Subcellular localization of the neuronal isoform of nitric oxide synthase in the rat brain: A critical evaluation

Rothe, Fritz; Canzler, U.; Wolf, Gerald

doi:10.1016/s0306-4522(97)00373-4

Cited by 61 publications

(33 citation statements)

References 46 publications

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“…Consequently, the NADPH-d reaction on the axolemma and different intracellular membranes can be considered to be specific. A similar ultrastructural localization of NADPH-d at membrane level has been reported in the nervous system of mammals (Wolf et al, 1992;Rothe et al, 1998), and crayfish (Johansson et al, 1996), supporting our observations. A negative aspect of the pre-embedding labeling of the enzyme reactions in electron microscopic preparations is that the formation of the polymer precipitate can mask the exact location of the target molecule.…”

Section: Specificity and Identity Of The Demonstration Of Nadph-d Andsupporting

confidence: 92%

“…3B). This is probably due to the crystallization of the polymer endproduct of the substrate BSPT at the site of NADPH oxidation, as it was suggested earlier (Rothe et al, 1998). NOS was detected at subcellular level in the cytoplasm of cortical NOS-IR neurons (Rothe et al, 1998), but was also found at the postsynaptic complex of glutamatergic synapses in the hippocampus (Szabadits et al, 2010).…”

Section: Specificity and Identity Of The Demonstration Of Nadph-d Andmentioning

confidence: 75%

“…From this point of view it is rather strange why only a single study has dealt until now with fine details of the localization of NOS (Johansson et al, 1996), in contrast to vertebrates (Rodrigo et al, 1997;Aoki et al, 1998;Rothe et al, 1998;Sancesario et al, 2000;Seress et al, 2005). At light microscopic level NOS was found mainly in sensory-and interneurons, and less in motor neurons of invertebrates, and it seemed that NO mainly acts in an anterograde way (Park et al, 1998;Wildemann and Bicker, 1999;Serfőző et al, 2008).…”

Section: In Invertebratesmentioning

confidence: 99%

“…It also concerns the immuno-electron microscopic visualization of NOS since ultrastructural and biochemical studies suggested that a significant amount and activity of NOS are of cytosolic location (Hecker et al, 1994;Huang et al, 1997;Zhou and Zhu, 2009). Hence the exact identity of the ultrastructural NADPH-d reaction product, which is only membrane-bound, remains for further discussion (Rothe et al, 1998).…”

Section: In Invertebratesmentioning

confidence: 99%

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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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Section: Specificity and Identity Of The Demonstration Of Nadph-d Andsupporting

confidence: 92%

Section: Specificity and Identity Of The Demonstration Of Nadph-d Andmentioning

confidence: 75%

Section: In Invertebratesmentioning

confidence: 99%

Section: In Invertebratesmentioning

confidence: 99%

See 2 more Smart Citations

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

Nacsa

Elekes

Serfözö

2015

Micron

View full text Add to dashboard Cite

show abstract

“…The observation of punctate fluorescence is in agreement with previous studies on NOS detection by immunohistochemistry and histochemistry (Gonzalez-Hernandez et al, 1996;Hecker et al, 1994). NOS is found within cells in association with subcellular organelles, especially with the endoplasmic reticulum and nuclear envelope and sometimes within the Golgi apparatus, and with the mitochondrial membrane (FaberZuschratter and Wolf, 1994;Hecker et al, 1994;Rothe et al, 1998). NO detection using DAF-2-DA has demonstrated punctate fluorescence in cultured neurons (Chen et al, 2001) and in brain slices (Brown et al, 1999;Buskila et al, 2005).…”

Section: Discussionsupporting

confidence: 90%

Simultaneous nitric oxide and dehydroascorbic acid imaging by combining diaminofluoresceins and diaminorhodamines

Rubakhin

Sweedler

2008

Journal of Neuroscience Methods

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Spatial measurements of nitric oxide (NO) production are important to understand the function and metabolism of this molecule. The reagent, 4,5-diaminofluorescein (DAF-2) and several structurally similar probes are widely used for detection and imaging of NO. However, DAF-2 also reacts with dehydroascorbic acid (DHA) in biological samples, with both products having nearly indistinguishable fluorescence spectra. Measurements using fluorimetry and fluorescence microscopy cannot easily differentiate NO-related fluorescent signals from DHA-related signals. While DAFs and the structurally related diaminorhodamines (DARs) both react with NO and DHA, they do so to different extents. We report a multiderivatization method to image NO and DHA simultaneously by using both DAF and DAR. Specifically, DAF-2 and DAR-4M are used to image NO and DHA concentrations; after reaction, the solutions are excited, at 495 nm to measure fluorescence emission from DAF-2, and at 560 nm to measure fluorescence emission from DAR-4M. Using the appropriate calibrations, images are created that depend either on the relative NO or the relative DHA concentration, even though each probe reacts to both compounds. The method has been validated by imaging NO production in both undifferentiated and differentiated pheochromocytoma cells.

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Immune stress activates putative nitric oxide-producing neurons in rat brain: Cumulative effects with restraint

1999

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Subcellular localization of the neuronal isoform of nitric oxide synthase in the rat brain: A critical evaluation

Cited by 61 publications

References 46 publications

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

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

Simultaneous nitric oxide and dehydroascorbic acid imaging by combining diaminofluoresceins and diaminorhodamines

Immune stress activates putative nitric oxide-producing neurons in rat brain: Cumulative effects with restraint

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