NADPH-diaphorase Research Articles

Neuroplastic and neuropathological changes in the central nervous system of the Gray mussel Crenomytilus grayanus (Dunker) under environmental stress

We studied here neuron ultrastructure, synaptic plasticity and subcellular localization of NADPH-diaphorase (NADPH-d), a cytochemical marker for nitric oxide syntase, in the pedal ganglia of the Gray mussel Crenomytilus grayanus sampled from the polluted and reference sites in Amursky Bay (Sea of Japan) at lower and higher water temperature (in the beginning and the end of August, respectively). At lower temperature, neuroplastic changes in mussel ganglia prevailed: a sharp increase in the number of cytosomes in NADPH-d-positive neurons and a sharp decrease in the number of mitochondria in both NADPH-d-positive and NADPH-d-negative neurons. At higher temperature, neurodegenerative changes prevailed: disruption of a part of NADPH-d-negative axons and interneuronal contacts, formation of concentric lamellar structures in the neuropils, and accumulation of autophagosomes in NADPH-d negative neurons. The results suggest that the stress-induced production of nitric oxide in cytosomes of mussel neurons and plasticity of gap junctions have a neuroprotective effect.

Distribution and phenotype of Phox2a‐containing neurons in the adult sprague‐dawley rat

Phox2a is a transcription factor that plays an essential role, with Phox2b, in the specification of the adrenergic and noradrenergic phenotype in developing brain. Localization of Phox2a in developing brainstem has demonstrated a high degree of correspondence between neurons expressing the transcription factor and those involved in the regulation of autonomic function. Although it is well established that the paralogous gene product Phox2b is widely expressed in adult brain, no study has mapped the distribution of Phox2a in the adult. The data reported here address that void. A well-characterized rabbit polyclonal antiserum was used for immunohistochemical localization of the transcription factor in adult rats. Sections through the rostrocaudal extent of brain were processed for dual immunocytochemical localization of Phox2a and catecholamine enzymes. Adjacent sections were used for dual localization of Phox2a and NADPH diaphorase, a marker of nitric oxide-containing neurons. The data demonstrate that Phox2a is present in all brainstem catecholamine neurons, in circumscribed populations of NADPH(+) neurons, and in a subset of neurons that influences sympathetic and parasympathetic outflow. In addition, Phox2a(+) neurons were observed within diencephalic and brainstem nuclei that regulate behavioral state. Considered with data demonstrating that Phox2a is part of the transcriptional complex that drives expression of dopamine-beta-hydroxylase and can also up-regulate expression of other genes, the data support the conclusion that Phox2a plays an important role in brainstem catecholamine neurotransmission and in the regulation of adaptive homeostatic functions in the adult nervous system.

Capillaries in the Brain Microcirculatory Bed in the Acute Period of Experimental Brain Injury

Objective: to provide a morphochemical evaluation of the capillaries in the brain microcirculatory bed of experimental animals in the acute period of brain injury (BI). Materials and methods. An experiment was carried out on 40 sexually mature Wister rats. Gradual BI was inflicted by a falling load blow on the right parietotemporal region, as described by T. F. Sokolova (1986). Brain magnetic resonance imaging was made in the animals an hour after injury infliction to define the extent of the damage and its site. Morphological studies of the brain were conducted 24 and 72 hours and 7 days after the injury. The capillaries were identified by the injection technique (Indian ink imbedding). The NO-producing function of endotheliocytes was evaluated using the NADPH-diaphorase histochemical technique. To study microcirculatory changes, the similar brain portions ipsilateral to the site of injury and in the intact hemisphere were compared in each animal. The changes in the diameter of capillaries, the volume density of the microcirculatory bed, the exchange surface area and activity of NADPH diaphorase in the capillary wall were analyzed. The findings were processed by the variation statistical method, by determining the arithmetic mean, the standard error of the arithmetic mean, and the test of significance. The findings give an insight into the mechanisms responsible for secondary ischemic lesions in the early period of brain injury. The NO-dependent capillary blood flow reduction leading to hypoxia may be one of the most important causes of secondary cerebral lesion. All changes in the dynamics of microvessels (their lumen and area) are in line with the activity of the enzyme. Conclusion. In severe BI, changes in the brain microcirculatory bed, its capillary link in particular, are manifested not only with in a traumatic injury focus, but also involve the brain as a whole. Key words: brain, brain njury, capillaries, nitric oxide (NO).

Neonatal Handling Impairs Spatial Memory and Leads to Altered Nitric Oxide Production and DNA Breaks in A Sex Specific Manner

Early life events lead to behavioral and neurochemical changes in adulthood. The aim of this study is to verify the effects of neonatal handling on spatial memory, nitric oxide (NO) production, antioxidant enzymatic activities and DNA breaks in the hippocampus of male and female adult rats. Litters of rats were non-handled or handled (10 min/day, days 1-10 after birth). In adulthood they were subjected to a Morris water maze or used for biochemical evaluations. Female handled rats showed impairment in spatial learning. They also showed decreased NO production, while no effects were observed in these parameters in male rats. No effects were observed on the number of hippocampal NADPH diaphorase positive cells. In the Comet Assay, male handled rats showed increased DNA breaks index when compared to non-handled ones. We conclude that neonatal handling impairs learning performance in a sex-specific manner, what may be related to NO decreased levels.

Age-related changes in nitric oxide synthase in the lateral geniculate nucleus of rats

Age-related changes in nitric oxide production in the visual system have not been well characterized. Therefore, we used staining and image-processing approaches to describe changes in levels of neuronal nitric oxide synthase (nNOS), the NADPH-diaphorase (NADPH-d) histochemical marker, and 3-nitrotyrosine in the lateral geniculate nucleus (LGN) of young and aged rats. The LGN plays an important role in the visual system, as it acts as a visual relay nucleus. Quantitative analysis of NADPH-d-positive and nNOS-immunoreactive neurons revealed significant optical density increases in the dorsal LGN and ventral LGN of aged rats; however, no significant changes were observed in the number of neurons with age. 3-Nitrotyrosine immunoreactivity was increased in the dorsal LGN and ventral LGN of aged rats. These results indicate that increased nitric oxide production and peroxynitrite may be associated with alterations in visual function during aging.

Nitroxidergic Nerve Fibers of Intracerebral Vessels

Light and electron histochemical methods were used to study the structure and distribution of neurons containing NADPH diaphorase and their processes in the parietal area of the cortex in rats. Most neurons were found to be characterized by tight associations with intracerebral vessels. The smallest distances between the axon plasmalemma and the smooth myocytes of intracerebral arteries in the cerebral cortex were at least 0.3-0.5 microm. Neuron bodies were located at functionally important locations of vessels (sites at which subsidiary vessels branched off, the origins of arterioles), and their processes accompanied vessels, densely entwining the vessels with their branches. Neurons whose dendrites contacted the bodies or process of above- or below-lying neurons often sent nerve conductors to arteries, veins, or capillaries. Thus, nitroxidergic neurons or groups of these neurons may monitor the state of the circulation at different points in the vascular bed, functioning as local nerve centers.

Nitric Oxide Synthase Mediates the Ability of Darbepoetin Alfa to Improve the Cognitive Performance of STOP Null Mice

STOP (stable tubule only polypeptide) null mice display neurochemical and behavioral abnormalities that resemble several well-recognized features of schizophrenia. Recent evidence suggests that the hematopoietic growth factor erythropoietin improves the cognitive performance of schizophrenics. The mechanism, however, by which erythropoietin is able to improve the cognition of schizophrenics is unclear. To address this question, we first determined whether acute administration of the erythropoietin analog known as darbepoetin alpha (D. alpha) improved performance deficits of STOP null mice in the novel objective recognition task (NORT). NORT performance of STOP null mice, but not wild-type littermates, was enhanced 3 h after a single injection of D. alpha (25 microg/kg, i.p.). Improved NORT performance was accompanied by elevated NADPH diaphorase staining in the ventral hippocampus as well as medial and cortical aspects of the amygdala, indicative of increased nitric oxide synthase (NOS) activity in these structures. NOS generates the intracellular messenger nitric oxide (NO) implicated in learning and memory. In keeping with this hypothesis, D. alpha significantly increased NO metabolite levels (nitrate and nitrite, NOx) in the hippocampus of both wild-type and STOP null mice. The NOS inhibitor, N (G)-nitro-L- arginine methyl ester (L-NAME; 25 mg/kg, i.p.), completely reversed the increase in hippocampal NOx levels produced by D. alpha. Moreover, L-NAME also inhibited the ability of D. alpha to improve the NORT performance of STOP null mice. Taken together, these observations suggest D. alpha enhances the NORT performance of STOP null mice by increasing production of NO.

NADPH-Diaphorase Activity and NO Synthase Expression in the Olfactory Epithelium of the Bovine

NADPH-diaphorase (NADPH-d) staining of the bovine olfactory epithelium was compared with the immunohistochemical localization of nitric oxide synthase (NOS), soluble guanylyl cyclase, and cGMP (cyclic guanosine 3',5'-monophosphate). Out of the three isoforms, only the inducible NOS (NOS-II) was found at the epithelial surface correlating with the strong labelling for NADPH-d. In contrast, light diaphorase staining associated with deeper epithelial regions did not coincide with any NOS immunoreactivity. As there is overlapping expression of NOS-II, soluble guanylyl cyclase and cGMP at the luminal surface morphologically occupied by dendritic knobs of olfactory receptor neurons and microvillar endings of supporting cells, the nitric oxide (NO)/cGMP pathway is likely to be involved in modulating the odour signals during olfactory transduction.

NADPH-d cells (mast cells) around and within the autonomic nerves of porcine renal hilus

The enzymehistochemical localization of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) reactive cells next and within the autonomic nerves in the porcine kidney's hilus, close to renal artery and vein as well as in its adventitia, was studied. It was established that the prevailing part of NADPH-d-reactive cells were distributed next to and within NADPH-d-reactive autonomic fibers around the renal vein. Comparatively small number of reactive cells was found around the renal artery, but single NADPH-d-reactive cells were found in the neighbouring autonomic nerves as well. The cells observed next to nerves had much more expressed reaction (granules) than those, located in the nerves. Toluidine blue staining on frozen and paraffine sections from the same areas showed well-expressed γ-ma metachromasia in mast cells with similar or identical localization. This indicated that observed reactive cells were mast cells. The obtained data convincingly showed that porcine NADPH-d cells may produce nitric oxide and in this way most probably they were involved in the function of autonomic nerves in the renal blood vessels (vasodilatation) and the kidney function, as well.

Regulation of cyclooxygenase activity in airway epithelium by endogenous nitric oxide

The role of nitric oxide (NO) in the activity of cyclooxygenase (COX) in cultured canine tracheal epithelium was studied. Tracheal epithelium spontaneously released prostaglandin E2 (PGE2), which is a product of COX. The release of PGE2 was increased by bradykinin and was decreased by two NO synthase inhibitors: NG-nitro-L-arginine methyl ester and NG-monomethyl-L-arginine. That decrease was reversed in the presence of L-arginine. Chrolpromadin, but not aminoguanidine, inhibited PGE2 production, which suggests that constitutive NO synthase is involved. Two stable NO donors, sodium nitroprusside and S-nitroso-N-acetyl DL-penicillamine, also increased the production of PGE2. These effects were abolished by coincubation with hemoglobin, which binds and inactivates NO, but not by methylene blue, an inhibitor of soluble guanylate cyclase. NADPH diaphorase histochemistry of cultured tracheal cells revealed activity in the periphery of the cytoplasm. These results suggest that, in cultured canine tracheal epithelium, NO directly interacts with COX to regulate PGE2 production.

Interstitial cells of Cajal in the cynomolgus monkey rectoanal region and their relationship to sympathetic and nitrergic nerves

The morphology of interstitial cells of Cajal (ICC) in the circular muscle layer of the cynomolgus monkey internal anal sphincter (IAS) and rectum and their relationship to sympathetic and nitrergic nerves were compared by dual-labeling immunohistochemistry. Contractile studies confirmed that nitrergic nerves participate in neural inhibition in both regions whereas sympathetic nerves serve as excitatory motor nerves only in the IAS. Muscle bundles extended from myenteric to submucosal edge in rectum but in the IAS bundles were further divided into "minibundles" each surrounded by connective tissue. Dual labeling of KIT and smooth muscle myosin revealed KIT-positive stellate-shaped ICC (ICC-IAS) within each minibundle. In the rectum intramuscular ICC (ICC-IM) were spindle shaped whereas stellate-shaped ICC were located at the myenteric surface (ICC-MY). ICC were absent from both the myenteric and submucosal surfaces of the IAS. Nitrergic nerves (identified with anti-neuronal nitric oxide synthase antibodies or NADPH diaphorase activity) and sympathetic nerves (identified with anti-tyrosine hydroxylase antibody) each formed a plexus at the myenteric surface of the rectum but not the IAS. Intramuscular neuronal nitric oxide synthase- and tyrosine hydroxylase-positive fibers were present in both regions but were only closely associated with ICC-IM in rectum. Minimal association was also noted between ICC-IAS and cells expressing the nonspecific neuronal marker PGP9.5. In conclusion, the morphology of rectal ICC-IM and ICC-MY is similar to that described elsewhere in the gastrointestinal tract whereas ICC-IAS are unique. The distribution of stellate-shaped ICC-IAS throughout the musculature and their absence from both the myenteric and submucosal surfaces suggest that ICC-IAS may serve as pacemaker cells in this muscle whereas their limited relationship to nerves suggests that they are not involved in neuromuscular transmission. Additionally, the presence of numerous minibundles, each containing both ICC-IAS and nerves, suggests that this muscle functions as a multiunit type muscle.

Vitamin E improves learning performance and changes the expression of nitric oxide-producing neurons in the brains of diabetic rats

We investigated the effects of chronic administration of vitamin E on nitric oxide (NO)-producing neurons in the brains of streptozotocin (STZ)-induced diabetic rats using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry. We further evaluated the effects of diabetes and vitamin E treatment on experimental anxiety and memory processes using the elevated plus maze (EPM) Trial 1/2 protocol. Wistar rats were divided into four groups: normoglycemics (N), normoglycemics treated with vitamin E (NVE), diabetics (D), and diabetics treated with vitamin E (DVE). Diabetes mellitus was induced by a single intraperitoneal injection of STZ (35mg/kg). Vitamin E (100mg/kg) or vehicle was administered orally by gavage (1ml/kg) once each day for 7 weeks. After behavioral testing, the dentate gyrus of the hippocampus (DG), striatum, paraventricular nucleus of the hypothalamus (PVN), supraoptic nucleus (SON), and dorsolateral periaqueductal grey (DLPAG) were analyzed for NADPH-d histochemistry. STZ-induced diabetic rats exhibited decreased locomotor activity and cognitive impairment compared with normoglycemic controls. The number of NADPH-d-positive neurons was increased in the DG, striatum, and DLPAG of diabetic rats. An increase in soma area was detected in all structures analyzed (DG, striatum, PVN, SON, and DLPAG) of STZ-induced diabetic animals. The present study showed that chronic administration of vitamin E ameliorates memory in STZ-induced diabetic rats and revealed that NOS-producing neurons have an increased soma area which can be restored, at least partially, by vitamin E treatment. These results suggest the potential use of vitamin E as an adjuvant therapy for the prevention and treatment of diabetic conditions.

Distribution of NADPH-diaphorase and AChE activity in the anterior leaflet of rat mitral valve

The mitral valve, as an active flap, forms the major part of the left ventricular inflow tract and therefore plays an important function in many aspects of left ventricular performance. The anterior leaflet of this valve is the largest and most ventrally placed of two leaflets that come together during ventricular systole to close the left atrioventricular orifice. Various neurotransmitters are responsible for different functions including controlling valve movement, inhibiting or causing the failure of impulse conduction in the valve and the sensation of pain. Nitric oxide acts as a gaseous free radical neurotransmitter, neuromediator and effective cardiovascular modulator. Acetyl-choline is known to function as a typical neurotransmitter. Histochemical methods for detection of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d), as an indirect nitric oxide-synthase marker, and method for detection of acetylcholinesterase (AChE) were used. Both methods were performed on the same valve sample. A widespread distribution of nerve fibres was observed in the anterior leaflet of the mitral valve. The fine NADPH-d positive (nitrergic) nerve fibres were identified in all zones of valve leaflet. AChE positive (cholinergic) nerve fibres were identified forming dense network and fibres organized in stripes. Endocardial cells and vessels manifested heavy NADPH-d activity. Our observations suggest a different arrangement of nitrergic and cholinergic nerve fibres in the anterior leaflet of the mitral valve. The presence of nitrergic and cholinergic activity confirms the involvement of both neurotransmitters in nerve plexuses and other structures of mitral valve.

An Arabidopsis mutant with high cyclic electron flow around photosystem I (hcef) involving the NADPH dehydrogenase complex.

Cyclic electron flow (CEFI) has been proposed to balance the chloroplast energy budget, but the pathway, mechanism, and physiological role remain unclear. We isolated a new class of mutant in Arabidopsis thaliana, hcef for high CEF1, which shows constitutively elevated CEF1. The first of these, hcef1, was mapped to chloroplast fructose-1,6-bisphosphatase. Crossing hcef1 with pgr5, which is deficient in the antimycin A-sensitive pathway for plastoquinone reduction, resulted in a double mutant that maintained the high CEF1 phenotype, implying that the PGR5-dependent pathway is not involved. By contrast, crossing hcef1 with crr2-2, deficient in thylakoid NADPH dehydrogenase (NDH) complex, results in a double mutant that is highly light sensitive and lacks elevated CEF1, suggesting that NDH plays a direct role in catalyzing or regulating CEF1. Additionally, the NdhI component of the NDH complex was highly expressed in hcef1, whereas other photosynthetic complexes, as well as PGR5, decreased. We propose that (1) NDH is specifically upregulated in hcef1, allowing for increased CEF1; (2) the hcef1 mutation imposes an elevated ATP demand that may trigger CEF1; and (3) alternative mechanisms for augmenting ATP cannot compensate for the loss of CEF1 through NDH.

A qualitative analysis of the regulation of cyclic electron flow around photosystem I from the post-illumination chlorophyll fluorescence transient in Arabidopsis: a new platform for the in vivo investigation of the chloroplast redox state

A transient in chlorophyll fluorescence after cessation of actinic light illumination, which has been ascribed to electron donation from stromal reductants to plastoquinone (PQ) by the NAD(P)H-dehydrogenase (NDH) complex, was investigated in Arabidopsis thaliana. The transient was absent in air in a mutant lacking the NDH complex (ndhM). However, in ndhM, the transient was detected in CO(2)-free air containing 2% O(2). To investigate the reason, ndhM was crossed with a pgr5 mutant impaired in ferredoxin (Fd)-dependent electron donation from NADPH to PQ, which is known to be redundant for NDH-dependent PQ reduction in the cyclic electron flow around photosystem I (PSI). In ndhM pgr5, the transient was absent even in CO(2)-free air with 2% O(2), demonstrating that the post-illumination transient can also be induced by the Fd- (or PGR5)-dependent PQ reduction. On the other hand, the transient increase in chlorophyll fluorescence was found to be enhanced in normal air in a mutant impaired in plastid fructose-1,6-bisphosphate aldolase (FBA) activity. The mutant, termed fba3-1, offers unique opportunities to examine the relative contribution of the two paths, i.e., the NDH- and Fd- (or PGR5)-dependent paths, on the PSI cyclic electron flow. Crossing fba3-1 with either ndhM or pgr5 and assessing the transient suggested that the main route for the PSI cyclic electron flow shifts from the NDH-dependent path to the Fd-dependent path in response to sink limitation of linear electron flow.

Biochemical changes in cavernosal tissue caused by single sided cavernosal nerve resection and the effects of alpha lipoic acid on these changes

ObjectiveOne of the most important complications of radical prostatectomy operation is erectile disfunction (ED). Oxidative stress patterns and apoptotic changes may happen in smooth muscles and endothelial cells of corpus cavernosum after neuropraxia or neurectomy. Alpha lipoic acid (ALA) shows its antioxidant properties by eliminating free radicals. In this experimental study we investigated the effects of ALA on rehabilitation of cavernosal tissue and nitric oxide synthase (NOS) containing nerve fibers on erectile tissue. Materials and methodsIn this study four groups were formed by inclusion of 63 adult fertile rats. Control group (n: 9), sham operation group (n: 18), 18 rats underwent unilateral neurectomy of a 5-mm. segment of the cavernous nerve (group DI) and another 18 rats group which ALA received after unilateral neurectomy (group DII). Assessments were done 3 weeks after neurectomy. ResultsWe assessed number of NOS containing nerve fibers via nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining. According to NADPH diaphorase staining group DII significantly recovered comparing group DI (48.89±19.00 and 17.22±6.67 respectively) (p<0.05). SOD activity is reduced in both; group DI and group DII (31.42±6.06 and 40.38±4.24). Nitrite+nitrate levels were elevated significantly in both group DI and group DII (0.52±0.05 and 0.44±0.02 micromole/gr wet tissue respectively) when compared with other groups (p<0.05). There is no statistical difference between results of group DI and Group DII (p>0.05). ConclusionThis study confirms that neurectomy caused decrease of intracavernous pressure and number of NOS fibers. Neurectomy and surgical trauma caused oxidative stress in rat corpus cavernosum. As a potent antioxidant ALA has positive effects on cavernosal tissue regeneration and rehabilitation by reducing oxidative stress. In this aspect, ALA may have a potential advantage in penile rehabilitation after radical prostatectomy.

Role of Sonography for Implantation and Sequential Evaluation of a VX2 Rabbit Liver Tumor Model

We investigated the role of sonography in the implantation process of a VX2 rabbit liver tumor model and sequential evaluation. Fifty rabbits were divided into 2 groups. Animals in group I underwent surgical implantation, whereas those in group II received percutaneous sonographically guided implantation. At 7, 14, 21, and 28 days after implantation, respectively, 5 rabbits in each group were examined with conventional, color Doppler (CD), contrast-enhanced (CE) pulse inversion harmonic (PIH), and CE CD sonography. Pathologic examination was performed with hematoxylin-eosin, nicotinamide adenine dinucleotide phosphate-diaphorase, and succinic dehydrogenase stains. Twenty-one rabbits with tumors survived in group I, and 22 with tumors survived in group II. The mean duration of implantation +/- SD in group II was 16.9 +/- 3.4 minutes, whereas that in group I was 21.5 +/- 4.1 minutes (P < .05). The tumor volume measured by conventional sonography increased from 0.28 +/- 0.14 cm(3) at 7 days to 16.49 +/- 5.50 cm(3) at 28 days in group I and from 0.31 +/- 0.19 to 19.79 +/- 4.70 cm(3) in group II, whereas no significant difference existed between the groups. On CD, CE PIH, and CE CD sonography, most tumors were hypervascular before 14 days and after 14 days had peripheral vessels and central hypovascular areas, which were shown as necrotic areas by pathologic examination. Sonographically guided implantation achieved a good success rate with convenient inoculation performance. Conventional gray scale, CD, CE PIH, and CE CD sonography were useful in sequential evaluation of tumor growth and characteristic vascularity.

Postnatal development of NADPH-d neurons in the enteric nervous system of the goat

The morphology and distribution of nitric oxide synthase (NOS)-expressing neurons in the myenteric plexus of small intestine of goat at different ages (2 weeks, and 2, 4, 6 and 12 months) were studied by using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry, which could stain NOS-expressing neurons as immunohistochemistry. The results showed that the NADPH-d neurons and their fibres formed a clear tertiary meshwork structure in the myenteric plexus. The shapes of stained neurons were diverse, and many neurons gathered to form numerous ganglia in variable size. During development, the density of NADPH-d neurons in the myenteric plexus of goat small intestine decreased from 27.07/mm2 at 2 weeks to 16.18/mm2 at 12 months. The total number calculated referred to the total serosal surface of small intestine, however, increased from 1.86×106 at 2 weeks to 5.84×106 at 6 months, before decreasing to 4.41×106 at 12 months. At different ages, the density of NADPH-d neurons was lowest, but the total number was highest, in the jejunum with respect to duodenum and ileum. The mean area of the perikaryon or nucleus was enlarged at first and then diminished as the goats growing. The nuclear:cytoplasmic size ratio in NADPH-d neurons at 2 weeks was much higher than that at 2, 4, 6 or 12 months (P<0.05), while no differences could be found among different ages (P>0.05).

Using a panel of immunomarkers to define homologies in mammalian brains

Brain mapping has relied on a small number of routine chemical stains for many decades. The advent of immunomarkers has had a major impact on the ability to define homologous nuclei from one species to another. The first atlas to present a panel of immunomarkers was that of Paxinos et al. (1999a,b) in the adult rat brain. The markers used were parvalbumin, calbindin, calretinin, SMI32, tyrosine hydroxylase, and NADPH diaphorase (plus nissl and acetylcholinesterase). The ‘signature’ of a nucleus of interest in a new species can be tested against the findings in the rat. Since the pattern of immunomarkers seems to be conserved in mammalian evolution, such extrapolations can be made with reasonable confidence. A marmoset brain stained with a comprehensive set of immunomarkers has recently been published on the internet (Tokuno et al., 2009) and we are in the process of defining nuclear homologies in this brain by comparison with the same markers in the rat. In this article, we present an example (mapping the amygdala in the marmoset) which demonstrates the application of this immunomarker panel in defining homologies. The technique is particularly valuable in situations where little data on hodology or electrophysiology are available.

Early observations of nitric oxide synthase in the intact brain and regenerating tentacle of the snail, Helix pomatia

Event Abstract Back to Event Early observations of nitric oxide synthase in the intact brain and regenerating tentacle of the snail, Helix pomatia Zoltán Serfözö1* and Károly Elekes1 1 Balaton Limnological Research Institute, HAS, Department of Experimental Zoology, Hungary In the present study, the regeneration dynamics of the Helix tentacle which is an established model for neuronal as well as organ regeneration was investigated by NADPH-diaphorase (NADPH-d) reaction which is the histochemical marker for nitric oxide synthase (NOS). The NOS activity and expression, as well as its possible impact on cysteine and tyrosine nitrosylation were also followed in intact brain and tentacle. In a period of 15 weeks, the terminal part of the right tentacle of ten animals was cut by every odd week; meanwhile the left tentacle of these groups remained intact for control. Neurons of the tentacular ganglion, their projections in the tentacular nerve innervating the lateral neuropil and the globuli cells of the procerebrum, showed NADPH-d reactivity. After 1 week of tentacle extirpation a faint NADPH-d reactivity was still seen in the distal part of injured axons of the tentacular nerve. At 10 weeks of regeneration NADPH-d reactivity reappeared in the newly forming tentacular ganglion, and a similar distribution to the control was obtained by 13 weeks. NOS activity of the brain was found 1422±42 dpm 14C citrulline/mg/ml protein/min/5 animals (n=4), whereas it was 2212±50 dpm in the tentacular ganglia. A fivefold dilution of the extracts resulted in an approx. five times higher activity in the brain (6691±193 dpm; n=4). L-NAME substrate analogue inhibitor proved not to be effective. From total brain extract a 140-150 kDa protein was detected by an anti-universal NOS antibody that was also labeled by an anti-S-nitrosocysteine antibody. In addition, cysteine nitrosylation was also found in other 5 proteins of 180, 170, 80, 50, and 40 kDa mw, respectively, whereas tyrosine nitration was not present in the intact Helix brain. These preliminary results show the presence of the NO signaling pathway in the snail CNS and tentacle and also suggest a possible involvement of NO in the tentacle regeneration. Supported by the OTKA PD75276 and K78224 grants. Conference: IBRO International Workshop 2010, Pécs, Hungary, 21 Jan - 23 Jan, 2010. Presentation Type: Poster Presentation Topic: Sensory and motor systems Citation: Serfözö Z and Elekes K (2010). Early observations of nitric oxide synthase in the intact brain and regenerating tentacle of the snail, Helix pomatia. Front. Neurosci. Conference Abstract: IBRO International Workshop 2010. doi: 10.3389/conf.fnins.2010.10.00267 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 06 May 2010; Published Online: 06 May 2010. * Correspondence: Zoltán Serfözö, Balaton Limnological Research Institute, HAS, Department of Experimental Zoology, Tihany, Hungary, serfozo@tres.blki.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Zoltán Serfözö Károly Elekes Google Zoltán Serfözö Károly Elekes Google Scholar Zoltán Serfözö Károly Elekes PubMed Zoltán Serfözö Károly Elekes Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.