Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “RADIOAUTOGRAPHY”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 775 records · Page 43Linked to original sources
PubMed1995

In vivo-synthesized radioactively labelled alpha-methyl serotonin as a selective tracer for visualization of brain serotonin neurons.

To investigate the use of alpha-[3H]methyl tryptophan (alpha-[3H]MTrp) as a tracer for the in vivo study of brain serotonergic neurons, we examined whether alpha-[3H]MTrp and its metabolite alpha-[3H]methyl serotonin (alpha-[3H]M5-HT) selectively label serotonergic neurons and whether once accumulated in these neurons, the radioactive metabolite behaves like endogenous serotonin. Rats received a systemic injection of 1-5 mCi of alpha-[3H]MTrp and 24 h later their brains were immediately removed or fixed by perfusion before removal. Tissue sections in which serotonergic neurons had been immunostained for 5-HT or its synthesizing enzyme, tryptophan hydroxylase, were processed for radioautography at the light and electron microscopic level. In another group of rats, the release of radioactivity from different brain areas was studied both under basal and depolarizing conditions. In the dorsal raphe nucleus, the light microscopic examination revealed almost complete colocalization between serotonergic neurons and those that accumulated radioactivity, with a heterogeneity in the content of alpha-[3H]M5-HT among the various cells. At the ultrastructural level, immunoidentified serotonergic perikarya and dendritic processes in the dorsal raphe nucleus, as well as nerve terminals in the cerebral cortex were also found to contain alpha-[3H]M5-HT. Under basal conditions, radioactivity was released from the brainstem raphe region and from projection areas such as the striatum and hippocampus. The basal output of alpha-[3H]M5-HT increased approximately twofold after a depolarizing 50 mM KCl solution was added to the perfusion fluid. These findings suggest that newly synthesized alpha-[3H]M5-HT can be released both at somatodendritic and terminal sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Quantification of free sphingosine in cultured cells by acylation with radioactive acetic anhydride.

A simple and sensitive method for quantification of sphingosine in cellular lipid extracts was developed. The assay is based on quantitative conversion of sphingosine to N-[3H]acetylated sphingosine ([3H]C2-ceramide) by N-acylation with [3H]acetic anhydride under certain conditions. Sphingosine was extracted from cultured cells with chloroform and methanol and then treated with base to remove interfering glycerolipids having reactive amino groups (e.g., phosphatidylethanolamine or phosphatidylserine). Sphingosine was acylated with [3H]acetic anhydride in the presence of 0.004 N NaOH. Acylation was complete in 1 h at 37 degrees C when sphingosine was present in the picomole range. After the acylation, samples were treated with NaOH to reduce background radioactivity by removing the remaining [3H]acetic anhydride and hydrolyzing any ester linkages formed during the acylation and resolved by thin-layer chromatography. [3H]C2-ceramide converted from sphingosine with the acylation was detected with radioautography and quantitated by scraping the corresponding band and counting its radioactivity with a scintillation counter. [3H]C2-ceramide formed was quantitatively measured. This assay allows quantification of sphingosine over a range of 10 to 1500 pmol. The amount of sphingosine in lipid extracts from cultured cells was proportional to the number of cells. Sphingosine levels in human gastric cancer KATO III cells, human promyelocytic leukemic HL60 cells, and human monoblastic U937 cells, determined by this method, were 26.6 +/- 2.2, 6.3 +/- 0.4, and 6.8 +/- 0.6 pmol per 10(6) cells, respectively. Our new procedure allows quantification of sphingosine levels present in the low picomole range in lipid extracts from biological samples.

Acetic Anhydrides↗
PubMed1994

Cholinergic neuronal differentiation factor (CDF)/leukemia inhibitory factor (LIF) binds to specific regions of the developing nervous system in vivo.

The cholinergic neuronal differentiation factor (CDF)/leukemia inhibitory factor (LIF) promotes survival and/or differentiation of specific neuronal populations in vitro. To investigate the spatial and temporal distribution of CDF/LIF binding sites in the developing rat nervous system, we localized binding of iodinated native rat CDF/LIF by radioautography. We find that specific CDF/LIF binding sites are present in distinct regions of the nervous system at defined developmental stages. CDF/LIF binding is detectable in all sensory ganglia examined, both neural crest- and placode-derived, and in all sympathetic and parasympathetic ganglia examined. In the central nervous system, in contrast, binding is restricted to specific regions in the motor, sensory, and limbic systems. These in vivo results not only support culture studies which suggest CDF/LIF involvement in development of specific populations of neurons, but also suggest new roles played by CDF/LIF.

Animals↗
PubMed2002

Electron microscopic investigation of nitrobenzene distribution and effect on plant root tip cells ultrastructure.

Electron microscopic radioautography demonstrated the penetration of [1-6(14)C]nitrobenzene in maize and soybean root tip cells: radioactive label was detected in cell wall, plasmalemma, nuclei, and cytoplasm. Among cytoplasmic organelles, the highest label was found in mitochondria and plastids. [1-6(14)C]nitrobenzene and/or products of its transformation accumulated in vacuoles. Study of the action of different concentrations of nitrobenzene on cell ultrastructural organization revealed the following picture. Nitrobenzene concentration up to 0.015 mM was harmless for plant cells. Increase of nitrobenzene concentration from 0.015 to 1.5 mM induced several pathological changes, up to the complete destruction of cells. The most damaged organelles were nuclei, mitochondria, and plastids. In the presence of 0.15 mM nitrobenzene the intensification of contacts among cell organelles, especially between endoplasmic reticulum and mitochondria/plastids, was observed. The data indicate some coordination between detoxication activity and energy metabolism during cell reaction to xenobiotic toxicity.

Carbon Radioisotopes↗
PubMed1998

Ligand-induced conformational changes in cyclic nucleotide phosphodiesterases and cyclic nucleotide-dependent protein kinases.

Three methods have been used to assess the conformational effects associated with ligand binding to two unrelated cyclic nucleotide receptor proteins: the cGMP-binding, cGMP-specific phosphodiesterase (cGB-PDE or PDE5A) and the cGMP-dependent protein kinase (PKG). The methods should be applicable to other proteins and to other types of modification such as phosphorylation. The procedures use either ion-exchange chromatography, size-exclusion chromatography, or native gel electrophoresis of these proteins in the absence and presence of regulatory ligands. Measurements from these respective approaches allow documentation of changes in the quaternary structure, surface electronegativity, and relative compactness (Stokes radius) of the protein molecule. The combined data allow the changes in protein conformation to be quantitated in terms of alterations in the axial ratio or length/width dimension of the molecule. The methods can be applied to partially purified proteins and to proteins that are available in limited quantities. Conformational changes due to stable modifications of proteins can be potentially examined in crude extracts of intact cells. Each of the methods can be tailored to optimize resolution of a particular protein under a variety of conditions. Activity measurements, Coomassie brilliant blue or silver staining of gels, radioautography, or Western blot analysis can be used for detection of the protein.

1-Methyl-3-isobutylxanthine↗
PubMed1999

Expression of melatonin receptors and 2-[125I]iodomelatonin binding sites in the pituitary of a teleost fish.

The mechanisms of the photoperiodic control of fish physiology (growth, reproduction) and behavior (locomotor activity) are far from being understood. We show here that 2-[125I]iodomelatonin binds specifically to membrane preparations from pike (Esox lucius, L.) pituitaries (KD: 556 pM; Bmax: 2.8 fmol/mg proteins). Radioautography indicated that the binding was restricted to a part of the pituitary only. Using polymerase chain reaction from pike genomic DNA we were able to isolate, subclone and sequence two fragments. The so-called P4 and P8 fragments displayed homology with, respectively, the Mel1a and Mel1b receptor subtypes. The P4 and P8, probes allowed detection of mRNAs corresponding to these receptors in different areas of the brain, including the pituitary. This is the first evidence that melatonin receptors and binding sites are expressed in the pituitary of a non-mammalian species. We suggest that in fish the melatonin-mediated photoperiodic control of neuroendocrine functions might involve a direct effect on the pituitary.

Amino Acid Sequence↗
PubMed1991

Serotonin: its role and receptors in enteric neurotransmission.

Enteric neural 5-HT receptors were analyzed and related to possible physiological actions of 5-HT. Receptors were identified electrophysiologically with intracellular microelectrodes and by studies of the binding of radioligands. Radioligand binding was assessed by rapid filtration and by radioautography. Three subtypes of 5-HT receptor, 5-HT1P, 5-HT3, and 5-HT1A, were identified. 5-HT1P receptors were found to mediate slow depolarizations of myenteric neurons that were associated with a decrease in membrane conductance. These responses were inhibited by 5-HTP-DP and by BRL 24924 and mimicked by 5- and 6-hydroxyindalpine. 5-HT1P receptors were labeled with high affinity by 3H-5-HT and were located on both submucosal and myenteric neurons and on processes of intrinsic neurons in the lamina propria. Serotonergic EPSPs were found to be mediated by 5-HT1P receptors; it is postulated that 5-HT1P receptors may be involved in initiation of the peristaltic reflex and in the regulation of gastic emptying. 5-HT3 receptors have been shown to be responsible for fast depolarizations of myenteric and submucosal neurons associated with a rise in membrane conductance. These responses are antagonized by ICS 205-930 and mimicked by 2-methyl-5-HT. 5-HT1A receptors have been reported by others to mediate hyperpolarizing responses of myenteric neurons associated with a rise in membrane conductance. Hyperpolarizing responses are also elicited by the 5-HT1A agonist, 8-OH-DPAT. No physiological role has yet been identified for 5-HT3 or 5-HT1A receptors in the ENS.

Animals↗
PubMed1992

Calories, fat, fibers, and cellular proliferation in Swiss Webster mice.

Increased cellular proliferation has been associated with the enhanced expression of several key stages in carcinogenesis. A standard protocol was used to investigate the effect of specific dietary regimens on cellular proliferation. Young adult Swiss Webster mice were fed for 30 days with modified AIN-76A semi-purified diets designed to illustrate the effects of the levels of dietary or calorie restriction, different fibers and bulking agents, and different fats on cellular proliferation. Female mice were used for the restriction and fat studies, males for the fiber and bulking agent studies. Vaginal smears were taken from females from treatment day 15, and the mice killed 2 days following the first estrus following 30 days feeding; males were killed on the 30th day. One hour before death, mice were injected ip with 0.25 micro Ci/g 3[H]-thymidine. Slides were prepared for radioautography and histopathology. Both dietary and calorie restriction led to reduced 3[H]-thymidine labeling indices in each of the seven tissues studied, the mammary gland being the most severely affected. Different fibers and bulking agents, in specific cases, reduced labeling in the duodenum but not to a consistent statistically significant extent in the colon or colo-rectal region. In the duodenum, oat bran and oat gum were the most effective while wood cellulose (alphacel) had no effect. Investigations on the effects of different fats is continuing. High levels of lard, menhaden oil, or cod liver oil as the fat component of the AIN-76A diet, led to much higher levels of labeled cells in the mammary gland or colo-rectal region than did fat components rich in vegetable oils. The labeling indices appeared to be inversely correlated with the level of linoleic acid in the diet, a presumption that has been confirmed by investigating a series of diets containing different levels of this acid. Anti-oxidants were not used in any of these fat-modified diets. The overall results obtained in these studies clearly indicate the utility of cellular proliferation studies in investigating the effects of dietary modifications.

Animals↗
PubMed1975

Observations of age and environmental influences on the thymus kept in tissue culture.

In tissue cultures of thymus of rats of various ages only the cells of Gomori- and PAS-positivity showed iodine incorporation as demonstrated by radioautography. Above other cells of the migration zone no grains indicating iodine accumulation were observable. The capacity of iodine accumulation depends on the age of the animal, namely it decreases with the ageing.

Aging↗
PubMed1990

From moclobemide to Ro 19-6327 and Ro 41-1049: the development of a new class of reversible, selective MAO-A and MAO-B inhibitors.

This study describes the serendipitous discovery of moclobemide, a short-acting MAO-A inhibitor which is in an advanced stage of clinical development as an antidepressant. The short duration of action of this MAO inhibitor containing a morpholine ring moiety is due to the complete reversibility (probably by metabolism of the inhibitory molecular species) of MAO-A inhibition. Since moclobemide is much more effective in vivo than expected from its in vitro activity, investigations to identify a possible metabolite(s) more active as MAO-A inhibitor than the parent compound were carried out. The study of the MAO inhibitory characteristics of several known and putative moclobemide metabolites did not allow the identification of a potent MAO-A inhibitor but led to the discovery of Ro 16-6491, a potent MAO-B inhibitor of novel chemical structure. Systematic chemical modification of the aromatic ring system of Ro 16-6491 finally provided Ro 19-6327 and Ro 41-1049 which are highly selective and reversible inhibitors of MAO-B and MAO-A, respectively. Tritiated derivatives of Ro 19-6327 and Ro 41-1049 were used in binding studies to elucidate their mechanisms of action and to study their cellular distribution by quantitative enzyme radioautography.

Animals↗
PubMed1990

Molecular neuroanatomy of MAO-A and MAO-B.

A selective, quantitative and high resolution technique (in vitro and in vivo enzyme radioautography) has been used to reveal the tissue distribution and abundance of MAO-A and MAO-B in the central nervous system and peripheral organs in the rat. The in vitro approach was also used to map the enzymes in human post-mortem brain. Furthermore, using in situ hybridization histochemistry, locus coeruleus and raphé neurons in the human brain were found to code for MAO-A and MAO-B respectively and not vice versa.

Animals↗
PubMed1994

Age-related changes on MAO in Bl/C57 mouse tissues: a quantitative radioautographic study.

The distribution of MAO-A and -B in brain and peripheral tissues of Bl/C57 mice and their changes during ageing were studied by quantitative enzyme radioautography with [3H]Ro41-1049 and [3H]Ro19-6327. In the brain, MAO-A decreased between weeks 4 and 8 and then remained unchanged until 25 months, whereas MAO-B increased for the whole period studied. Heart also showed a continuous increase in MAO-B, but not MAO-A, with ageing, and liver showed a decrease in MAO-B in the older animals. These results show marked species differences in the distribution and age-related changes of MAO and might help to elucidate the high sensitivity of Bl/C57 mouse of MPTP, which increases with age.

Aging↗
PubMed1995

Structure of the embryonic primate spinal cord at the closure of the first reflex arc.

Early development of the spinal cord was studied in macaque monkey embryos, using light- and electron microscopy, Golgi impregnation and [3H]thymidine radioautography. All neurons engaged in the first reflex arc are generated before E27 in the 165 day gestation period. The earliest-generated cells differentiate into motoneurons while the commissural and association neurons are generated later. Synaptogenesis starts at E27 in the basal plate, and 2 days later in the alar plate. The first synapses appear as symmetrical junctions situated on the neuronal perikarya and proximal dendrites. Closure of the first spinal reflex are is established within 2 days and follows an antidromic pattern as related to the physiological spread of nerve impulses: synapses on motoneuronal somata and primary dendrites in the basal plate appear first and are followed by synapses in the alar plate, between dorsal root axon collaterals and somata of borderline (commissural/association) neurons. Commissural axons grow towards the floor plate, cross the midline and proceed caudo-rostrally, while association fibers remain ipsilateral. The first wave of apoptosis (programmed cell death) thins out dense populations of nerve and glial cells by E30. Some of the early-generated borderine cells that form commissural and association interneurons, seem to play the role of transient target cells and die once the definitive axonal pathways are established. Since transient cells form numerous synapses, deprivation from the afferent impulses is not a likely cause of their elimination. The present results indicate that the initial developmental events, including formation of the first reflex arc in the primate spinal cord, occur considerably earlier in respect to birth than in other mammals, but that the schedule of cellular events and cellular mechanisms seem to be the same.

Animals↗
PubMed1978

High resolution radioautographic study of newly formed protein in striated muscle with emphasis on red and white fibres.

The elaboration and distribution of newly formed proteins in the striated muscle of 21-day-old mice were investigated by quantitative radioautography at intervals between 2 and 240 min after intravenous injection of tritiated leucine. In radioautographs, the localization and the relative label concentration were comparatively estimated for the different components of mitochondria-rich fibres, in particular of red fibres, from the tibialis anterior muscle and of mitochondria-poor fibres from the oesophageal muscle. As early as 2 min after injection, radioactivity was detected over the nucleus, the polysome-rich sarcoplasm, the A and I bands, the Z lines, and the mitochondria in the two fibre types. Label localization did not change with time. The relative label concentration increased similarly in the polysome-rich sarcoplasm and the A and I bands of both fibre types within 30 min after injection, a confirmation that biosynthesis of myofibrillar proteins takes place rapidly. In each case, concentration was higher in the Z lines than in the I bands, and higher in the I bands than in the A bands, thus showing "in vivo" that the rates of synthesis of sarcomere protein components are not uniform. However, the relative label concentration was found to be higher in the Z lines of mitochondria-poor fibres than of mitochondria-rich fibres: this suggests that a higher synthetic rate of Z line protein, and probably of alpha actinin, is characteristic of the first type. Inversely, the concentration was higher in the mitochondria of mitochondria-rich fibres. This lead to the belief that high rate of protein synthesis in these organelles may account for the high rate of label incorporation into this type of fibre.

Animals↗
PubMed1986

The dopaminergic innervation of the goldfish pituitary. An immunocytochemical study at the electron-microscope level using antibodies against dopamine.

The dopaminergic innervation of the goldfish pituitary gland was studied by immunocytochemistry at the electron-microscope level using highly specific antibodies against dopamine coupled to bovine serum albumin with glutaraldehyde. A satisfactory preservation of the tissue was achieved after immersion in 5% glutaraldehyde in phosphate buffer containing sodium metabisulfite to prevent oxidation of the endogenous dopamine. The immunocytochemical procedure was performed on Vibratome sections using the preembedding method. Immunoreactivity was restricted to part of the neurosecretory type-B fibers (diameter of the secretory vesicles lower than 100 nm) in which it was found to occupy the whole cytoplasm. Labeled fibers were observed within the neurohypophysis in the different parts of the gland and in the adenohypophyseal tissue where immunoreactive profiles were detected in close apposition to the different cell types. These data are in agreement with previous results obtained by means of radioautography and further support a role for dopamine in the neuroendocrine regulation of pituitary functions in teleosts.

Animals↗
PubMed1984

Distribution of [3H]noradrenaline and [3H]serotonin in photophores of porichthys notatus. An electron-microscopic radioautographic analysis.

Photophores of Porichthys notatus were examined by electron-microscopic radioautography following incubation in tritiated noradrenaline ( [3H]NA) or serotonin ( [3H]5-HT). Nerve varicosities surrounding the photocytes were found to accumulate [3H]NA but not [3H]5-HT, providing compelling evidence for the catecholaminergic nature of the monoaminergic innervation of photophores. The photocytes themselves appeared selectively labelled with both tracers, but the intensity of labelling after [3H]5-HT incubation was considerably greater than after [3H]NA. Stereological sampling of organelle content in photocytes showed ultrastructural differences between [3H]NA- and [3H]5-HT-labelled cells, probably related to light emission induced by NA. The main changes noted after incubation with [3H]NA were mitochondrial swelling and disorganization, increased coalescence of photocytic vesicles and extrusion of vesicular material into the extracellular matrix. With respect to the subcellular localization of [3H]NA and [3H]5-HT within the photocytes, statistical analysis of the distribution of silver grains disclosed a preferential affinity of both labels for appositional zones between mitochondria and coalescent vesicles. Moreover, in the case of 5-HT, selective affinity was also exhibited by sites comprising vesicular membrane and adjacent cytoplasm, suggesting binding of this biogenic amine to the entire membrane of photocytic vesicles.

Animals↗
PubMed1984

Homing of germinal-center cells into germinal centers of lymph node via afferent lymphatics. An autoradiographic study in rabbits.

Affinity of lymphoid cells for the microenvironment of germinal centers (GC), as detectable in transfer experiments by rapid homing in spleen GC from the blood, is a capacity expressed by only a subset of lymphoid cells, in particular by those constituting a GC. However, when introduced into the blood stream, these cells do not home into GC of lymph nodes and gut-associated lymphoid tissues. To investigate further this homing inability for high endothelial venule (HEV)-containing lymphoid tissues, GC cells isolated from donor rabbit appendix were labeled in vitro with 3H-leucine and injected into an afferent lymph vessel of recipient popliteal lymph nodes. Draining lymph nodes were removed 15 min to 24 h after cell administration and prepared for radioautography. For reference, the migration of cells isolated from Peyer's patches and thoracic duct lymph was also studied. By use of appendix GC cells, large numbers of labeled cells were found to migrate into GCs of the outer cortex centripetally, i.e., from the subcapsular sinus through the lymphocyte corona into the GC proper. The same was observed for cells from Peyer's patches, although in smaller numbers. Thoracic duct lymphocytes were only localized in the lymphocyte corona and the deep cortex. Thus, appendix GC cells and a subpopulation of cells from Peyer's patches can reach lymph node GC, but only when administered intralymphatically.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Synthesis and migration of proteins and glycoproteins in juxtaglomerular cells of sodium-deficient rats. An ultrastructural radioautographic study.

Sections of juxtaglomerular cells from sodium-deficient rats were subjected to radioautography after a single intravenous injection of L-tyrosine-3,5 3H or of L-fucose 3H to identify the sites of synthesis and to follow the migration of newly-formed proteins and glycoproteins. As early as 2 min after injection of L-tyrosine 3H, the label was highest in the rough endoplasmic reticulum (RER), suggesting that cisternal ribosomes are sites of protein synthesis. By 60 min, much of the label had migrated from the RER to the Golgi complex. Some radioactivity was already present over specific granules by 2 min but a peak was reached at 4 h. The label over myofilaments was evident at all time intervals, indicating a certain incorporation of tyrosine into their contractile and/or structural proteins. The label over the cell surface peaked at 4 h. After injection of L-fucose 3H, there was an early and important relative specific radioactivity in the Golgi complex at 5 min with a peak at 20 min and a decrease thereafter. The label increased slightly but steadily in secretory granules and cell surface to reach maxima at 4 h. A low level of radioactivity was recorded in mitochondria at all time intervals. After injection of both fucose 3H and tyrosine 3H, the label was detected at relatively low levels in the cytosol. These results suggest that renin, as the major secretory glycoprotein of juxtaglomerular cells, is synthetized in the RER, packaged in the Golgi complex and found relatively rapidly in newly-formed secretory granules. Part of the fucose and tyrosine labels is also associated with the thick cell coat of these cells.

Animals↗