Demonstration of double-labelled branched neurons in the CNS of the rat by retrograde axonal transport of iron-dextran complex and HRP. An application to epoxy-embedded material.
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Biomedical subjects
Publications and source records attributed to B Berger.
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While instituting a new program in fetoscopy, the authors explored the effects of sedation with meperidine (Demerol) and promethazine HCl (Phenergan) during fetoscopy conducted on 10 mothers at 14 to 18 weeks' gestation. Each received 100 mg of meperidine and 50 mg of promethazine HCl approximately 30 minutes before the procedure. In 3 instances, a repeat half-dose injection was made at 30 minutes for adequate analgesia. Sex of the fetus was determined accurately in 9 instances. In 9 of 10 cases, visualization was satisfactory; technical problems in 1 case precluded visualization. Complications included 2 superficial fetal limb hematomas and 2 instances of minimal fetal bleeding from an unknown site. Three patients required perforation of anterior placentas. Amniotic fluid was invariably clear but became progressively turbid. Sedation allowed a longer period for clear amniotic fluid by decreasing fetal movements progressively over the first hour; fetal activity showed a prompt return thereafter. The clarity of the amniotic fluid was adversely influenced by external manipulation of the fetus. The authors conclude that sedation has obvious benefits for the mother and may facilitate evaluation of the fetus during fetoscopy.
A new radioautographic technique for the visualization of central dopaminergic fibers is described. This technique is based upon in vitro uptake of triatiated norepinephrine or tritiated dopamine in the presence of monoamine oxidase inhibitor and specific uptake inhibitor in noradrenergic fibers. After a short step in glutaraldehyde, the use of osmium vapours as a fixative, instead of osmic acid immersion, allows a satisfactory retention of triatiated amines within dopaminergic fibers.
The retrograde axonal transport of an iron-dextran complex was observed in neurons of the substantia nigra and of the intralaminar nuclei of the thalamus, after previous injection into the striatum. The histochemical demonstration of iron is simple and rapid, and can be combined with that of horseradish peroxidase, under precise conditions in the sequence of reactions. The iron-dextran complex revealed to be a valuable material for neuronal connectivity studies in the central nervous system.
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The retrograde axonal transport of an iron-dextran complex leads to a labelling of neural cell bodies in the central nervous system (CNS) of the rat. This tracer and horseradish peroxidase (HRP) can both be demonstrated histochemically in same cell bodies of intralaminar thalamic neurons in the central lateral nucleus, after injection of iron-dextran in the striatum and injection of HRP in the motor cortex. This is made possible by processing the sections first for HRP and then for ferric ions by Perl's reaction. This method allows an accurate demonstration of divergent axonal projections and is compatible with cytoarchitectonic studies on the same sections.
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Long-term survival of mesencephalic and striatal cells from mouse embryos in dissociated primary cultures is described. Catecholaminergic neurons in mesencephalic culutres were identified histochemically and by measuring [3H]dopamine uptake and synthesis from [3H]tyrosine. According to experiments using specific inhibitors of catecholamine uptake, at least two-thirds of the catecholaminergic neurons are dopaminergic. These neurons differentiated whether or not striatal target cells were present, but striatal cells stimulated the development of the dopaminergic neurons. [3H]Dopamine uptake was increased by at least 2-fold regardless of the age of the cocultures (4-15 days). Enhanced [3H]dopamine synthesis was also observed (at least 2-fold) at later times (12-15 days).
A pigmented fundus lesion in a 47-year-old woman gave a positive 32P test result and angiography showed a separate tumor circulation suggesting a diagnosis of choroidal melanoma. Examination after local resection of the tumor revealed massive retinal gliosis, a rare condition not normally considered in the differential diagnosis of choroidal melanoma. We review the clincial and histologic features that differentiate massive retinal gliosis from choroidal melanoma and astrocytic hamartoma.
We treated a patient with a subacute subdural hematoma who manifested only a complete third cranial nerve paralysis and decreased visual acuity. The gravity of delayed diagnosis in such a situation should prompt the ophthalmologist to consider subdural hematoma as the underlying cause in cases of third nerve paralysis.
The dissecting procedure and tissue culture technique on collagen coated coverslips in Leighton tubes, of newborn Mouse nucleus locus coeruleus and the regular histofluorescence demonstration of catecholaminergic fibers in the outgrowth zone are described. This simple method allows 10 day--long experiments .
An in vitro method was developed to separately visualize dopaminergic, noradrenergic and serotoninergic terminals in the cerebral, hippocampal and cerebellar cortices of the rat, by means of the glyoxylic histochemical fluorescence method. Animals were pretreated with alpha-methylparatyrosine to deplete catecholamine stores. Thin vibratome sections were made and incubated in the presence of various exogenous amines and inhibitors of the catecholaminergic and serotoninergic transport systems. Experimental conditions insuring the best specificity were determined and the validity of the combined pharmacological and histochemical approach was further tested by using animals in which the cortical dopaminergic, noradrenergic or serotoninergic innervations were destroyed. Under the experimental conditions used, norepinephrine as well as alpha-methyl-norepinephrine were taken up in only noradrenergic and dopaminergic terminals. A separate visualization of the two systems was obtained by using specific uptake inhibitors. Dopamine was taken up not only in catecholaminergic but also in serotominergic terminals. The uptake of DA in serotoninergic fibers was inhibited by a specific inhibitor of the serotoninergic transport system or by the presence of serotonine in the incubating medium. The signification and the implications of thes results are discussed.
The demonstration of fluorescent catecholamines and horseradish peroxidase (HRP) in the same neuron has been achieved in the Rat in two ways: by submitting vibratome sections to a modified glyoxylic acid fluorescence method followed by the usual procedure to reveal HRP; or by combining the last procedure with the cryostat technique of Chiba et coll. After HRP injection into the striatum or the nucleus accumbens of the Rat, non-fluorescent HRP labelled neurons were observed in the substantia nigra and the ventral tegmental area respectively, in addition to the HRP labelled fluorescent dopaminergic neurons.
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This report documents the second case of an ovarian pregnancy with a Cu-7 intrauterine contraceptive device. All of Spiegelberg's criteria were satisfied. A summary of the literature is presented, revealing 33 ovarian pregnancies in users of an IUD. The overall frequency of primary ovarian pregnancy is 1:40,000 deliveries. There is a ratio of one ovarian pregnancy to nine ectopic pregnancies among IUD users, compared to 1:150 to 1:200 in the general population.
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After the destruction of the noradrenergic ascending pathways, the localization of frontal cortical fields receiving fibers from the dopaminergic mesocortical system has been studied in rats, using a glyoxylic-paraformaldehyde method. The dopaminergic innervation was distributed in two main areas. The area of highest density was a medial field which spread in the medial cortex anterior and dorsal to the genu of the corpus callosum. It did not reach the shoulder region except in the foremost part of the frontal lobe where dopaminergic fibers were scattered in the whole cortex, including the molecular layer. A deep sulcal field was situated between the dorsal bank of the rhinal sulcus and the lateral cortex above it. In addition, a moderately dense band of dopaminergic fibers was observed between the corpus callosum and the anterior commissura, beside the accumbens nucleus. Similar data were obtained with dopamine uptake experiments on reserpine-treated but otherwise normal animals. The frontal areas receiving dopaminergic innervation coincide strikingly with the 'prefrontal cortex' as defined by neuroanatomical studies, which is assumed to be more or less equivalent to the prefrontal cortex of primates and derives direct projections from the amygdala. The functional implications of these findings are discussed.