Abstract:
:Nitric oxide (NO) production in the sensory neurons of the rad nodose ganglion was studied by examining the distribuiotn of NO synthase (NOS) by use of NADPH diaphorase (NADPHD) histochemistry and immunohistochemistry ofr the presence of isoformes of NOS: neuronal (nNOS), endothelial (eNOS) and the inducible isoform (iNOS). Distribution and changes in NO production during acute hypoxia were studied in vital vibratome sections with the fluorescent marker for NO, diaminotriazolofluorescein (DAF-2T). Furthermore, changes in reactive oxygen species (ROS) in vibratome slices were examined utilizing 2',7'-dichlorofluorescein (DCF). By use of these histochemical methods, a positive NADPH reaction and positive immunoreactivity for eNOS were noted in all neurons observed. While for nNOS immunoreactivity, both strongly positive cells but also many negative cells are seen., no iNOS immunoreactive cells were observed. In vital vibratome slices, a dot-like distribution of fluorescence for DAF-2T, indicating production of NO, was observed in the nodose ganglion cells. Neurons exposed to hypoxia showed stronger DAF-2T fluorescence than cells exposed to normoxia, indicating an increased production of NO during hypoxia. When Ca(2+) was removed from the incubation buffer, the intensity of fluorescence for DAF-2T decreased but did not disappear completely. Using a photoconversion technique, DAF-2T was localized in the inner membrane of mitochondria in the ganglion cells by electron microscopy. The level of DCF signals for detection of ROS was higher in neurons incubated in the normoxic medium than those incubated under conditions of hypoxia. Nerve cells exposed to hypoxia followed by reoxygenation (3 min in normoxic conditions) showed higher fluorescence for DCF than those exposed to normoxia. The results of the present study demonstrate clearly that the basal production of NO in viscerosensory neurons is increased during hypoxia and is due to the isoform eNOS rather than nNOS, moreover, that ROS is augmented by reoxygenation but not during hypoxia.
journal_name
Brain Resjournal_title
Brain researchauthors
Yamamoto Y,Henrich M,Snipes RL,Kummer Wdoi
10.1016/s0006-8993(02)03826-xsubject
Has Abstractpub_date
2003-01-24 00:00:00pages
1-9issue
1eissn
0006-8993issn
1872-6240pii
S000689930203826Xjournal_volume
961pub_type
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