SEARCH · PubMed Health

Results for “RADIOAUTOGRAPHY”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

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

At least 1,009 records · Page 56Linked to original sources

PubMed1992

Transcobalamin II--cobalamin binding sites are present on rabbit germ cells.

Specific binding sites for rabbit transcobalamin II have been found on isolated adult rabbit germ cells. Scatchard analysis revealed a single class of binding sites for [57Co]cyanocobalamin-transcobalamin II with an association constant (Ka) of 1.3 x 10(10) M-1 and 700 sites per cell. Binding was reversible, saturable and calcium dependent. Electron microscope radioautography following incubation with iodinated transcobalamin II at 4 degrees C led to a detectable labeling mainly restricted to the plasma membrane.

Animals↗

PubMed1990

Ultrastructural localization of oxytocin mRNA in the rat hypothalamus by in situ hybridization using a synthetic oligonucleotide.

In situ hybridization (ISH) was used to study at the electron microscope level, the subcellular localization of oxytocin (OT) mRNA in the rat hypothalamic magnocellular neurons. Rat brains were fixed with paraformaldehyde and glutaraldehyde and vibratome slices were incubated with a 25-base synthetic oligonucleotide complementary to OT mRNA and labelled at the 3'-end with [3H]dCTP. Hybridized slices were embedded in Epon after post-fixation with osmium tetroxide and cut into ultrathin sections that were processed for ultrastructural radioautography. OT mRNA was observed in magnocellular neurons of supra-optic and paraventricular nuclei in the vibratome sections. On ultrathin sections, the cytological preservation appeared to be satisfactory. Except for a few silver grains over the nucleus, sometimes close to its membrane, most grains were localized over the cytoplasm of some magnocellular neurons, where they frequently overlapped the endoplasmic reticulum. To decrease exposure time, ISH was also performed with OT probes labelled with a long tritiated tail. In this case, clusters of silver grains occurred over the cell nuclei not only in magnocellular neurons but also in non-secretory neurons and even in glial cells. However, an excess of poly C added to the hybridization buffer strongly decreased this non-cytoplasmic labelling. In conclusion, the results obtained with the short-tailed oligonucleotides demonstrate that these synthetic oligonucleotides have possible applications for the ultrastructural localization of mRNAs and constitute a powerful tool for the dynamic study of cellular mRNA processing in several physiological and experimental conditions.

Animals↗

PubMed1996

Detection of MAO-A and MAO-B mRNAs in monkey brainstem by cross-hybridization with human oligonucleotide probes.

The distribution of mRNAs encoding the isoenzymes monoamine oxidase A and B (MAO-A and MAO-B) in monkey locus coeruleus and dorsal raphe nucleus was studied by in situ hybridization histochemistry using 35S-labelled oligodeoxynucleotide probes complementary to cloned human sequences. MAO-A mRNA was highly expressed in noradrenergic neurons of the locus coeruleus while MAO-B mRNA was abundantly and exclusively localized in serotoninergic neurons of the raphe. However, upon emulsion radioautography raphe neurons showed a level of MAO-A mRNA signal noticeably above the background. Our results indicate the utility of human MAO oligodeoxynucleotide probes to identify homologous species of transcripts in the brain of a non-human primate. They also suggest the coexistence of the isoenzymes in raphe neurons as well as the potential role of MAO-A in metabolizing serotonin in vivo.

Animals↗

PubMedDate not supplied

Radioautographic localization of somatostatin-14 and somatostatin-28 binding sites in the rat brain.

Somatostatin-14 (S14) and its precursor, somatostatin-28 (S28), are widely distributed throughout the rat brain, suggesting that they could act as neurotransmitter or neuromodulator in the central nervous system. The present study was undertaken to study the localization of S14 and S28 receptors in the rat brain determined by "in vitro" radioautography. The study performed on slide mounted frozen brain section with iodinated S14 and S28 analogs revealed an identical distribution of binding sites for the two forms of somatostatin. A good correlation could be observed between receptor distribution and immunohistologically localized neuropeptides except for striatum and hypothalamus. However, receptors were not detectable in the hypothalamus and were found in low concentration in the caudate-putamen nucleus, two regions containing high amounts of S28 and S14, suggesting a high occupancy of receptors in these areas by endogenous peptides or an inverse correlation between receptor and peptide concentrations.

Animals↗

PubMed1984

Synaptic input to vasopressin neurons of the paraventricular nucleus (PVN).

Following injections of horseradish peroxidase into the PVN, retrogradely filled cells were found in regions of the limbic system known to contain glucocorticoid concentrating neurons [4, 31, 44]. To determine if these regions which include the lateral septum, medial amygdala and ventral subiculum have a monosynaptic input to vasopressin neurons we developed a double label ultrastructural technique [20] to simultaneously visualize immunoreactive neuropeptide and anterogradely transported HRP. Following injections of tracer into all three of these regions, HRP labeled fibers were seen at the light microscopic level to form a halo in the perinuclear, cell poor zone around the PVN. Ultrastructural examination of this area resulted in the discovery of a small number of limbic system synapses on vasopressin dendrites. These synapses were most numerous in the ventral and medial portion of the cell poor zone. A similar pattern of innervation was seen for the supraoptic and suprachiasmatic nucleic which also contain vasopressin cells whose dendrites extend beyond the nuclear boundaries. In a similar fashion we were interested in determining the distribution of noradrenergic terminals on vasopressin neurons in the various subnuclei of the PVN. We have combined immunocytochemistry for vasopressin with radioautography for 3H-norepinephrine (NE) at the ultrastructural level. NE terminals were numerous in the periventricular zone, innervating both vasopressin containing dendrite and non-immunoreactive dendrites and cell bodies. The vasopressin dendrites could originate from cells either resident in the periventricular zone or from cells situated in more lateral subnuclei. In the main, lateral magnocellular region, noradrenergic terminals were very few in number and innervated almost exclusively non-vasopressin containing structures. These studies demonstrate the need for ultrastructural analysis of synaptic input to neurosecretory cells.

Animals↗

PubMedDate not supplied

Anatomical substrate for neurotensin-acetylcholine interactions in the rat basal forebrain.

We have previously shown by combined radioautography and acetylcholinesterase histochemistry that the distribution of 125I-neurotensin (NT) binding sites was in register with that of cholinergic neurons in the rat nucleus basalis magnocellularis (NBM). The present study utilized three experimental approaches to elaborate on the type and cellular localization of NT binding sites in the NBM. Competition studies using levocabastine, a selective blocker of the low affinity NT binding component, revealed that most of the 125I-NT binding sites labeled in the NBM are of the levocabastine-insensitive high affinity type, known to correspond to the physiologically active receptor. Ibotenic acid-induced lesions of the NBM produced a marked reduction in both cholinesterase reactivity and cellular 125I-NT binding suggesting that most of the labeled sites are associated with the cholinergic neurons themselves rather than with an afferent input to those cells. Finally, examination of the high resolution radioautographic distribution of 125I-NT binding sites in semithin sections revealed that a proportion of 125I-NT-labeled receptors is associated with the plasma membrane of magnocellular perikarya and proximal processes, thereby providing an anatomical substrate for a local action of NT in the NBM.

Acetylcholine↗

PubMed1994

VIP binding sites in adult rat hypothalamus: nuclear distribution and daily variations.

Vasoactive intestinal peptide (VIP) neurons are present in the suprachiasmatic nucleus (SCN) and project to various hypothalamic nuclei. Several results suggest that hypothalamic VIP is involved in the mediation of synchronizing information. [125I]Vasoactive intestinal peptide binding has been reported to be especially high in the hypothalamus. We show here by radioautography by dipping that [125I]VIP binding is high and homogeneously distributed in the SCN. The correspondence between VIP binding and terminals densities supports the hypothesis that VIP acts through synapses in the SCN. Moderate and fairly homogeneous VIP binding is also found in the paraventricular nucleus (PaVN), whereas VIP terminals are reported to be restricted to the sub-paraventricular area. This suggests that VIP could diffuse and thus function as a paracrine neuromodulator in the PaVN. [125I]Vasoactive intestinal peptide binding is stable along the nycthemer in the suprachiasmatic, peri-, and paraventricular nuclei as well as in all other areas investigated.

Animals↗

PubMedDate not supplied

Differential age-related changes of MAO-A and MAO-B in mouse brain and peripheral organs.

Distribution and age-related changes of MAO in BL/C57 mouse were studied by quantitative enzyme radioautography with [3H]Ro41-1049 and [3H]Ro19-6327. In the brain, MAO-A was highest in locus coeruleus and interpeduncular nucleus, and MAO-B in raphe nuclei, paraventricular thalamic nucleus, and ependyma of ventricles. Extremely high MAO-B levels were also measured in the choroid plexus in contrast to the very low MAO-B levels in rat choroid plexus. With aging, brain MAO-A showed a clear decrease between 4 and 9 weeks, followed by no change between 9 weeks and 19 months, and a slight increase between 19 and 25 months. On the other hand, all brain structures showed age-related increases in MAO-B. Peripheral organs showed different patterns of MAO age-related changes. Particularly interesting was the marked MAO-B increase in heart, parallel to the MAO-A increase in rat heart. Also of interest is the decrease of liver MAO-B in old animals, which, together with the increase of MAO-B in the brain, might underlie the high sensitivity of old BL/C57 mice to MPTP.

Aging↗

PubMed1996

Kinetics of internalization and subcellular binding sites for T3 in mouse liver.

The intracellular fate of radiolabeled T3 taken up by mice hepatocytes in vivo was determined at specific time intervals (2-120 min) after injection by quantitative electron microscopic radioautography. Injection of a 200-fold excess of unlabeled T3 together with [125I]-T3 resulted in a more than 90% inhibition of radioactivity detected in hepatocytes. A simple grain density (GD) analysis of radioautograms revealed that a specific labeling (GD > 1) was displayed by only five cell compartments: the plasma membrane, lipid droplets, mitochondria, nuclear envelope and nuclear matrix whereas other compartments were not labeled. Labeled compartments showed distinct changes in the pattern of labeling over time: the plasma membrane was labeled only 2 min after T3 injection, whereas labeling of the nuclear envelope was high at 2 min, decreased at 15 min and progressively increased to maximal measured levels at 120 min. After a lag time of 30 min, nuclear matrix labeling increased progressively with time. Mitochondrial labeling was found to be specific at any time point studied but showed no change over time. These ultrastructural data have been confirmed in vitro by the interaction of T3 with plasma membrane, nuclear membrane, nuclear matrix and mitochondria by real-time biospecific interaction analysis in a BIAcore system. These results demonstrate that T3 binds to hepatocytes before internalization, is transported both to mitochondria and to the nuclear envelope and translocated into the nuclear matrix.

Animals↗

PubMed1985

Cell proliferation kinetics in the gastric remnant.

Biopsy specimens of the gastroenterostomal area in 60 long-standing asymptomatic postgastrectomy patients and of the mid-corpus region in five normal controls were cultured in [3H]-TdR. Using radioautography, thymidine incorporation rate (TIR) and cell position of the labelled cells were scored. In the normal gastric mucosa cell proliferation was limited to the progenitor cell region. TIR of the stomal area was higher compared to the normal mucosa. In addition, an upward shift of the proliferative compartment towards the luminal surface was observed. In severe dysplasia an expanded pool of DNA-synthesizing cells was present in the uppermost layers of the gastric mucosa, but even in the surrounding non-dysplastic mucosa an upward shift of labelled cells could be demonstrated. These proliferative changes are considered to be compatible with increased cancer risk. The sequence of proliferative events suggests that early cancer formation in the gastric remnant takes place in the superficial mucosa and therefore is easily amenable to endoscopic biopsy.

Cell Division↗

PubMedDate not supplied

Histopathological and radioautographical studies on the forestomach of F344 rats treated with butylated hydroxyanisole and related chemicals.

Butylated hydroxyanisole, when fed to male Fischer 344 rats for periods of 9 days or more, led to forestomach epithelial cell necrosis and regeneration. Both the induced proliferation and the histopathological changes were considerably more prevalent in the prefundic region of the forestomach than in the mid-region. Using [Me-3H]thymidine, a specific DNA precursor, and radioautography it was shown that the effect of the antioxidant was apparently threshold at 0.25% in the diet after both 9 days and 3 months of treatment and that the proliferation was dependent on the continuous presence of the antioxidant in the diet at 3 and 6 months. Some other phenols and acids were investigated after 9-27 days' feeding; most induced some degree of epithelial cell proliferation in the rat forestomach, although some had a greater effect on the mid-region than on the prefundic region. These observations are discussed in terms of the likely relevance of butylated hydroxyanisole-induced forestomach tumours to the possibility that the antioxidant may lead to cancer in humans exposed to lower levels in their diet.

Administration, Oral↗

PubMed1994

Effect of addition of dried healthy or diseased parsnip root tissue to a modified AIN-76A diet on cell proliferation and histopathology in the liver, oesophagus and forestomach of male Swiss Webster mice.

Umbelliferous crop plants, including the parsnip (Pastinaca sativa L.), elaborate enhanced levels of furocoumarins, including psoralens, when subjected to biotic or abiotic stress. These furocoumarins are recognized to lead to phototoxicity. In this study, the effect of these agents, which are present in diseased parsnip root tissue, on the liver and two tissues on the route of entry to the body (the oesophagus and forestomach) were investigated. Young male Swiss Webster mice were fed for approximately 30 days with modified AIN-76A diets containing 32.5% dried healthy, 32.5% apparently healthy or 32.5% fungicide-treated parsnip root tissue, and 8, 16 or 32.5% dried diseased (Phoma complanata-infected) parsnip root tissue. As controls, three modified AIN-76A diets differing in their edible starch-to-sucrose ratios (C1-C3) were administered for an equal time. Dried healthy parsnip root tissue, compared with controls, did not significantly affect any of the indices of cellular proliferation or histopathological parameters that were assessed. Histopathological examination of the oesophagus and forestomach demonstrated no significant changes as a result of feeding any of the diets containing parsnip tissue. In the liver, the highest level (but neither of the two lower levels) of dried diseased parsnip root tissue led to swelling of the cytoplasm in cells surrounding the central vein of hepatic lobules, with consequent compression of the peripheral cells. Using [3H]thymidine radioautography, a dose-related increase in cell labelling with the level of diseased parsnip root tissue was demonstrated in the liver. Compared with control diet C2 only, the extent of [3H]thymidine labelling in the liver was increased in mice receiving apparently healthy parsnip tissue; a slight, not statistically significant, increase was also noted with fungicide-treated parsnip tissue. Increased [3H]thymidine labelling with the feeding of diseased parsnip tissue was also found in the greater curvature of the forestomach and the region of the oesophageal-forestomach junction, but not at the glandular junction of the forestomach nor in the mid-oesophagus.

Animal Feed↗

PubMed1984

Short-term pathological and proliferative effects of butylated hydroxyanisole and other phenolic antioxidants in the forestomach of Fischer 344 rats.

Food grade butylated hydroxyanisole (BHA) when incorporated in the diet and fed to male Fischer 344 rats for 9 or 27 days induced proliferative squamous epithelial changes in the lesser curvature of the forestomach proximate to the glandular stomach. These changes were assessed histopathologically and by [methyl-3H]thymidine radioautography. It was shown that BHA mixed dry into powdered diet, incorporated into the diet in corn oil, or in a pelleted diet, induced similar effects. When levels of 2%, 1%, 0.5%, 0.25%, 0.1% and 0% BHA were incorporated in rat diet for 9 days, the proliferative effect appeared to show a no effect level at 0.25% based on the [methyl-3H]thymidine-labelling index. Other food use antioxidants, namely butylated hydroxytoluene or tertiary butylhydroquinone, induced a lesser response than BHA at the maximum dose employed in the study. Propyl gallate was without effect. Propyl-4-hydroxybenzoate, a food use phenol, on the other hand, induced a less pronounced response than BHA but was more effective than the other antioxidants. Because increased cellular proliferation often provides an optimal milieu for tumor formation, it is suggested that these observations may be relevant to rat forestomach tumors induced by BHA.

Animals↗

PubMedDate not supplied

Uptake of biogenic amines by thyroid glands.

The biogenic amines, serotonin (5-HT) and norepinephrine, activate follicular cells and are endogenous thyroid constitutents. Both amines are taken up by throid lobes incubated in vitro. Uptake of 5-HT is temperature and concentration dependent, is inhibited by chlorimipramine more than by desmethylimipramine, and although it is inhibited by norepinephrine, 5-HT uptake is unaffected by chemical sympathetomy with 6-hydroxydopamine. Electron microscopic radioautography reveals that follicular cells are responsible for uptake of 5-HT and adrenergic nerves for uptake of norepinephrine. These data are consistent with the hypothesis that biogenic amines are intrathyroidal transmitters acting on follicular cells, and that uptake mechanisms may contribute to their inactivation.

Animals↗

PubMed1978

Radioautographic study of the rat brain, uterus and vagina after [3H]R-5020 injection.

Localization of [3H]R-5020, a potent synthetic progestin, was examined by radioautography in the central nervous system and the genital tract of estrogen-primed, ovariectomized rat. In the brain, a concentration of radioactivity was found in the nuclei of neurons localized in different specific areas: the preoptic region and the medio-basal hypothalamus. In the anterior pituitary, cells which concentrated the hormone were found. The uterine tissues (uterine horns and cervix) presented a nuclear concentration of radioactivity in stromal and muscle cells but little or no radioactivity in luminal and glandualr epithelia. In the vagina of the same animals, radioactivity was observed in the connective tissue cells, muscle cells and epithelial cells. Hormonal specificity and saturability of radioactivity binding in these target tissues were established by competition with an excess of unlabeled hormones. In the presence of an excess of unlabeled progesterone the nuclear concentration of radioactivity was diminished in all areas, while no inhibition of radioactivity incorporation was observed with unlabeled corticosterone. These results suggest the presence of specific progesterone receptors in the hypothalamus, the pituitary and the genital tract tissues of the rat.

Animals↗

PubMed1986

Distribution of epidermal growth factor receptors in the rat ovary.

The distribution of epidermal growth factor (EGF) was studied in the ovary using light microscope radioautography which was performed at different time intervals (2-60 min) after intravenous (i.v.) injection of [125I]EGF into adult rat at random stages of the estrous cycle and also after topical localization of iodinated EGF on slide-mounted frozen ovarian sections. After i.v. injection of label, the labeling was mostly observed in the theca interna cells of secondary, preovulatory and atretic follicles and luteal cells of corpus luteum. Primordial and primary follicles did not show any significant labeling. The time-course study performed on luteal cells showed that, 2 min after injection, most silver grains were found at the periphery of the cells. At the 10 min time interval, silver grains were found both at the periphery and over the cytoplasm of these cells. The number of grains was very reduced over the cytoplasm at the 60 min time interval. In the in vitro study, a positive radioautographic reaction was seen in the same cellular elements as found in vivo, with the additional labeling of the granulosa cells of growing and preovulatory follicles. Control experiments indicated that the radioautographic labeling was due to specific interaction of [125I]EGF with its receptor. These results clearly indicate that EGF binding sites are present in luteal, thecal and granulosa cells, and provide support for the inhibitory and stimulatory actions of EGF on different parameters of ovarian cells.

Animals↗

PubMed1979

High resolution radioautographic identification of [3H]GABA labeled neurons in the rat nucleus raphe dorsalis.

Following intraventricular administration of [3H]GABA into the lateral ventricle of the rat radioautography demonstrates the presence of labeled cell bodies and fibers within the nucleus raphe dorsalis (NRD). Pharmacological controls suggest that labeled perikarya whose diameter does not exceed 10 micrometer and which are not probably more than ten in the entire NRD could be GABAergic. The effects of 5,7-dihydroxytryptamine (5,7-DHT) pretreatment support the hypothesis that labeled fibers could belong at least to two populations: ones being probably serotonergic and the others being possibly GABAergic. These results are in good agreement with the existence of an intrinsic GABAergic system within the rat NRD.

Animals↗

PubMed1979

Evidence for a multiple innervation of subcommissural ependymocytes in the rat.

Glandular ependymocytes of the rat subcommissural organ (SCO) receive a multiple innervation: a serotonergic afference identified by radioautography and at least one other input of a hitherto unknown transmitter content. Both types of terminals make well differentiated axo-glandular synapses with SCO-ependymocytes, but display different morphological characteristics. Serotonergic terminals being considered as inhibitory, it is suggested that the other type of terminal is excitatory.

Animals↗