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Neuropeptide Y in submucosal ganglia: regional differences in the innervation of guinea-pig large intestine.

Since information about possible regional differences in the innervation of the guinea-pig large intestine is incomplete, a comparative study was made of the occurrence of neurones and nerve fibres of the submucosa showing immunoreactivity (IR) to neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). In addition, a quantitative analysis was made of submucosal neurones in regions of guinea-pig large intestine selected for probable differences in their function. There were two principal findings: First, the density of NPY-IR neurone somata was high in the ascending colon (mean +/- SEM 3148 +/- 464 neurones/cm2; n = 5 animals) and progressively declined in an anal direction, the descending colon having 348 +/- 125 neurones/cm2 (in the same 5 animals); immunoreactive cell bodies were rare in the rectum. The reduced density was also reflected in a fall in the number of NPY-IR neurones/ganglion from 3.0 +/- 0.3 in the ascending colon to 0.5 +/- 0.2 in the descending colon. Second, varicose NPY-IR intraganglionic fibres were a conspicuous feature of the duodenum, caecum, transverse colon, descending colon and rectum, but not of the ileum, ascending colon or distal spiral. Moreover, in the descending colon and rectum the fibres were arranged in a loose 'cobweb' structure around non-NPY-IR neurone somata; in the caecum, there was an apparent paucity of NPY-IR somata but the exceptionally dense intraganglionic varicose fibre network may have obscured NPY-IR somata. In all regions, fibre baskets were rare. In the ascending colon, only 25 +/- 5% of ganglia (compared to 92 +/- 2% of ganglia in the descending colon) showed any intraganglionic nerve fibres; furthermore, when they occurred, these were not of the 'cobweb' type but, rather, they gave the ganglia a speckled appearance. In very immature fetuses at a stage of development when no neuropeptide somata could be found in either the myenteric or submucosal plexuses, many NPY-IR nerve fibres were present in the submucosa with a distribution similar to that of adult guinea pigs. With respect to the density of VIP-IR neurones in the large intestine, there was only a 40% reduction in the number of neurones/cm2 from proximal to distal colon, in contrast to the corresponding 90% reduction in the density of NPY-IR neurones. The number of VIP-IR neurones/ganglion (6.4) and the proportion of ganglia with VIP-IR fibres (> 90%) were constant. It is concluded that the striking regional dissimilarities in (i) the occurrence of NPY-IR neurone somata and (ii) in the disposition of intraganglionic NPY-IR nerve fibres indicate potentially important regional differences in the functions of neuropeptide Y as an antisecretory peptide in the local regulation of chloride transport in the mucosa and as a modulator of ganglionic transmission, respectively.

Animals↗

Age-dependent stimulation by atrium explants or nerve growth factor of nerve fibre outgrowth from cocultured embryonic rat sympathetic ganglia.

Whole sympathetic superior cervical ganglia of 14- and 15-day-old rat embryos were cultured in a collagen gel medium for 24 h with or without explants of heart atrium from the same animals or from newborn rats. The extent of nerve fibre outgrowth was estimated by counting the number of nerve fibres crossing each arc of a sector drawn in the microscope ocular. Only few nerve fibres extended from the ganglia of the 14-day-old embryos cultured in the pure control medium. Addition of exogenous nerve growth factor did not stimulate the poor growth. While the growth of nerve fibres in the presence of atrium explants of newborn rats did not significantly differ from that in cultures without any target tissue, the presence of atrium explants of the same, 14-day-old, embryo resulted in clearly enhanced fibre outgrowth from the ganglia, which was inhibited by antiserum to the nerve growth factor. The ganglia of 15-day-old embryos cultured without target tissue in the control medium produced only sparse fibre outgrowth which was excessively increased by exogenous nerve growth factor. Coculture with atrium explants of 15-day-old embryos or newborn rats produced an increased overall nerve fibre growth, as well as predominance of those fibres growing toward the target tissue. The growth-promoting effect of the atrium of 15-day-old embryos was only partially prevented by antiserum to the nerve growth factor, while that elicited by newborn rat atrium was totally inhibited.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Evidence for a rostrocaudal organization in dorsal root ganglia during development as demonstrated by intra-uterine WGA-HRP injections into the hindlimb of rat fetuses.

The development of the sensory innervation of the rat hindlimb was studied with special attention to the dorsal root ganglia and the lumbar plexus. Injections of wheat germ agglutinin-horseradish peroxidase were made into the hindlimb of 30 rat fetuses of gestational ages ranging from embryonic day 15-18. Additionally wheat germ agglutinin-horseradish peroxidase was applied to the sciatic nerves of 8 neonatal rats and 3 adults. The saphenous nerves of 2 neonatal rats were labeled. Injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the hindlimb of the fetuses result in completely and partially labeled dorsal root ganglia. Partial labeling always concerns the rostral or caudal part of a dorsal root ganglion. The associated dorsal roots of partially labeled dorsal root ganglia are also partially labeled in a corresponding rostrocaudal fashion. Reconstructions of the labeled nerves following injections into the hindlimb suggested that the somata of the sensory neurons of a particular nerve can be restricted to the rostral or caudal half of a dorsal root ganglion. For example: the rostral half of the fourth lumbar dorsal root ganglion belongs to the femoral nerve and its caudal half to the sciatic nerve. The results of application of wheat germ agglutinin-horseradish peroxidase to the central ends of the cut sciatic and saphenous nerves in neonatal rats confirmed these observations. So the rostrocaudal organization in the dorsal root ganglia stems from the distribution pattern of the peripheral nerves.

Animals↗

Calcium-binding proteins in primate basal ganglia.

This paper describes the distribution of the calcium-binding proteins calbindin-D28k. Parvalbumin and calretinin in primate basal ganglia. The data derive from immunocytochemical studies undertaken in squirrel monkeys (Saimiri sciureus) and in normal human individuals. In the striatum, calbindin labels medium-sized spiny projection neurons whereas parvalbumin and calretinin mark two separate classes of aspiny interneurons. The striatal matrix compartment is markedly enriched with calbindin while striatal patches (striosomes) display a calretinin-rich neuropil. In the pallidum, virtually all neurons contain parvalbumin but none express calbindin. Calretinin occurs only in a small subpopulation of both large and small pallidal neurons. In the subthalamic nucleus, there exists a multitude of parvalbumun-positive cells and fibers but the number of calretinin and calbindin-positive neuronal elements is small. In the substantia nigra/ventral tegmental area complex, calbindin and calretinin occur principally in dopaminergic neurons of the dorsal tier of the pars compacta and in those of the ventral tegmental area. Parvalbumin is strictly confined to the GABAergic neurons of the pars reticulata and lateralis. Calbindin-rich fibers abound in the pars reticulata and lateralis, while calretinin-positive axons are confined to the pars compacta. These results indicate that calbindin and parvalbumin are distributed according to a strikingly complementary pattern in primate basal ganglia. Calretinin is less ubiquitous but occurs in all basal ganglia components where it labels distinct subsets of neurons. Such highly specific patterns of distribution indicate that calbindin, parvalbumin and calretinin may work in synergy within primate basal ganglia.

Animals↗

Effect of thyrotropin releasing hormone on the accumulation of cAMP by parietal ganglia of a gastropod.

1. We had previously shown that thyrotropin releasing hormone (TRH) can stimulate the in vitro accumulation of cAMP by the parietal ganglia of the pond snail Lymnaea emarginata (Grimm-Jørgensen, 1980). 2. The mechanism by which TRH affects cAMP metabolism in parietal ganglia was further studied. 3. The TRH-induced accumulation of cAMP is preceded by a lag period and is of long duration. 4. TRH does not stimulate basal or guanylylimidodiphosphate-stimulated adenylate cyclase activity and is unable to cause an increase in cAMP accumulation when the incubation is carried out in the presence of a phosphodiesterase inhibitor. 5. This finding is compatible with the hypothesis that TRH may cause an increase in cAMP accumulation by means of decreasing phosphodiesterase activity. 6. When the ganglia were incubated with 3H-TRH and the localization of the labeled TRH examined by autoradiographic techniques, reduced silver grains were present only over glial and connective tissue elements. 7. The observed effect of TRH on the cAMP metabolism in parietal ganglia may be due to its action on non-neuronal cells.

1-Methyl-3-isobutylxanthine↗

Selective localization of alpha-fetoprotein and serum albumin within the sensory ganglia cells of developing chicken.

The presence of alpha-fetoprotein (AFP) and serum albumin (SA) has been described within developing brain cells of mammals and chicken. In order to avoid the influence of the choroid plexuses in the transport, via the cerebrospinal fluid, of these proteins to brain cells, we studied the presence of AFP and SA within chicken sensory ganglion cells during ontogeny using the indirect immunoperoxidase technique. Our results show that before day 9 of egg incubation, both AFP and SA can be detected within the trigeminal and spinal ganglia cells. Henceforth, the labeling for AFP starts decreasing in the spinal ganglia but not in the trigeminal ganglion. The labeling for SA remains constant in both structures. Differences between AFP and SA staining in the spinal ganglia are maximal on day 11 of incubation, when AFP is no longer detected. It is concluded that at this time, the uptake of AFP by spinal ganglia cells is switched-off while SA is still taken up. As the blood concentration of both proteins is similar at this time, the finding reported here gives support to the advanced hypothesis suggesting the presence of specific receptors for AFP and/or SA in embryonic neural cells.

Animals↗

Coexistence of substance P- and methionine5-enkephalin-like immunoreactivity in nerve cells of the myenteric ganglia in the cat ileum.

Intramural ganglia of colchicine-injected cat ileum were examined immunohistochemically using antisera to substance P (SP), vasoactive intestinal polypeptide (VIP) and methionine5-enkephalin (ENK). SP- and ENK-like immunoreactive nerve cells were preferentially located in the myenteric ganglia, whereas VIP-like immunoreactive nerve cells were distributed in both myenteric and submucous ganglia. A small number of nerve cells in the myenteric ganglia showed immunoreactivities to both ENK and SP antisera, but not to VIP antiserum. These findings provide supportive evidence for the coexistence of SP and ENK in a small population of neurones in the cat myenteric plexus.

Animals↗

Nicotinic transmission in sympathetic ganglia: blockade by the snake venom neurotoxin kappa-bungarotoxin.

Kappa-bungarotoxin, a snake venom alpha-neurotoxin, blocks nicotinic transmission in chick lumbar sympathetic ganglia as well as in the rat superior cervical ganglion. Intracellular and extracellular recordings support the conclusion that the toxin has a specific action at the postsynaptic nicotinic receptors on ganglia neurons. The affinity of kappa-bungarotoxin for receptors in chick ganglia appears to be higher than that for receptors in the rat ganglia. Kappa-bungarotoxin can thus be used as a ligand for studying nicotinic receptors in a variety of neuronal preparations.

Animals↗

The subserosal ganglia of the human taenia.

Specimens of the taenia from the sigmoid colon of female patients undergoing surgery for carcinoma of the rectum were studied histochemically and immunohistochemically for acetylcholinesterase (AChE) and for vasoactive intestinal polypeptide (VIP)-, substance P (SP)-, somatostatin (SOM)-, neuropeptide Y (NPY)-, calcitonin gene-related peptide (CGRP)- and Met-enkephalin (mENK)-immunoreactivity. Autonomic ganglia were observed on the serosal surface of the longitudinal muscle of the taenia. The subserosal ganglia contained SP-, mENK-, NPY-, SOM-, but not CGRP- or VIP-immunoreactive nerve fibres. In addition, they contained SP-, mENK- and NPY-, but not CGRP-, SOM- and VIP-immunoreactive nerve cell bodies (although CGRP- and VIP-immunoreactive nerve fibres were observed in the longitudinal muscle of the taenia). AChE-activity was found both in nerve fibres and nerve cell bodies in these ganglia. The greatest numbers of nerve cell bodies contained AChE, followed in decreasing order by SP, mENK and NPY. The possible function of the subserosal ganglia of the human taenia is discussed.

Acetylcholinesterase↗

Neuropeptide Y mRNA regulation in rat sympathetic ganglia: effect of reserpine.

To study the mechanism underlying trans-synaptic neuropeptide regulation, mRNA levels of neuropeptide Y were examined in the rat stellate and superior cervical ganglia using a specific neuropeptide Y cRNA probe. Basal levels of neuropeptide Y mRNA were detectable in total RNA extracts from single ganglia. Reserpine induced a large rise in ganglion neuropeptide Y mRNA. Decentralization prevented the increase of neuropeptide Y mRNA content in the ganglia. This suggests that the reserpine induced increase in neuropeptide Y mRNA was dependent on transsynaptic stimulation. Consequently, neuropeptide mRNA levels in sympathetic ganglia may be under the control of preganglionic impulse flow.

Autonomic Fibers, Preganglionic↗

Postnatal ontogeny of the neurotrophin receptors trk and trkB mRNA in rat sensory and sympathetic ganglia.

Using reverse transcription followed by polymerase chain reaction, we examined the expression of mRNA for the tyrosine kinase receptors trk and trkB in rat sensory and sympathetic ganglia during postnatal development. While the levels of both trk and trkB mRNA in the dorsal root ganglia (DRG) decreased two-fold, they increased by seven and two times, respectively, in superior cervical ganglia. The developmentally regulated and tissue-specific expression of trk and trkB genes suggest that peripheral ganglia differ in their responsiveness to neurotrophins in neonatal and adult rats. We found that the temporal pattern of trk expression in DRG neurons correlates with the observed age-dependent ability of nerve growth factor to induce the biosynthesis of the neuropeptide substance P.

Animals↗

Distribution of substance P-like immunoreactivity in the spinal cord and dorsal root ganglia of the human foetus and infant.

Using the indirect immunofluorescence method, the distribution of substance P-like-immunoreactivity was studied in spinal cord and dorsal root ganglia of 25 human foetuses ranging from 12 to 29 weeks of gestational age. The spinal cord and dorsal root ganglia of three infants (1 day-, 2 and 4 month-old) were also investigated as a post-natal reference. On the whole, the substance P distribution patterns seen in infants were already visible throughout most of foetal life. The highest density of substance P-like-immunoreactive fibres was localized over the superficial layers of the dorsal grey horn. Punctiform immunofluorescence was often found over the white matter especially in the funiculi dorsalis et lateralis. In the ventral horn, substance P immunoreactive fibres were few and far between in the grey matter and were only detected from foetal stage 16 weeks. In addition, longitudino-frontal sections through the dorsal regions revealed repetitive arrangements of substance P-like-immunoreactive fibres along the whole spinal cord. In dorsal root ganglia only a few immunoreactive cells were observed. These findings demonstrate the wide and early occurrence of substance P-like-immunoreactivity in the human foetus spinal cord and dorsal root ganglia. They suggest that the development of the substance P neuronal system begins early in ontogenesis and is regionally differentiated.

Fetus↗

Extent of survival and vascularization of adult superior cervical sympathetic or nodose ganglia transplanted into the septal nuclei or choroid fissure of adult rats.

Adult superior cervical sympathetic ganglia were auto-transplanted, and adult nodose ganglia were homografted into the septal nuclei or the choroid fissure of adult Wistar rats. At times from 4 h to 9 weeks after operation, the distribution of surviving transplanted neurons was compared with the development of the transplant vascularization, as visualized by transcardial Indian ink filling of the host vascular system. Within 24 h, the ganglionic neurons and Schwann cells of the interior of the transplants in both sites were necrotic. The surviving neurons and Schwann cells formed a shell, occupying those areas of the transplant periphery which were in direct contact with the host circulation. Occasional ink-filled vessels were evident at this time in transplants in the choroid fissure, but there were none in the septal nuclei, where vessels did not appear until the next day. Blood vessels reached the centre of the ganglia by 3-4 days in the choroid fissure and one week in the septal nuclei, the finest diameter capillaries forming last. At longer survivals there was a slow loss of neurons, notable between 1 and 2 months, and leading progressively (especially in the septal transplantation site) to the disappearance of all but a very small number of ganglionic neurons. The general findings were similar for both types of ganglion, and in both sites, but the initial cell loss was much greater for both types of ganglia in the septum (over 90%) as compared with about a 50% loss in the choroid fissure. The initial rapid cell loss was probably a result of ischaemia. The subsequent, slow progressive loss may be associated with failure to make or receive neuronal connections, or the absence of appropriate growth factors.

Animals↗

Neurokinin A in capsaicin-sensitive neurons of the guinea-pig inferior mesenteric ganglia: an additional putative mediator for the non-cholinergic excitatory postsynaptic potential.

The presence of neurokinin-A-like immunoreactivity in guinea-pig inferior mesenteric ganglia was detected by radioimmunoassay procedures. Pretreating the animals with capsaicin 7 days prior to experimentations reduced the mean content of neurokinin-A-like immunoreactivity by 85% from its control value of 150 +/- 31.3 fmol per ganglion. High-performance liquid chromatography revealed that neurokinin-A-like immunoreactivity was heterogenous as in addition to neurokinin A, peaks corresponding to the amphibian tachykinin eledoisin and to neuropeptide K were detected, and they too were depleted by capsaicin. Electrophysiological studies showed that neurokinin A applied either by superfusion or by pressure ejection evoked a slow depolarization in the majority of inferior mesenteric ganglia neurons in vitro. Neurokinin-A-evoked depolarizations in the majority of cells tested were associated with a small increase in membrane input resistance. However, the responses were increased by membrane hyperpolarization: the extrapolated mean equilibrium potential of neurokinin-A-induced depolarization was -36 mV. Removal of extracellular sodium but not chloride ions suppressed the neurokinin-A-induced depolarization. The slow depolarization elicited either by exogenously applied substance P or by repetitive stimulation of hypogastric nerves was reversibly eliminated in the presence of neurokinin A. Collectively, our studies suggest that neurokinin-A-like immunoreactivity may coexist with substance-P-like immunoreactivity in capsaicin-sensitive fibers in the guinea-pig prevertebral ganglia and that the similarity of the actions of neurokinin A on the one hand and substance P on the other raises the possibility that non-cholinergic excitatory potentials elicited in the inferior mesenteric ganglia may be generated by not one but a number of closely related tachykinins.

Animals↗

Quantitative histochemical analysis of cytochrome oxidase in rat dorsal root ganglia and its co-localization with carbonic anhydrase.

A quantitative histochemical method was developed and standardized and then used to characterize the heterogeneity of cytochrome oxidase activity among primary afferent neuronal cell bodies in dorsal root ganglia of rat. In addition, the relationship between cytochrome oxidase and carbonic anhydrase activities in these neurons was determine. In tests of the procedure, the density of cytochrome oxidase reaction product evaluated repeatedly in individual neurons within sections of ganglia was found to increase linearly over incubation periods of up to 6 h. The heterogeneity in cytochrome oxidase activity in ganglia was not simply a reflection of the heterogeneity in ganglion cell sizes. On the whole, each class of ganglion cell exhibited the full range of staining densities encountered but intense staining was observed in many more large type A cells than small type B cells. The latter, together with their termination fields within the substantia gelatinosa of the spinal cord, were lightly stained. A significant positive correlation was found between neuronal size and staining density (r = 0.43). However, the large scatter in the plot of these two variables suggests that the expression of cytochrome oxidase in sensory neurons is governed to a considerable extent by properties of these neurons that are unrelated to their size. Analysis of cytochrome oxidase and carbonic anhydrase activities in the same ganglion cells revealed that all neurons with dense staining for the oxidase were anhydrase positive. Conversely, however, some intensely anhydrase-positive cells exhibited only light staining for cytochrome oxidase. The heterogeneity of cytochrome oxidase activity among neurons in dorsal root ganglia may be related to the steady state electrophysiological activity of distinct populations of sensory neurons which in turn may be related to the specific sensory modalities these populations transmit. The observation that some neurons with the greatest abundance of carbonic anhydrase do not contain high or even moderate levels of cytochrome oxidase suggests some degree of dissociation between the functional requirement for carbonic anhydrase in sensory neurons and the rate of energy expenditure in these cells.

Animals↗

Atrial natriuretic factor constitutes an intrinsic functional unit within superior cervical ganglia of the rat.

The molecular forms of atrial natriuretic factor were studied in the sympathetic ganglia of the rat. The peptide atrial natriuretic factor was also tested for its ability to induce intracellular changes in ganglionic elements. Chromatographic evaluation of extracted ganglionic atrial natriuretic factor revealed the presence of proatrial natriuretic factor together with lower molecular weight peptides. Atrial natriuretic factor induced a maximal six-fold increase of cGMP accumulation within ganglia in vitro, most probably in principal ganglionic cells. Its effect on cGMP was not mediated by acetylcholine or any other neurotransmitter because it persisted after muscarinic receptor blockade and in a calcium-free medium and was not affected by ganglia decentralization. Thus, atrial natriuretic factor appears to be produced by a structural neural component of ganglia (in preganglionic cholinergic neurons or small intensely fluorescent cells?) and has receptors at sites different from its source. It is suggested that atrial natriuretic factor may be locally involved in the process of neurotransmission and may be yet another peptide neurotransmitter and/or neuromodulator.

Animals↗

Serotonin-immunoreactive varicosities in the cell body region and neural sheath of the snail, Helix pomatia, ganglia: an electron microscopic immunocytochemical study.

The distribution and connections of serotonin-immunoreactive fibers in the cell body region and neural sheath of the central ganglia of the snail, Helix pomatia, have been examined. The cell body region of the ganglia is supplied by an extremely dense network of varicose serotonin-immunoreactive fibers which surround neuronal perikarya in the ganglia. Immunoreactive processes also run to the neural sheath of both the ganglia and the peripheral nerve roots, forming a dense network. Electron microscopy revealed five different connections of serotonin-immunoreactive varicosities, according to their target: (i) non-specialized contacts with neuronal perikarya; (ii) non-specialized contacts with axon processes on the surface of the peripheral nerve roots; (iii) non-specialized neuromuscular connections with smooth muscle fibers in the neural sheath; (iv) varicosities engulfed by glial processes in both the cell body region and neural sheath; (v) varicosities embedded in the connective tissue elements of the sheath either partly or completely free of glial processes. In all cases of appositions no membrane specializations could be observed on either site of the contacts. These observations provide morphological evidence for non-synaptic regulatory actions of serotonin-containing neurons in Helix central nervous system: (i) modulation of the activity of neuronal perikarya; (ii) involvement in neuromuscular regulation; (iii) neurohormonal modulation of peripheral processes by release through the neural sheath.

Animals↗

Excitatory amino acid binding sites in the basal ganglia of the rat: a quantitative autoradiographic study.

Quantitative receptor autoradiography was used to determine the distribution of excitatory amino acid binding sites in the basal ganglia of rat brain. alpha-Amino-3-hydroxy-5-methylisoxazole-4-propionic acid, N-methyl-D-aspartate, kainate, quisqualate-sensitive metabotropic and non-N-methyl-D-aspartate, non-kainate, non-quisqualate glutamate binding sites had their highest density in striatum, nucleus accumbens, and olfactory tubercle. Kainate binding was higher in the lateral striatum but there was no medial-lateral striatal gradient for other binding sites. N-Methyl-D-aspartate and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid binding sites were most dense in the nucleus accumbens and olfactory tubercle. There was no dorsal-ventral gradient within the striatal complex for the other binding sites. Other regions of the basal ganglia had lower densities of ligand binding. To compare binding site density within non-striatal regions, binding for each ligand was normalized to the striatal binding density. When compared to the striatal complex, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid and metabotropic binding sites had higher relative density in the globus pallidus, ventral pallidum, and subthalamic nucleus than other binding sites. Metabotropic binding also had a high relative density in the substantia nigra. Non-N-methyl-D-aspartate, non-kainate, non-quisqualate glutamate binding sites had a high relative density in globus pallidus, ventral pallidum, and substantia nigra. N-Methyl-D-aspartate binding sites had a low relative density in pallidum, subthalamic nucleus, substantia nigra and ventral tegmental area. Our data indicate heterogeneous distribution of excitatory amino acid binding sites within rat basal ganglia and suggest that the character of excitatory amino acid-mediated neurotransmission within the basal ganglia is also heterogeneous.

Animals↗