Horseradish Peroxidase (Total 56051 Papers Found)

The present study examines the spatial distribution within rat locus coeruleus of neurons projecting to particular brain regions. In order to accurately recreate, in digital and pictorial formats, the spatial distribution of neurons for the entire nucleus locus coeruleus, three-dimensional reconstructions were created which specified the location of each individual Nissl-stained locus coeruleus cell in each of five nuclei. Dynamic computerized displays were visually analyzed and statistically co ...
Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria (blue-green algae), and microcystin-leucine-arginine (MC-LR) is among the most frequent and most toxic microcystin congeners. In this study, a free amino group was introduced to MC-LR at its seventh amino acid residue with 2-mercaptoethylamine, and the product aminoethyl-MC-LR was coupled to bovine serum albumin (BSA) and horseradish peroxidise (HRP) by glutaraldehyde to be complete anti ...
Whether the increased capillary permeability characteristic of diabetes extends to the blood-brain barrier is presently unclear. We have examined in streptozotocin-diabetic rats the permeability of the blood-brain barrier at the level of 12 discrete brain regions employing 3 intravenous tracers of different molecular weight: sucrose, inulin and horseradish peroxidase. In animals killed 5 min after tracer injection both the sucrose and the inulin spaces were similar to controls. In order to asses ...
The location, number and size of the motoneurons innervating the ischiocavernosus muscle, identified by means of horseradish-peroxidase (HRP) retrograde transport, were studied (1) in adult untreated male rats, (2) in adult male rats castrated before puberty, and (3) in adult male rats castrated before puberty and injected with testosterone from the day of castration. After injection of HRP into the ischiocavernosus muscle, labeled motoneurons were found in the dorsolateral and dorsomedial colum ...
There is increasing evidence that glial cells respond to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), and astrocytes have been shown to possess GABAA receptors both in vivo and in vitro. A recent study by Sakatani et al. (Proc. R. Soc. Lond. B247, 155 (1992)) demonstrated the transient expression of functional GABAA receptors in the developing rat optic nerve, but axonal and glial components of the response were not distinguished. To help address this problem, we have determin ...
Intracellular recordings were made from ventral anterior and ventral lateral (VA-VL) thalamic neurons in the cat. VA-VL neurons were tested for responsiveness to activation of cortical, pallidal and cerebellar afferents, and were identified morphologically by intracellular injection of HRP. Orthodromic activation of cortical and pallidal afferents produced primarily an initial inhibition (due in part to oligosynaptic circuitry) while activation of cerebellar afferents produced an initial excitat ...
The afferents to the periaqueductal gray in the rabbit have been described following hydraulic pressure injection of horseradish peroxidase at various sites throughout this structure. Every third section was reacted with tetramethylbenzidine, for the localization of afferent neurons. At the site of the deposit alternate sections were reacted with tetramethylbenzidine, Hanker-Yates reagent, or diaminobenzidine, for comparative assessment of the injection site. A large number of retrogradely label ...
An electron microscopic study on the synaptic connections between neurons of ventrolateral nucleus of thalamus (VL) and pyramidal tract neurons (PTNs) in cat motor cortex was conducted by means of the anterograde degenerating procedure coupled with horseradish peroxidase (HRP) intracellular staining. Following VL lesions, a large majority of the degenerating terminals were found to terminate on dendritic spines and a few on the dendritic shaft. An asymmetric type synapse formed by a VL degenerat ...
Membrane protein isolation is a challenging problem. In fact especially their extraction from the respective membrane is difficult and often goes along with losses in yield. Usually expensive detergents are needed to extract the target protein from the membrane. Therefore finding an efficient overexpression and extraction method and an alternative to detergents is desirable. In this study we describe a new and fast method to express, extract and purify an engineered variant of the FhuA protein ( ...
In order to determine whether neurones in the parvicellular reticular formation are in direct synaptic contact with motor neurones innervating masticatory muscles, a combined retrograde and anterograde transport study was carried out in the rat at both light and electron microscopic levels. The animals received injections of the retrograde tracers wheat germ agglutinin conjugated to horseradish peroxidase or cholera toxin B conjugated to horseradish peroxidase into the masticatory muscles and of ...
  • Helmut Flemm,
  • Zeitschrift für Zellforschung und Mikroskopische Anatomie
  • 2004
1. The findings of other investigators with respect to the perineurium being an efficient diffusion barrier are confirmed. This also holds true for a protein of small molecular weight and size. 2. By means of endoneurial or epineurial application the substance never reaches the third perineurial cell layer. Pinocytotic vesicles account for the transport of peroxidase through the cytoplasm of the cells. 3. The perineural cell membranes and the interconnecting tight junctions form a barrier for ho ...
The axial ligand effect on reactivity of heme enzymes is explored by means of density functional theoretical calculations of the oxidation reactions of propene by a model compound I species of horseradish peroxidase (HRP). The results are assessed vis-à-vis those of cytochrome P450 compound I. It is shown that the two enzymatic species perform C=C epoxidation and C–H hydroxylation in a multistate reactivity scenario with FeIII and FeIV electromeric situations and two different spin states, do ...
Monoclonal antibodies to human IgM were produced by fusing the Sp 2/0-Ag 14 line of mouse myeloma cells with spleen cells from BALB/c mice immunized with purified human IgM. From 6 clones which secreted antibody to human IgM, the one which produced the highest levels of antibody and grew relatively rapidly was selected for expansion and production of immune reagents for viral IgM antibody assays. Mouse ascitic fluids produced with this clone of cells had antibody titers for human IgM of 1 X 10(- ...
A new chemiluminescence technique has been assessed for the detection of reactive oxygen species generated by purified populations of human sperm. This revised protocol involves the use of horse-radish peroxidase (HRP) in combination with a luminol analogue, 7-dimethyl amino-naphthalin-1,2-dicarbonic acid hydrazide (DNDH), that exhibits two-three times the quantal efficiency of luminol itself. The chemiluminescent signal generated with these reagents was significantly (P less than 0.001) greater ...
A unique approach of developing a bar code version of lateral-flow enzymatic-based assay for the semi-quantification of hydrogen peroxide is described. The proposed assay system is mainly composed of a goat anti-mouse IgG-horseradish peroxidase conjugate (Gt anti-M IgG-HRP)-coated nitrocellulose (NC) membrane and a peroxidase substrate pad. Unlike the bar code immunochromatographic assay which depends on the stepwise capture of analyte, the principle of enzyme-based bar code lateral-flow assay i ...
Topical drug delivery is preferred in the eye to avoid under or over medication and undesired side effects of systemic administration. In order to maintain adequate drug levels in ocular tissues, frequent drug installation is necessary in vision threatening conditions like glaucoma, corneal ulcers due to microbial infections, etc. Only a part of the installed drug reaches the aqueous humor and the rest of it is drained by the nasolacrimal duct. Positively charged liposomes have been found to enh ...
A procedure for fabricating an enzyme electrode has been described based on the effective immobilization of horseradish peroxidase (HRP) to a nano-scaled particulate gold (nano-Au) monolayer modified chitosan-entrapped carbon paste electrode (CCPE). The high affinity of chitosan entrapped in CCPE for nano-Au associated with its amino groups has been utilized to realize the use of nano-Au as an intermediator to retain high bioactivity of the enzyme. Hydrogen peroxide (H(2)O(2)) was determined in ...
Oxidation of cytochrome c peroxidase with hydrogen peroxide to form the initial oxidized intermediate, cytochrome c peroxidase compound I, drastically alters the proton hyperfine nmr spectrum. In contrast to studies of horseradish peroxidase, where the spectrum of horseradish peroxidase compound I is similar to that of the native protein, cytochrome c peroxidase compound I exhibits only broad resonances near 17 and 30 ppm from 2,2-dimethyl-2-silapentane-5-sulfonate. No unique resonances attribut ...
Immunology laboratories widely rely on the enzyme linked immunosorbent assay (ELISA) for protein detection and identifi cation. [ 1 , 2 ] A typical 96-well plate sandwich ELISA product provides inside each well an adsorbed primary antibody that binds its specifi c antigen in the fi rst step of the assay. Subsequently, a biotinylated antibody is added and binds to the antigen via a second recognition site. Streptavidin-linked horseradish peroxidase is then introduced, binds to the biotin moiety a ...
The apical cytoplasm of several absorbing epithelia contains well-developed apical tubules (AT) which contribute to membrane recycling from endocytic vacuoles to the apical cell membrane. In this study, we examined three-dimensional structures of the AT in rat kidney proximal tubule cells by transmission and scanning electron microscopy. In thin sections, the AT appeared as straight tubules with a rather constant diameter (70–90 nm), but others were curved and, occasionally, branching. No AT ...
The vagal innervation of the proximal gastrointestinal (GI) tract is lateralized. To determine whether this pattern is specified as early as the perinatal period, neonatal rat pups were given unilateral cervical vagotomies. Separate groups received (1) transections below the left nodose ganglion, (2) left cervical resections that included removal of the nodose ganglion, or (3) sham surgeries. At 4 months of age, each animal's vagal afferent projections from the unoperated side were mapped by inj ...
Enzyme immunoassay for determination of total immunoglobulin E in children and adolescents based on combination of two monoclonal antibodies with high affinity and specificity to different epitopes of IgE molecule has been developed. Monoclonal antibodies IgE/11-4 are used for sensitization of a solid phase (96-well plate), mcAB IgE/11-1 conjugated with horseradish peroxidase are used as a developing reagent. The following analytical characteristics of a test-system are established: range of ass ...
For immunohistological analysis, simultaneous detection of multiple cellular epitopes, as compared to single staining of serial sections, is sometimes needed. Therefore, immunoenzyme triple-staining protocols were tested with polyclonal and monoclonal antibodies on tissue sections and cytospin preparations. Various immunoconjugates were used in different combinations of methods, of which not all proved to be suitable. Of the tested protocols, one yielded superior results for both monoclonal and ...
We investigated whether vacuolated cervical anterior horn cells of the wobbler mouse maintain axons to the periphery, and if these morphologically abnormal neurons are capable of supporting axonal regeneration. Using retrograde axonal transport, we applied horseradish peroxidase (HRP) to peripheral nerves or muscles and with electron microscopy sought evidence for perikaryal labeling in vacuolated neurons in 23 wobbler mice. When HRP was injected into forelimb muscles, 12 of 36 vacuolated neuron ...
Amplification of immunological signals with catalyzed reporter deposition (CARD) allows improved detection of scarce tissue antigens in light and electron microscopy. The technique takes advantage of the oxidation ability of horseradish peroxidase (HRP), in the presence of hydrogen peroxide, to yield the accumulation of one of its specific reporter-tagged substrates. This immunocytochemical approach continues to be improved by the introduction of new reporter molecules tagged to tyramine or to o ...
The position of the motornuclei projecting through the median (Mn) and ulnar (Ul) nerves to the cat distal forelimb has been investigated. Horseradish peroxidase (HRP) and fluorescent (Fl) compounds have been used as retrograde tracers. They were either injected into forelimb muscles or applied to the proximal end of transected forelimb nerves. Limb muscles that were not investigated were carefully denervated. The position and the architecture of the individual motornuclei were traced with HRP. ...
In guinea-pigs given 400 micrograms per kg of tunicamycin, the toxin responsible for the disease known as annual ryegrass toxicity, alterations in blood-brain barrier permeability were assessed with vascular tracers of differing particle size. The toxin caused loss of integrity of the barrier to the smallest diameter tracer, horseradish peroxidase, but the cerebral endothelium was able to retain larger particles of ferritin and colloidal gold in the circulation. The effect of tunicamycin on cult ...
Inputs to the nucleus interpositus anterior (NIA) of the cat from precerebellar nuclei which are thought to give rise to mossy fibers were studied using electrophysiological and anatomical techniques. Stimulation of one of these precerebellar nuclei, the lateral reticular nucleus (LRN) evoked monosynaptic EPSPs in NIA neurons. These EPSPs were followed by polysynaptic IPSPs and late depolarization mediated by the response of the cerebellar cortex. Similar responses were occasionally seen followi ...
Measurement of temperature within the cerebral cortex, hypothalamus, cerebellum and medulla of rats sham-, heat- or microwave-exposed revealed the presence of a thermal gradient within the brain. In all groups, cerebral cortex and the cerebellum were cooler than the deeper hypothalamus and medulla. Exposure to 2450 MHz CW microwaves or ambient heat (42 +/- 2 degrees C) resulted in measurable elevation of regional brain temperature, but without alteration of temperature gradients normally observe ...
Mouse peritoneal macrophages were cultured in vitro, exposed to exogenous tracers, and examined by transmission electron microscopy. Native, anionic ferritin and horseradish peroxidase did not bind to the cell surface and were exclusively found in endocytic vesicles and lysosomes after uptake into the cells. Cationized ferritin (CF) bound to the plasma membrane and was then rapidly internalized. Intracellularly, CF first reached the lysosomes, but was later also transferred to and passed through ...
Projection neurons in the rat hypothalamic paraventricular nucleus (pv) were identified using the retrograde transport of horseradish peroxidase (HRP) following injections of this tracer into the neurohypophysis or jugular vein. The objective was to determine if parvocellular pv neurons might be labelled by either injection in addition to the well known magnocellular contingent. Injections confined to the neurohypophysis resulted in the labelling of only magnocellular pv neurons, while intrajugu ...
Two new substrate chromogens for alkaline phosphatase (ALP) detection have been recently synthesized for use in molecular biology research, salmon and magenta phosphate. We show here that these two chromogens have advantageous characteristics for immunocytochemistry as well. Their relatively delicate pink- and magenta-colored products do not mask the colors produced by other staining procedures. In addition, the reaction products of these substrates are insoluble in water, ethanol, and xylene, p ...
A sensitive staining procedure for glucose oxidase (GOD) as marker in immunohistology is described. The cytochemical procedure involves a two-step enzyme method in which GOD and horseradish peroxidase (HRP) are coimmobilized onto the same cellular sites by immunological bridging or by the principle of avidin-biotin interaction. In this coupled enzyme technique, H2O2 generated during GOD reaction is the substrate for HRP and is utilized for the oxidation of chromogens such as 3,3′-diaminobenzid ...
We have investigated the developmental changes of intrahemispheric neuronal connections of the areas 17 & 18 ocular dominance columns in monocularly deprived cats. Single cortical columns were microiontophoretically injected with horseradish peroxidase and 3D reconstruction of retrogradely labelled cells' region was done. Ocular dominance of injected columns and their coordinates in the visual field map were determined. In area 17 it was shown that for non-deprived eye the connections of columns ...
Morphological and physiological studies were undertaken in the cat lumbosacral spinal cord in order to examine individual last-order excitatory interneurons in the disynaptic pathway between low threshold superficial peroneal (SP) afferents and flexor digitorum longus (FDL) motoneurons. Last-order interneurons projecting to the FDL motor nucleus were visualized with activity-dependent trans-synaptic transport of lectin-conjugated horseradish peroxidase (WGA-HRP). The location of lumbar interneur ...
The binding of a number of ligands to the heme protein indolamine 2,3-dioxygenase has been examined with UV-visible absorption and with natural and magnetic circular dichroism spectroscopy. Relatively large ligands (e.g., norharman) which do not readily form complexes with myoglobin and horseradish peroxidase (HRP) can bind to the dioxygenase. Except for only a few cases (e.g., 4-phenylimidazole) for the ferric dioxygenase, a direct competition for the enzyme rarely occurs between the substrate ...
This study investigated the effect of early life stress on blood-brain barrier functional development in an altricial mammal, the Wistar rat. Forced swimming between gestational days 10-20 was used as a stress procedure in pregnant rats. After delivery, half of the control litters underwent 180 min maternal separation from postnatal day 2-20. Controls were kept without any stress manipulations. At sacrifice between postnatal days 1-30 subjects were given intracardiac Evans blue or the lectin whe ...
In horseradish peroxidase (HRP), hydrogen bonding of Asn-70 to His-42 enhances the catalytic activity of the histidine, and mutation of His-42 to a neutral residue greatly decreases peroxidase activity. The N70D/H42A HRP mutant is compared here to the previously characterized H42A mutant to determine if the Asp-70 substitution can rescue the catalytic activity. The N70D/H42A and H42A mutants give Compound I species with a high ratio of H2O2 at the low rates of 37 and 81 M-1 s-1 at 4 degrees C, r ...
Correlation of the main peripheral branches of the facial nerve with morphological subdivisions of the facial nucleus was examined in the guinea pig by the retrograde horseradish peroxidase method. The facial nucleus of the guinea pig was divided cytoarchitectonically into the dorsolateral, lateral, intermediate, medio-intermediate, medial, and ventromedial divisions; the ventromedial division was further divided into the major, dorsal and lateral parts. Six main branches of the facial nerve wer ...
Embryonic neural tissues were taken from the ventral mesencephalon, the striatal anlage, the cerebellum, or the dorsal telencephalon. Prior to grafting these tissues were dissociated. This provided the opportunity to generate experimental cografts and allowed the introduction of neuronal tracers. Projections from grafts that contained dopaminergic neurons of the mesencephalon were identified immunocytochemically with antisera to tyrosine hydroxylase. Acetylcholinesterase histochemistry was emplo ...
The intracellular distribution of horseradish peroxidase (HRP) transported intra-axonally from the mesenteric nerve to the wall of the small intestine has been examined electron microscopically. After 2 days, some of the neurons of the myenteric and submucosal plexuses showed an accumulation of HRP. The HRP reaction product consisted of fine, discrete cytoplasmic granules and it was located in lysosome-like dense bodies with a 400-500 nm of diameter. The morphological features of the labelled ne ...
Sensory neocortex of the tree shrew was divided into three main areas: the visual field, the auditory field, and the somatic field which includes motor cortex. Cortical cells which project to the tectum were identified by injecting HRP into superficial or deep layers of the superior colliculus and into various parts of the inferior colliculus. The main result is that these descending projections are well organized according to their origin in the three main sensory fields of the cortex. (1) Audi ...
Multilayer assemblies were prepared by alternating adsorption of monolayers of monoclonal antibody against horse radish peroxidase (anti-HRP) and dextran sulfate (DS) on solid supports at acid pH. After crosslinking with glutaraldehyde, DS was washed out of the film with buffered physiological saline, while the antibody remained immobilised on the support. Assembly was monitored in situ on germanium supports by infrared multi-internal reflection spectroscopy. The binding capacity of the immobili ...
  • Merav Jacobson,
  • The Journal of neuroscience : the official journal of the Society for Neuroscience
  • 1981
Clonal origins of Rohon-Beard neurons in Xenopus were determined quantitatively by injecting horseradish peroxidase into individual blastomeres at the 16-cell stage and later counting labeled and unlabeled Rohon-Beard neurons. Two different patterns of cleavage were selected. In pattern X, all Rohon-Beard neurons originated from three blastomeres (V1.1, V1.2, and V2.2) on each side; in pattern Y, all Rohon Beard neurons originated from two blastomeres (V1.2 and V2.2) on each side. Counts of Roho ...
We examined abdominal vagal afferents (n = 33) and the distributions of their intraganglionic laminar endings (IGLEs) in the duodenum. Rats (male, Sprague-Dawley) received a partial subdiaphragmatic vagotomy that spared a single branch. Wheat germ agglutinin-horseradish peroxidase (0.5-1.0 μl) was injected into the nodose ganglion ipsilateral to the vagotomized side. We observed that the hepatic branch does not project to the stomach, that the accessory celiac and celiac branches course along t ...
It was shown by the method of retrograde axonal transport of horseradish peroxidase that the posterolateral thalamic nucleus (NPL) in rats receives considerable ascending projections from the superior colliculus (SC), the dorsal part of the lateral geniculate body (LGB), and the pretectal region (PT) and smaller projections from n. ventralis posterior (VP) and n. ventralis lateralis (VL) of the thalamus, the ventral part of LGB, the zona incerta, and anterior hypothalamus. The most marked descen ...