Biomedical subjects

H H Schmidt

Publications and source records attributed to H H Schmidt.

At least 109 records · Page 6Linked to original sources

PubMed1995

Nitric oxide nerves in the uterus are parasympathetic, sensory, and contain neuropeptides.

Nitric oxide (NO) is synthesized in neurons and is a potent relaxor of vascular and nonvascular smooth muscle. The uterus contains abundant NO-synthesizing nerves which could be autonomic and/or sensory. This study was undertaken to determine: 1) the source(s) of NO-synthesizing nerves in the rat uterus and 2) what other neuropeptides or transmitter markers might coexist with NO in these nerves. Retrograde axonal tracing, utilizing Fluorogold injected into the uterine cervix, was employed for identifying sources of uterine-projecting neurons. NO-synthesizing nerves were visualized by staining for nicotinamide adenine dinucleotide phosphate (reduced)-diaphorase (NADPH-d) and immunostaining with an antibody against neuronal/type I NO synthase (NOS). NADPH-d-positive perikarya and terminal fibers were NOS-immunoreactive (-I). Some NOS-I/NADPH-d-positive nerves in the uterus are parasympathetic and originate from neurons in the pelvic paracervical ganglia (PG) and some are sensory and originate from neurons in thoracic, lumbar, and sacral dorsal root ganglia. No evidence for NOS-I/NADPH-d-positive sympathetic nerves in the uterus was obtained. Furthermore, double immunostaining revealed that in parasympathetic neurons, NOS-I/NADPH-d-reactivity coexists with vasoactive intestinal polypeptide, neuropeptide Y, and acetylcholinesterase and in sensory nerves, NOS-I/NADPH-d-reactivity coexists with calcitonin gene-related peptide and substance P. In addition, tyrosine hydroxylase(TH)-I neurons of the PG do not contain NOS-I/NADPH-d-reactivity, but some TH-I neurons are apposed by NOS-I varicosities. These results suggest NO-synthesizing nerves in the uterus are autonomic and sensory, and could play significant roles, possibly in conjunction with other putative transmitter agents, in the control of uterine myometrium and vasculature.

Amino Acid Oxidoreductases↗

PubMed1995

Colocalization of neuronal nitric oxide synthase with GABA in rat cuneate nucleus.

Pre- and post-embedding immunocytochemistry were employed in this electron microscopic investigation of cuneate neurons that are enriched in GABA and in nitric oxide synthase, the enzyme responsible for the synthesis of nitric oxide. GABAergic neurons are local circuit interneurons; 10-20% of them also contain nitric oxide synthase. These are among the smallest GABA-positive perikarya. We describe a network of processes in the rat cuneate nucleus that are immunopositive for nitric oxide synthase. Axon terminals positive for nitric oxide synthase are small and make synapses mainly onto dendrites; they make only occasional axo-axonic contacts. Double-labelling immunocytochemistry verified that the large majority of terminals positive for nitric oxide synthase also contained GABA. However, most GABA-positive profiles were negative for nitric oxide synthase and GABA-positive terminals that are negative for nitric oxide synthase frequently made axo-axonic contacts. These results suggest that nitric oxide synthase is within a specialized subpopulation of interneurons in the cuneate nucleus.

Amino Acid Oxidoreductases↗

PubMed1995

Hodgkin's disease developing after spontaneous remission of chronic lymphocytic leukemia.

We present a 71-year-old patient with chronic lymphocytic leukemia diagnosed 27 years ago. Initially, the disease was staged as Rai II and the patient suffered from secondary immunoglobulin deficiency. Nevertheless, no treatment was necessary at that time. Because of disease progression a single course of chemotherapy was given in 1984. During the following year there was a constant decline of the WBC, accompanied by normalization of the immunoglobulins; both have remained stable ever since that time. However, there was still residual bone marrow infiltration, indicating persisting CLL. In 1993 cervical lymphadenopathy occurred with acute onset. A diagnostic lymphadenectomy revealed Hodgkin's disease of the nodular-sclerosing subtype. The patient was staged as II-III according to the Ann Arbor Classification and underwent radiation therapy. Cytogenetic examination of the bone marrow revealed a normal karyotype with an inversion of chromosome 9. This case demonstrates the rate coincidence of two lymphoproliferative disorders in the same patient. The clinical course and the immunologic findings of this patient are presented, together with a review of the literature.

Adult↗

PubMed1995

Evidence for bidirectional changes in nitric oxide synthase activity in the rat striatum after excitotoxically (quinolinic acid) induced degeneration.

Nitric oxide, a gaseous inter- and intracellular messenger, is thought to mediate neurotoxicity via excitatory amino acid receptors which may contribute to the pathogenesis of a variety of neuronal diseases. Excitotoxin lesions induced by quinolinic acid were made unilaterally in the rat striatum to study biochemically, light- and electron microscopically the possible involvement of the nitric oxide synthesizing enzyme nitric oxide synthase in degeneration processes. 5 days after quinolinic acid injection nitric oxide synthase activity in the striatum was elevated to 196.5% (P < 0.005% as compared to controls). There was no requirement of Ca2+ for the enzyme activity measured indicating that the elevation is due to the inducible isoform of nitric oxide synthase. Parallel to the depletion of neurons by quinolinic acid a massive gliosis was seen. Whereas quiescent astroglial cells in the normal striatum did not show any light microscopically detectable nicotinamide adenine dinucleotide phosphate diaphorase reaction, reactive astroglia revealed a substantial labeling distributed over the cell body and their stellar processes. Within the lesion and, particularly, close to the needle tract the number of microglia/macrophages labeled by isolectin B4 increased dramatically. Reactive microglial cells macrophages, situated along the needle tract and characterized by a pseudopodic or a globular shape, contained highest staining activity. At the ultrastructural level only disintegrated, if any, neuronal perikarya were seen five days after quinolinic acid injection while numerous reactive glial cells were observed. Reactive astroglia showed nicotinamide adenine dinucleotide phosphate diaphorase activity by displaying a substantial labeling of the nuclear envelope and endoplasmic membranes. Occasionally stained mitochondria were encountered. Globular-shaped (ameboidal) microglia near the needle tract were rich in phagocytotic debris and, apart from formazan-positive endomembranes, their plasmalemma was often nicotinamide adenine dinucleotide phosphate diaphorase stained. Additionally, in those cells regions of highly electron-dense puncta were seen which differ sharply from other cytoplasmic areas. Such sand-like accumulations of nicotinamide adenine dinucleotide phosphate diaphorase positive grains have never been observed in other cell types, indicating a special type of nitric oxide synthase representation, possibly that of the inducible isoform.

Amino Acid Oxidoreductases↗

PubMed1995

Aldehyde fixation differentially affects distribution of diaphorase activity but not of nitric oxide synthase immunoreactivity in rat brain.

The effect of aldehyde fixation on NADPH- and NADH-dependent diaphorase (d) histochemistry and nitric oxide synthase (NOS) immunocytochemistry in the brain was investigated by comparing the distribution of these enzymes in in situ nitrocellulose blots of unfixed brain sections with that in aldehyde-fixed brain sections. Substitution of NADPH by NADH yielded no gross differences in cellular distribution in the native blot, whereas in fixed sections NADH produced nonspecific staining of the entire section. In the in situ blot NADPHd histochemistry therefore visualized general nitroblue tetrazolium reductase (NBTr) activity, which was particularly strong in hippocampal pyramidal neurons and cerebellar Purkinje cells. Aldehyde fixation abolished the anatomical pattern of general NBTr activity and changed the histochemical distribution in that of the NADPHd activity associated with the distribution of NOS-I immunoreactivity (ir). Fixation intensified NADPHd histochem- ical staining in specific neurons, resulting in outstanding, Golgi-like staining of these neurons in several brain regions, whereas the general NBTr activity in pyramidal and Purkinje cells disappeared. In contrast to the histochemical diaphorase distribution, the distribution of NOS-I ir on blots and in aldehyde-fixed brain sections was similar. No NOS was observed in hippocampal pyramidal and cerebellar Purkinje neurons. In regions like cerebral and cerebellar cortex and striatum the applied anti NOS-I serum had a higher affinity for the native protein. It is concluded that aldehydes, rather than to progressively suppress NOS-unrelated enzymes, differentially elicit NADPHd activity in some groups of neurons while leaving NOS-ir unaffected.

Animals↗

PubMed1995

Induction and variants of neuronal nitric oxide synthase type I during synaptogenesis.

In the adult central nervous system, nitric oxide (NO) is formed from L-arginine by the so-called constitutive or type I NO synthase (NOS-I155). However, expression of NOS-I155 immunoreactivity and activity was low or not detectable in developing mouse and rat brain. NOS-I155 was sharply induced coincident with the onset of synaptogenesis in specific brain regions. This was followed by a second phase in which total NOS-I155 expression decreased both in specific cell populations and in the total synaptosomal subcellular fraction.Furthermore, two putative variants of NOS-I were transiently observed: an NOS-I-immunoreactive protein with increased electrophoretic mobility (NOS-I144) and a transient hypersensitivity of NOS-I155 to the competitive substrate inhibitor N omega-nitro-L-arginine. It is concluded that NOS-I expression is not constitutive but locally induced. In the central nervous system, this regionally specific, biphasic pattern of postnatal NOS-I induction is consistent with a role for NO in synaptogenesis and synaptic plasticity.

Amino Acid Oxidoreductases↗

PubMed1995

Synaptic interactions between primary afferent terminals and GABA and nitric oxide-synthesizing neurons in superficial laminae of the rat spinal cord.

The superficial laminae (I and II) of the spinal dorsal horn receive small caliber primary afferent fibers responsive to noxious stimulation, and contain local circuit neurons that modulate afferent input. Many of these neurons are GABAergic; about a third of these also synthesize nitric oxide. We identified three main morphological types of primary afferent terminals in superficial laminae after injections of a tracer selective for small caliber afferents into the sciatic nerve of rats. The relative densities of the three types varied through the dorsoventral extent of laminae I and II. Synaptic contacts of each type with GABA- and nitric oxide synthase (NOS)-containing dendrites and axon terminals were determined by preembedding and postembedding immunocytochemistry. Nonglomerular primary afferent terminals, likely to originate from peptidergic unmyelinated fibers, were not seen in synaptic contact with either GABA- or NOS-containing neurons. Primary afferent terminals at the center of type 1 glomeruli (C1) and at the center of type 2 glomeruli (C2) are likely to originate from unmyelinated and small myelinated fibers, respectively. GABAergic terminals contacted more C2 than C1 terminals, suggesting more effective presynaptic inhibition of C2 terminals. Many GABAergic terminals were also positive for NOS, but all GABAergic terminals presynaptic to primary afferent terminals were negative for NOS. Only C2 terminals established frequent synapses with NOS-positive dendrites. (ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Oxidoreductases↗

PubMed1994

Postnatal development of nitric oxide synthase type 1 expression in the lumbar spinal cord of the rat: a comparison with the induction of c-fos in response to peripheral application of mustard oil.

The distribution of the neuronal isoform of the enzyme nitric oxide synthase (type 1) has been investigated in the lumbar spinal cords of neonatal rats (2-20 days old). Large multipolar neurones were present from day 2 around the central canal, in a band across the neck of the dorsal horn and at the thoracic level in the intermediolateral cell column, whereas staining was absent from laminae II. By 20 days the laminae II staining was similar to that found in the adult. NOS expression in lamina II paralleled the development of c-fos expression in this lamina in response to peripheral application of mustard oil.

Administration, Topical↗

PubMed1994

[Fish eye disease].

A 67-year-old man complained of impaired vision at night for several months. He was known to have arcus senilis (arcus lipoides corneae) since aged 21 years. For the last 20 years both corneas had become progressively more cloudy. His mother was said to have had similar eye changes. Ophthalmological examination discovered no abnormality other than marked corneal dystrophy with hardly separable arcus senilis. General physical examination was normal. Total cholesterol and triglyceride levels in serum were within normal limits, but serum concentration of high density lipoprotein (HDL) cholesterol (8 mg/dl) was reduced as were, within the high density lipoprotein fraction, the concentrations of triglyceride (4 mg/dl), phospholipids (38 mg/dl), the proportion of cholesterol esters (31%), and the cholesterol-esterification rate (51 nmol/ml . h). The HDL-associated activity of lecithin-cholesterol-acetyltransferase activity was scarcely measurable (0.9 nmol/ml . h). The signs in this case (cloudy cornea, marked decrease in serum HDL cholesterol concentration without premature arteriosclerosis) are typical of fish eye disease.

Aged↗

PubMed1994

NO at work.

Explore the source record for details and available documents.

Animals↗

PubMed1994

Upregulation of neuronal nitric oxide synthase and mRNA, and selective sparing of nitric oxide synthase-containing neurons after focal cerebral ischemia in rat.

Nitric oxide synthase-containing neurons are presumed to be resistant to neurodegeneration and neurotoxicity, however this resistance has not been demonstrated after focal cerebral ischemia. We therefore measured the temporal profile of neuronal nitric oxide synthase (NOS-I) mRNA and immunoreactivity and NADPH-diaphorase reactivity over a one week period after permanent middle cerebral artery (MCA) occlusion in 48 male Wistar rats and compared these data to ischemic cell damage as evaluated on hematoxylin and eosin (H & E) stained sections by light microscopy. NOS-I mRNA increased as early as 15 min after MCA occlusion in the ipsilateral striatum and maximal expression of NOS-I was found in the ipsilateral cortex and striatum 1 h after MCA occlusion. The numbers of NOS-I-containing neurons in the ipsilateral cortex and striatum were significantly greater (P < 0.05) than NOS-I-containing neurons in the contralateral hemisphere at 2-48 h after the onset of ischemia. The number of NOS-I-containing neurons peaked at 4 h after MCA occlusion. Neurons exhibited shrinkage or were swollen at 1 to 4 h after MCA occlusion. At 24-48 h after ischemia, neurons in the ischemic lesion appeared to be eosinophilic or ghost like on H & E stained sections. However, some of these neurons retained morphological integrity on the NOS-I immunohistochemical sections. At 168 h after ischemia, all neurons within the lesion appeared necrotic on H & E stained sections; however, scatterred neurons expressed NOS-I and NADPH-diaphorase. The rapid upregulation of NOS-I and mRNA in the ischemic lesion suggests that NOS-I is involved in focal cerebral ischemic injury; the expression of NOS-I by neurons that retain their morphological structure in the area of the infarct suggests that NOS-I-containing neurons are more resistant to the ischemic insult. Our data also indicate a close association of NOS-I immunoreactivity and NADPH-diaphorase reactivity in ischemic brain.

Amino Acid Oxidoreductases↗

PubMed1994

Functional coupling of a Ca2+/calmodulin-dependent nitric oxide synthase and a soluble guanylyl cyclase in vertebrate photoreceptor cells.

Electrophysiological recordings on retinal rod cells, horizontal cells and on-bipolar cells indicate that exogenous nitric oxide (NO) has neuromodulatory effects in the vertebrate retina. We report here endogenous NO formation in mammalian photoreceptor cells. Photoreceptor NO synthase resembled the neuronal NOS type I from mammalian brain. NOS activity utilized the substrate L-arginine (Km = 4 microM) and the cofactors NADPH, FAD, FMN and tetrahydrobiopterin. The activity showed a complete dependence on the free calcium concentration ([Ca2+]) and was mediated by calmodulin. NO synthase activity was sufficient to activate an endogenous soluble guanylyl cyclase that copurified in photoreceptor preparations. This functional coupling was strictly controlled by the free [Ca2+] (EC50 = 0.84 microM). Activation of the soluble guanylyl cyclase by endogenous NO was up to 100% of the maximal activation of this enzyme observed with the exogenous NO donor compound sodium nitroprusside. This NO/cGMP pathway was predominantly localized in inner and not in outer segments of photoreceptors. Immunocytochemically, we localized NO synthase type I mainly in the ellipsoid region of the inner segments and a soluble guanylyl cyclase in cell bodies of cone photoreceptor cells. We conclude that in photoreceptors endogenous NO is functionally coupled to a soluble guanylyl cyclase and suggest that it has a neuromodulatory role in visual transduction and in synaptic transmission in the outer retina.

Amino Acid Oxidoreductases↗

PubMed1994

Co-localization of nitric oxide synthase and NGF receptor in neurons in the medial septal and diagonal band nuclei of the rat.

The relationship of neurons that express nitric oxide synthase type I (NOS-I), the synthetic enzyme of the free radical nitric oxide (NO), with cells that contain the low affinity p75 nerve growth factor receptor (NGFr) was examined in the basal forebrain region of the rat using double immunohistofluorescence. NOS and NGFr were found to be co-localized in a neuronal population that displayed a selective distribution. Double immunopositive neurons were evident in the medial septum and in rostral levels of the diagonal band nuclei where the vast majority (more than 80%) of NOS-immunoreactive cells were also NGFr-positive; the two cell populations were separate at more caudal levels of the basal forebrain. These findings support the chemical heterogeneity of septal and basal forebrain neurons which contain NGFr and indicate that in the rat a subset of neurons of the septum-diagonal band complex may release NO.

Amino Acid Oxidoreductases↗

PubMed1994

The extravascular contractile system in the human placenta. Morphological and immunocytochemical investigations.

In the human placenta, besides the fetal blood vessel system a second extravascular contractile system exists. It is localized in the chorionic plate and runs in a longitudinal direction and adjacent to fetal blood vessels into the stem villi, where it forms perivascular contractile sheaths. Characteristically, cells of the extravascular contractile system are extremely long and spindle-shaped and give rise to fine cell processes, by which they obviously contact each other or insert into the basement membrane of the trophoblast. They show immunoreactivity with desmin, vimentin, alpha-actin, myosin, nitric oxide synthase type I (brain form) and dipeptidyl peptidase IV. The ultrastructure suggests that cells of the extravascular contractile system are related to smooth muscle cells, including subpopulations with myofibroblastic features. In stem villi a few cells are nitric oxide synthase type I immunoreactive. These cells are thought to be specialized smooth-muscle-like cells of the extravascular contractile system or cells of the extravascular contractile system related to paraneurons that generate nitric oxide, which, in turn, may modulate the tone of perivascular contractile sheaths. The high dipeptidyl peptidase IV activity suggests that modulation of the extravascular contractile system may also occur by substance P.

Actins↗

PubMed1994

Expression of nitric oxide synthase immunoreactivity by interstitial cells of the canine proximal colon.

A subpopulation of interstitial cells (ICs) are interposed between nerve terminals and smooth muscle cells in the gastrointestinal tract and may participate in neuromuscular transmission. These cells appear to be targets for NO released from enteric inhibitory nerves and respond to exogenous NO with: (i) an elevation in cGMP levels; (ii) an increase in intracellular Ca2+; (iii) and release of a diffusible substance that has tentatively been identified as NO. For the latter to be possible, ICs must express a constitutive isoform of NOS. This study characterized the expression of NOS-like immunoreactivity (NOS-LI) in ICs of the canine colon using 3 antibodies raised against the 2 known constitutive forms of NOS (i.e., neural (nNOS) and endothelial (eNOS) isoforms). Antibodies raised against cNOS and an antibody raised against rat cerebellar nNOS labeled ICs along the submucosal surface of the circular muscle layer (IC-SM), along the surface of septa that separate the circular muscle into fiber bundles (IC-SM), and in the myenteric region between the circular and longitudinal muscle layers (IC-MY). Another antibody raised against rat cerebellar nNOS failed to label ICs. Cultured IC-SM also expressed NOS-LI, suggesting that this feature of the IC phenotype survives culture conditions. Arteriolar endothelial cells in the canine colon were labeled with the same 2 antibodies that labeled ICs, suggesting there are significant structural similarities between NO synthases in ICs and endothelial cells. The data suggest that IC-SM and IC-MY express a constitutive form of NOS. Synthesis of NO by ICs may influence electrical rhythmicity and may serve to amplify and even propagate enteric inhibitory neurotransmission.

Amino Acid Oxidoreductases↗

PubMed1994

Subpopulations of GABAergic neurons in laminae I-III of rat spinal dorsal horn defined by coexistence with classical transmitters, peptides, nitric oxide synthase or parvalbumin.

GABAergic neurons in laminae I-III of the spinal dorsal horn may contain one or more of the following compounds: glycine, acetylcholine, neuropeptide Y, enkephalin, nitric oxide synthase or parvalbumin. Although the pattern of co-localization of some of these compounds is understood, it is not known which types of GABAergic neurons contain parvalbumin, or whether nitric oxide synthase coexists with peptides, acetylcholine or parvalbumin in any of these neurons, and in this study we have used immunocytochemistry and enzyme histochemistry to resolve these issues. Parvalbumin-immunoreactivity was restricted to those GABA-immunoreactive neurons that also showed glycine-immunoreactivity and was not co-localized with neuropeptide Y-immunoreactivity or NADPH diaphorase activity. By combining NADPH diaphorase histochemistry with immunocytochemistry with an antiserum to nitric oxide synthase, we were able to show that NADPH diaphorase activity was a reliable marker for nitric oxide synthase in the spinal cord. Neurons that possess GABA- but not glycine-immunoreactivity may contain neuropeptide Y, enkephalin, acetylcholine or NADPH diaphorase, and all of the cholinergic neurons appear to contain NADPH diaphorase. By combining immunofluorescent detection of neuropeptide Y or enkephalin with NADPH diaphorase histochemistry, we showed that peptide-immunoreactivity did not coexist with NADPH diaphorase. This suggests that neither of these peptides coexists with nitric oxide synthase or with acetylcholine in neurons in the superficial dorsal horn. Several phenotypically distinct groups of GABA-immunoreactive neuron can therefore be identified in laminae I-III of the dorsal horn, and these may represent different functional types of inhibitory neuron.

Acetylcholine↗

PubMed1994

Nitric oxide synthase expression reveals compartments of cerebellar granule cells and suggests a role for mossy fibers in their development.

The developmental expression and cellular distribution of nitric oxide synthase was investigated in the murine cerebellum and in cerebellar neurons developing under controlled in vitro conditions. Cerebellar granule cells expressed nitric oxide synthase only after migration to the internal granule cell layer. Initially, the nascent internal granule cell layer throughout the cerebellum stained uniformly for nitric oxide synthase, but during the second postnatal week, a pattern emerged consisting of clusters of heavily stained granule cells separated by areas of unstained granule cells. This pattern persisted into adulthood. There was a close temporal correlation between innervation of the granule cell layer by mossy fibers and the emergence of granule cell compartments as defined by levels of nitric oxide synthase expression. Granule cells in dissociated cultures derived from cerebellar anlagen prior to mossy fiber innervation also express nitric oxide synthase. The time-course of nitric oxide expression was independent of electrical activity of the neuronal network forming in vitro. However, suppression of spontaneous electrical activity resulted in enhanced nitric oxide synthase expression. These findings indicate that granule cell precursors are endowed with an intrinsic program which regulates nitric oxide synthase induction and which is executed independently of correct positional cues. The data also suggest that electrical activity of ingrowing mossy fibers down regulates nitric oxide synthase expression and plays an important role in the generation of granule cell compartments. These compartments may contribute to the functional organization of the cerebellar cortex.

Afferent Pathways↗

PubMed1994

Type I nitric oxide synthase fully accounts for NADPH-diaphorase in rat striatum, but not cortex.

The novel gaseous neuromediator nitric oxide is thought to play an important role in development and plasticity. Despite this, gene-knockout mice lacking neuronal (Type I) nitric oxide synthase exhibit relatively normal brain development and behavior. The nervous system of these mice (especially the forebrain) retains some calcium-dependent nitric oxide synthesis, presumably reflecting other isozymes. Type I nitric oxide synthase has NADPH-dependent diaphorase activity. However, this stain also recognizes other isozymes, and it remains controversial whether all diaphorase-positive neurons contain Type I nitric oxide synthase. To assess whether neurons containing another isoform of nitric oxide synthase may be present in the forebrain of normal rodents, we studied co-localization of diaphorase staining with immunocytochemistry for Type I nitric oxide synthase. Co-localization was complete in the striatum, but some neurons deep in cortex were diaphorase-positive and immunonegative, and therefore may contain a splice variant or novel isozyme of nitric oxide synthase.

Amino Acid Oxidoreductases↗