Selective labeling of vagal sensory nerve fibers in the lower esophageal sphincter with anterogradely transported WGA-HRP

Clerc, Nadine; Condamin, Madeleine

doi:10.1016/0006-8993(87)91464-8

Cited by 31 publications

(21 citation statements)

References 31 publications

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“…The highly arborizing terminal endings at the myenteric ganglia of the esophagus, which have been called intraganglionic laminar endings (IGLEs) (Rodrigo et al 1975;Berthoud et al 1997), were marked by intense fluorescence (Figure 6). In addition, fibers were also observed in the submucous and mucous layer in the cervical and lower part of the esophagus, as described in previous studies (Clerc and Condamin 1987;Neuhuber 1987) (Figure 3). Unspecific staining was occasionally found in erythrocytes and macrophages.…”

Section: Peripheral Projections Of the Nodose Afferentssupporting

confidence: 83%

“…1994;Berthoud et al 1997). Vagal afferents were also observed in the submucosa and mucosa of the esophagus in rat and cat, some penetrating deeply into the epithelium (Clerc and Condamin 1987;Neuhuber 1987;Neuhuber and Clerc 1990). These different types of vagal afferents were also detected by tyramide-biotin amplification and subsequent incubation with streptavidin-FITC.…”

Section: Discussionmentioning

confidence: 95%

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Tyramide Amplification Allows Anterograde Tracing by Horseradish Peroxidase-conjugated Lectins in Conjunction with Simultaneous Immunohistochemistry

Kressel

1998

J Histochem Cytochem.

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SUMMARY Current protocols for a combined approach of anterograde tracing with carbocyanine dyes or horseradish peroxidase (HRP) conjugates and immunohistochemistry represent a compromise between sensitive detection of the tracer and the immunohistochemical procedure. Therefore, it was investigated whether the use of tyramide amplification allows sensitive anterograde tracing with wheat-germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) in conjunction with simultaneous immunohistochemistry. Vagal afferents were anterogradely labeled by injection of WGA-HRP into the nodose ganglion of rats. By use of tyramide-biotin amplification, a dense fiber plexus of vagal afferents was visualized centrally in the nucleus of the solitary tract and in retrogradely labeled neurons in the dorsal vagal nucleus. In the esophagus and duodenum, large-and small-caliber vagal fibers and terminals could be demonstrated comparably to conventional tracing techniques using carbocyanine dyes or WGA-HRP and TMB histochemistry. Combination with immunohistochemistry could easily be done, requiring only one more incubation step, and did not result in loss of sensitivity of the tracing. With this method and confocal microscopy, the presence of Ca binding proteins in vagal afferent terminals could be demonstrated. Tyramide amplification allows sensitive anterograde tracing with low background staining in conjunction with immunohistochemistry of intra-axonal markers.

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Section: Peripheral Projections Of the Nodose Afferentssupporting

confidence: 83%

Section: Discussionmentioning

confidence: 95%

Tyramide Amplification Allows Anterograde Tracing by Horseradish Peroxidase-conjugated Lectins in Conjunction with Simultaneous Immunohistochemistry

Kressel

1998

J Histochem Cytochem.

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“…One previous study in the cat esophagus tried to identify vagal afferent nerve endings employing injection of horseradish peroxidase in the nodose ganglion and subsequent light microscopy. They indicated that numerous afferent fibers were running through the muscle layers, but it could not be established whether nerve endings were present within the muscle layers (10). ICC were not identified in that study.…”

mentioning

confidence: 75%

“…Interestingly, despite gap junction contacts between degenerating ICC and smooth muscle cells, the latter remained undamaged in many of the animals, suggesting that the trophic influence is not essential for smooth muscle cells. It is likely that the ICC are associated with vagal afferent sensory fibers since injection in the nodose ganglion labeled these neural structures and not motor fibers (10). Furthermore, Fox and coworkers (20,49) showed an intimate relationship between IMAs and ICC-IM in the fundus.…”

Section: Discussionmentioning

confidence: 99%

Survival dependency of intramuscular ICC on vagal afferent nerves in the cat esophagus

Huizinga¹,

Reed²,

Berezin³

et al. 2008

American Journal of Physiology-Regulatory, Integrative and Comp

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cells of Cajal (ICC) have been proposed as stretch receptors for vagal afferent nerves in the stomach based on immunohistochemical studies. The aim of the present study was to use electron microscopy and the anterograde degeneration technique to investigate ultrastructural features and survival dependency of ICC associated with vagal afferent innervation of the cat esophagus. This is the first report on the ultrastructural characteristics of ICC in the cat esophagus. Intramuscular ICC (ICC-IM) were identified throughout the musculature, whereas ICC in the myenteric plexus were rare. ICC-IM were particularly numerous in septa aligned with smooth muscle bundles. They were in synapse-like contact with nerve varicosities and in gap junction contact with smooth muscle cells. Smooth muscle cells also made contact with ICC through peg and socket junctions. Precision damage through small-volume injection of saline in the center of the nodose ganglion from the lateral side, known to selectively affect sensory nerves, was followed within 24 h by degeneration of a subset of nerve varicosities associated with ICC-IM, as well as degeneration of the associated ICC-IM. Smooth muscle cells were not affected. Nerves of Auerbachs plexus and associated ICC were not affected. In summary, ICC-IM aligning the esophageal muscle bundles form specialized synapse-like contacts with vagal afferent nerves as well as gap junction and peg-and-socket contacts with smooth muscle cells. This is consistent with a role of ICC-IM as stretch receptors associated with vagal afferent nerves; the ICC-vagal nerve interaction appears essential for the survival of the ICC. interstitial cells of Cajal; esophagus; stretch receptor; vagus; afferent innervation; intramuscular interstitial cells of Cajal FOR NORMAL ESOPHAGEAL function, the vagus nerves provide the major sensory and motor innervation to both striated and smooth muscle portions of the esophagus. Sensory pathways from the esophagus have been studied both electrophysiologically (40, 41) and by nerve tracing techniques (4,11,28). However, the precise location and structure of the sensory receptor(s) involved in control of esophageal motor function remain controversial. It is likely that more than one structurally and functionally distinct type of mechanoreceptor exists within the esophagus (4, 5, 8). As early as 1929, intraganglionic laminar nerve endings (IGLE) were considered candidates for esophageal stretch receptors (34). In an elegant study by Berthoud and Powley (4), intramuscular vagal afferents were shown to be positioned and structured as likely candidates for the gastric stretch receptor. Intramuscular arrays (IMA) enter circular or longitudinal muscle layers, run parallel to respective muscle fibers, and bifurcate to create an array of parallel terminal elements in stomach and sphincters (28). In the stomach, these terminal elements consist of interconnected parallel axonal telodendria that are arrayed in close proximity to one another and that form appositions with interstitial cells ...

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“…From the absence of any labeled fibers in animals with cervical vagal transection immediately following the nodosal injection, we can also rule out any nonvagal transport of the dye, such as through the circulation. Although tritiated leucine (Sato and Koyano 1987) and W G A -H R P (Neuhuber 1987;Clerc and Condamin 1987) have been successfully used to label vagal afferents selectively, DiI offers distinct advantages. Tissue processing is extremely simple and takes little time, and fixed tissue can be stored for months or years without a reduction in label.…”

Section: Methodological Considerationsmentioning

confidence: 99%

Vagal sensors in the rat duodenal mucosa: distribution and structure as revealed by in vivo DiI-tracing

Kressel²,

et al. 1995

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Results from functional studies point to the importance of chemoreceptive endings in the duodenum innervated by vagal afferents in the regulation of gastrointestinal functions such as gastric emptying and acid secretion, as well as in the process of satiation. In order to visualize the vagal sensory innervation of this gut segment, vagal afferents were selectively labeled in vivo by injecting the lipophilic carbocyanine dye DiI into either the left or the right nodose ganglion of young adult rats. Thick cryostat sections or whole-mounted peels of muscularis externa or submucosa of formalin-fixed tissue were analyzed with conventional and/or confocal microscopy. In the mucosa, many DiI-labeled vagal afferent fibers were found with terminal arborizations mainly between the crypts and the villous lamina propria. In both areas, vagal terminal branches came in close contact with the basal lamina, but did not appear to penetrate it so as to make direct contact with epithelial cells. Labeled vagal afferent fibers in the villous and cryptic lamina propria were found to be in intimate anatomical contact with fibrocyte-like cells that may belong to the class of interstitial cells of Cajal, and with small granular cells that might be granulocytes or histiocytes. Although our analysis was not quantitative, and considering that labeling was unilateral and not complete, it appears that the overall density of vagal afferent mucosal innervation was variable; many villi showed no evidence for innervation while other areas had quite dense networks of arborizing terminal fibers in several neighboring villi. Analysis of separate whole-mounted muscularis externa and submucosa peels revealed the presence of large bundles of labeled afferent fibers running within the myenteric plexus along the mesenteric attachment primarily in an aboral direction, with individual fibers turning towards the antimesenteric pole, and either penetrating into the submucosa or forming the characteristic intraganglionic laminar endings (IGLEs). Although the possibility of individual fibers issuing collaterals to myenteric IGLEs and at the same time to mucosal terminals was not demonstrated, it cannot be ruled out. These anatomical findings are discussed in the context of absorptive mechanisms for the different macronutrients and the implication of enteroendocrine cells such as CCK-containing cells that may function as intestinal "taste cells".

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Selective labeling of vagal sensory nerve fibers in the lower esophageal sphincter with anterogradely transported WGA-HRP

Cited by 31 publications

References 31 publications

Tyramide Amplification Allows Anterograde Tracing by Horseradish Peroxidase-conjugated Lectins in Conjunction with Simultaneous Immunohistochemistry

Tyramide Amplification Allows Anterograde Tracing by Horseradish Peroxidase-conjugated Lectins in Conjunction with Simultaneous Immunohistochemistry

Survival dependency of intramuscular ICC on vagal afferent nerves in the cat esophagus

Vagal sensors in the rat duodenal mucosa: distribution and structure as revealed by in vivo DiI-tracing

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