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  • 1
    Electronic Resource
    Electronic Resource
    The distributions and coexistence of peptides in nerve fibres of large bowel affected by Hirschsprung's disease (1991)
    Springer
    Pediatric surgery international 6 (1991), S. 322-332 
    Details
    ISSN: 1437-9813
    Keywords: Neuropeptides ; Coexistence ; Hirschsprung's disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The distributions of nerve fibres immunoreactive for the peptides calcitonin gene-related peptide (CGRP), enkephalin (ENK), neuropeptide Y (NPY), somatostatin (SOM), substance P (SP), and vasoactive intestinal peptide (VIP) and the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) were studied in healthy colon and samples of ganglionic and aganglionic colon from cases of proven Hirschsprung's disease. Studies of coexistence of reactivities in nerve fibres were performed to predict the possible origins of fibres that are found in the aganglionic bowel, e. g., from sensory or sympathetic ganglia. The muscularis externa of the ganglionic colon contained many nerve fibres immunoreactive for ENK, SP, and VIP, fewer for NPY, and only rare fibres reactive for CGRP, SOM, or TH. In ganglionic colon reactivities for SP and ENK coexisted in nerve fibres in the muscularis externa but in aganglionic colon no ENK immunoreactivity was found and most SP fibres were double-labelled with CGRP reactivity, indicating their probable sensory nature. Abnormally increased numbers of somatostatin-reactive fibres and noradrenergic fibres (marked by TH) were noted in the external muscle, but no coexistence was seen between these reactivities and only a small proportion of the noradrenergic fibres in the muscle showed NPY reactivity although almost all around blood vessels did. Many fibres in the diseased segment had coexistence of NPY and VIP reactivities; these may arise from more orally located intrinsic cell bodies or from pelvic parasympathetic ganglia. In the mucosa of aganglionic colon there was a striking lack of SP-reactive fibres while other fibre types were often normal in number. It is concluded that nerve fibres from sensory ganglia, sympathetic ganglia, nerve cells located more oral in the ganglionated part, and possibly from pelvic parasympathetic ganglia invade the aganglionic bowel in Hirschsprung's disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00178648
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  • 2
    Electronic Resource
    Electronic Resource
    Charybdotoxin and iberiotoxin but not apamin abolish the slow after-hyperpolarization in myenteric plexus neurons (1994)
    Springer
    Pflügers Archiv 428 (1994), S. 300-306 
    Details
    ISSN: 1432-2013
    Keywords: Potassium channels ; Enteric nervous system ; After-hyperpolarization ; Toxins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Myenteric neurons of guinea-pig ileum were studied with intracellular microelectrodes. The specific toxins charybdotoxin, iberiotoxin and apamin were used to characterize the prolonged after-hyperpolarizations of AH neurons in this preparation. Charybdotoxin and iberiotoxin blocked prolonged after-hyperpolarizations in 23 of 24 AH neurons, but apamin had no effect on 5 of 5 AH neurons. Abolition of the after-hyperpolarizations was accompanied by depolarization and increases in input resistances of those AH neurons affected, but the shapes of action potentials were unchanged. The excitability of the AH neurons was enhanced as shown by an increase in the number of action potentials evoked by a 500-ms depolarizing current pulse or by a train of 15 ms depolarizing current pulses (10Hz). The other class of myenteric neurons, S neurons, was also investigated. The 19 S neurons studied fired action potentials only at the start of a 500 ms depolarization, but the toxins had no effect on this behaviour or on their other properties. Intracellular injection of Neurobiotin into the neurons studied and subsequent immunohistochemical staining to localise the calcium-binding protein, calretinin, indicated that all major classes of S neurons were included in the sample. Thus, the prolonged after-hyperpolarizations in AH neurons may be due to opening of a large-conductance (BK) calcium-dependent potassium channel, but similar channels play little or no role in regulation of the excitability of S neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00724511
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization of 5-HT receptors mediating contraction and relaxation of the longitudinal muscle of guinea-pig distal colon in vitro (1994)
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 349 (1994), S. 455-462 
    Details
    ISSN: 1432-1912
    Keywords: 5-HT receptors ; Guinea-pig colon ; Longitudinal muscle ; Tachykinins ; Enteric neurons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A range of agonists and antagonists were used to characterize the receptors through which 5-hydroxytryptamine (5-HT) contracts and relaxes the longitudinal muscle of segments of guinea-pig distal colon, in vitro. 5-HT contracted the longitudinal muscle over the concentration range 10−9 to 10−4 mol/l. The 5-HT3 receptor agonist, 2-methyl-5-HT, produced concentration dependent contractions over the range 10−6 to 10−4 mol/l. 5-methoxytryptamine, an agonist at 5-HT4 receptors, caused contractions over a concentration range of 10−8 to 10−4 mol/l. The 5-HT4 antagonist, SDZ 205-557 (5 × 10−7 mol/l) substantially suppressed the responses to low concentrations of 5-HT and to 5-methoxytryptamine, but had no effect on the responses to higher concentrations of 5-HT. In contrast, the 5-HT3 antagonist, granisetron (10−6 mol/l), blocked the effect of 2-methyl-5-HT and substantially depressed responses to high concentrations of 5-HT, but had no effect on lower concentrations of 5-HT. Granisetron produced a small reduction in the response to 5-methoxytryptamine. Tetrodotoxin (TTX) (3 × 10−7 mol/l) almost abolished the response to 5-methoxytryptamine and markedly suppressed the response to 2-methyl-5-HT, but the responses to 5-HT were only partially reduced. The 5-HT, antagonist, methiothepin 10−6 mol/l. depressed the response to 5-HT 10−7 to 10−4 mol/l. and blocked its TTX insensitive component. The 5-HT2 antagonist, ketanserin, in concentrations up to 10−5 mol/l, had no effect on the contractions evoked by 5-HT. The response to 5-HT was substantially depressed by hyoscine (3 × 10−6 mol/l. The tachykinin antagonist, spantide 10−5 mol/l. depressed the response to 5-HT but to a lesser extent than hyoscine. Spantide and hyoscine combined completely blocked the contractile responses to 5-HT Responses to 2-methyl-5-HT were partially suppressed by hyoscine (3 x 10−6 mol/l. and spantide (10−5 mol/l) and completely blocked when both byoscine and spantide were present. Contractions evoked by 5-methoxytryptamine were partially blocked by hyoscine (3 × 10−6 mol/l) and were unaffected by spantide (10−5 mol/l), but a combination of hyoscine and spantide completely blocked such responses. When the excitatory transmission was blocked with hyoscine (3 × 10−6 mol/l) and spantide 10−5 mol/l) and the tone of the muscle raised, an inhibitory response to 5-HT was revealed that had a threshold concentration between 10−7 mol/l) and 3 × 10−7 mol/l, and a maximum effect at 10−4 mol/l. It was blocked by TTX (3 × 10−7 mol/l) and granisetron 10−6 mol/l. while N-nitro-l-arginine (NOLA) (10−4 mol/l) and SDZ 205-557 (5 × 10−7 mol/l) had no effect. Apamin A 10−6 mol/l. partially suppressed this response. It is concluded that 5-HT3, 5-HT4 and 5-HT1-like receptors mediate contraction of the longitudinal muscle of the distal colon. The 5-HT3 and 5-HT4 receptors are located on the excitatory motor neurons innervating the longitudinal muscle and the 5-HT1-like receptor is located on the muscle. 5-HT3 receptors are also found on inhibitory neurons to the muscle.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00169133
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  • 4
    Electronic Resource
    Electronic Resource
    Co-localization of nitric oxide synthase immunoreactivity and NADPH diaphorase staining in neurons of the guinea-pig intestine (1992)
    Springer
    Histochemistry and cell biology 97 (1992), S. 375-378 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal nitric oxide synthase (NOS), an enzyme capable of synthesizing nitric oxide, appears to be identical to neuronal NADPH diaphorase. The correlation was examined between NOS immunoreactivity and NADPH diaphorase staining in neurons of the ileum and colon of the guinea-pig. There was a one-to-one correlation between NOS immunoreactivity and NADPH diaphorase staining in all neurons examined; even the relative staining intensities obtained were similar with each technique. To determine whether pharmacological methods could be employed to demonstrate that NADPH diaphorase staining was due to the presence of NOS, tissue was pre-treated with NG-nitro-l-arginine, a NOS inhibitor, or l-arginine, a natural substrate of NOS. In these experiments on unfixed tissue, it was necessary to use dimethyl thiazolyl tetrazolium instead of nitroblue tetrazolium as the substrate for the NADPH diaphorase histochemical reaction. Neither treatment caused a significant decrease in the level of NADPH diaphorase staining, implying that arginine and NADPH interact at different sites on the enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00270041
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  • 5
    Electronic Resource
    Electronic Resource
    Origins of synaptic inputs to calretinin immunoreactive neurons in the guinea-pig small intestine (1993)
    Springer
    Journal of neurocytology 22 (1993), S. 531-546 
    Details
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Calretinin immunoreactivity is almost completely confined to two classes of neuron in the myenteric plexus of the guinea-pig small intestine, longitudinal muscle motor neurons and ascending interneurons. Nerve cell bodies of the two classes can be readily identified by their sizes and positions in ganglia. The motor neurons, which are small Dogiel type I neurons, are about 20% and the interneurons, which are medium-sized Dogiel type I neurons, are about 5% of myenteric neurons. In the present work, we have also discovered a minor population (0.1%) of small filamentous neurons. In unoperated regions of intestine, at the light microscopic level, numerous calretinin immunoreactive nerve fibres were found in the tertiary plexus that innervates the longitudinal muscle and a medium density of varicose fibres formed pericellular endings in the myenteric ganglia. After double myotomy operations, in areas of plexus 0.5 to 1.5 mm wide which were isolated from ascending and descending inputs, calretinin-immunoreactive fibres of the tertiary plexus were unchanged, but the periceliular endings in the ganglia disappeared. Both the ascending interneurons and the longitudinal muscle motor neurons received ultrastructurally identified synapses and close axonal contacts that were calretinin-immunoreactive. These were counted in semi-serial sections from normal intestine and from regions between myotomy operations. In unoperated intestine, the proportions of calretinin-immunoreactive synapses on small, calretinin-immunoreactive, Dogiel type I nerve cells and small filamentous nerve cells were 30% and 0.1% respectively and on medium-sized Dogiel type I cells the proportion was 28%. Electron microscopy revealed an almost complete loss of immunoreactive inputs to the small Dogiel type I cells between double myotomies, but the number of unreactive inputs was the same as in normal intestine. This work demonstrates that the ascending calretinin-immunoreactive interneurons connect with one another to form ascending chains in the myenteric plexus and that they also provide about 1/3 of the inputs received by calretinin-immunoreactive longitudinal muscle motor neurons. Many of the remaining inputs to these motor neurons are local; we have deduced that these are mainly from primary sensory neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01189041
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  • 6
    Electronic Resource
    Electronic Resource
    Ultrastructure and synaptology of neurons immunoreactive for gamma-aminobutyric acid in the myenteric plexus of the guinea pig small intestine (1990)
    Springer
    Journal of neurocytology 19 (1990), S. 539-549 
    Details
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immunoreactivity for gamma-aminobutyric acid is located in one morphologically-defined class of nerve cell body in the myenteric plexus of the guinea pig small intestine. These are a subgroup of the Dogiel type I nerve cells, characterized by their lamellar dendrites, about 1 μm thick and flattened in the plane of the myenteric plexus, and one (or rarely two) long axonal process that extends to either the longitudinal or the circular muscle. At an ultrastructural level the dendrites were characterized by their open cytoplasm in which were scattered granular vesicles, pale mitochondria, Golgi apparatus and endoplasmic reticulum. A large proportion of the dendritic surface was in direct contact with the extra-ganglionic space. In the cell body region, which was away from the ganglion surface, the nucleus was surrounded by a thin rim of cytoplasm. The cytoplasmic features are quite distinct from those of Dogiel type II neurons but they were shared by many other non-immunoreactive neurons. Synaptic inputs, which were all non-immunoreactive, were found on the dendrites, cell bodies, axon hillocks and axons of the gamma-aminobutyric acid-immunoreactive neurons. The predominant vesicle type in the presynaptic elements was the small clear vesicle, 40–60 nm in diameter. Based on two gamma-aminobutyric acid-immunoreactive cells that were examined in serial section, about 40–50% of synapses are dendritic, 20–25% are somatic, and 30–35% are on the axon hillock or first 50–70 μm of the axon. No synapses formed by immunoreactive varicosities were found on non-immunoreactive neurons or in the neuropil of the myenteric ganglia. Moreover, the lamellar dendrites or soma of gamma-aminobutyric acid neurons were never presynaptic elements forming relationships with other elements in the ganglia. It is concluded that the gamma-aminobutyric acid reactive Dogiel type I neurons are motor neurons providing inputs to the circular and longitudinal muscle layers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01257242
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  • 7
    Electronic Resource
    Electronic Resource
    Light- and electron-microscopic immunochemical analysis of nerve fibre types innervating the taenia of the guinea-pig caecum (1992)
    Springer
    Cell & tissue research 270 (1992), S. 125-137 
    Details
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Caecum ; Neurochemistry ; Neuropeptides ; Nitric oxide synthase ; Chemical coding ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present work was undertaken to determine by immunocytochemical methods which of the putative enteric neurotransmitters are contained in axons supplying the guinea-pig taenia coli and what proportion of axons is accounted for by the presence of these substances. Numerous fibres displayed immunoreactivity for dynorphin (DYN), enkephalin (ENK), γ-aminobutyric acid (GABA), nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP), but, in contrast to other gut regions, fibres showing immunoreactivity for gastrin-releasing peptide, galanin and neuropeptide Y were rare in the taenia. Fibres reactive for calbindin, calcitonin gene-related peptide, cholecystokinin, 5-hydroxytryptamine and somatostatin were also rare. Tyrosine hydroxylase-like immunoreactivity (TH-LI) was present in numerous fibres that disappeared after extrinsic denervation, a procedure that did not detectably affect any of the other major groups of fibres. Simultaneous staining of extrinsically denervated preparations revealed that SP-LI and VIP-LI were located in separate fibres, and ultrastructural studies showed these to be 58% and 33% of intrinsic fibres supplying the muscle. Immunoreactivity for the general marker, neuron-specific enolase, was located in 95–98% of axons. ENK-LI and DYN-LI were in the same axons, and similar proportions of the fibres with either SP-LI or VIP-LI, about 85%, contained immunoreactivity for ENK and DYN. All VIP-LI fibres, but no SP-LI fibres, were reactive for NOS. The results imply that the taenia of the guinea-pig caecum is innervated by two major groups of enteric neurons: (i) excitatory neurons that contain ACh, SP, other tachykinins, and, in most cases, DYN-LI and ENK-LI; and (ii) inhibitory neurons that contain NOS-LI, VIP-LI, in most cases, the two opioids and, quite probably, ATP as a transmitter. GABA-LI is contained in a smaller population of intrinsic axons. Even though the taenia represents one of the simplest tissues for examining transmission from enteric neurons to intestinal muscle, it shares some of the complexity of other regions, in that four major axon types supply the muscle and both the enteric excitatory and enteric inhibitory neurons contain multiple transmitters.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00381887
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  • 8
    Electronic Resource
    Electronic Resource
    Total numbers of neurons in myenteric ganglia of the guinea-pig small intestine (1993)
    Springer
    Cell & tissue research 272 (1993), S. 197-200 
    Details
    ISSN: 1432-0878
    Keywords: Myenteric plexus ; Intestine, small ; Neurons ; Enteric nervous system ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Two techniques that are thought to stain all of the neurons in the myenteric ganglia of the intestine are NADH diaphorase histochemistry and immunhistochemistry using a “nerve cell body” antiserum. However, this assumption has never been directly verified. In the present study myenteric ganglia of the guinea-pig ileum were prepared as whole-mounts and stained with either of these techniques. All nerve cells that could be identified in the whole-mounts were counted. The whole-mounts were then embedded flat in resin and serially sectioned at 1 μm. Nerve cells were identified and counted from the serial sections, and the data compared to those obtained from the whole-mounts. NADH diaphorase histochemistry did not reveal all the neurons at incubation times that gave selective staining. In contrast, “nerve cell body” antiserum stained the entire neuronal population. To determine the total number of nerve cell bodies/ganglion and the proportion of nerve cell bodies with calbindin immunoreactivity, whole-mounts that had been processed for calbindin immunohistochemistry were serially sectioned and reconstructed. The total number of neurons per myenteric ganglion was 105±10 (SE). Calbindin-immunoreactive neurons comprised about 20% of the myenteric neurons, which is considerably less than previous estimates, because previously the total population has been underestimated. The spatial density of myenteric neurons in the undistended ileum of the guinea-pig is 17300 nerve cells/cm2.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00323587
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  • 9
    Electronic Resource
    Electronic Resource
    Evidence that myenteric neurons of the gastric corpus project to both the mucosa and the external muscle: myectomy operations on the canine stomach (1991)
    Springer
    Cell & tissue research 266 (1991), S. 475-481 
    Details
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Stomach ; Vasoactive intestinal peptide ; Galanin ; Gastrin-releasing peptide ; Substance P-Dog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of nerve cell bodies and fibres in the canine stomach was investigated using antibodies to the general neuronal marker, neuron-specific enolase. Prominent ganglia containing many reactive nerve cells were found in the myenteric plexus of the gastric corpus and antrum. Nerve cells were absent from the submucosa of the corpus and were extremely rare in the antrum. Renoval of areas of longitudinal muscle and myenteric plexus from the corpus (myectomy), with 7 days allowed for axon degeneration, resulted in the loss of fibres reactive for galanin, gastrin-releasing peptide, substance P and vasoactive intestinal peptide from both the circular muscle and mucosa in the area covered by the lesion. Combined vagotomy and sympathetic denervation did not significantly affect these fibres, but did cause fibres reactive for calcitonin gene-related peptide to degenerate. It is concluded that the myenteric plexus of the gastric corpus, like the myenteric plexus of the small intestine and colon, is the source of nerve fibres innervating the circular muscle, but, in contrast to other regions of the gastrointestinal tract, myenteric ganglia, not submucous ganglia, are the major, or sole, source of the intrinsic innervation of the mucosa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318588
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  • 10
    Electronic Resource
    Electronic Resource
    Distribution of enteric nerve cells projecting to the superior and inferior mesenteric ganglia of the guinea-pig (1993)
    Springer
    Cell & tissue research 271 (1993), S. 333-339 
    Details
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Prevertebral ganglia ; Retrograde tracing ; Calbindin ; Vasoactive intestinal peptide (VIP) ; Intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Retrograde tracing, using Fast Blue dye, was employed to determine the distribution of enteric nerve cells that project to the superior mesenteric and inferior mesenteric ganglia of the guinea-pig. Retrogradely labelled neurons were found in the myenteric but not submucous ganglia. When the superior mesenteric ganglion was injected, labelled neurons were found in low frequencies (less than 5 nerve cell bodies/cm2) in the duodenum, jejunum, ileum, caecum and proximal colon. The distal colon was analysed in five segments of equal length (1–5; oral to anal). Segment 1 had about 4 labelled nerve cells/cm2, whereas segments 2 to 5 displayed an average of about 25 nerve cells/cm2. The rectum contained about 36 labelled neurons/cm2. After injection of the inferior mesenteric ganglia with Fast Blue, no labelled neurons were found in the duodenum, jejunum, ileum or caecum. No labelled cells were observed in the gallbladder. A small number of labelled cells occurred in the proximal colon and in segment 1 of the distal colon. The frequency of labelled cells increased markedly in the more anal regions of the distal colon, and reached a peak in the rectum (138 cells/cm2). Both nerve lesions and immersion of the cut nerve in Fast Blue solution showed that the superior mesenteric nerve carries the axons of neurons located in the middle distal colon to the superior mesenteric ganglion. Almost half of the neurons in the rectum that project to the inferior mesenteric ganglia do so via the hypogastric nerves. Of neurons that projected to the inferior or superior mesenteric ganglia from the colon or rectum, similar proportions (about 75–80%) showed immunoreactivity for calbindin or VIP. For each of the prevertebral ganglia (coeliac, superior mesenteric and inferior mesenteric) the great majority of peripheral inputs arise from the large intestine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318620
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