Immunohistochemical visualization of the enteric nervous system using antibodies against Protein gene product (PGP) 9.5

Krammer, Heinz-Jürgen; Karahan, S. Tuna; Rumpel, Elisabeth; Klinger, Matthias; Kühnel, Wolfgang

doi:10.1016/s0940-9602(11)80029-4

Cited by 72 publications

(47 citation statements)

References 9 publications

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“…NSE immunoreactivity was used as a nonspecific marker for cell bodies because NSE antibodies have been a useful tool for locating enteric ganglia and nerve fibers in the intestine, stomach, and gallbladder (Scheuermann et al, 1989;Furness et al, 1990aFurness et al, ,b, 1991Talmage et al, 1992), although it has not been conclusively established that NSE immunoreactivity is a marker for all enteric neurons. Therefore, we also used an antibody against protein gene product 9.5 (PGP), which appears to be a reliable marker for visualizing the general pattern of innervation in the heart and has been reported to stain enteric neurons (Gulbenkian et al, 1987;Horsch et al, 1993;Krammer et al, 1993). We found that the PGP antibody always colabeled NSE-immunoreactive cell bodies and vice versa, which suggests that the vast majority of, if not all, gastric enteric cell bodies were accounted for.…”

Section: Neurochemical Codingmentioning

confidence: 93%

Neurochemical coding of enteric neurons in the guinea pig stomach

Schemann

Schaaf

Mäder

1995

J of Comparative Neurology

156

147

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The aim of this study was to investigate the neurochemical coding of myenteric neurons in the guinea pig gastric corpus by using immunohistochemical methods. Antibodies and antisera against calbindin (CALB), calretinin (CALRET), choline acetyltransferase (ChAT), calcitonin gene-related peptide (CGRP), dopamine beta-hydroxylase (DBH), beta-endorphin (ENK), neuropeptide Y (NPY), neuron-specific enolase (NSE), nitric oxide synthase (NOS), protein gene product 9.5 (PGP), parvalbumin (PARV), serotonin (5-HT), somatostatin (SOM), substance P (SP), tyrosine hydroxylase (TH), and vasoactive intestinal peptide (VIP) were used. Double- and triple-labeling studies revealed colocalization of certain transmitters and enabled the identification of distinct subpopulations of gastric enteric neurons. NPY/VIP/NOS/ENK were present in 28% of all neurons, whereas 11% had NPY/VIP/DBH/ChAT; NOS-only neurons made up 2% of the population. The combination SP/ChAT/ENK occurred in 21% of the population, whereas SP/ChAT/ENK/CALRET and SP/CHAT/SOM/ +/- CALRET was identified in 5% and 6% of all cells, respectively. 5-HT-containing neurons comprised 2% of all cells and could be further classified by the presence of additional antigens as 5-HT/SP/(ChAT) or 5-HT/VIP/(ChAT). Approximately 21% of all neurons contained only ChAT with no additional antigen present and are referred to as ChAT/-. Gastric myenteric ganglion cells were not immunoreactive for CALB, PARV, CGRP, or TH. The results of this study indicate that gastric myenteric neurons can be characterized on the basis of different chemical coding. Neurochemical coding of corpus myenteric neurons revealed some similarities and significant differences in comparison with other regions of the gut. These differences might reflect adaptation of enteric nerves according to regional specialization and the distinct functions of the proximal stomach as a gastric reservoir.

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Section: Neurochemical Codingmentioning

confidence: 93%

Neurochemical coding of enteric neurons in the guinea pig stomach

Schemann

Schaaf

Mäder

1995

J of Comparative Neurology

156

147

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“…PCR for LCMcaptured samples and control tissues from cerebral cortex and left heart ventricle were processed as described above. PCR for protein gene product 9.5 (PGP 9.5), a pan-neuronal marker (36), was carried out to ascertain the neuronal identity of captured myenteric cells. Negative control contained all reagents, except that 1 l H 2O was substituted for reverse transcriptase in the RT reaction to exclude the possibility of genomic or other DNA contamination.…”

Section: Ucns and Crf Receptor Gene Expression In The Gc Of Naïve Ratsmentioning

confidence: 99%

“…The glandular middle portion of the mucosa expressed the highest levels of three Ucns, and Ucn 1 and Ucn 2 mRNA were also found prominently in muscle layers while Ucn 3 mRNA was also remarkably expressed in the neck zone. Furthermore, we characterized the mRNA expression of Ucns in laser-captured GC myenteric neurons identified by their prominent coexpression with the neuronal marker PGP 9.5 (36). The sequencing of PCR products ascertains that their identities matched to the known sequences of rat Ucn 1, Ucn 2, and Ucn 3.…”

Section: Distribution Of Ucns In the Gc Of Naïve Ratsmentioning

confidence: 99%

Urocortins and CRF type 2 receptor isoforms expression in the rat stomach are regulated by endotoxin: role in the modulation of delayed gastric emptying

Yuan

Taché

2012

American Journal of Physiology-Gastrointestinal and Liver Physi

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Peripheral activation of corticotropin-releasing factor receptor type 2 (CRF2) by urocortin 1, 2, or 3 (Ucns) exerts powerful effects on gastric function; however, little is known about their expression and regulation in the stomach. We investigated the expression of Ucns and CRF2 isoforms by RT-PCR in the gastric corpus (GC) mucosa and submucosa plus muscle (S+M) or laser captured layers in naive rats, their regulations by lipopolysaccharide (LPS, 100 μg/kg ip) over 24 h, and the effect of the CRF2 antagonist astresssin2-B (100 μg/kg sc) on LPS-induced delayed gastric emptying (GE) 2-h postinjection. Transcripts of Ucns and CRF2b, the most common wild-type CRF2 isoform in the periphery, were expressed in all layers, including myenteric neurons. LPS increased Ucn mRNA levels significantly in both mucosa and S+M, reaching a maximal response at 6 h postinjection and returning to basal levels at 24 h except for Ucn 1 in S+M. By contrast, CRF2b mRNA level was significantly decreased in the mucosa and M+S with a nadir at 6 h. In addition, CRF2a, reportedly only found in the brain, and the novel splice variant CRF2a-3 were also detected in the GC, antrum, and pylorus. LPS reciprocally regulated these variants with a decrease of CRF2a and an increase of CRF2a-3 in the GC 6 h postinjection. Astressin2-B exacerbated LPS-delayed GE (42–73%, P < 0.001). These data indicate that Ucn and CRF2 isoforms are widely distributed throughout the rat stomach and inversely regulated by immune stress. The CRF2 signaling system may act to counteract the early gastric motor alterations to endotoxemia.

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“…Thereafter, the sections were rinsed in PBS and incubated with rabbit antiprotein gene product (PGP) 9.5 (1:40 for 48 h at RT; 7863-0504; Biogenesis, UK). PGP 9.5 was used as a marker of fibers and neurons of the enteric nervous system (Krammer et al, 1993).…”

Section: Double Immunostainingmentioning

confidence: 99%

Distribution of ghrelin peptide in the gastrointestinal tract of stomachless and stomach‐containing teleosts

Arcamone

Neglia

Gargiulo

et al. 2009

Microscopy Res & Technique

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The occurrence and localization of ghrelin peptide in the gastrointestinal tract of Carassius auratus and Dicentrarchus labrax, two fish species that exhibit different feeding behavior, different habitats, and different anatomical organizations of the gastroenteric tract, were examined by immunohistochemical methods and western blotting analysis. All of the gastrointestinal segments studied displayed immunohistochemical localizations of ghrelin peptide. Numerous single or clustered immunoreactive cells were found along the gastric folds, particularly in the pyloric region of Dicentrarcus labrax, whereas scattered ghrelin immunoreactive cells were observed in the intestinal epithelium of both fish species. Double immunolabeling PGP 9.5/ghrelin demonstrated the localization of ghrelin peptide also in nerve fibers and neuronal cells of the submucosal and myenteric plexuses, often in association with vascular structures. Western blotting analysis confirmed the presence of ghrelin peptide in the gatrointestinal tract of both species studied, whose molecular weight was similar to that of the corresponding mammalian prepro-ghrelin. The findings could support the hypothesis that this peptide is an important appetite regulator in fish and could confirm the presence of the ghrelin peptide, starting from its precursor proteins, in the gastrointestinal tract of the goldfish and the sea bass.

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Immunohistochemical visualization of the enteric nervous system using antibodies against Protein gene product (PGP) 9.5

Cited by 72 publications

References 9 publications

Neurochemical coding of enteric neurons in the guinea pig stomach

Neurochemical coding of enteric neurons in the guinea pig stomach

Urocortins and CRF type 2 receptor isoforms expression in the rat stomach are regulated by endotoxin: role in the modulation of delayed gastric emptying

Distribution of ghrelin peptide in the gastrointestinal tract of stomachless and stomach‐containing teleosts

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