Quantification of neurons in the myenteric plexus: an evaluation of putative pan-neuronal markers

Phillips, Robert; Hargrave, Sara L.; Rhodes, Brie S.; Zopf, David; Powley, Terry L.

doi:10.1016/j.jneumeth.2003.10.004

Cited by 122 publications

(134 citation statements)

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“…First, in the present experiment, the stomach had the lowest density of alpha-synuclein-positive myenteric neurons in the proximal GI tract, with higher densities found in the small intestine (i.e., jejunum > duodenum > stomach). Indeed, the proportion of gastric myenteric neurons that are alpha-synuclein-positive is quite low: comparisons of the counts of alpha-synucleinpositive neurons in the present experiment with estimates of the total population of gastric and intestinal myenteric neurons in our earlier experiments using the pan-neuronal marker Cuprolinic Blue (Phillips et al, 2003(Phillips et al, , 2004a suggests that, at most, less than 3% of all gastric myenteric neurons express alpha-synuclein. In contrast, moving distally through the small intestine, the proportion of alpha-synuclein neurons increases to 6% of the neurons in the proximal duodenum, 13% of the neurons in the distal duodenum, and 22% of the neurons in the jejunum.…”

Section: Alpha-synuclein-positive Pathways Innervating the Gutmentioning

confidence: 38%

“…Whole mounts from an additional four unoperated control rats were permanently labeled with DAB to visualize the alpha-synuclein innervation of the GI tract, with the GI tissue from one of the four controls counterstained with 0.5% Cuprolinic Blue (quinolinic phthalocyanine; Polysciences, Inc., Warrington, PA) to label the entire population of myenteric neurons (Phillips et al, 2004a). Specificity of the staining was assessed for each rat by omitting the primary antibody in randomly chosen jejunal whole mounts.…”

Section: Permanent Immunohistochemistrymentioning

confidence: 99%

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Alpha-synuclein-immunopositive myenteric neurons and vagal preganglionic terminals: Autonomic pathway implicated in Parkinson's disease?

et al. 2008

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The protein alpha-synuclein is implicated in the development of Parkinson's disease. The molecule forms Lewy body aggregates that are hallmarks of the disease, has been associated with the spread of neuropathology from the peripheral to the central nervous system, and appears to be involved with the autonomic disorders responsible for the gastrointestinal (GI) symptoms of individuals afflicted with Parkinson's. To characterize the normative expression of alpha-synuclein in the innervation of the GI tract, we examined both the postganglionic neurons and the preganglionic projections by which the disease is postulated to retrogradely invade the CNS. Specifically, in Fischer 344 and Sprague Dawley rats, immunohistochemistry in conjunction with injections of the tracer DextranTexas Red was used to determine, respectively, the expression of alpha-synuclein in the myenteric plexus and in the vagal terminals. Alpha-synuclein is expressed in a subpopulation of myenteric neurons, with the proportion of positive somata increasing from the stomach (~3%) through duodenum (proximal, ~6%; distal, ~13%) to jejunum (~22%). Alpha-synuclein is co-expressed with the nitrergic enzyme NOS or the cholinergic markers calbindin and calretinin in regionally specific patterns: ~90% of forestomach neurons positive for alpha-synuclein express NOS, whereas ~92% of corpus-antrum neurons positive for alpha-synuclein express cholinergic markers. Vagal afferent endings in the myenteric plexus and the GI smooth muscle do not express alpha-synuclein, whereas, virtually all vagal preganglionic projections to the gut express alpha-synuclein, both in axons and in terminal varicosites in apposition with myenteric neurons. Vagotomy eliminates most, but not all, alpha-synuclein-positive neurites in the plexus. Some vagal preganglionic efferents expressing alphasynuclein form varicose terminal rings around myenteric plexus neurons that are also positive for the protein, thus providing a candidate alpha-synuclein-expressing pathway for the retrograde transport of putative Parkinson's pathogens or toxins from the ENS to the CNS.

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Section: Alpha-synuclein-positive Pathways Innervating the Gutmentioning

confidence: 38%

Section: Permanent Immunohistochemistrymentioning

confidence: 99%

Alpha-synuclein-immunopositive myenteric neurons and vagal preganglionic terminals: Autonomic pathway implicated in Parkinson's disease?

et al. 2008

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“…We can justify the controversial results for the NADPH-dp neurons in Group D, C, CS and DS when compared to other reports: the diabetes maintenance period as well as the neurochemical phenotype of the stained neurons was not similar. Enhanced changes in the neuronal density can be better observed when a generalized neuronal population is stained regardless of their neurochemical phenotype, as occurs, for example, with the use techniques such as the cuprolinic blue (Phillips et al 2004), myosin V (Zanoni et al 2003), NADH-diaphorase (Sant'Ana et al 2001, Young et al 1993, Clebis et al 2004, , Zanoni et al 2007, Silva et al 2008, methylene blue (Hernandes et al 2000), among others, which stain the whole neuronal population, including the nitrergic neurons. When analyzing specific populations, such as NADPH-dp neurons, more specific results were obtained.…”

Section: Discussionmentioning

confidence: 99%

Effects of ascorbic acid supplementation in ileum myenteric neurons of streptozotocin-induced diabetic rats

et al. 2009

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295Pesq. Vet. Bras. 29(4): 295-302, abril 2009 RESUMO.-[Efeitos da suplementação do ácido ascórbico nos neurônios mientéricos do íleo de ratos diabéticos induzidos por estreptozootocina.] A exacerbação do estresse oxidativo e da via do poliol que comprometem o plexo mioentérico são alterações metabóli-cas características do diabetes. O ácido ascórbico (AA) é antioxidante e inibidor da aldose redutase, podendo atuar como neuroprotetor. Verificaram-se os efeitos da suplementação com AA sobre o número e a área do perfil do corpo celular (PC) de neurônios mioentéricos em ratos diabéticos induzidos por estreptozootocina. Formaram-se quatro grupos com cinco animais cada: normoglicêmico (C); diabético (D); diabético tratado com AA (DS); e normoglicêmico tratado com AA (CS). Três vezes por semana administrou-se 50mg de AA para cada animal (grupos DS e CS). Após 90 dias e eutanásia com tiopental, o íleo foi coletado e processado para a técnica da NADPHdiaforase. Não se observaram diferenças (P>0,05) na densidade neuronal entre os grupos. A área do PC foi menor (P<0,05) para os grupos DS e CS, com incidência maior de neurônios com área do PC superior a 200μm 2 para os grupos C e D. Concluiu-se que o AA não influenciou a densidade neuronal do íleo, mas foi neuroprotetor The exacerbation of the oxidative stress and of the polyol pathway which impair damage myenteric plexus are metabolic characteristics of diabetes. The ascorbic acid (AA) is an antioxidant and an aldose reductase inhibitor, which may act as neuroprotector. The effects of AA supplementation on the density and cellular body profile area (CP) of myenteric neurons in STZ-induced diabetes in rats were assessed. Four groups with five animals each were formed: normoglycemic (C); diabetic (D); AA-treated diabetic (DS) and AA-treated normoglycemic (CS). Dosagen of 50mg of AA were given, three times a week, for each animal (group DS and CS). Ninety days later and after euthanasia, the ileum was collected and processed for the NADPH-diaphorase technique. There were no differences (P>0.05) in the neuronal density among the groups. The CP area was lower (P<0.05) in the DS and CS groups, with a higher incidence of neurons with a CP area exceeding 200μm 2 for groups C and D. The AA had no influence on the neuronal density in the ileum but had a neuroprotective effect, preventing the increase in the CP area and allowing a higher number of neurons with a CP area with less than 200μm 2 .INDEX TERMS: Vitamin C, diabetes, intestine, myenteric plexus.

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“…The immunohistochemical localization of these substances has been important for the analysis of the neuronal circuitry of the ENS [5,6,40]. It has been reported that the neuronal loss in the gastrointestinal tract of the rat starts in early adulthood and continues in a roughly linear manner over the rodent's lifespan [36][37][38][39].…”

Section: Discussionmentioning

confidence: 99%

Age-Related Changes of Calbindin D-28k-Immunoreactive Neurons in the Myenteric Plexus of Gerbil Duodenum

Choi

Lee

Chung

et al. 2008

The Journal of Veterinary Medical Science

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ABSTRACT. We examined the age-related changes of calbindin D-28k (CB)-immunoreactive neurons and overall populations of neurons in the myenteric plexus of gerbil duodenum using whole mount preparations and immunohistochemistry. The circumference of duodenum increased age-dependently. CB-immunoreactive neurons were observed in all groups, and most of them had the Dogiel type II morphology. The fully developed cobweb-like structures were observed in the myenteric plexus of duodenum at postnatal month (PM) 3 to 24. Although the highest numbers of CB-immunoreactive neurons and overall population were observed in PM 1.5, it is related with significant increase of the size of circumference between PM 1.5 to PM 3. CB-immunoreactive neurons were slightly decreased with age between PM 3 to PM 24. We have also found that whole numbers of myenteric neurons were also significantly decreased in PM 24 group. These results suggest that loss of overall numbers of myenteric neurons and CB-immunoreactive neurons may be related with age-related neurodegeneration and functional loss of duodenum in the gerbil. KEY WORDS: aging, calbindin, duodenum, gerbil, myenteric plexus.J. Vet. Med. Sci. 70(4): 343-348, 2008 The autonomic nervous system is divided into two divisions, e.g. parasympathetic nervous system and sympathetic nervous system. In recent decade, the enteric nervous system (ENS) is the largest division of the autonomic nervous system, because it can perform its function independently from the central nervous system [7,20,44]. It consists of local nerve networks embedded in the gut wall and is subdivided into two ganglionated plexuses, i.e., the submucosal and myenteric plexuses [40,44]. The ENS mediates complex reflex activities involving intestinal motility, mucosal transportation and blood flow [7]. In electrophysiological studies, the majority of the enteric neurons could be placed in one of two classes: synaptic neurons or after-hyperpolarizing (AH) neurons [21,22,26,29,34].The myenteric plexus lies between the longitudinal and circular muscular layers and extends entire length of the gut. It has been known that the myenteric plexus primarily provides motor innervation to the two muscular layers and secremotor innervation to the mucosa [15,44].Calbindin D-28k (CB), one of the calcium binding proteins, plays an important role as an intracellular facilitator of calcium diffusion or as a modulator of neurotransmission [3,4,16,17]. In neurons, action potentials are also carried in part by inward calcium current [22,26,34]. It has been reported that AH neurons were sensory neurons because these neurons lacked fast excitatory synaptic inputs [21,22,26,29,34]. More recently, with the help of the combination of the electrophysiological and immunohistochemical methods, it has been identified that the AH neurons had Dogiel type II morphology [14,18,26].Although the distribution of CB-containing neuronal system has been reported upon the forebrain [11,23,24], comprehensive knowledge about the maturation of CB neurons in myenteri...

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Quantification of neurons in the myenteric plexus: an evaluation of putative pan-neuronal markers

Cited by 122 publications

References 25 publications

Alpha-synuclein-immunopositive myenteric neurons and vagal preganglionic terminals: Autonomic pathway implicated in Parkinson's disease?

Alpha-synuclein-immunopositive myenteric neurons and vagal preganglionic terminals: Autonomic pathway implicated in Parkinson's disease?

Effects of ascorbic acid supplementation in ileum myenteric neurons of streptozotocin-induced diabetic rats

Age-Related Changes of Calbindin D-28k-Immunoreactive Neurons in the Myenteric Plexus of Gerbil Duodenum

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