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Michael Schemann

Publications and source records attributed to Michael Schemann.

17 recordsLinked to original sources
PubMed2006

Hydrogen sulfide is a novel prosecretory neuromodulator in the Guinea-pig and human colon.

BACKGROUND & AIMS: Hydrogen sulfide (H(2)S) has been suggested as a novel gasomediator. We explored its unknown neuromodulatory role in human and guinea-pig colon. METHODS: We used immunohistochemistry to detect H(2)S-producing enzymes cystathionine gamma-lyase (CSE) and cystathionine beta-synthase (CBS) in enteric neurons, Ussing chambers to measure mucosal ion secretion, and neuroimaging with voltage- and Ca(++)-sensitive dyes to record H(2)S effects on guinea-pig and human enteric neurons. RESULTS: More than 90% of guinea-pig and human submucous and myenteric neurons were colabeled for CSE and CBS. Myenteric interstitial cells of Cajal were CSE-immunoreactive. The exogenous H(2)S donor NaHS (0.2-2.5 mmol/L) concentration-dependently increased chloride secretion in human and guinea-pig submucosa/mucosa preparations, but not in the colonic epithelial cell line T84. The secretory response was reduced significantly by tetrodotoxin (0.5 micromol/L), capsaicin desensitization (10 micromol/L), and the transient receptor potentials vanilloid receptor 1 antagonist capsazepine (10 micromol/L). The endogenous H(2)S donor L-cysteine also induced secretion that was diminished significantly by capsaicin desensitization, the CBS inhibitor amino-oxyacetic acid, and the CSE inhibitor propargylglycine. NaHS increased spike discharge in 23% of guinea-pig and 36% of human submucous neurons, but had no effect on Ca(++) mobilization in cultured guinea-pig enteric neurons. This excitatory response was reduced significantly by capsaicin desensitization and capsazepine, but not by glibenclamide (10 micromol/L). CONCLUSIONS: The presence of H(2)S-producing enzymes in human and guinea-pig enteric neurons, the excitatory action on enteric neurons, and the prosecretory effects of NaHS suggest H(2)S as a novel gut-signaling molecule. Its action mainly involves transient receptor potentials vanilloid receptor 1 receptors on extrinsic afferent terminals, which in turn activate enteric neurons.

Adult↗
PubMed2006

Role of enteric glia in intestinal physiology: effects of the gliotoxin fluorocitrate on motor and secretory function.

The role of enteric glia in gastrointestinal physiology remains largely unexplored. We examined the actions of the gliotoxin fluorocitrate (FC) on intestinal motility, secretion, and inflammation after assessing its efficacy and specificity in vitro. FC (100 microM) caused a significant decrease in the phosphorylation of the glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diaz-4-yl)amino]-2-deoxyglucose in enteric glial cultures and a reduction in glial uptake of the fluorescent dipeptide Ala-Lys-7-amino-4-methylcoumarin-3-acetic acid in both the ileum and colon. Dipeptide uptake by resident murine macrophages or guinea pig myenteric neurons was unaffected by FC. Incubation of isolated guinea pig ileal segments with FC caused a specific and significant increase in glial expression of the phosphorylated form of ERK-1/2. Disruption of enteric glial function with FC in mice reduced small intestinal motility in vitro, including a significant decrease in basal tone and the amplitude of contractility in response to electrical field stimulation. Mice treated with 10 or 20 micromol/kg FC twice daily for 7 days demonstrated a concentration-dependent decrease in small intestinal transit. In contrast, no changes in colonic transit or ion transport in vitro were observed. There were no changes in glial or neuronal morphology, any signs of inflammation in the FC-treated mice, or any change in the number of myenteric nitric oxide synthase-expressing neurons. We conclude that FC treatment causes enteric glial dysfunction, without causing intestinal inflammation. Our data suggest that enteric glia are involved in the modulation of enteric neural circuits underlying the regulation of intestinal motility.

4-Chloro-7-nitrobenzofurazan↗
PubMed2006

Enteric nervous system.

PURPOSE OF REVIEW: Our aim was to provide a synopsis of how the field of enteric neurobiology has advanced during the past year. RECENT FINDINGS: With such a large number of studies to choose from and given our emphasis in last year's issue on developmental aspects of the enteric nervous system, we have focused on several key themes reflecting the current interest in the way the enteric nervous system is altered in disease. SUMMARY: The new basic science information gathered during the past year provides insight into pathophysiological processes and will pave the way for improved understanding of both organic and 'functional' gastrointestinal disorders.

Animals↗
PubMed2005

Functional expression of the peptide transporter PEPT2 in the mammalian enteric nervous system.

The peptide transporter PEPT2 mediates transmembrane uptake of small peptides. So far, its expression has not been evidenced in the gastrointestinal tract. We have investigated peptide transport activity in the neuromuscular layers of the gastrointestinal tract by using the fluorescent tracer-dipeptide beta-Ala-Lys-Nepsilon-7-amino-4-methyl-coumarin-3-acetic acid (Ala-Lys-AMCA). Whole-mount preparations from mouse, rat, and guinea pig stomach and small and large intestine were incubated with Ala-Lys-AMCA in the presence or absence of the uptake-inhibitors L-histidine, D-phenylalanyl-L-alanine (D-Phe-Ala), glycyl-L-sarcosine (Gly-Sar), glycyl-L-glutamine (Gly-Gln), benzylpenicillin, and cefadroxil. Fluorescence microscopy revealed that Ala-Lys-AMCA specifically accumulated in both ganglionic layers of the enteric nervous system (ENS) in all regions and species studied. This could be inhibited by Gly-Sar, D-Phe-Ala, Gly-Gln, and cefadroxil, but not by free histidine and benzylpenicillin, indicating uptake via PEPT2. Accordingly, dipeptide uptake was completely abolished in PEPT2-deficient mice. Reverse transcriptase-polymerase chain reaction analysis detected a PEPT2-specific transcript in extracts from the ganglionic ENS layers of mouse small and large intestine, further proving that enteric dipeptide transport activity is specifically mediated via PEPT2. The cellular site of dipeptide uptake was immunohistochemically localized to enteric glial cells and tissue-resident macrophages. In addition, dipeptide uptake occurred in a neurochemically defined subset of neurons in the guinea pig ENS. Our results constitute the first functional evidence for dipeptide transport activity in the ENS. PEPT2-mediated dipeptide transport in enteric glia could contribute to the clearance of neuropeptides in the ENS. In addition, the fluorophore-coupled dipeptide uptake via PEPT2 is a novel vital marker for glial cells in the ENS.

Animals↗
PubMed2005

Serotonin excites neurons in the human submucous plexus via 5-HT3 receptors.

BACKGROUND & AIMS: Serotonin (5-hydroxytryptamine [5-HT]) is a key signaling molecule in the gut. Recently, the neural 5-HT3 receptor received a lot of attention as a possible target in functional bowel diseases. Yet, the 5-HT3 receptor-mediated changes in properties of human enteric neurons is unknown. METHODS: We used a fast imaging technique in combination with the potentiometric dye 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl]vinyl]pyridinium betaine to monitor directly the membrane potential changes in neurons of human submucous plexus from surgical specimens of 21 patients. An Ussing chamber technique was used to study 5-HT3 receptor involvement in chloride secretion. RESULTS: Local microejection of 5-HT directly onto ganglion cells resulted in a transient excitation of enteric neurons characterized by increased spike discharge. This response was mimicked by the 5-HT3 receptor agonist, 2-methyl-5-HT, and blocked by the 5-HT3 receptor antagonist, tropisetron. The proportions of 5-HT-responsive nerve cells per ganglion ranged from 25.5% +/- 18.4% in the duodenum to 54.2% +/- 46.9% in the colon. Interestingly, 2-methyl-5-HT did not evoke chloride secretion in the human intestine but it did in the guinea-pig intestine. Specific 5-HT3A and 5-HT3B receptor subunit immunoreactivity as well as 5-HT3A and 5-HT3B receptor-specific messenger RNA were detected in the tissue samples. Based on co-labeling with the pan-neuronal marker HuC/D we conclude that submucous nerve cells potentially express heteromeric 5-HT3A/B receptors. CONCLUSIONS: We show that 5-HT excited human enteric neurons via 5-HT3 receptors, which may comprise both 5-HT3A and 5-HT3B receptor subunits.

Action Potentials↗
PubMed2005

Enteric nervous system.

PURPOSE OF THE REVIEW: The purpose of this review is to provide a synopsis of how the field of enteric neurobiology has advanced during the past 2 years. RECENT FINDINGS: With more than 500 studies from which to choose, the authors have focused on several themes that illustrate recent progress. There has been an explosion of interest in the development of the enteric nervous system driven by the need to understand development abnormalities, particularly in Hirschsprung disease, and fueled by technical advances for investigating how neural crest-derived cells migrate, proliferate, and differentiate into enteric neurons and glia. The use of neural stem cells as a therapeutic strategy aimed at repopulating regions of bowel, where enteric neurones are reduced or absent, is on the horizon. Enteric reflexes involve interactions between sensory neurons, interneurons, and motor neurons. Recent findings suggest this distinction may be blurred, with neurons having multifunctional properties, perhaps because enteric neurons, unlike their central nervous system counterparts, are directly exposed to mechanical forces that they regulate. Another topic the authors have highlighted is pharmacology, with new tools for investigating ion channels, receptors, and transporters, leading to an expanding list of molecular mechanisms that regulate neuronal excitability. Long-term alterations in the expression of these molecules during disease or injury may underlie many gastrointestinal disorders that currently have unknown etiology. The authors finish with a look to the future and what may be the subject of this review next time. SUMMARY: Basic science information gathered during the past 2 years provides insight into pathophysiologic processes and will pave the wave for improved understanding of both organic and 'functional' gastrointestinal disorders.

Animals↗
PubMed2005

Control of gastrointestinal motility by the "gut brain"--the enteric nervous system.

The enteric nervous system (ENS) as the "brain of the gut" is pivotal for normal muscle activity in the gut. Neuronal circuits within the ENS are designed to control gut motility independent of central inputs. To fulfill this task the ENS contains all necessary elements for coding mechanical and chemical stimuli, interneuronal communication and efferent output to the muscle. This review provides a summary of the ENS circuits that control muscle activity, the main transmitters and neuromodulators involved and the functional implications for the normal and diseased gut.

Digestive System↗
PubMed2004

Calbindin-immunoreactive neurones in the ovine rumen.

In small laboratory animals, such as guinea pigs, immunoreactivity for the calcium-binding protein calbindin (CALB) can be used to distinguish functionally different classes of myenteric neurones. The rumen of sheep is a highly specialized gastrointestinal region, and the control of its functions requires specific intrinsic innervation patterns. The aim of this study was to neurochemically identify and characterize CALB-positive myenteric neurones of the ovine rumen. Therefore, we performed quadruple immunohistochemistry against CALB, substance P (SP), vasoactive intestinal peptide (VIP), and nitric oxide synthase (NOS) using whole-mount preparations of the ruminal myenteric plexus. On average, 3 +/- 2 and 1 +/- 0.4 myenteric neurones/ganglion were CALB-immunoreactive in suckling lambs and adult sheep, respectively. These neurones had Dogiel type-I morphology. Most of them (89.2% +/- 8.7% and 71.7% +/-44.8% in suckling lambs and adult sheep, respectively) did not colocalize any of the other antigenes. Since it has been shown in previous studies that ruminal myenteric neurones are immunoreactive for either choline acetyltransferase (ChAT) or NOS, we defined neurones which were CALB-positive and NOS-negative as CALB/ChAT. The other CALB-positive neurones were encoded CALB/NOS/+/-VIP (10.3% +/- 9.3% and 26.7% +/- 46.2% in suckling lambs and adult sheep, respectively) or CALB/ChAT/SP (0.5% +/- 1.0% and 1.7% +/- 1.9% in suckling lambs and adult sheep, respectively). We used cryostat sections of the ruminal wall to analyze the projections of the CALB-positive neurones. CALB-immunoreactive somata were exclusively located within the myenteric plexus. CALB-immunoreactive nerve fibers were found primarily in the lamina propria of the ruminal papillae. We conclude that CALB-positive myenteric neurones within the ovine rumen project to the epithelium; however, their functional role remains to be investigated.

Animals↗
PubMed2004

Intrinsic innervation patterns of the smooth muscle in the rumen and reticulum of lambs.

The rumen and reticulum of sheep serve as a fermentation chamber. Both compartments exhibit specific motility patterns. With developmental changes, the size of the reticulorumen dramatically increases when newborn lambs mature to adult sheep. This makes it possible to investigate the intrinsic innervation of the reticuloruminal muscles in lambs by taking the entire reticulum and rumen into account. The aim of the study was to analyse the projections and neurochemistry of myenteric neurons in the rumen and reticulum, which project to the inner or outer muscle layer, respectively. Therefore, we applied retrograde tracing with the fluorescent dye 1,1'-didodecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI) and subsequent immunohistochemical detection of choline acetyltransferase (ChAT), substance P (SP) and vasoactive intestinal peptide (VIP). In both compartments innervation of both the inner and the outer muscle layer consisted mainly of cholinergic neurons (65-70%). The majority of them co-localized SP. The non-cholinergic neurons projecting to the muscle expressed immunoreactivity for VIP. Polarized innervation of the muscle layers was found neither in the rumen nor in the reticulum. Consequently, intrinsic innervation patterns for the smooth muscle layers in the rumen and reticulum differ from all gastrointestinal regions examined thus far.

Animals↗
PubMed2003

Virokinin, a bioactive peptide of the tachykinin family, is released from the fusion protein of bovine respiratory syncytial virus.

Tachykinins, an evolutionary conserved family of peptide hormones in both invertebrates and vertebrates, are produced by neuronal cells as inactive preprotachykinins that are post-translationally processed into different neuropeptides such as substance P, neurokinin A, and neurokinin B. We show here that furin-mediated cleavage of the bovine respiratory syncytial virus fusion protein results in the release of a peptide that is converted into a biologically active tachykinin (virokinin) by additional post-translational modifications. An antibody directed to substance P cross-reacted with the C terminus of mature virokinin that contains a classical tachykinin motif. The cellular enzymes involved in the C-terminal maturation of virokinin were found to be present in many established cell lines. Virokinin is secreted by virus-infected cells and was found to act on the tachykinin receptor 1 (TACR1), leading to rapid desensitization of this G protein-coupled receptor as shown by TACR1-green fluorescent protein conjugate translocation from the cell surface to endosomes and by co-internalization of the receptor with beta-arrestin 1-green fluorescent protein conjugates. In vitro experiments with isolated circular muscle from guinea pig stomach indicated that virokinin is capable of inducing smooth muscle contraction by acting on the tachykinin receptor 3. Tachykinins and their cognate receptors are present in the mammalian respiratory tract, where they have potent effects on local inflammatory and immune processes. The viral tachykinin-like peptide represents a novel form of molecular mimicry, which may benefit the virus by affecting the host immune response.

Animals↗
PubMed2003

Reticular groove and reticulum are innervated by myenteric neurons with different neurochemical codes.

The reticulum and the reticular groove are functional distinct compartments within the ovine forestomach. While the reticulum takes part in various motor functions, such as mixing, retaining, and rejecting the forestomach ingesta, the reticular groove serves mainly as a bypass between the esophagus and the abomasum. To accomplish these different tasks, the compartments develop specific motility patterns that are controlled by intrinsic neural circuits. In this study the intrinsic innervation by myenteric neurons was analyzed by quadruple immunohistochemistry against cholineacetyl transferase (ChAT), nitric oxide synthase (NOS), substance P (SP), and vasoactive intestinal peptide (VIP). Four neurochemically different subpopulations of myenteric neurons were found in the reticulum and the floor of the reticular groove: ChAT/-, ChAT/SP, NOS/-, and NOS/VIP. The neuronal proportions were calculated relative to all myenteric neurons. Neurons of the reticulum were mostly immunoreactive for ChAT (89% +/- 3%), whereas neurons adjacent to the reticular groove predominantly expressed a nitrergic phenotype (62% +/- 4%). ChAT-positive neurons were also immunoreactive for SP (ChAT/SP: 64% +/- 3% reticulum; 25% +/- 1% reticular groove) or were purely cholinergic (ChAT/-: 25% +/- 4% reticulum; 13% +/- 3% reticular groove). NOS-positive neurons colocalized VIP (NOS/VIP: 10% +/- 3% reticulum; 46% +/- 1% reticular groove) or none of the other neurotransmitters (NOS/-: 1% +/- 1% reticulum; 17% +/- 3% reticular groove). Analysis of the soma sizes revealed that in both compartments the nitrergic neurons were significantly larger than the cholinergic neurons. It is suggested that the specific neurochemical code in combination with a specific morphology leads to a precise regulation of the specialized tasks of the reticulum and reticular groove by subpopulations of myenteric neurons.

Animals↗
PubMed2003

Age-associated plasticity in the intrinsic innervation of the ovine rumen.

The rumen of adult sheep functions as a large fermentation chamber. In the newborn suckling ruminant, the rumen is bypassed and milk enters the abomasum directly. It was the aim of our study to investigate whether the transmitter content of intrinsic nerves changes with the developmental stage. The neurochemical code of myenteric neurons in the rumen from suckling lambs, fattened lambs and adult sheep was determined by using quadruple immunohistochemistry against choline-acetyltransferase (ChAT), nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP). Three neurochemically distinct subpopulations were identified within the rumen. They expressed the code ChAT/-, ChAT/SP and NOS/VIP. The number of ChAT/SP neurons did not change during development. It was 62% in the newborn lamb and remained stable in fattened lambs (63%) and adult sheep (63%). By contrast, the number of ChAT/- neurons decreased significantly from 20% in suckling lambs to 11% and 7% in fattened lambs and adult sheep, respectively. Simultaneously, the proportion of NOS/VIP neurons increased from 16% in suckling lambs to 29% in adult sheep. The increase in the proportion of NOS/VIP immunoreactive neurons indicates an adaptation to large volumes of ingesta at the beginning of roughage intake and rumination. We conclude that the age-associated changes in neurochemical code of myenteric neurons in the forestomach are related to the adaption of the rumen to different functional properties during development.

Aging↗
PubMed2002

Ruminal muscle of sheep is innervated by non-polarized pathways of cholinergic and nitrergic myenteric neurones.

The motility patterns of the reticulorumen evoke mainly mixing of the ingesta. So far unknown, intrinsic neural circuits of the enteric nervous system are involved in the control of these motility patterns. The aim of the study was to characterize neurochemically sheep ruminal myenteric neurones, in particular the neural pathways innervating the ruminal muscle layers. Cell bodies within the myenteric plexus projecting to the longitudinal or circular muscle layer were retrogradely labelled by direct application of the fluorescent tracer 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) onto the circular or longitudinal muscle. The neurochemical code of myenteric neurones was identified by their immunoreactivity for choline acetyltransferase (ChAT), nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP). According to their neurochemical code, ruminal myenteric neurones were divided into three populations: ChAT/SP (68% of all myenteric neurones), NOS/VIP (26% of all myenteric neurones) and ChAT/- (5% of all myenteric neurones). Application of DiI onto the circular or longitudinal muscle revealed on average 64 or 44 labelled cell bodies in the myenteric plexus, respectively. DiI-labelled neurones expressed the code ChAT/SP or NOS/VIP. In the pathways to circular or longitudinal muscle, ChAT/SP-positive neurones outnumbered NOS/VIP-immunoreactive neurones by 5:1 and 2:1. Pathways to the circular or longitudinal muscle did not exhibit any pronounced polarized innervation patterns. This study demonstrated specific projections of myenteric neurones to the ruminal muscle. Neurones expressing the code ChAT/SP might function as excitatory muscle motor neurones, whereas NOS/VIP neurones are likely to act as inhibitory muscle motor neurones.

Animals↗
PubMed2002

Cholinergic and noncholinergic innervation of the smooth muscle layers in the bovine abomasum.

The intrinsic innervation of muscle layers in the mammalian gastrointestinal tract has been mainly studied in nonruminants. The aim of this study was to identify intrinsic motor neurones in the bovine abomasum that innervate the circular and longitudinal muscles. Circular (CMN) and longitudinal muscle motor neurones (LMN) were selectively labeled by application of the retrograde tracer 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) onto the muscle layers. The transmitter phenotype was determined by immunohistochemical detection of choline acetyltransferase (ChAT), nitric oxide synthase (NOS), and neurone-specific enolase (NSE). On average, the myenteric ganglia contained 61 +/- 19 NSE-positive cell bodies, of which 89% were ChAT-positive and 10% were NOS-positive. Only 0.7% of NSE-positive neurones (41 of 5,777) contained both ChAT and NOS. Application of DiI onto the circular and longitudinal muscles revealed on average 60 +/- 27 (n = 4) and 68 +/- 36 (n = 4), respectively, labeled cell bodies in the myenteric plexus. For the circular and longitudinal muscles the proportions of ascending to descending neurones were 76 : 24% and 54 : 46%, respectively. While most ascending CMN were ChAT-positive (96%), 51% of the descending CMN were ChAT-negative. All ascending and 95% of descending LMN were ChAT-positive. It was concluded that cholinergic excitatory innervation is predominant in both muscle layers of the abomasum. Whereas the circular muscle receives cholinergic excitatory and nitrergic inhibitory innervation, the longitudinal muscle is only innervated by cholinergic pathways. This innervation pattern is different from that in gastric muscle layers in monogastric animals.

Abomasum↗
PubMed2002

Cutting-edge technology. III. Imaging and the gastrointestinal tract: mapping the human enteric nervous system.

Monitoring membrane potentials by multisite optical recording techniques using voltage-sensitive dyes is ideal for direct analysis of network signaling. We applied this technology to monitor fast and slow excitability changes in the enteric nervous system and in hundreds of neurons simultaneously at cellular and subcellular resolution. This imaging technique presents a powerful tool to study activity patterns in enteric pathways and to assess differential activation of nerves in the gut to a number of stimuli that modulate neuronal activity directly or through synaptic mechanisms. The optical mapping made it possible to record from tissues such as human enteric nerves, which were, until now, inaccessible by other techniques.

Digestive System↗