Architecture and Chemical Coding of the Inner and Outer Submucous Plexus in the Colon of Piglets

Petto, Carola; Gäbel, Gotthold; Pfannkuche, Helga

doi:10.1371/journal.pone.0133350

Cited by 12 publications

(27 citation statements)

References 61 publications

(71 reference statements)

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“…Then, the following primary antibodies were used: rabbit polyclonal anti-P2X2 antibody (ab48864, RRID: AB_881833, Abcam, diluted 1:400), mouse monoclonal anti-Calbindin D-28k (CB300, RRID: AB_10000347, Swant® Swiss Antibodies, diluted 1:2500), Chicken polyclonal anti-Neurofilament 200 (NFH, RRID: AB: AB_2313552, Aves Labs, INC., diluted 1:10000), Chicken polyclonal anti-TH (THY, RRID: AB_100113440, Aves Labs, INC., diluted 1:100), Rat monoclonal Anti-Substance P (SP, ab7340, RRID: AB_305866, Abcam, diluted 1:1000) and rabbit polyclonal Anti-Neuropeptide Y (22940, RRID: AB_2307354, ImmunoStar Inc, diluted 1:1000). The specificity of the primary antibodies has been published elsewhere 71 – 75 . For slices treated with mouse monoclonal CB300 antibody, an additional incubation with unconjugated Goat anti-mouse Fab fragment (10 ug/ml) in blocking solution was applied for 1 h at RT.…”

Section: Methodsmentioning

confidence: 99%

Selective tracking of FFAR3-expressing neurons supports receptor coupling to N-type calcium channels in mouse sympathetic neurons

Colina

Puhl

Ikeda

2018

Sci Rep

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Activation of short-chain free fatty acid receptors 3 (FFAR3) has been suggested to promote sympathetic outflow in postganglionic sympathetic neurons or hamper it by a negative coupling to N-type calcium (CaV2.2) channels. Heterogeneity of FFAR3 expression in sympathetic neurons, however, renders single neurons studies extremely time-consuming in wild-type mice. Previous studies demonstrated large variability of the degree of CaV2.2 channel inhibition by FFAR3 in a global population of rat sympathetic neurons. Therefore, we focused on a small subpopulation of mouse sympathetic neurons using an FFAR3 antibody and an Ffar3 reporter mouse to perform immunofluorescent and electrophysiological studies. Whole-cell patch-clamp recordings of identified FFAR3-expressing neurons from reporter mice revealed a 2.5-fold decrease in the CaV2.2-FFAR3 inhibitory coupling variability and 1.5-fold increase in the mean ICa2+ inhibition, when compared with unlabeled neurons from wild-type mice. Further, we found that the ablation of Ffar3 gene expression in two knockout mouse models led to a complete loss-of-function. Subpopulations of sympathetic neurons are associated with discrete functional pathways. However, little is known about the neural pathways of the FFAR3-expressing subpopulation. Our data indicate that FFAR3 is expressed primarily in neurons with a vasoconstrictor phenotype. Thus, fine-tuning of chemically-coded neurotransmitters may accomplish an adequate outcome.

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Section: Methodsmentioning

confidence: 99%

Selective tracking of FFAR3-expressing neurons supports receptor coupling to N-type calcium channels in mouse sympathetic neurons

Colina

Puhl

Ikeda

2018

Sci Rep

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“…The neurons not preferentially approached by capillaries, among them the morphologically striking, nondendritic type II neurons, were shown to be nonnitrergic but cholinergic [Brehmer et al, 2004;Wolf et al, 2007]. It can be assumed that additional neuroactive substances are colocalized with nNOS in the respective myenteric and submucosal type III neurons [Timmermans et al, 2001;Kapp et al, 2006;Wojtkiewicz et al, 2012;Petto et al, 2015]. The further deciphering of the chemical code of porcine type III neurons including quantitative data will be performed in a future project.…”

Section: Enteric Neurons Approached By Capillariesmentioning

confidence: 98%

Neurovascular Interface in Porcine Small Intestine: Specific for Nitrergic rather than Nonnitrergic Neurons

Jabari

Neuhuber²,

Brehmer³

2016

Cells Tissues Organs

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In the 1970s, by using classic histological methods, close topographical relationships between special areas of enteric ganglia and capillaries were shown in the pig. In this study, by application of double and triple immunohistochemistry, we confirmed this neurovascular interface and demonstrated that these zones are mainly confined to nitrergic neurons in the myenteric and the external submucosal plexus. In the upper small intestine of the pig, the respective neurons display type III morphology, i.e. they have long, slender and branched dendrites and a single axon. In another set of experiments, we prepared specimens for electron-microscopical analysis of these zones. Both ganglia and capillaries display continuous basement membranes, the smallest distances between them being 1,000 nm at the myenteric and 300 nm at the external submucosal level. The capillary endothelium was mostly continuous but, at the external submucosal level, scattered fenestrations were observed. This particular neurovascular relationship suggests that nitrergic neurons may require a greater amount of oxygen and/or nutrients. In guinea pig and mouse, previous ischemia/reperfusion experiments showed that nitrergic neurons are selectively damaged. Thus, a preferential blood supply of enteric nitrergic neurons may indicate that these neurons are more vulnerable in ischemia.

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“…With this study, we confirmed our hypothesis on the presence of MEN in the SMP of the porcine and human colon. So far, studies on SMP of the pig focused on the electrophysiology of neurons as revealed by intracellular recordings 37 or the expression of transmitter and neuropeptides 8,[38][39][40][41][42] . Although the pig is frequently advocated as a model for humans [31][32][33][34][35][36] , there are surprisingly few original studies which back up this notion.…”

Section: Discussionmentioning

confidence: 99%

“…Immunohistochemical staining of submucosal MEN of the porcine proximal and distal colon for ChAT and SP immunoreactivity showed no significant differences between the intestinal regions investigated regardless of whether tensile or compressive forces were applied. A study performed in the porcine colon revealed that the proportion of enteric ISMP neurons immunoreactive for ChAT was 43%, 66% of those also expressed SP, and no neurons solely expressed SP 8 . Our immunohistochemical staining of MEN mainly reflected these data, and this agrees with our previous results in the guinea pig ileum and stomach, where MEN also did not show a particular neurochemical code 14 , 16 .…”

Section: Discussionmentioning

confidence: 99%

“…Our immunohistochemical staining of MEN mainly reflected these data, and this agrees with our previous results in the guinea pig ileum and stomach, where MEN also did not show a particular neurochemical code 14 , 16 . Due to the fact that the majority of ISMP neurons project into the mucosal epithelium 8 , 9 , we could speculate that those cholinergic MEN are most likely to be involved in the regulation of secretory functions.…”

Section: Discussionmentioning

confidence: 99%

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Compression and stretch sensitive submucosal neurons of the porcine and human colon

Filzmayer

Elfers

Michel

et al. 2020

Sci Rep

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The pig is commonly believed to be a relevant model for human gut functions-however, there are only a few comparative studies and none on neural control mechanisms. To address this lack we identified as one central aspect mechanosensitive enteric neurons (MEN) in porcine and human colon. We used neuroimaging techniques to record responses to tensile or compressive forces in submucous neurons. Compression and stretch caused Ca-transients and immediate spike discharge in 5-11% of porcine and 15-24% of human enteric neurons. The majority of these MEN exclusively responded to either stimulus quality but about 9% responded to both. Most of the MEN expressed choline acetyltransferase and substance P; nitric oxide synthase-positive MEN primarily occurred in distal colon. The findings reveal common features of MEN in human and pig colon which we interpret as a result of species-independent evolutionary conservation rather than a specific functional proximity between the two species. The enteric nervous system (ENS), which is integrated into the wall of the gastrointestinal tract (GIT) from the oesophagus to the anal sphincter, enables the GIT to generate reflex activity independent from central influences 1-3. The ENS is organized in two complex neuronal networks called the submucosal plexus (SMP) and the myenteric plexus (MP). In both, the pig and human the SMP consists of two layers interconnected by interganglionic nerve fiber tracts 4,5. The SMP regulates mostly epithelial functions, such as secretion and absorption, as well as blood flow, cell proliferation and immune responses. The distinct parts of the SMP are called inner SMP (ISMP) and outer SMP (OSMP) with the ISMP being located closely to the lamina muscularis mucosae, whereas the OSMP is located on the luminal side of the circular muscle layer 6. In the porcine and human colon mainly neurons in the ISMP predominantly project to the mucosa 7-9 , and hence likely regulate epithelial functions 10. Enteric neurons can be activated by various stimuli, including chemical stimuli as well as mechanical distortion 11-17. During muscle contraction and relaxation enteric neurons are constantly exposed to distorting forces 16,18. Remarkably, even enteric neurons classically defined as interneurons or motoneurons are mechanosensitive, suggesting that mechanosensitive enteric neurons (MEN) are multifunctional 14,15,19. With intracellular recording techniques it has been shown that MEN in the MP respond to distension as well as to mucosal distortion 20-22. An important step forward was the use of imaging techniques, as they allowed to record simultaneously from a larger set of neurons and were also a prerequisite to study enteric neurons in larger animals 14-17,23,24. Using von Frey hair probing and intraganglionic volume injection, compression sensitive MEN have been identified in the MP of the guinea pig gastric corpus, ileum and colon 14,16,23 as well as in mouse ileum and colon 15. Furthermore, von Frey hair probing was used to identify compression sensitive MEN in isol...

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Architecture and Chemical Coding of the Inner and Outer Submucous Plexus in the Colon of Piglets

Cited by 12 publications

References 61 publications

Selective tracking of FFAR3-expressing neurons supports receptor coupling to N-type calcium channels in mouse sympathetic neurons

Selective tracking of FFAR3-expressing neurons supports receptor coupling to N-type calcium channels in mouse sympathetic neurons

Neurovascular Interface in Porcine Small Intestine: Specific for Nitrergic rather than Nonnitrergic Neurons

Compression and stretch sensitive submucosal neurons of the porcine and human colon

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