Demonstration of a unique population of neurons with NADPH-diaphorase histochemistry

Scherer-Singler, U.; Vincent, Steven R.; Kimura, Hiroshi; McGeer, Edith G.

doi:10.1016/0165-0270(83)90085-7

Cited by 667 publications

(281 citation statements)

References 9 publications

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“…The ascending colon of four animals of each group were washed and filled with phosphate buffer (PBS; pH 7.4), fixed in 4% paraformaldehyde (Merk, Darmstad, Germany) prepared in 0.1M phosphate buffer (PBS; pH 7.4) for 30 minutes, immersed in 0.3% Triton X-100 ® (Sigma, St .Louis, USA) dissolved in saline phosphate buffer (PBS, pH 7.4) for 10 minutes and then washed ten times (10 minutes each) in PBS and immersed in the incubation medium for the neuronal evidenciation of the NADPH-diaphorase, during two hours 19 . This medium contained in each dl: 25 mg of NBT; 50 mg of β-NADPH (Sigma, Steinheim, Germany), 0.3% Triton X-100 in 0.1M Tris-HCl buffer, pH 6.0 (GibcoBRL, N.Y. USA).…”

Section: Methodsmentioning

confidence: 99%

Effects of protein and vitamin B deficiency on blood parameters and myenteric neurons of the colon of rats

Sant’Ana

Molinari

Miranda-Neto

2001

Arq. Neuro-Psiquiatr.

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-The aims of this work were to evaluate the effects of the deficient ingestion of protein and vitamin B on the biochemical and hematologic parameters and on the NADH-and NADPH-diaphorase positive myenteric neurons. The control animals (n=10) received commercial chow and the experimental rats (n=10) received chow with protein level reduced to 8% during 120 days. At the time of killing blood was collected for assessment of the blood and hematologic parameters and the ascending colon for quantitative analysis of the neurons of the myenteric plexus. It was observed that the reduction of the protein level to 8% coupled to the reduction of the levels of vitamin B in adult rats neither led to qualitative or quantitative changes on red or white blood cells, nor decreased globulin levels, induced the formation of edema or gave rise to clinical signs typical of protein or vitamin B deficiency. On the other hand, the experimental protocol led to less weight gain, change on the body composition with fat deposition; decrease of the values of serum total protein and albumin; reduction of the area of colon and density of nitrergic and NADH-diaphorase myenteric neurons inferior to the expected.KEY WORDS: enteric neurons, proteic desnutrition, ascending colon, hematology, vitamin B.Efeitos da deficiência de proteínas e vitamina B sobre parâmetros sanguíneos e neurônios mioentéricos do colo de ratos adultos RESUMO -Os objetivos deste trabalho foram avaliar as repercussões da ingestão deficiente de proteínas e vitaminas do complexo B sobre parâmetros bioquímicos e hematológicos e neurônios mientéricos NADH e NADPH-diaforase positivos. Os animais controle (n=10) receberam ração comercial e os animais experimentais (n=10) ração com teor protéico reduzido para 8% durante 120 dias. Durante o sacrifício, coletou-se o sangue dos animais para avaliação de parâmetros sanguíneos e hematológicos e o colo ascendente para a análise quantitativa dos neurônios do plexo mioentérico. Observou-se que a redução do teor de proteínas na dieta para 8% associado a redução no teor de vitaminas do complexo B em ratos adultos, não causa alterações qualitativas e quantitativas das células sanguíneas das séries vermelha e branca, não leva a redução do valor da globulina nem a formação de edema ou ao surgimento de sinais clínicos característicos das carências de proteínas e vitaminas do complexo B. Por outro lado causa: menor ganho de peso corporal; alteração da composição corporal com acúmulo de gordura; redução nos valores de proteína total e albumina; redução na área do colo e evidenciação de densidade de neurônios nitrérgicos e NADH-diaforase positivos inferior ao esperado.PALAVRAS CHAVE: neurônios entéricos, desnutrição protéica, colo ascendente, hematologia, vitamina B. The utilization of animal models in studies of food restriction with known and controlled diets has as an advantage the improved knowledge about the many aspects of the human desnutrition 1 . Research on desnutrition aimsat simulating natural conditions, which involve deficienc...

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Section: Methodsmentioning

confidence: 99%

Effects of protein and vitamin B deficiency on blood parameters and myenteric neurons of the colon of rats

Sant’Ana

Molinari

Miranda-Neto

2001

Arq. Neuro-Psiquiatr.

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“…For light microscopy the method of Scherer-Singler et al (1983), for electron microscopy the method of Wolf et al (1992) was applied with slight modification. Samples for light microscopy were incubated in 1 mM reduced NADPH, 0.2 mM nitro blue tetrazolium chloride (NBT), and 0.1% Triton-X 100 (TX) containing Tris-HCl buffer (0.1 M, pH 8.1) in dark at room temperature until the blue formazan precipitate appeared (approx.…”

Section: Nadph-diaphorase Histochemistrymentioning

confidence: 99%

Ultrastructural localization of NADPH diaphorase and nitric oxide synthase in the neuropils of the snail CNS

Nacsa

Elekes

Serfözö

2015

Micron

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This manuscript is contextually identical with the following published paper:For the first time, we have shown that NADPH-diaphorase reactivity and NOS immunoreactivity are bound to transmitter vesicle membranes in varicosities of ganglionic neuropil of an invertebrate. The presence of NOS in certain synaptic configurations of the snail CNS suggests a role of NO in fast and target specific interneuronal signaling processes. 3 ABSTRACTComparative studies on the nervous system revealed that nitric oxide (NO) retains its function through the evolution. In vertebrates NO can act in different ways: it is released solely or as a co-transmitter, released from presynaptic or postsynaptic site, spreads as a volumetric signal or targets synaptic proteins. In invertebrates, however, the possible sites of NO release have not yet been identified. Therefore, in the present study, the subcellular distribution of the NO synthase (NOS) was examined in the central nervous system (CNS) of two gastropod species, the terrestrial snail, Helix pomatia and the pond snail, Lymnaea stagnalis, which are model species in comparative neurobiology. For the visualization of NOS NADPH-diaphorase histochemistry and an immunohistochemical procedure using a universal anti-NOS antibody were applied. At light microscopic level both techniques labeled identical structures in sensory tracts ramifying in the neuropils of central ganglia and cell bodies of the Lymnaea and Helix CNS. At ultrastructural level NADPH-d reactive/NOS-immunoreactive materials were localized on the nuclear envelope and membrane segments of the rough and smooth endoplasmic reticulum, as well as the cell membrane and axolemma of positive perikarya. NADPH-d reactive and NOS-immunoreactive varicosities connected to neighboring neurons with both unspecialized and specialized synaptic contacts. In the varicosities, the majority of the NADPH-d reactive/NOS-immunoreactive membrane segments were detected in round and pleomorph agranular vesicles of small size (50-200 nm). However, only a small portion (16%) of the vesicles displayed the NADPH-d reactivity/NOSimmunoreactivity. No evidence for the postsynaptic location of NOS was found. Our results suggest that the localization of NADPH-diaphorase and NOS is identical in the snail nervous system. In contrast to vertebrates, however, NO seems to act exclusively in an anterograde way possibly released from membrane segments of the presynaptic transmitter vesicle surface. Based on the subcellular distribution of NOS, NO could be both a volume and a synaptic mediator, in addition NO may function as a co-transmitter.

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“…This medium contained in 100 ml: 25 mg of NBT; 50 mg of b-NADPH (Sigma, Steinheim, Germany), 03% Triton X-100 in Tris HCl buffer (GibcoBRL, New York, USA) 0.1 M (pH 6.0). After incubation the segments were opened at the mesocolic insertion, washed three times in PBS (five min each) and immersed in 5% paraformaldehyde solution 25 .…”

Section: Methodsmentioning

confidence: 99%

Effect of protein and vitamin B deficiency on the morpho-quantitative aspects of the myenteric plexus of the descending colon of adult rats

Araújo

Sant’Ana

Molinari

et al. 2003

Arq. Neuro-Psiquiatr.

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We carried out this work with the purpose of studying the effects of protein and vitamin B deficiency on the morphologic and quantitative aspects of the myenteric plexus of the descending colon of adult Rattus norvegicus. Twenty-eight rats were divided in two groups, one of them receiving chow with 22% protein level (control) and the other fed with chow having 8% protein level without vitamin B supplementation, during 120 days. Whole-mounts of the descending colon were prepared and stained with Giemsa, NADH-diaphorase and NADPH-diaphorase. The undernourished rats had a body weight 11.84% less than the control group. Relative to the controls, the experimental group had a colonic area 48% smaller, 51.9% less Giemsa-stained neurons, 28.3% less NADH-diaphorase positive neurons and 24.2% less NADPH-diaphorase positive neurons.

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Demonstration of a unique population of neurons with NADPH-diaphorase histochemistry

Cited by 667 publications

References 9 publications

Effects of protein and vitamin B deficiency on blood parameters and myenteric neurons of the colon of rats

Effects of protein and vitamin B deficiency on blood parameters and myenteric neurons of the colon of rats

Ultrastructural localization of NADPH diaphorase and nitric oxide synthase in the neuropils of the snail CNS

Effect of protein and vitamin B deficiency on the morpho-quantitative aspects of the myenteric plexus of the descending colon of adult rats

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