salivary glands

Summary The distribution of formaldehyde-induced fluorescence and acetylcholine-esterase (AChE) activity was histochemically investigated in certain salivary glands of the cow (submandibular gland), guinea pig and hamster (submandibular and sublingual glands). Adrenergic nerves occur around the secretory acini of the bovine, guinea pig and hamster submandibular glands, as well as around those of the hamster sublingual gland. The mucous secretory acini of the guinea pig sublingual gland, however, seem to be devoid of adrenergic nerve supply. Except in the sublingual gland of the hamster, no adrenergic nerves occur in relation to duct cells.The pattern of AChE activity is similar to that of adrenergic nerves. Thus, AChE-positive nerves form a network around secretory acini of all the five glands examined. Furthermore, AChE activity was also observed in nerve fibres in close proximity to striated duct cells.Both adrenergic and AChE-containing fibres were observed around blood vessels of different sizes. Ganglionic cells are occasionally to be seen; they all display AChE-activity. No adrenergic ganglionic cells were observed in any of the glands examined.All glands were also studied in the electron microscope. Interest was focussed on the fine structure of the autonomic nerves with special reference to their contents and type of storage vesicles.The content of noradrenaline was chemically determined in each type of salivary gland studied.This work was supported by grants from the University of Umeå and from the Swedish Society for Medical Research and was also carried out within a research organization supported by the Swedish Medical Research Council (projects B73-04X-712-08C and B73-04X-56-09C). The authors are indebted to Miss Kristina Karlsson and Miss Marianne Borg for valuable technical assistance.     

Adrenergic neurons in teleost retina

Summary The adrenergic retinal neurons of perch and trout were studied with the fluorescence microscopical method of Falck and Hillarp. Pilot studies were also performed on pike, plaice, cod, eel, goldfish, cunner, black moor, cichlid and carp. Only minor differences were noted between the species.Adrenergic varicose terminals occur in three sublayers of the inner plexiform layer. The layer adjacent to the ganglion cells is the most elaborate. Adrenergic perikarya occur in the innermost cell rows of the inner nuclear layer, sending branches to all sublayers of the inner plexiform layer. Adrenergic perikarya also occur among the ganglion cells, sending their branches to the innermost sublayer of adrenergic fibres in the inner plexiform layer. Weakly fluorescent adrenergic fibres can be seen running through the entire depth of the inner nuclear layer. They originate from the adrenergic perikarya of the inner nuclear layer, and they end in an elaborate plexus of adrenergic terminals among the horizontal cells. Either of the horizontal cell types can be in contact with adrenergic terminals, but the middle horizontal cells have the greatest density about them, being surrounded by baskets of adrenergic terminals of presumably synaptic character. It cannot be excluded that some horizontal cells contain a catecholamine.Microspectrofluometry revealed dopamine in the perch and trout retinal neurons.The research reported in this document has been sponsored by USPHS Grant No. 06092 and by a Research Professorship from Research to Prevent Blindness, Inc. to A.M.L. and by the Swedish Medical Research Council (B69-14X-712-04C and B68-14X-2321-01).     

Potentiation by cocaine of relaxations of the guinea-pig colon caused by noradrenaline and by stimulation of adrenergic nerves.

The distal colon of the guinea-pig is relaxed by noradrenaline, by isoprenaline and by the stimulation of fibres running with the colonic nerves or intramurally. The relaxations in response to stimulation of the colonic nerves have a guanethidine-sensitive (adrenergic) and a guanethidine-insensitive (non-adrenergic) component. Cocaine causes a three-fold sensitization of the muscle to noradrenaline but no sensitization to isoprenaline. Cocaine increases the duration, but does not affect the amplitude, of the relaxation observed when adrenergic nerves are stimulated, and affects neither duration nor amplitude of the non-adrenergic response. The adrenergic nerve terminals lie in Auerbach's plexus, not in the longitudinal muscle. It is concluded that the sensitization to noradrenaline and the increases in durations of responses to adrenergic nerve stimulation are due to inhibition of catecholamine uptake into adrenergic nerves by cocaine. It appears that, even where the neuromuscular separation is large as it is in the colon, the concentration of exogenous noradrenaline at the receptors can be decreased by neuronal uptake, and the uptake mechanism can modify responses to nerve stimulation in vitro.     

Development of the ocular adrenergic nerve supply in man and guinea-pig

Summary The development of adrenergic nerves to the anterior eye segment was studied in human and guinea-pig embryos. Adrenergic terminals had already appeared in the earliest human embryos available (4–6 cm). They first appeared mainly in nerve trunks in the primitive chorioid, especially in the region of the developing ciliary body. Adrenergic nerves then grow into different structures of the eye as these develop, but typical terminals in contact with effector cells appeared late during the development, about the 25–30 cm stage. No adrenergic nerves were observed in the chamber angle. Corneal adrenergic nerves (also intraepithelial terminals) appeared much more frequently in embryos than in adults. No adrenergic neurons were observed in the retina. In the guinea-pig, the first adrenergic fibres were observed at about gestation day 35. The general principle of the development was very similar to that of the humans. At gestation day 45 to 50, the supply of adrenergic fibres was essentially that of the adult animal, except that the corneal adrenergic fibres were increasing until just before birth and that the adrenergic terminals of the chamber angle appeared shortly before term.This work was supported by grants from the Association for the Aid of Crippled Children, H. Hiertas Stiftelse, and the Swedish Medical Research Council (Project no. B71-14X-2321-05B).     

Uptake of3H-adrenaline and14C-noradrenaline into neuronal and extraneuronal tissue compartments in the perfused gas gland of the swimbladder of the Atlantic cod,Gadus morhua

Summary The neuronal and extraneuronal accumulation of radiolabelledl-adrenaline andl-noradrenaline was studied in the gas gland of the swimbladder of the Atlantic cod,Gadus morhua. Both amines are taken up into the tissue compartments, and a preference for noradrenaline for both uptake processes was demonstrated. A relatively high neuronal accumulation compared to earlier results (cod spleen; Ungell and Nilsson 1984) was seen and this is probably due to the more dense innervation of the swimbladder gas gland. A higher extraneuronal accumulation may be due to the presence of arterial smooth muscle.It is concluded that the adrenergic nerves of the swimbladder gas gland of the cod preferentially accumulate noradrenaline, a situation similar to that in mammals.Abbreviations A/NA adrenaline/noradrenaline ratio - PBA phenoxybenzamine - PNMT phenylethanolamine-N-methyl tranferase     

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.     

Evidence for the existence of monoamine neurons in the central nervous system

Summary With the help of the highly specific and sensitive fluorescence method of Falck and Hillarp together with the histochemical and pharmacological criteria for the specificity of the fluorescence reaction convincing evidence has been obtained that the fine, varicose nerve fibres observed in a vast number of regions in the mammalian central nervous system (mouse, hamster, rat, guineapig, rabbit, cat), which exhibit a green or yellow fluorescence, contain primary catecholamines and 5-HT respectively. Strong support has been given for the view that CA fibres showing a rapid recovery after administration of -MMT contain DA, while those showing a slow recovery contain NA.There is little doubt that the monoamine-containing fibres in the brain represent the terminal ramifications of axons belonging to specific monoamine neurons and that they are true synaptic terminals. They seem to make their contacts via the varicosities which have extremely high concentrations of amines and in all probability represent the presynaptic structures, specialized for synthesis, storage and release of the amines. The central monoamine terminals thus have the same characteristic appearance as the adrenergic synaptic terminals in the peripheral nervous system.All the data strongly support the view that the specific central neurons giving rise to the terminals are monoaminergic, i.e. function by releasing their amines from the synaptic terminals. Consequently, DA, NA and 5-HT seem to be central neurotransmitters.Not only the median eminence but also the nuc. caudatus putamen, tuberculum olfactorium, nuc. accumbens and the small circumscribed areas medial to nuc. accumbens contain very fine (partly sublightmicroscopical) CA terminals. These areas react to treatment with reserpine, nialamide-dopa and -MMT in the same way and since the nuc. caudatus putamen and tuberculum olfactorium are known to have a high DA content it seems likely that abundant DA terminals are accumulated in these special areas.The Following Abbreviations are Used CA Catecholamine - DA Dopamine - dopa 3.4-Dihydroxy-phenylalanin - NA Noradrenaline - A Adrenaline - 5-HT 5-Hydroxytryptamine - -MMT -Methyl-meta-tyrosine - MAO Monoamine oxidase For generous supplies of drugs the author is indebted to the following companies: Swedish Ciba, Stockholm, Sweden (reserpine); Swedish Pfizer, Stockholm, Sweden (nialamide); Abbott Research Laboratories, Chicago, USA. (MO 911). This study has been supported by a Public Health Service Grant (NB 02854-04) from the National Institute of Neurological Diseases and Blindness and by grants from the Knut and Alice Wallenberg Foundation, and the Swedish Medical Research Council.     

Fluorescence studies on neural structures and endocrine cells in the pancreas of the cat

Summary With the use of the Falck-Hillarp histochemical technique for the detection of monoamines, nerve fibre fluorescence is observed throughout the tail of the pancreas of the cat and the arrangement and distribution of the nerve fibres can be studied in both the exocrine and endocrine tissue. In the exocrine pancreas, adrenergic nerve fibres innervate arterioles, larger veins and major pancreatic ducts. Adrenergic nerve fibres also appear to terminate on the non-adrenergic nerve cell bodies of the intrapancreatic ganglia. In the islets of Langerhans, adrenergic nerve fibres innervate both the endocrine cells and blood vessels. Some of the islet cells exhibit fluorescence with the Falck-Hillarp technique and these cells have been identified as alpha cells. In animals treated with reserpine, the fluorescence in nerve fibres and in alpha cells is absent.The author wishes to thank ProfessorG. C. Schofield and Dr.G. C. Smith for their encouragement and valuable criticism during the course of this study. The assistance of MissJ. Bennett and MissW. Kemp and the photographic help of Mr.J. S. Simmons, F.R.P.S., are gratefully acknowledged. The diagram was drawn by MissS. Flett.     

NEUROCHEMICAL PROPERTIES OF ADRENERGIC NERVES REGENERATED AFTER 6-HYDROXYDOPAMINE

Abstract— The uptake-storage properties and synthesis of noradrenaline, and fluorescence morphology of adrenergic nerves which have been allowed to regenerate for 4 weeks after a chemical sympathectomy produced by 6-hydroxydopamine have been investigated in mouse iris and atrium. The regenerated nerve terminals displayed a lower formaldehyde-induced fluorescence intensity whereas the non-terminal axons exhibited a stronger fluorescence intensity and a more beaded appearance compared with mature nerves. The endogenous noradrenaline concentration after 6-hydroxydopamine was 30% in iris and 45% in atrium compared to control values. Recovery of [³H]noradrenaline uptake was found to be more rapid than that of endogenous noradrenaline concentration after the 6-hydroxydopamine treatment. [³H]Noradrenaline uptake in regenerating and adult mature nerves both obeyed Michaelis-Menten kinetics having identical K_m values. There was a close correlation between [³H]noradrenaline uptake and nerve density of adrenergic nerves regenerated after 6-hydroxydopamine. These results show that [³H]noradrenaline uptake is a better index for the number of regenerated nerve terminals than is the endogenous noradrenaline concentration. The retention of [³H]noradrenaline taken up and accumulated in vitro was about the same in regenerated and mature nerves, although a slight tendency to less effective retention was observed in the regenerated nerves. Subcellular distribution studies showed that relatively less [³H]noradrenaline was recovered in the microsomal fraction after 6-hydroxydopamine treatment. The formation of ¹⁴C-labelled catecholamines from [¹⁴C]DOPA was higher in regenerating nerves than indicated by the endogenous noradrenaline concentration but lower than that indicated by the [³H]noradrenaline. It is concluded that the regenerating nerves contain less endogenous noradrenaline than adult mature nerves and that the uptake mechanism develops promptly, whereas the development of the storage mechanism lags behind.     

Fluorescence-microscopical demonstration of a population of gastro-intestinal nerve fibres with a selective affinity for Quinacrine

Summary A population of nerve fibres in the gastro-intestinal tract of mice showing a high affinity for quinacrine was revealed by fluorescence microscopy. Similar results were obtained in rats and guinea pigs. Whole-mounts of sheets of the smooth muscle layer following incubation in 10^-6-10^-7 M quinacrine for 15–60 min revealed fine fluorescent varicose nerve fibers in the myenteric plexus of Auerbach both around nerve cell bodies and in the interconnecting strands. Many fibers were also present between the strands of the plexus, especially running parallel to the circular muscle layer. Such fibers were not seen in similarly quinacrine-incubated irides. A proportion of the cell bodies in Auerbach's plexus also showed quinacrine accumulation. These cells were apparently smaller neurons, sometimes with fluorescent processes. Intraperitoneal injections of quinacrine failed to demonstrate nerve fibers, but some cell bodies in Auerbach's plexus were positive. Subsequent paraformaldehyde treatment for monoamine visualization showed persistent adrenergic nerve terminals in the intestine and iris. These nerves seemed to be fewer and had a more yellow fluorescence than normally. The identity of the quinacrine-positive fibers is discussed with respect to recent suggestions that purinergic, substance P, enkephalin, and somatosin-containing nerves, in addition to adrenergic and cholinergic nerves, are present in the gut wall.Supported by the Swedish Medical Research Council (04X-03185). Magnus Bergvalls Stiftelse and Karolinska Institutets Fonder. For generous gifts of Mepacrine we thank Winthrop, Skärholmen, Stockholm, Sweden. The skilful technical assistance of Miss Gerd Boetius and Miss Maud Eriksson is gratefully acknowledged     

Fluoreszenzmikroskopische und elektronenmikroskopische Untersuchungen an regenerierenden adrenergischen Nervenfasern

Zusammenfassung Die Regeneration peripherer adrenergischer Nervenfasern wurde fluoreszenzmikroskopisch nach einer von Falck u. Mitarb. angegebenen Methode, ferner elektronenmikroskopisch untersucht. Schon 24 Std nach einer Durchschneidung des N. ischiadicus (Kaninchen) schwellen die Axone unmittelbar am Schnittrand des zentralen Stumpfes an. Diese Anschwellungen weisen einen großen Gehalt an Catecholaminen auf. Die primären Axonanschwellungen fallen der initialen Degeneration anheim. Die nachfolgende Axonaussprossung beginnt mit der Bildung von Wachstumsendkolben, die sich von der Trümmerzone des zentralen Stumpfes aus in die Nervennarbe vorschieben. Die Wachstumsendkolben adrenergischer Fasern sind elektronenmikroskopisch durch das vesicular erweiterte neuroendoplasmatische Reticulum und durch eine große Zahl von Catecholaminkörpern gekennzeichnet. Es wird angenommen, daß die Catecholaminkörper der adrenergischen Regenerate überwiegend Noradrenalin enthalten, das im Bereich der Wachstumsendkolben vermehrt an das Grundplasma und das Interstitium abgegeben wird. Die Feinstruktur der adrenergischen Wachstumsendkolben hat große Ähnlichkeit mit normalen adrenergischen Nervenfaserendigungen.

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Summary The regeneration of peripheral adrenergic nerve fibres is investigated with the method described by Falck et al. and by means of electron microscopy. Within 24 hours after the severance of the N. ischiadicus (rabbit) a swelling of the axons, which are situated in the immediate vicinity of the sectioned area in the central stump, is observed. These swellings have a high content of catechol amines. These primary axon swellings undergo degeneration. The subsequent proliferation of axons starts with a formation of end-bulbs, which grow from the debris-zone of the central stump into the scar of the nerve. The end bulbs of adrenergic fibres are electron microscopically distinguishable through their neuroendoplasmic reticulum expanded to form vesicles and a large number of catecholamine-bodies. It is suggested that the cate-cholamine-bodies in the adrenergic regeneration tissue contain predominantly noradrenaline, which in the region of the end-bulbs is secreted predominantly into the interstitial ground substance and the interstitium. The fine structure of the adrenergic end-bulbs has a similar appearance to that of normal adrenergic nerve fibre endings.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft     

Extrinsic adrenergic innervation of the extrahepatic biliary duct system in guinea-pigs,cats and rhesus monkeys

Summary The localization of catecholamines has been investigated in the extrahepatic biliary duct system of cats, guinea-pigs and rhesus monkeys. In fluorimetric determinations noradrenaline was found to be the main primary catecholamine present in the biliary tract of rhesus monkeys. There exist regional differences in the noradrenaline content: Fairly low amounts were detected in the lower fundus of the gall-bladder (0.28 g/g). Increasing concentrations were measured in the corpus vesicae felleae (0.35 g/g), reaching a maximum level in the collum vesicae (0.49 g/g) and the ductus cysticus (0.50 g/g). The noradrenaline content of the choledochus and the choledocho-duodenal junction including Oddi's sphincter was much lower: 0,27 and 0,25 g/g respectively. The noradrenaline level in the small intestine of the rhesus monkey amounted to less than half the concentration found in the biliary ducts. Neither dopamine nor adrenaline have been detected. Fluorescence microscopical analysis reveals the presence of adrenergic nerves in the bile ducts which correspond to the measured noradrenaline concentrations: All parts of the biliary duct system in the different species investigated contain an elaborate perivascular adventitial plexus and adrenergic fibres confined to adventitial non-adrenergic ganglia. In guinea-pigs adrenergically innervated ganglia extend into the smooth muscle layer. The smooth muscle layer of the gall-bladder and the terminal choledochus in cats and rhesus monkeys is penetrated by a wide-meshed adrenergic ground plexus. This plexus was absent in guinea-pigs. The smooth musculature of the sphincter Oddi lacks a specialized adrenergic nerve supply in all species investigated. Finally, bound to the arterial vascular bed inside the propria in all parts of the biliary tract from all species investigated a prominent perivascular plexus is present. It is concluded that the smooth musculature of the gall-bladder and the terminal choledochus (the sphincter region excluded) in cats and monkeys receives 1. a direct sympathetic noradrenergic inhibitory innervation and 2. an indirect sympathetic noradrenergic inhibitory innervation which acts on intrinsic excitatory neurons and is present in all species investigated. The functional significance of the direct and indirect inhibitory innervation to the smooth musculature of the gall-bladder is discussed in detail.Dedicated to Professor Bengt Falck.Supported by the Deutsche Forschungsgemeinschaft and Joachim-Jungius-Gesellschaft zur Förderung der Wissenschaften, Hamburg.     

6-AMINODOPAMINE-INDUCED DEGENERATION OF CATECHOLAMINE NEURONS

the effects of 6-aminodopamine on central and peripheral catecholamine neurons using fluorescence histochemical and isotope techniques have been investigated. Systematic administration of 6-aminodopamine (20 mg/kg intraveneously) produced a rapid (within 1 h) and long-lasting depletion of endogenous noradrenaline in adrenergic nerves of mouse atrium and iris with a concomitant loss of [³H]noradrenaline uptake. The effects were dosedependent. Accumulations of noradrenaline in non-terminal axons were observed histochemically, indicating that 6-aminodopamine induces neuronal damage. Desipramine completely blocked the 6-aminodopamine induced noradrenaline depletion and reduction in [³H]noradrenaline uptake, indicating that 6-aminodopamine has to be taken up by the axonal ‘membrane pump’ to produce its effects. Themonoamine oxidase inhibitor, nialamide, potentiated the effect of 6-aminodopamine on [³H]noradrenaline uptake. 6-Aminodopamine did not affect the cell bodies of the adrenergic neurons and there was a reappearance of adrenergic nerves and recovery of [³H]noradrenaline uptake. 6-Aminodopamine does not seem to pass the blood-brain barrier after systemic injection. Intraventricular injection of 6-aminodopamine in rats led to a considerable reduction in endogenous whole brain noradrenaline and [³H]noradrenaline uptake in slices from cerebral cortex and hypothalamus. Similar, but less pronounced effects were observed on dopamine neurons in the caudate nucleus. Histochemically, pronounced accumulations of transmitter were observed in the axons of the catecholamine neurons. The results obtained favour the view that 6-aminodopamine is able to produce an acute and selective degeneration of catecholamine neurons similar to that seen after the neurotoxicagent, 6-hydroxydopamine. Both compounds seemed to be approximately equally potent in their neurotoxicity, although 6-aminodopamine seemed to be more generally toxic.     

Adrenergic nerves to the dermal melanophores of the rainbow trout,Salmo Gairdneri

Summary The stellate processes and cell bodies of the dermal melanophores in the rainbow trout are intimately enclosed by a plexus of thin varicose nerves which display a specific catecholamine fluorescence. The nerves contain probably small amounts of noradrenaline and have the ability to take up and concentrate this amine. Denervation of the skin leading to dispersion of the melanophores causes the nerves to disappear. The findings leave little doubt that the dermal chromatic motor nerves are adrenergic.This study was supported by a grant from the Swedish Research Council for Natural Sciences (99-35) and was carried out within a research organization sponsored by the Swedish Medical Research Council (Projects No. B69-14X-56-05C and No. B69-14X-712-04C).     

Combined fluorescence histochemistry and 3h-noradrenaline measurements of adrenergic nerves

Summary A method is described which combines the histochemical fluorescence technique of Falck and Hillarp with isotope measurements in the same pieces of tissue. Tissue pieces incubated in isotope solutions were treated for fluorescence microscopy and examined. They were then removed from the microscopical slides, and the radioactivity determined. It was shown that NA¹ content and estimated fluorescence intensity were well correlated. The procedure devised is of special value when isotope measurements are needed of structures which can be safely identified only in the fluorescence microscope, and it has been used for quantitative estimations of adrenergic innervation.Abbreviations used Na noradrenaline - cpm counts per minute - dpm desintegrations per minute