Cytophotometric study of catecholamines and energy metabolism enzymes in rat celiac plexus neurons under cold and emotional stress

Catecholamine concentration and energy metabolism enzymes were investigated using histochemical and computer analysis techniques in celiac ganglia neurons under cold and emotional stress. Acute short-term cooling was found to lead to intensified catecholamine fluorescence as well as succinate and lactate dehydrogenase activity in celiac ganglia neurons, whereas short-lasting emotional stress produced no changes on fluorescence or energy metabolism enzymes.Institute of Physiology, Academy of Sciences of the Belorussian SSR, Minsk. Translated from Neirofiziologiya, Vol. 20, No. 6, pp. 743–749, November–December, 1988.     

Parasympathetic and sympathetic postganglionic synapses in ureterovesical autonomic pathways

Summary Histochemical techniques for acetylcholinesterase and catecholamine show that ureterovesical ganglia of both cat and dog contain dense intraganglionic cholinergic and adrenergic plexuses. Ramifications of both plexuses surround most cholinergic and adrenergic ganglion cell bodies as pericellular synaptic plexuses. Similar pericellular plexuses exist around extraganglionic cholinergic and adrenergic ganglion cells. Both adrenergic and cholinergic synaptic fibers persist in denervated pregnaglionic nerve-free specimens, indicating that cholinergic synaptic fibers are postganglionic parasympathetic in nature. The presence of adrenergic (postganglionic sympathetic) and postganglionic parasympathetic synapses around cell bodies in ureterovesical ganglia provides a morphologic basis for the sympathoinhibitory and muscarinic parasympatho-excitatory phenomena described in these ganglia.     

Microfluorimetric study of adrenergic structures in nerve plexuses of the cat pelvic cavity following decentralization and in the normal

Microfluorimetric techniques were used to investigate catecholamine concentration in small, intensely fluorescent cells and adrenergic fibers of the cat pelvic plexus ganglia and intramural ganglia of the urinary bladder and rectum in the control and following sympathetic and parasympathetic denervation. The cells examined could be divided between catecholamine- and serotonin-containing types. Parasympathetic denervation brought about an increase in the number of cells displaying serotinergic fluorescence and heightened fluorescence in the adrenergic fibers of the pelvic plexus ganglia and intramural ganglia of the urinary bladder, without affecting degree of fluorescence in those of the rectal intramural ganglia. Sympathetic denervation failed to change fluorescence level in the cells and adrenergic fibers in pelvic plexus and urinary bladder ganglia but caused the almost complete disappearance of the adrenergic fibers in the rectal intramural ganglia.Institute of Physiology, Academy of Sciences of the Belorussian SSR, Minsk. Translated from Neirofiziologiya, Vol. 18, No. 4, pp. 496–502, July–August, 1986.     

Structure and functional changes in the gastric intramural nervous plexus in emotionally stressed rats exposed to low doses of long-term ionizing irradiation and lead

Changes in the catecholamine content in adrenergic fibres, acetylcholinesterase activity, and in the energy metabolism enzymes lactate dehydrogenase and succinate dehydrogenase in neurons of the gastric intramural plexus during emotional stress in rats a day after combined exposure to prolonged (30 days) ionizing radiation in a total dose 1.0 Gy and 0.6 mg/kg lead were studied. A decrease in catecholamines in adrenergic fibres and acetylcholinesterase and lactate dehydrogenase activity in neurons was observed. An enhanced sensitivity of the gastric intramural plexus after the prolonged exposure to small doses of ionizing radiation and lead in conditions of emotional stress was suggested.     

Fluorescence microscopic study of the architecture and structure of an adrenergic network in the plexus myentericus (Auerbach), plexus submucosus externus (Schabadasch) and plexus submucosus internus (Meissner) of the porcine small intestine

The distribution of adrenergic fibres in the ganglionated plexuses of the porcine small intestine has been made on air-dried stretch preparations using the glyoxylic acid fluorescence method. Adrenergic fluorescent fibres occur in the ganglia and internodal strands of the three fundamental ganglionated plexuses: the myenteric plexus (Auerbach) and the two superimposed meshworks of the plexus submucosus , i.e. the plexus submucosus externus ( Schabadasch ) and the plexus submucosus internus (Meissner). The plexus Auerbach consists of densely glyoxylic acid induced fluorescent (GIF) elongated ganglia with in general a longitudinal axis running parallel to the circular muscle layer and large dense interconnecting fibre tracts with primary, secondary and tertiary subdivisions. In the ganglia, the fibres are varicose, forming large fluorescent 'baskets' which might be related to the occurrence of well defined enteric neurones. The plexus Schabadasch can be distinguished from the plexus Meissner by its size, strongly fluorescent ganglia and broad densely fluorescent internodal strands. The pattern of fluorescing ring-like formations at the margin and out of the nodes, clearly present in the Auerbach and Schabadasch plexuses, completely lack in the plexus Meissner, the latter being narrow-meshed with smaller fluorescent 'baskets', indicating that the corresponding neurones are smaller in size. In the ganglionic nodes of all three plexuses the axons display comparatively more varicosities than in the fibre tracts. Each of the three main ganglionated enteric plexuses are quite different with regard to the pattern of the adrenergic network both in the ganglia and in the strands.     

Morphofunctional changes in the adrenergic innervation of the major cerebral arteries during bilateral electrostimulation of the locus coeruleus

The intramural adrenergic nervous apparatus of cerebral arteries was studied in adult rabbits after 3-10 sessions of electrical stimulation of locus coeruleus. The activity of nerve structures was determined by estimating the density of adrenergic perivascular plexuses and by semi-quantitative cytophotometry of changes in the catecholamine content of nerve varicosities. The stimulation was followed by a 28.2 +/- 1.5% increase in adrenergic innervation density. while catecholamine content in perivascular nerve structures displayed a tendency to decrease. The problem of central effects on cerebral blood flow autoregulation is discussed.     

Adrenergic innervation of the gut musculature in vertebrates

Summary The adrenergic innervation of the gut musculature has been compared in various vertebrates (two teleost fish, an amphibian, a reptile and a mammal) by the fluorescent histochemical localization of certain monoamines. Very few, if any, adrenergic nerves occur within the longitudinal gut muscle of any of these animals, except for the taenia coli of the guinea-pig caecum. In contrast, the circular smooth muscle coat is supplied by varicose adrenergic nerves. These nerve fibres are particularly numerous in the toad large intestine, guinea-pig caecum, and throughout the eel gut, but are generally sparse or absent from the musculature of the stomach and small intestine of the trout, toad, lizard and guinea-pig. The extent of adrenergic innervation of the muscle has been discussed in relation to the physiology of the different muscle coats and to the general structure of the enteric plexuses in the vertebrate gut.     

Changes in the level of eosinophils, corticosterone and catecholamine metabolism in the dynamics of emotional-painful stress

A correlation has been established between changes in blood eosinophil content, corticosterone level and catecholamine metabolism during emotional painful stress. The authors assume that the differences in the functions of the adrenergic and pituitary-adrenal systems permit an objective determination of the parameters of stress formation in extremal exposures.     

The autonomic innervation of Rana ridibunda intestine

1. The innervation of Rana ridibunda intestine has been studied by the following methods: (a) demonstration of cholinesterase activity; (b) FIF method for catecholamines; (c) immunohistochemistry for VIP, SP and SOM, and (d) conventional electron microscopy. 2. The intrinsic intestinal innervation is represented by cholinergic-, VIPergic-, SP- and SOM-like plexuses. The intestinal adrenergic component is of extrinsic origin. 3. The intestinal peptidergic innervation is the most developed, the large intestine being the portion where the studied peptidergic plexuses are more widely distributed. 4. Against a poorly developed cholinergic/adrenergic innervation, it seems that there is a predominant peptidergic innervation in the amphibians intestine wall. 5. Taking into account that amphibians sacral parasympathetic as well as sympathetic innervation development are limited, it could be considered that in vertebrates the intestinal peptidergic innervation is phylogenetically earlier and hence better developed.     

Morphofunctional restructuring of the nervous system of the lymphoid organs during electrostimulation of the hypothalamus

The intramural nervous apparatus of rabbit lymphoid organs was examined after 15 and 30 sessions of electric stimulation of the posterior hypothalamus. The function of nerve cells was evaluated by cytophotometry according to the content of catecholamines and acetylcholinesterase (ACE) activity. In the mesenteric lymph nodes, stimulation led to a decrease in the activity of both adrenergic and ACE-containing nerve cells. The spleen demonstrated a reduction in the catecholamine content in the perivascular nerve plexuses, with a simultaneous rise of the catecholamine content in the fibers located in the parenchyma, which evidences inhibition of the neurogenous mechanisms of vasomotor control and possible selective activation of the sympathetic mechanisms involved in the regulation of the immunogenesis. Thymic preparations impregnated with silver salts manifested high argyrophilia of the nerve cells, characteristic of the reactive phase of the destructive process. The problems of the hypothalamic effects on the blood supply and functions of the immune system organs are discussed.     

Increased ovarian follicular angiogenesis and dynamic changes of follicular vascular plexuses induced by equine chorionic gonadotropin in the gilt

Follicular angiogenesis and capillary degeneration are crucial ovarian processes in folliculogenesis. The present study was conducted to assess the changes in population of follicular vascular plexuses with different capillary status in prepubertal gilts 72 h after equine chorionic gonadotropin (eCG) (1,250 IU) treatment, using combined vascular corrosion casting and scanning electron microscopy. Follicular fluid concentrations of estradiol, progesterone and vascular endothelial growth factor (VEGF) were determined to confirm the follicular status. Based on the proliferative or degenerative characteristics of their capillaries, follicles were classified into three categories: active angiogenesis, low angiogenesis and degeneration. Irrespective of exogenous gonadotropin treatment in vivo, small follicular vascular plexuses (<4 mm in diameter) exhibited all three conditions in casted ovaries, while medium (4–5 mm) and large (>5 mm) plexuses showed only active angiogenesis or degeneration. eCG treatment significantly increased the population of large, but decreased that of small follicular plexuses. Most large follicular vascular plexuses showed active angiogenesis with higher follicular fluid estradiol:progesterone ratios and VEGF concentration. eCG also increased the percentage of medium follicular plexuses with active angiogenesis. The populations of small follicular plexuses with active angiogenesis were higher in controls, but decreased after eCG treatment. However, treatment of gilts with the gonadotropin increased the percentage of small plexuses (<1.0 mm) with low angiogenesis and those (1–3.9 mm) with extensive capillary degeneration. These findings are consistent with the hypothesis that angiogenesis is involved in selection and growth of small follicles in gilts under the regulation of gonadotropin.This work was supported by grants from the Program for Promotion of Basic Research Activities for Innovative Biosciences and Research for the Future Program, the Japan Society for the Promotion of Science (JSPS-RFTF97L00904) and the Canadian Institutes of Health Research (MOP-15691). J.Y.J. is a recipient of a CIHR-STIRRHS Postdoctoral Fellowship. A preliminary report of this work was presented at the 49th Annual Meeting of the Canadian Fertility and Andrology Society, 5–8 November 2003, Victoria, British Columbia, Canada     

Innervation of the mucosa of the small intestine of laboratory animals. I. Architecture, light microscopic structure and histochemical differentiation

1. The small intestine mucosa has a dense and differentiated innervation. The most dense nerve plexuses are contained in the villi. Each tubular gland is surrounded by a rope-ladder-like plexus, mostly developed in the lower and upper third. 2. According to our observations most of the axons in the mucosa are cholinergic. Cholinergic and adrenergic axons take part in the innervation of the glands. Moreover enterochromaffine cells can be innervated for instance sympathetically. Intraepithelial axons could not be found.     

Effect of intravascular laser irradiation of blood on the status of the adrenergic and cholinergic structures of the small intestine in experimental peritonitis

In the experiment performed on 60 mongrel dogs the effect of intravascular laser radiation of blood (ILRB) to adrenergic and cholinergic fibers of the small intestine has been studied at treatment of experimental generalized suppurative peritonitis. At treatment of the experimental peritonitis by means of traditional method only one month after its termination a positive reaction to acetylcholinesterase (AChE) is noted, structural organization of cholinergic fibers corresponds to the norm, and density of their plexuses increases. Application of ILRB at treatment of the experimental peritonitis facilitates to increasing density of the cholinergic fibers already on the 21st day after termination of the treatment; their AChE increases essentially, their structural organization corresponds to the norm. In cytoplasm of neurons of the muscular-intestinal plexus a positive reaction to AChE is revealed; this demonstrates their increased functional activity. The small intestine adrenergic apparatus at peritonitis undergoes less manifested structural-chemical alterations. After termination of treatment by means of traditional methods it corresponds to the norm on the 7th day, and after ILBR treatment----on the 3d day. Thus, application of ILBR in treatment of the experimental peritonitis enhances the development of regenerative processes in adrenal and cholinergic structures of the small intestine, facilitates a more manifested demonstration of compensatory-reparative possibilities of the organism.     

Adrenergic terminal structures in the mesentery of mammals

This work is devoted to the study of adrenergic terminal structures in the mesentery of mammals (cat, dog). The investigation was performed with the Falck-Hillarp method of catecholamine fluorescence microscopy on total stretch mesentery preparations. The investigation showed that richly developed perivascular plexus constitute the basis of the adrenergic innervation system of the mesentery. In numerous points of these plexuses, single adrenergic fibers or polyaxonal structures are observed to issue into nonvascular areas of the mesentery where after repeated dichotomic division they pass into the preterminal and terminal parts. Being constructed on the principle of extended or restrained arborizations, these innervating structures have a morphological similarity with free sensory nerve endings. In this connection, the question of the possible existence of the sensory (afferent) links in the catecholamine-containing vegetative nerve plexuses is discussed.     

Auerbach's plexus of mammals and man: Electron microscopic identification of three different types of neuronal processes in myenteric ganglia of the large intestine from rhesus monkeys,guinea-pigs and man

Summary Ganglia from Auerbach's plexus of the large intestine (caecum, appendix vermiformis, colon transversum and rectum) in man, rhesus monkey and guinea-pig are composed of nerve cells and their processes, typical Schwann cells and a vast neuropil. The neuropil consists of dendrites and axons of intrinsic nerve cell perikarya and axons of extrinsic neurons. Axonal profiles in large nerve fibre bundles are of uniform size and appearance, embedded in infoldings of Schwann cell cytoplasm and contain occasional large granular vesicles, mitochondria and neurotubules. Preterminal axons widen into vesicle filled varicosities, some of which establish synaptic contact with intrinsic nerve cell bodies.At least three different types of neuronal processes can be distinguished in the myenteric neuropil according to the size, appearance and commutual proportion of vesicles present in axonal varicosities, and their ability to accumulate exogenous 5- and 6-hydroxydopamine and 5-hydroxydopa: 1. Axonal enlargements containing a major population of small electron lucent synaptic vesicles (350–600 Å in diameter) together with a small number of membrane-bound, opaque granules (800–1,100 Å). These profiles have been identified as cholinergic axons. The boutons establish synaptic contacts with dendritic processes of intrinsic nerve cell bodies; membrane specializations are found at the preand postsynaptic sites. 2. Axonal beads of sometimes very large diameter, containing an approximately equal amount of large granular vesicles (850–1,600 Å) and small, electron lucent or faintly opaque vesicles (400–600 Å). The granular core of the large vesicles is of medium electron density and may either fill the entire vesicle or is separated from the limiting membrane by a more or less clear interspace. The fibres probably belong to intrinsic neurons, and because of the similarity of the large, membrane-bound vesicles with neurosecretory elementary granules, they have been designated p-type fibres (polypeptide fibres). The granular core of the vesicles in these fibres becomes more electron dense after treatment with 5-OH-dopa. The accumulation of an amine precursor analogue in combination with a possible storage of a polypeptide substance (or an ATP-like substance) resembles the situation in several diffusely distributed endocrine cell systems. 3. Varicosities of axons equipped with small (400–600 Å) empty or sometimes granular vesicles, medium sized (500–900 Å) vesicles with highly electron dense cores and occasional large (900–1,300 Å) granular vesicles. Pretreatment with 5-OH-dopamine increases the electron density in almost all medium-sized granular vesicles and some of the large granular vesicles; an osmiophilic core develops in some small vesicles. 6-hydroxydopamine results in degenerative changes in the varicosities of this type of neurons. Concomitantly, both catecholamine analogues markedly reduce neuronal noradrenaline in the large intestine, as demonstrated by fluorescence histochemistry and in fluorimetric determinations. The ultrastructural features of these varicosities and their reaction to 5- and 6-OH-dopamine indicate that they belong to adrenergic, sympathetic nerves. No membrane specializations could be detected at sites of close contact of the adrenergic boutons with dendrites and cell bodies of intrinsic nerve cells.Supported by grants from the Deutsche Forschungsgemeinschaft.Supported by a grant from Albert Pahlsson's Foundation, Sweden. The work was carried out within a research organization sponsored by the Swedish Medical Research Council (projects No. B70-14X-1007-05B, B70-14X-712-05, and B70-14X-56-06).     

Histochemical and chemical studies on pre- and postnatal development of the different systems of “short” and “long” adrenergic neurons in peripheral organs of the rat

Summary The adrenergic innervation in the submaxillary gland, heart, kidney, small intestine, and accessory male genital organs and the development of the adrenal chromaffin cells and the sympathetic ganglia were studied in the rat from 15 days post coitum to 16 days post partum using the fluorescence histochemical method of Falck and Hillarp. The postnatal development of the noradrenaline concentrations in the heart and vas deferens was followed by fluorometric determinations.At about 15 days post coitum, the anlagen of the sympathetic chains were well visible in the form of two dorsal segmented columns of small branching sympathicoblasts exhibiting an intense catecholamine fluorescence. In the midline, ventrally to these two anlagen, another column of sympathicoblasts developed; this seemed to give rise to the prevertebral ganglia and to the short adrenergic neurons supplying the internal genital organs. At the level of the adrenal anlagen, small intensely fluorescent chromaffin cells were collected in two bilateral groups which became enclosed by adreno-cortical cells. This enclosure was, however, not complete even at two weeks post partum.Bundles of growing sympathetic nerves were visible in the periphery of the various organs studied at 19–21 days post coitum. A terminal innervation of the organs suggestive of a functional transmitter mechanism did not start to establish until at or immediately after birth. The final pattern of innervation was usually reached at about one week post partum, and the following development proceeded largely in the form of a quantitative increase in the number of nerves participating in the innervation apparatus. The adult level of noradrenaline in the heart and vas deferens was reached three to five weeks after birth. The small intestine was an exception in that the final pattern of innervation in the wall was attained immediately after birth.There was no overt difference in the rate of development of the terminal sympathetic innervation in organs supplied by short adrenergic neurons (accessory male genital organs) compared to the innervation of the submaxillary gland, heart and kidney, which receive classical long adrenergic neurons.The work was supported by a grant from the Association for the Aid of Crippled Children, New York, and was carried out within a research organization sponsored by the Swedish Medical Research Council (grants No. B71-14X-56-07A and B71-14X-712-06A).     

Age and changes in the nerve supply of the superior and inferior venae cavae in man

The superior and inferior venae cavae have been studied in 51 human being at various age, beginning from newborns using histochemical methods. After birth concentration of neural plexuses in the venae cavae increases up to the period of puberty, and then after 40years of age, certain rarefication of adrenergic and then cholinergic neural plexuses is noted. Choline- and adrenergic innervation is revealed in myocardial fibers penetrating the walls of the superior and inferior venae cavae.     

Fluorescence histochemical study on regional differences in the sympathetic nerve supply of the choroid plexus from various laboratory animals

Summary The adrenergic nerve supply of the choroid plexus in all four ventricles was studied by the Falck-Hillarp histofluorescence technique in nine different species, and the noradrenaline concentration in whole plexus tissue was determined by a radioenzymatic method. The nerve density was usually in the order: third > lateral > fourth ventricular plexuses. Plexuses of the pig and cat possessed the largest number of nerves; the innervation was intermediary in the baboon, guinea-pig, rat, rabbit and hamster, whereas only few fluorescent nerves were found in the cow and mouse plexuses. Sympathetic denervation showed an ipsilateral supply from the superior cervical ganglia to the lateral plexuses and a mixed contribution to the midline plexuses. The total noradrenaline concentration varied between 0.10 and 0.73 ng per mg protein.     

Intrinsic and extrinsic innervation of the amphibians esophagic myenteric plexuses

The innervation of Rana ridibunda esophagus myenteric plexuses has been studied by the following methods: demonstration of cholinesterase activity; FIF method for catecholamines; immunohistochemistry for VIP, SP and SOM, and conventional electron microscopy. The cholinergic innervation is important in the esophagus wall where, in addition to the well known extrinsic component, there is a rich intrinsic plexus with cells and fibres widely distributed. The esophagus, together with the intestine, are the Rana gut portions where the adrenergic component is more broadly expressed. The adrenergic innervation seems to be almost entirely of extrinsic origin. We have shown that, for the tested peptides, there is an intrinsic innervation represented by VIP, SP and SOM like plexuses. We do not discard nonetheless an extrinsic component. The ultrastructure reveals the morphological characteristics of the enteric neurons as well as the fine inter-relationships between the nervous elements and the functional components of the esophagic wall.     

Axoplasmic flow of adrenaline and monoamine oxidase in amphibian sympathetic nerves

Summary The accumulation of both A and MAO proximal to a ligature on toad spinal nerves has been shown to occur at a slower rate than in mammals. As in mammals, there are two components of axonal transport in amphibian nerves, with the accumulation of A reaching a peak at between 4 and 7 days (cf. 2–4 days for NA in mammals), while MAO accumulation does not reach its maximum before 9 days (cf. 7 days in mammals). No accumulation occurs after sympathectomy, providing evidence for localization of MAO within amphibian sympathetic adrenergic nerves. Distal accumulation of MAO occurs in toad sympathetic nerves; this has not been reported to occur in mammalian nerves. Distal accumulation reaches a peak at 2–4 days, which suggests either a fast retrograde flow of MAO or that induction of MAO is occurring. These results are discussed in relation to differences between mammalian and amphibian sympathetic nerves and to the events occurring following ligation of these nerves.We wish to thank Judy Lenk, Vivienne Einhorn and Barbara Peachey for their assistance with the initial MAO histochemical work. This work was supported by grants from the National Heart Foundation of Australia and the Australian Research Grants Committee.