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PubMed1992

Light and electron microscopic radioautography of DNA synthesis in the endometria of pregnant-ovariectomized mice during activation of implantation window.

Pregnant mice were ovariectomized at pre-implantation stage and exogenous nidatory estradiol was administered to evaluate the DNA synthesis of the endometrial cells during activation of uterine receptivity for blastocyst implantation. After 0, 3, 6, 12 and 18 hrs. of estradiol treatment, the animals received 3H-thymidine injection, sacrificed 1 hr. later, and the uteri were prepared for light and electron microscopic radioautography. At time 0, no labelled stromal or epithelial cells was found in the endometrium. According to the time-lapse after estradiol induction, a gradual increase of labelled stromal and endothelial cells was seen in the endometrium. The highest labeling index was observed at the antimesometrial side of the implantation sites and the lowest value was found at the interimplantation site. The cells found at mesometrial side of the implantation site showed an intermediate labeling index. Eighteen hrs. after estradiol treatment, the labelled stromal cells found near the implantation chamber resembled the morphology of decidual cells while those labelled cells localized at the interimplantation sites were similar to the fibroblast. The uterine luminal epithelial cells showed low DNA synthesis after estradiol treatment resulting in only a few labelled cells at the interimplantation sites and no labelled cells at the implantation sites. A similar labeling pattern was seen in the glandular epithelium. The distribution of labelled cells seen among the regions of pregnant endometrium under estradiol effect suggest that DNA synthesis related to uterine activation for blastocyst implantation is a focal reaction, where the luminal epithelium does nt proliferate while the stromal and endothelial cells around the conceptus increase the DNA synthesis to prepare the endometrial decidualization.

Animals↗
PubMed1978

[Sleep and monoamines: differential radioautography of central neurons after systematic injection of tritiated 5-hydroxytryptophane (5 HTP 3H) or of dihydroxyphenylalanine (DOPA 3H)].

In normal cats 90 min. after the intravenous injection of 5 mg/kg of 5 HTP and 3H 5 HTP or DOPA and 3H DOPA a few reactive nerve cell bodies were identified by radioautography. On the contrary, 48 hrs. after p-chlorophenylalanin or subtotal lesion of raphe nuclei, these tracers showed numerous reactive neurons inside or outside the central monoaminergic systems.

5-Hydroxytryptophan↗
PubMed1992

Quantitative enzyme radioautography with 3H-Ro 41-1049 and 3H-Ro 19-6327 in vitro: localization and abundance of MAO-A and MAO-B in rat CNS, peripheral organs, and human brain.

Monoamine oxidases A and B (MAO-A and MAO-B) oxidatively deaminate neurotransmitter and xenobiotic amines. Since the cellular localization of the isoenzymes in the CNS and peripheral organs determines to a large extent which substrate has access to which isoenzyme, knowledge of their tissue distribution and cellular localization is essential. Here we describe how reversible and selective inhibitors of MAO-A and MAO-B [Ro 41-1049 and Ro 19-6327 (lazabemide), respectively] can be used, as tritiated radioligands, to map the distribution and abundance of the enzymes in microscopic regions of the rat CNS and peripheral organs, and human brain by quantitative enzyme radioautography. The in vitro binding characteristics of both radiolabeled inhibitors revealed them to be selective, high-affinity ligands for the respective enzymes. KD and Bmax values for 3H-Ro 41-1049 in rat cerebral cortex were 10.7 nM and 7.38 pmol/mg protein, respectively, and for 3H-Ro 19-6327 were 18.4 nM and 3.45 pmol/mg protein, respectively. In accordance with their potencies as enzyme inhibitors, binding to MAO-A and MAO-B was competitively inhibited by clorgyline (IC50 = 1.4 nM) and L-deprenyl (selegiline; IC50 = 8.0 nM), respectively. The capacities of various rat and human tissues to bind the radioligands correlated extremely well with their corresponding enzyme activities. As revealed by the respective binding assays, the distribution and abundance of MAO-A and MAO-B in the tissues investigated differed markedly. MAO-A was most abundant in the locus coeruleus, paraventricular thalamus, bed nucleus of the stria terminalis, median habenular nucleus, ventromedial hypothalamus, raphe nuclei, solitary tract nucleus, inferior olives, interpeduncular nucleus, claustrum, and numerous peripheral tissues, including liver, vas deferens, heart, superior cervical ganglion, and exocrine and endocrine pancreas. In contrast, MAO-B was most abundant in the ependyma, circumventricular organs, olfactory nerve layer, periventricular hypothalamus, cingulum, hippocampal formation, raphe nuclei, paraventricular thalamus, mammillary nuclei, cerebellar Bergmann glia cells, liver, posterior pituitary, renal tubules, and endocrine pancreas. The cellular localization of the isoenzymes in both rat and human brain differs markedly and does not reflect the distribution of the presumed natural substrates, for example, absence of MAO-A in serotoninergic neurons. Indeed, the present evidence suggests that, whereas MAO-A is found in noradrenergic and adrenergic neurons, MAO-B occurs in astrocytes, serotoninergic neurons, as well as ventricular cells, including most circumventricular organs. The physiological roles of the enzymes are discussed in the light of these findings, some of which were unexpected.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Localization of the anti-allergic agent tranilast in the urinary bladder of rats as demonstrated by light microscopic radioautography.

Tranilast (N-(3,4-dimethoxycinnamoyl)anthranilic acid), an anti-allergic drug, has been developed and used for the treatment of anaphylactic hypersensitivity such as bronchial asthma in Japan. Recently, many cases with eosinophilic cystitis associated with administration of this drug have been reported. In a series of studies to elucidate the pathoetiology of this unusual cystitis, the distribution of tranilast in urinary bladder of rats was investigated. Light microscopic radioautography using tritiated tranilast showed that this drug was localized specifically in the transitional epithelial cells of the mucosa and the endothelial cells of venous vessels for a fairly long time after an oral administration. These results should be taken into consideration in further studies of this drug-associated cystitis.

Animals↗
PubMed1991

Protein synthesis in the livers of aging mice studied by electron microscopic radioautography.

The application of 3H-leucine results in labeling of the liver cells of mice in which protein is synthesized at various ages of the animals. Quantitative changes of protein synthesis in the hepatocytes of aging mice were studied by electron microscopic radioautography. The silver grains in the hepatocytes were mainly located over the rough surfaced endoplasmic reticulum, mitochondria, Golgi apparatus, cytoplasmic matrix, and a few over the nuclei. The number of silver grains in the cytoplasm and nuclei of the hepatocytes gradually increased after birth, reached the maximum at 1 month after birth, thereafter it continued to decrease with aging until the 24th month. The number of silver grains in the hepatocyte cytoplasm was more than that in nuclei at various ages. The number of silver grains in the rough surfaced endoplasmic reticulum and mitochondria gradually increased from embryo to 1 month after birth, thereafter it continued to decrease with aging until the 24th month. The number of silver grains in the Golgi apparatus showed almost no change from fetal stage to 6 months after birth, thereafter it continued to decrease with aging until the 24th month. The number of silver grains in the cytoplasmic matrix gradually increased from fetal stage to 2 months after birth, then decreased with aging until the 24th month. These changes reflect the quantity of protein synthesized in each cell organelle at various ages of animals.

Aging↗
PubMed1991

[Electron microscope-radioautography study of inveterate non-healing wounds].

Morphological and functional peculiarities of long non-healing wound granulation tissue cells have been studied. The cell functional and proliferating activities were defined by electron-microscopic radioautography of 3H-uridine and 3H-thymidine incorporation. There are few vessels and the lumens are narrow. The walls of the most of the vessels are in the state of destruction. There are multiple macrophages with phagosomes filled with detritus. Fibroblasts have endoplasmic reticulum with widened small canals. Not multiple collagen fibers have broken periodic striation.

Adult↗
PubMed1990

Time-related changes in the distribution of 45Ca in the developing enamel of rat incisors as revealed by radioautography.

The distribution and movement of calcium through the enamel organ and into the forming enamel was examined by means of quantitative 45Ca radioautography in rat incisors. Dislocation of radiocalcium in the specimen was minimized during histologic and radioautographic processing by using rapid freeze/freeze-substitution and dry emulsion coating methods. At 30 sec. after the 45Ca injection, distinct peaks of radioactivity occurred in the connective tissue immediately adjacent to the enamel organ and the infranuclear compartment of secretory ameloblasts. An intense labeling also occurred in the superficial layers of the forming enamel extending 15 microns below the surface. The grain density in the distal cytoplasm of secretory ameloblasts increased at later time periods, whereas all other regions of the enamel organ showed a considerable decrease in radioactivity. The radioactivity in the infranuclear compartment of ameloblasts with numerous mitochondria remained relatively high at 2 min. but was abolished by 10 min. after the injection. The grain density in the enamel matrix became much stronger but the labeled regions only extended to 20, 30, and 40 microns below the surface at 2, 10, and 60 min. after the injection, respectively. The application of wet emulsion over similarly prepared sections caused a severe dislocation of radiocalcium in the specimens. These data confirmed the rapid penetration of systemically administered radiocalcium into the surface layers of forming enamel and its slow diffusion to the deeper layers. The time-related changes in relative grain densities at various regions of the ameloblasts support the coexistence of a relatively slow transcellular pathway for calcium through the secretory ameloblast layer.

Ameloblasts↗
PubMed1988

Direct functional assessment of human osteoblasts by radioautography: methodology and application in end stage renal disease.

Fragments approximately 1 mm3 of freshly obtained iliac crest cancellous bone were incubated for 30 minutes at 37 degrees C in Krebs buffer containing 3H-leucine or 3H-proline followed by radioautography. Grains per osteoblast, reflective of protein synthetic activity, were counted in normal controls and patients with end stage renal disease with or without significant aluminum storage in their bones. Compared with control the end stage renal disease patients without significant aluminum storage had more large osteoblasts with increased grain counts which were ascribed to secondary hyperparathyroidism. Cells from patients with greater than 30% surface aluminum had more small osteoblasts with grain counts intermediate between control and less than 2% aluminum, ascribed to an inhibitory effect of aluminum on the secondary hyperparathyroidism. Cell activity did not depend on location next to a tetracycline label or osteoid and it was suggested that all bone lining cells could be considered as osteoblasts. Aluminum storage was associated with impaired mineralisation in the presence of normal or increased osteoblastic protein synthetic activity suggesting that in these patients the mineralisation defect was not primarily due to an abnormality in osteoblast function.

Aluminum↗
PubMed1979

[Thyroid incorporation of radioactive iodine in neotenic tritons (Triturus helveticus Raz.). In vivo studies by high resolution radioautography. Comparison of iodine incorporation in adults and larvae].

In vivo kinetics of radioiodine incorporation have been established for 70 newts, collected from Triturus helveticus Raz. populations where spontaneous neoteny occurs. Fifty of these newts which appeared to present four degrees of neoteny (more or less metamorphosed), have been studied. Main results obtained were: 1) The intensity of the iodine incorporation varies according to the degree of neoteny. The higher is this one, the lower is the intensity. 2) In general, iodine uptake rates are more or less similar to these of adults, but more often the evolution of the 125I uptake approaches a plateau-curve. From thyroid radioautographies on some animals (17 neotenic and 2 adult newts) for which in vivo iodine kinetics have been examined and on 9 normal larvae in premetamorphosis or in metamorphosis, we have noticed in those neotenic thyroids, a very small number of silver grains on apical vesicles and colloidal lumina when the degree of neoteny is the highest. This number of marked apical vesicles and colloid is more important in the neotenics which have lost some larval characters. Silver grains on colloid droplets indicating thyroid hormone excretion are inexistent in the most larval neotenics, more numerous in most metamorphosed neotenics. But they always are less numerous than in metamorphosing larvae. For a few animals, the fixation curve is very high, probably because of a stimulation of thyroid function, induced by the test and not because of a goitrous tendancy, as radioautographic observation of their thyroid shows. Thyroid function and its relation with the origin of this total and accidental neoteny are discussed.

Aging↗
PubMed1982

[Ultrastructural localization of dopaminergic receptors in the rat corpus striatum by radioautography using tritiated domperidone].

An attempt to localize dopaminergic receptors in the rat neostriatum by high-resolution radioautography was realized using intracerebral injections of the new ligand (antagonist) 3H-domperidone. In tissue regions located far from the injection site, the weaker diffuse radioautographic reaction permitted us to observe the existence of clusters of silver grains over some cerebral structures. The specificity of this type of labelling was tested using intraperitoneal injections of large amounts of haloperidol in order to block the studied receptors. Thus, we observed specific labelling over some nerve terminals as well as a low number of synaptic contact of the asymmetric type (however some synapses of the symmetric type were also labelled). These result agree with our previous work (1), and confirm the existence of dopaminergic synapses in the rat caudate-putamen.

Animals↗
PubMedDate not supplied

[Mast cell origin studied by H3-thymidine radioautography].

Reproduction of mast cells has been investigated by means of H3-thymidine radioautography in 26 mature mice of CC-57 White line and in 40 new-born nonlinear albino rats under normal conditions and experimental cold effect. It is established that differentiated mast cells incorporate H3-thymidine and can enter the phase of DNA synthesis of a proliferative cycle.

Animals↗
PubMed1984

Differences in d-[3H]lysergic acid diethylamide binding in mouse cortex and hippocampus in vivo and in vitro revealed by radioautography and rapid filtration studies.

The localization of d-[3H]lysergic acid diethylamide ([3H]LSD) binding sites in mouse brain was compared in vivo and in vitro. Radioautography of brain sections incubated with 6 nM [3H]LSD in vitro revealed substantial specific binding in cortex (CTX), especially in layers III to IV and anterior cingulate gyrus, and in areas CA1 and dentate gyrus of hippocampus (HIP). In sections of brains from mice that received 100 nmol of [3H]LSD per kg and were killed 10, 15 or 30 min later, specific [3H]LSD binding in CTX had a pattern of distribution similar to that observed in vitro. In contrast, the pattern of specific [3H]LSD binding in HIP in vivo differed from the results obtained in vitro, in that it was sparse and lacked differential subregional distribution. The low specific [3H]LSD binding in vivo in HIP but not in CTX was confirmed by homogenate filtration studies of brain areas from mice that received 100 nmol of [3H]LSD per kg. The levels of free [3H]LSD, obtained after correction for time-dependent metabolism of [3H]LSD, did not vary among regions, but [3H]LSD specifically bound in HIP was 30 to 50% of that in CTX. In contrast, steady-state binding studies in vitro in membrane preparations from CTX and HIP demonstrated a similar density and affinity of [3H]LSD binding sites in the two regions. Comparison of [3H]LSD binding characteristics in vivo and in vitro suggests possible mechanisms causing the lower specific binding in HIP in vivo, including modulation of the binding sites that differ in CTX and HIP.

Animals↗
PubMed1983

Dividing and newly produced cells in the corpus callosum of adult mouse cerebrum as detected by light microscopic radioautography.

New cell production in the corpus callosum and subependyma of 4 month old mice was analysed by light microscopic radioautography after 3H-thymidine injections. In the subependymal region of the lateral ventricle, about 10% of cells were labeled in mice examined 2 h after 3H-thymidine, and 40 to 50% of cells were labeled after 7 d of isotope infusion. In corpus callosum of mice 2 h after precursor injection, the few cells (0.1 to 0.2%) that were labeled had the appearance of "immature cells", and were presumably incompletely-differentiated neuroglial precursor cells which were preparing to divide. After 7 d of continuous 3H-thymidine administration, more labeled neuroglia (about 2%) were detected in corpus callosum; these newlyproduced cells included several astrocytes and some oligodendrocytes, as well as immature cells. Since the immature cells were the most frequently observed type of dividing cell within the normal adult corpus callosum, it is probable that the new astrocytes and oligodendrocytes were the products of divisions of their respective precursor cells.

Animals↗
PubMed1982

The fate of newly synthesized proteins in nerve and glial cells following nerve injury as shown by electron microscope radioautography.

Following peripheral crush of hypoglossal nerve the fate of newly synthesizing proteins in corresponding nerve and glial cells was traced out using light and high resolution radioautography after intraventricular injection of 3H-leucine. Radioautographic analysis was applied on the normal and chromatolytic neurons and their glial surroundings 45 min after injection of labelled precursor. Both chromatolytic neurons and surrounding glia showed higher incorporation of labelled proteins than those of control nerve and glial cells. Relative specific radioactivity (RSR) of ergastoplasm and lysosomes were increased in the chromatolytic neurons, whereas RSR of nuclei, mitochondria and Golgi apparatus were decreased. Contrary to neurons, RSR of corresponding compartments in experimental glial cells except lysosomes showed reciprocal values in comparison with control glial cells. The higher RSR of mitochondria and Golgi apparatus in experimental glial cells depended mostly on the astrocytes, whereas those of nuclei and lysosomes were related first of all with microglial cells. These findings are correlated with present concepts of the complex interdependency between neurons and glia in which mutual regulatory controls and influences are exerted.

Animals↗
PubMed1981

[3H]glucosamine and [3H]proline radioautography of embryonic mouse dental basement membrane.

[3H]proline and [3H]glucosamine radioautography was performed to analyze the labeling pattern of mouse embryonic dental basement membrane before and during odontoblast terminal differentiation. Sixteen- and eighteen-day-old first lower molars and trypsin-isolated enamel organs, as well as EDTA-isolated dental papillae, were used. Continuous labeling for 12 to 24 hr was required with [3H]proline to obtain a clear labeling of epithelial-mesenchymal junction in intact tooth germs or accumulation of surface label in trypsin-isolated enamel organs. With [3H]glucosamine, after 6-hr labeling, the epithelial-mesenchymal junction was heavily labeled and the trypsin-isolated enamel organs accumulated substantial amounts of surface label, corresponding to the redeposited basement membrane. At Day 16 stage, these labels always had a uniform distribution and decreased during chase without any redistribution. At Day 18 stage, when the terminal differentiation of odontoblasts occurred the label accumulated in a unique pattern: much more label was at the epithelial surface corresponding to the top of the cusps than in the apical parts. During chase and only in intact tooth germs epithelial surfaces which had labeled poorly during pulse became labeled, but those labeling heavily during pulse lost label. This pattern existed only in the presence of mesenchyme. EDTA treatment of [3H]glucosamine-labeled teeth enabled us to obtain isolated dental papillae with surface label. Distribution of this label was exactly the same as that for the epithelial-mesenchymal junction of intact teeth. During chase, these dental papillae completely lost the surface label. The mesenchyme seen to control the synthesis and/or the degradation of epithelially derived [3H]glucosamine-labeled material.

Animals↗
PubMed1995

Nucleic acid synthesis in the developing mouse ovary, uterus and oviduct studied by light and electron microscopic radioautography.

The DNA and RNA synthesis in the developing mouse ovary and oviduct were studied by 3H-thymidine and 3H-uridine radioautography. The results showed that both DNA and RNA syntheses were active in all surface epithelial cells, in stromal and follicular cells of the ovaries between postnatal days 1 to 7; then they decreased significantly from day 14 on. However, the DNA synthesis in the epithelial and in the stromal cells of the uteri and oviducts was active at days 1 and 3 and decreased from day 7. The RNA synthesis in the uteri and oviducts was active at postnatal day 1, increased from postnatal day 1 to day 14, decreasing between days 30 to 60. An unparalleled alternation of the DNA and RNA synthesis was shown between the ovary and uterus or oviduct. After postnatal day 3, the RNA synthesis was more important in the uterus and oviduct than in the ovary.

Animals↗
PubMed1995

Study on the aging changes of DNA and protein synthesis of bipolar and photoreceptor cells of mouse retina by light and electron microscopic radioautography.

The DNA and protein synthesis of the bipolar cells and photoreceptor cells of aging mouse retina from fetal to postnatal 1 year were studied by light and electron microscopic radioautography. Meanwhile the ultrastructure of these cells was observed by electron microscopy. The bipolar and photoreceptor cells incorporated 3H-thymidine from embryonic day 19 until 1 week after birth, while 3H-leucine from embryonic day 19 to postnatal 1 year. The 3H-thymidine labelled cells had undeveloped cell organelles in their cytoplasm. On the contrary, the unlabelled cells had well developed cell organelles, mainly mitochondria and endoplasmic reticulum in their cytoplasm. The mitochondria became elongated when the outer segments were well developed. The protein synthesis of the bipolar cells and photoreceptor cells as demonstrated with 3H-leucine labeling was vigorous at embryonic day 19 and early postnatal days when the inner and outer segments were formed. The peak of this phenomenon was observed at 1 day after birth. The activity of protein synthesis was also detected at the adult stage. However there was no significant fluctuation in any stages from 14 days after birth to 1 year.

Aging↗
PubMed1993

Study on the macromolecular synthesis in aging mouse seminiferous tubules by light and electron microscopic radioautography.

Morphologic alternation and the changes of DNA, RNA and protein syntheses of the seminiferous tubules of aging mice were studied by light and electron microscopic radioautography. The morphology varied characteristically during the developmental periods. At embryonic and neonatal stages, the labelled myoid cells and Sertoli cells with 3H-thymidine were well observed, while at adult period, the labeling indices of both cells decreased to low levels. In contrast to this, the radioautograms showed that the gonocytes did not incorporate the labelled thymidine at embryonic day 19. As development proceeded to the 4th day after birth, the activity of DNA synthesis of gonocytes started. The labeling indices of spermatogonia reached the first peak at 3 weeks and were nearly constant thereafter. On the other hand, the activity of RNA synthesis of the cells in seminiferous tubules was not prosperous at embryonic day 19 and the neonatal stage. It was vigorous at adult and maintained a relatively high level at old period. The aging change of protein synthesis was detected in all kinds of the cells one hour after the 3H-leucine labeling. These results indicated that the old mice still kept adequate spermatogenesis. The Sertoli and myoid cells may play an important role in spermatogenesis in the whole life.

Aging↗