Colocalization of nitric oxide synthase and NADPH-diaphorase in cultured myenteric neurons

Saffrey, M. Jill; Hassall, C. J. S.; Hoyle, Charles H.V.; Belai, Abebech; Moss, Jonathan; Schmidt, Hanna; Förstermann, Ulrich; Murad, Ferid; Burnstock, Geoffrey

doi:10.1097/00001756-199204000-00011

Cited by 90 publications

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“…The tissue was first incubated with potassium phosphate-buffered saline (KPBS) plus 0.4% Triton X-100 for 60 min and then incubated for 24-48 hr at 4°C with a rabbit polyclonal antibody (I :4SOO; #676 I-9, Abbott Laboratories, Abbott Park, IL) to rat cerebellar NOS. The rabbit polyclonal antibody to rat cerebellar NOS is specific for NOS isoform I from the brain and does not cross-react with NOS isoforms II or III from induced macrophages or endothelial cells, respectively (Schmidt et al, 1992). After thorough washing, sections were exposed to biotinylated anti-rabbit IgG followed by streptavidinhorseradish peroxidase (I :300).…”

Section: Drgmentioning

confidence: 99%

“…In these ganglia, an average of 37.0 2 4.0% (L6) and 20.6 + 2.2% (Sl), respectively, of FG-labeled pelvic afferent neurons were NOS-IR compared to 1.1 i 0.5% (L6) and 2.5 t 1.4% (Sl) Nitric oxide (NO) has been implicated as a neurotransmitter or neuromodulator at various sites in the mammalian nervous system (Scherer-Singler et al, 1983;Bredt et al, 1990;Pitkanen and Amaral, 1991;Anderson, 1992;Blottner and Baumgarten, 1992;Schmidt et al, 1992;Valtschanoff et al, 1992b). Neurons containing NO have been identified histochemically by the presence of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity or immunocytochemically with an antibody for NO synthase (NOS), the enzyme responsible for NO synthesis (Bredt et al, 1991;Dawson et al, 1991;Vincent and Kimura, 1992).…”

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confidence: 99%

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Increased expression of neuronal nitric oxide synthase (NOS) in visceral neurons after nerve injury

1995

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The distribution of nitric oxide synthase immunoreactivity (NOS-IR) and the changes in this distribution after peripheral axotomy were examined in lumbosacral afferent and preganglionic neurons (PGNs) innervating the pelvic viscera of the male rat. The visceral neurons in L6-S1 and L1-L2 dorsal root ganglia (DRG) and in the spinal cord were identified by retrograde axonal transport following injection of Fluorogold (FG) into the major pelvic ganglion (MPG). Axotomy was performed by removing the MPG on one side 2–4 weeks prior to sacrificing the animals. A differential distribution of NOS-IR was detected in DRG cells at different segmental levels of control animals. Significantly greater numbers of NOS-IR cells were present in thoracic (T8, T10, T12; 30–44 cell profiles/section) and rostral lumbar DRGs (L1-L2; 3–15 NOS-IR cell profiles/section) compared to caudal lumbosacral (L5-S1) DRGs (0.2–0.7 cell profiles/section). A significant increase in the number of NOS-IR cells was detected in the L6-S1 DRG (p < or = 0.001; 11 NOS-IR cell profiles/section) but not in the L2 or L5 DRG ipsilateral to axotomy. In these ganglia, an average of 37.0 +/- 4.0% (L6) and 20.6 +/- 2.2% (S1), respectively, of FG-labeled pelvic afferent neurons were NOS-IR compared to 1.1 +/- 0.5% (L6) and 2.5 +/- 1.4% (S1) contralateral to the axotomy. Following axotomy, a significantly greater percentage of dye-labeled pelvic visceral afferents in the L1 and L2 DRG also exhibited NOS-IR in comparison to the contralateral side. Following axotomy, NOS-IR fibers were detected along the lateral edge of the dorsal horn extending from Lissauer's tract to the region of the sacral parasympathetic nucleus (SPN) on the ipsilateral side of the L6 and S1 spinal segments. These NOS-IR fibers were not detected in adjacent spinal segments (L4, L5, or S2). Axotomy also changed the numbers of NADPH-d-positive and NOS-IR cells in the region of the SPN in the L6 spinal segment. Contralateral to the axotomy 38.3 +/- 4.0% of PGNs in the L6 spinal segment were colabeled with NOS-IR; however, ipsilateral to axotomy, a significantly greater percentage (61.0 +/- 3.0%; p < or = 0.01) of PGNs exhibited NOS-IR. Axotomy did not alter the distribution of PGNs in the S1 segment exhibiting NOS-IR. These results indicate that NOS-IR in visceral afferent and PGNs is plastic and can be upregulated by peripheral nerve injury.

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Section: Drgmentioning

confidence: 99%

mentioning

confidence: 99%

Increased expression of neuronal nitric oxide synthase (NOS) in visceral neurons after nerve injury

1995

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“…In all double-labelling experiments, NADPH-diaphorase histochemical staining was performed prior to immunolabelling, as the latter tended to mask the diaphorase activity SAFFREY et al, 1992).…”

Section: Resultsmentioning

confidence: 99%

“…NOS in neurons was localized using specific antisera (6761-9 and 6763-5) raised against NOS purified from the rat cerebellum (courtesy of U. FORSTERMANN). Other studies in our laboratory have shown that these antisera give specific labelling for NOS in other autonomic ganglia HASSALL et al, 1992;SAFFREY et al, 1992). For double labelling, NADPH-diaphorase staining was carried out prior to immunolabelling.…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…In the autonomic nervous system, pharmacological and electrophysiological evidence suggests that NO is an important mediator of some non-adrenergic, non-cholinergic (NANC) inhibitory responses (BAND, 1992). Subpopulations of neurons which express nitric oxide synthase (NOS) activity have been localized in the rat enteric ganglia (BREDT, HWANG and SNYDER, 1990;DAWSON et al, 1991;BELAI et al, 1992), guinea-pig enteric ganglia (COSTA et al, 1992;SAFFREY et al, 1992;YOUNG et al, 1992), guinea-pig cardiac ganglia and rat adrenal gland (AFEWORK et al, 1992). In all these autonomic ganglia, NOS-immunoreactivity has been found to be colocalized with NADPH-diaphorase activity.…”

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NADPH-diaphorase is Colocalized with Nitric Oxide Synthase and Vasoactive Intestinal Polypeptide in Rat Pancreatic Neurons in Culture.

Tay

Moules

1994

Archives of Histology and Cytology

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Summary. NADPH-diaphorase activity together with nitric oxide synthase-and vasoactive intestinal polypeptide-immunoreactivities were examined in the neurons of the rat pancreas in culture. Nitric oxide synthase (NOS)-and vasoactive intestinal polypeptide (VIP)-immunoreactivities were localized in a subpopulation of neurons which were also NADPH-diaphorase positive. All neurons expressing colocalized activities for NOS/ VIP and NADPH-diaphorase were solitary cells in culture. Each of these double-labelled neurons was characterized by a round or oval cell body, with short and long processes emanating from it. Some of these processes traversed several micrometres before terminating on the exocrine acinar and endocrine cells. The present study provides evidence that nitric oxide may act as a regulatory agent in the inhibitory neural control of the secretory functions in the pancreas, thereby maintaining the milieu interieur of the organism.

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Distribution and colocalization of NADPH‐diaphorase activity, nitric oxide synthase immunoreactivity, and VIP immunoreactivity in the newly hatched chicken gut

1995

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Colocalization of nitric oxide synthase and NADPH-diaphorase in cultured myenteric neurons

Cited by 90 publications

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Increased expression of neuronal nitric oxide synthase (NOS) in visceral neurons after nerve injury

Increased expression of neuronal nitric oxide synthase (NOS) in visceral neurons after nerve injury

NADPH-diaphorase is Colocalized with Nitric Oxide Synthase and Vasoactive Intestinal Polypeptide in Rat Pancreatic Neurons in Culture.

Distribution and colocalization of NADPH‐diaphorase activity, nitric oxide synthase immunoreactivity, and VIP immunoreactivity in the newly hatched chicken gut

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