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W Kummer

Publications and source records attributed to W Kummer.

At least 109 records · Page 6Linked to original sources

Neurotensin-like immunoreactivity in presumptive baroreceptor neurons innervating the guinea pig carotid sinus.

This study was carried out to determine the ultrastructure and origin of previously described neurotensin-like immunoreactive (NT-LI) nerve fibres in the wall of the carotid sinus of the guinea pig. In our degeneration experiments, these NT-LI axons were unaffected by surgical sympathectomy but disappeared in response to transection of the carotid sinus nerve, thus suggesting a sensory origin. This assumption could be supported by combined retrograde tracing and immunohistochemistry. Primary afferent neurons of the sensory glossopharyngeal (petrosal) ganglion projecting to the carotid sinus were identified by injecting the retrograde fluorescent tracer, Fluoro-Gold, into the sinus wall, and some of these neurons displayed NT-LI. Within the carotid sinus, the terminals of these NT-LI sensory neurons exhibited ultrastructural features characteristic of baroreceptor endings, i.e. axonal swellings filled with mitochondria and closely associated to elastin. However, many endings also fulfilling the ultrastructural criteria for baroreceptors were devoid of immunolabelling. Thus, we conclude that the NT-LI terminals constitute a subgroup rather than the entire population of baroreceptor endings within the guinea pig carotid sinus. With respect to the established pharmacological effects of NT in guinea pig, we propose an involvement of NT-LI fibres in the modulation of baroreception at the peripheral level.

Animals↗

Expression of c-JUN, JUN B and JUN D proteins in rat nervous system following transection of vagus nerve and cervical sympathetic trunk.

Expression of c-JUN, JUN B and JUN D proteins was investigated in axotomized neurons following transection of the vagus nerve and the cervical sympathetic trunk in the rat. Vagotomy induced the expression of c-JUN and JUN D in the nodose ganglion, dorsal motor nucleus of the vagus nerve and nucleus ambiguus, whereas JUN B was not expressed in these areas, c-JUN and JUN D appeared after 10 h in the nodose ganglion and after 24 h in the dorsal motor nucleus of the vagus nerve with a maximum of immunoreactivity after 48 h. The c-JUN protein remained expressed at an increased level up to 100 days, whereas the immunoreactivity of JUN D declined after five days. Crush of the vagus nerve initially evoked an intense expression of c-JUN and JUN D, but in the course of regeneration the expression of c-JUN and JUN D had returned to more basal levels after 100 days. Similar to vagotomy, application of colchicine and vinblastine on to the intact vagus nerve induced expression of c-JUN and JUN D. On the other hand, application of lidocaine prior to vagotomy did not prevent the expression of these proteins. Transection of the cervical sympathetic trunk induced expression of c-JUN and JUN D, but not of JUN B, in the preganglionic sympathetic neurons of thoracic spinal cord. In these neurons, expression of c-JUN was still enhanced after 60 days whereas JUN D had returned to basal level. One hour after vagotomy, c-JUN and JUN B were transynaptically expressed in the area of central termination of sensory vagal neurons and declined within 10 h to basal levels. JUN D showed a late onset of expression, it appeared after 5 h and persisted for 60 days in this area. We postulate that the expression of c-JUN and JUN D in axotomized neurons is induced by deprivation of a target-derived suppressor.

Animals↗

[Peculiarities of occupational health vibration measurements of manually controlled banging machines].

Specific problems of the measurement of mechanical vibrations on the grips of hand-held tools are discussed in this paper. Especially the measurement on percussive-typ tools, e.g. pneumatic hammers, percussive drills and others, is focussed. The possibilities of affecting the measuring results because of mistakes in the measurement procedure and because of technological peculiarities of the working process are explained and hints for minimizing these influences are given.

Bone Diseases↗

Vasoactive intestinal polypeptide receptor-like immunoreactivity in human sweat glands.

Human sweat and sebaceous glands were studied immunohistochemically with a monoclonal antibody recognizing a vasoactive intestinal polypeptide (VIP) receptor. The staining pattern correlated well with the known distribution of sympathetic VIP-containing nerves in the human skin. The luminal cell layer of the sweat gland ducts and some acinar cells of the secretory coil of eccrine sweat glands were the major sites of VIP receptor-like immunoreactivity. From these findings and the known pharmacological actions of VIP it is concluded that a major role of VIP released from sympathetic nerves in the skin is to regulate chloride reabsorption from the primary sweat at the ductal segment.

Humans↗

Immunohistochemistry of the guinea-pig trachea using an anti-idiotypic antibody recognizing substance P receptors.

The airways receive a dense innervation from sensory neurons containing substance P (SP). An anti-SP anti-idiotypic antibody (anti-Id ab) recognizing SP receptors was previously characterized pharmacologically and proved to be useful in immunohistochemistry of the central nervous system. This antibody was used to localize SP binding sites in the guinea-pig trachea by immunohistochemistry. Immunolabelling was considered as specific when it could be prevented by a) preabsorption of the anti-Id ab with a C-terminal specific monoclonal anti-SP antibody, and b) preincubation of the tissue sections with either of the tachykinins, substance P and neurokinin A, in the presence of the inhibitor of neutral endopeptidase, phosphoramidon, and addition of these compounds into the antibody incubation medium. Moreover, immunofluorescence was absent when the acetone-fixed of fresh frozen sections were exposed to the detergent Tween 20 prior to immunohistochemistry, which points to a membrane localization of the detected tissue antigen, as expected for SP receptors. Compared with previous reports on autoradiographic localization of SP receptors in the guinea-pig trachea, the present immunohistochemical approach proved to be superior in enabling discrimination of labelled elements: Trachealis muscle, cylindrical epithelial cells and some roundish, singly lying cells in the epithelium and subepithelial lamina propria displayed specific immunofluorescence. These morphological findings match well with the known pharmacological actions of SP on the guinea-pig trachea.

Animals↗

Neuronal pathways in the guinea-pig lumbar sympathetic ganglia as revealed by immunohistochemistry.

Tyrosine hydroxylase (TH)- and peptide-immunoreactivity of postganglionic neurons and of nerve fibres in guinea pig lumbar paravertebral sympathetic ganglia 2-4 after transection of the communicating rami and the visceral branches, respectively, were investigated by single- and double-labelling techniques. Six subpopulations of postganglionic neurons were discriminated immunohistochemically: two cell types, which were immunoreactive to only one of the applied antisera - TH, and vasoactive intestinal polypeptide (VIP); and four cell types in which immunoreactivity was colocalized - TH/neuropeptide Y (NPY), NPY/VIP, dynorphin/alpha-neoendorphin and dynorphin (alpha-neoendorphin)/NPY. Small intensely fluorescent (SIF) cells dependent on their location exhibited differential immunobehaviour to NPY-/dynorphin-(alpha-neoendorphin-) and TH-antisera. Immunoreactivity to substance P (SP), calcitonin gene-related peptide (CGRP), met-enkephalin-arg-phe (MEAP) and leu-enkephalin was present in nerve fibres but not in postganglionic neurons with frequent colocalization of SP/CGRP- and MEAP/leu-enkephalin- and, sometimes leu-enkephalin/SP- and dynorphin/SP-immunoreactivity. TH-immunoreactive intraganglionic nerve fibres were numerically more increased after cutting the visceral branches, than after transection of the communicating rami. Vice versa, NPY-, VIP-, dynorphin- and alpha-neoendorphin-immunoreactive nerve fibres were particularly increased in number after cutting the communicating rami. Many but not all of the nerve fibres exhibited colocalization of two of these peptides. SP-, CGRP-, and enkephalin-immunoreactive nerve fibres were not visibly affected by cutting the visceral branches but virtually disappeared after lesioning the communicating rami.

Animals↗

Catecholamines and catecholamine-synthesizing enzymes in guinea-pig sensory ganglia.

Cranial and spinal sensory ganglia of the guinea-pig were investigated by means of histochemistry and biochemistry for the presence of catecholamines and catecholamine-synthesizing enzymes. Sensory neurons exhibiting immunoreactivity to the rate-limiting enzyme of catecholamine synthesis, tyrosine hydroxylase (TH), were detected by immunohistochemistry in lumbo-sacral dorsal root ganglia, the nodose ganglion and the petrosal/jugular ganglion complex. The carotid body was identified as a target of TH-like-immunoreactive (TH-LI) neurons by the use of combined retrograde tracing and immunohistochemistry. Double-labelling immunofluorescence revealed that most TH-LI neurons also contained somatostatin-LI, but TH-LI did not coexist with either calcitonin gene-related peptide- or substance P-LI. TH-LI neurons did not react with antibodies to other enzymes involved in catecholamine synthesis, i.e., aromatic amino acid decarboxylase (AADC), dopamine-beta-hydroxylase (D beta H), and phenylethanolamine-N-methyl-transferase (PNMT). Petrosal neurons as well as their endings in the carotid body lacked dopamine- and L-DOPA-LI. Sensory neurons did not display glyoxylic acid-induced catecholamine fluorescence. Ganglia containing TH-LI neurons were kept in short-term organ culture after crushing their roots and the exiting nerve in order to enrich intra-axonal transmitter content at the ganglionic side of the crush. However, even under these conditions, catecholamine fluorescence was not detected in axons projecting peripherally or centrally from the ganglia. Sympathetic noradrenergic nerves entered the ganglia and terminated within them. Accordingly, biochemical analyses of guinea-pig sensory ganglia revealed noradrenaline but no dopamine. In conclusion, catecholamines within guinea-pig sensory ganglia are confined to sympathetic nerves, which fulfill presently unknown functions. The TH-LI neurons themselves, however, lack any additional sign of catecholamine synthesis, and the presence of enzymatically active TH within these neurons is questionable.

Animals↗

Simultaneous immunohistochemical demonstration of vasoactive intestinal polypeptide and its receptor in human colon.

The present study reports an immunohistochemical approach for localizing the immunoreactivity of vasoactive intestinal polypeptide (VIP) receptor in the human colon, by using a monoclonal antibody which recognizes the VIP-receptor of a human colonic adenocarcinoma cell line. Simultaneous demonstration of immunoreactive VIP-receptor of a human colonic adenocarcinoma cell line. Simultaneous demonstration of immunoreactive VIP-receptor and VIP was achieved by a double-labelling procedure employing immunogold silver staining for VIP-receptor, and a biotinylated secondary antibody followed by streptavidin-Texas Red, to visualize VIP. The immunoreactive VIP receptor was found at two locations receiving dense VIP innervation: myenteric ganglia and mucosal epithelium. Epithelial cells displayed intense labelling at the basolateral membrane, which confirmed earlier binding studies on fractionated membranes. A small number of enteroendocrine cells was also recognized by the VIP-receptor antibody. Smooth muscle and cells of the immune system were not stained by the monoclonal antibody, indicating that it recognized an epitope not common to VIP-receptors of all locations. Thus, the immunohistochemical approach of VIP-receptor localization differs from autoradiography in (a) precise cellular localization, (b) possibility of simultaneous demonstration of receptor and ligand immunoreactivity, and (c) selectivity to a certain receptor population which, however, is presently not fully characterized.

Antibodies, Monoclonal↗

Three types of neurochemically defined autonomic fibres innervate the carotid baroreceptor and chemoreceptor regions in the guinea-pig.

The innervation of the carotid body, carotid sinus, and neighbouring arteries (common carotid artery; external carotid artery; occipital artery; ascending pharyngeal artery) was investigated in guinea-pigs by means of glyoxylic acid-induced catecholamine-fluorescence and immunohistochemistry using a variety of antisera against neuropeptides and tyrosine hydroxylase (TH). Fibres displaying catecholamine-fluorescence, TH- and neuropeptide Y-like immunoreactivity (NPY-LI) were less numerous in the carotid sinus than in all other arterial segments. Vasoactive intestinal polypeptide (VIP)-LI axons were almost lacking in the common carotid, external carotid and occipital arteries, consistently found in the carotid sinus, and more numerous in the ascending pharyngeal artery. Catecholaminergic, TH-, NPY- and VIP-LI fibres were observed deep in the media of the carotid sinus, where the baroreceptor terminals are located. In contrast, they did not enter the media in the adjacent arterial segments. All these fibres disappeared following excision of the superior cervical ganglion, but were unaffected by combined transection of the carotid sinus nerve and resection of the no-dose ganglion, suggesting a sympathetic origin. Double-staining immunofluorescence revealed at least three types of autonomic, presumably sympathetic fibres in the carotid sinus: 1) TH+/NPY+, 2) NPY+/VIP+, and 3) VIP+ fibres. This points to a non-noradrenergic efferent innervation of the carotid sinus in addition to the hitherto known noradrenergic sympathetic fibres. The three populations of autonomic fibres seen in the carotid sinus were also observed in the carotid body, but the paucity of NPY+/VIP+ double-labelled fibres raises doubt as to the functional significance of this particular fibre type in modulating arterial chemoreception. The multiplicity of neurochemically defined autonomic nerves to the carotid baro- and chemoreceptor regions probably reflects functionally separate pathways that are differently regulated and exert different effects.

Animals↗

Immunohistochemical distribution and colocalization of regulatory peptides in the carotid body.

Current investigations on the immunohistochemical occurrence and co-occurrence of biogenic polypeptides in the mammalian carotid body were reviewed and extended by our own recent findings. The family of chromogranins and related peptides in glomus cells appears to have a widespread interspecies distribution, whereas other peptides investigated occur in a species-specific pattern. Immunoreactivity to antisera against opioids, which derive from the proenkephalin sequence, appears to be present in glomus cells of the rabbit, cat, dog, and a shrew. Conversely, glomus cells of pig and guinea pig predominantly are immunoreactive to cleavage products of prodynorphin, which co-occur in some cells with substance P and met-enkephalin-arg-phe, respectively. In the rat and Callithrix jacchus, opioid immunoreactivity is present in nerve fibres but not in glomus cells. Immunoreactivity to other peptides, such as neurotensin, cholecystokinin, neuropeptide Y, and galanin, is found only in one or two particular species. Neurotensin immunolabelling occurs in beagle dog glomus cells, which are known to lack substance P. Cholecystokinin immunoreactivity is present in glomus cells of dog and Callithrix, and co-exists with chromogranin A, neuropeptide Y, and substance P. Substance P appears to exist in both carotid body glomus cells and nerve fibres. Substance P immunoreactivity is present in glomus cells of all species investigated, except dog. Coexistence of substance P and calcitonin gene-related peptide (CGRP) is demonstrated in nerve fibres of the guinea pig carotid body, which originate in the petrosal and jugular ganglia. Other peptides visualized immunohistochemically in mammalian carotid body nerve fibres are vasoactive intestinal peptide and neuropeptide Y. The functional significance of the various peptides present in the carotid body is discussed.

Animals↗

Vagal paraganglia of the rat.

Paraganglia are associated with every branch of the rat vagus nerve except the pharyngeal branch. Some of the paraganglia closely resemble the glomus caroticum, whereas others appear like small, intensely fluorescent (SIF) cells of autonomic ganglia. The paraganglionic cells of SIF cell-like bodies (SLB) store catecholamines (the most abundant is probably noradrenaline) and in some cases neurotensin. The innervation pattern of SLB is variable and their physiological role remains unclear. Paraganglionic cells of glomus-like bodies (GLB) predominantly store dopamine and probably also to a lesser extent noradrenaline. These putative chemoreceptor organs receive sensory innervation from nodose ganglion neurons as revealed by degeneration experiments and by anterograde neuronal tracing. Substance P- and calcitonin gene-related peptide-immunoreactive fibres seen in the region of vascular entry into the GLB may account for some of these sensory fibres, but the peptide/classical transmitter stored in sensory terminals synapsing on paraganglionic cells is unknown. Ultrastructural immunocytochemistry revealed vasoactive intestinal polypeptide (VIP)-immunoreactive fibres lying in the interstitial space between paraganglionic cells and large capillaries. These fibres may originate from VIP-immunoreactive neurons, being frequently attached to GLB. The major difference between GLB and the glomus caroticum concerns their blood supply and related innervation: Arteries and arterioles do not penetrate into GLB and, accordingly, noradrenaline- and neuropeptide Y-containing nerve fibres are lacking within GLB. This peculiar arrangement of paraganglionic parenchyma and arterial blood supply may be one of the reasons for the different physiological properties of vagal and carotid arterial chemoreceptors.

Amines↗

Morphology and immunocytochemistry of two endocrine cell types in the guinea-pig esophageal epithelium.

Two types of endocrine cells in the basal layer of the keratinized stratified squamous epithelium in the guinea-pig esophagus were studied with immunohistochemistry by means of a streptavidin-biotin-bridge technique and by the immunofluorescence double-labelling technique. Cell-type I exhibited immunoreactivity to chromogranin A and to nucleosidetriphosphate-adenosinediphosphate-(ADP)-phosphotransfera se. Ultrastructurally, this cell type was characterized by small cytoplasmic dense-core vesicles in which immunoreactive product was localized. Cell-type II contained large membrane-limited granules, which were moderately electron dense. These granules displayed somatostatin immunoreactivity. Both cell types were located in close vicinity to non-myelinated nerve fibers and small blood vessels. The results provide evidence that, independent from the type of lining epithelium, the gastro-enteropancreatic system in guinea-pig extends to the lower portion of the esophagus.

Animals↗

Ultrastructure of calcitonin gene-related peptide- and substance P-like immunoreactive nerve fibres in the carotid body and carotid sinus of the guinea pig.

Previous studies have demonstrated that substance P- (SP) and calcitonin gene-related peptide-like immunoreactivities (CGRP-LI) coexist in sensory nerve fibres in the guinea-pig carotid body and carotid sinus. In the present study the ultrastructure of these nerve fibres was investigated by means of single- and double-labelling immunocytochemistry. In both, carotid body and carotid sinus immunoreactive fibres were unmyelinated axons of small diameter (0.12-0.56 microns). At the subcellular level, SP- and CGRP-LI were colocalized in intra-axonal dense core vesicles, suggesting corelease and simultaneous action of these two compounds. SP/CGRP-LI nerve fibres within the carotid body were mainly found in the interparenchymal connective tissue, but also occurred in relationship to blood vessels and nests of glomus cells. Neither in the carotid body not in the carotid sinus, SP/CGRP-LI axons corresponded to the large terminals which are generally considered to represent the main chemoreceptor and baroreceptor endings, respectively. Thus, SP/CGRP-LI fibres either belong to the chemo- and baroreceptors of the C-fibre class or constitute a fibre population not directly involved in conduction of baro- and chemoreflexes.

Animals↗

Peptidergic innervation of arterial chemoreceptors.

The peptidergic innervation of arterial chemoreceptor organs (the rat carotid body and vagal paraganglia; guinea pig carotid body) was studied immunohistochemically. Five different populations of nerve fibres in the guinea pig carotid body could be discriminated according to their origin and their chemical coding. The innervation pattern of the rat carotid body differed in some aspects. Comparison of the rat carotid body and vagal paraganglia suggested that autonomic neuropeptide Y-like immunoreactive fibres act primarily via vascular mechanisms rather than directly on the chemoreceptor tissue. Sensory fibres were shown to contain immunoreactivities for substance P, calcitonin gene-related peptide (rat and guinea pig) and somatostatin (guinea pig). The functional role of the identified peptide-containing sensory fibres remains to be established.

Animals↗

Neuropeptide distribution in the cervico-thoracic paravertebral ganglia of the cat with particular reference to calcitonin gene-related peptide immunoreactivity.

Paraffin sections of cervical and upper thoracic paravertebral ganglia of the cat were investigated by immunohistochemistry using antisera directed against calcitonin gene-related peptide (CGRP). The relationships of CGRP-immunoreactive structures to those exhibiting immunoreactivity to antisera against other regulatory peptides and dopamine-beta-hydroxylase (DBH), respectively, were studied in consecutive sections. Singly scattered CGRP-immunoreactive neuronal perikarya were observed in the superior and middle cervical ganglia as well as in the stellate ganglion. These neurons also displayed immunoreactivity to vasoactive intestinal polypeptide (VIP), and some additionally exhibited faint substance-P immunoreactivity. DBH- and neuropeptide Y-immunoreactive ganglion cells were not identical with CGRP-immunoreactive neuronal cell bodies. According to the immunoreactive properties of varicosities, which abut on CGRP/VIP-immunoreactive perikarya, three types of CGRP/VIP-immunoreactive ganglion cells could be distinguished: (1) CGRP/VIP-immunoreactive neurons being surrounded by somatostatin-immunoreactive nerve fibers, (2) neurons being approached by both DBH- and met-enkephalin-immunoreactive varicosities, and (3) neurons receiving both DBH- and neurotensin-immunoreactive fibers. The stellate and upper thoracic ganglia harbored clusters of intensely VIP-immunoreactive somata, which lacked CGRP-immunoreactivity. Fine somatostatin-immunoreactive and coarse CGRP-immunoreactive fibers were distributed within these clusters, whereas patches of neurotensin-immunoreactive fibers were complementarily arranged. At all segmental levels investigated, a few postganglionic neurons were approached by both CGRP-immunoreactive and substance P-immunoreactive varicosities, but lacked a VIP-immunoreactive innervation. Therefore, CGRP/substance P-immunoreactive fiber baskets appeared rather to be of extraganglionic origin than to emerge from intraganglionic CGRP/VIP/SP neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Retrograde neuronal labelling and double-staining immunohistochemistry of tachykinin- and calcitonin gene-related peptide-immunoreactive pathways in the carotid sinus nerve of the guinea pig.

The origin of tachykinin- and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibres in the guinea pig carotid body and carotid sinus was determined by retrograde labelling of the carotid sinus nerve with Fluoro-gold and immunohistochemical double staining with fluorescein- and rhodamine-conjugated second antisera. Fluoro-gold-labelled perikarya with characteristic features of primary sensory neurones were numerous in the glossopharyngeal (petrosal) ganglion and occurred rarely in the closely attached superior vagal (jugular) ganglion. An efferent pathway from the brainstem could not be detected. Co-existence of tachykinin- and CGRP-LI was observed in 25-47% of labelled sensory neurones; less than 1% of Fluoro-gold-containing perikarya were exclusively stained by CGRP antiserum. Co-existence of tachykinin- and CGRP-LI was also demonstrated in nerve fibres of the carotid body and carotid sinus. Somatostatin-, cholecystokinin- and dynorphin-LI did not co-exist with tachykinin-LI in these fibres. Thus, tachykinin/CGRP-LI fibres in the carotid presso- and chemoreceptive areas exhibit a peptide pattern being generally characteristic for sensory fibres supplying great vessels in the guinea pig. In view of the present findings doubt is raised as to a primary involvement of these fibres in presso- or chemoreception, although a modulatory influence on these specific functions appears to be likely.

Animals↗