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Biomedical subjects

A Minelli

Publications and source records attributed to A Minelli.

At least 73 records · Page 4Linked to original sources
PubMed1994

Cellular localization and laminar distribution of AMPA glutamate receptor subunits mRNAs and proteins in the rat cerebral cortex.

The cellular and laminar distributions of the alpha-amino-3-hydroxy-5- methyl-4-isoxazole propionate (AMPA) receptor subunits GluR1-4 have been investigated in the cerebral cortex of adult rats by in situ hybridization with 35S-labeled cRNA probes and by immunocytochemistry with subunit-specific antibodies. In sections incubated with the GluR1-4 antisense probes, specific hybridization signal was observed in many but not all cortical cells. Experiments with in situ hybridization and antibodies to glial fibrillary acidic protein (GFAP) showed that percentages of GFAP-immunoreactive cells labeled by the GluR1-4 probes were 20%, 9.4%, 8.2%, and 57.3%, respectively. A semiquantitative evaluation revealed that about 56% of cortical neurons contained the GluR1 subunit, 80% the GluR2, 63% the GluR3, and 44% the GluR4. The number of grains associated with every neuron was determined from sections exposed for 15 days, the background level was subtracted, and labeled neurons were divided into four groups: A (< or = 10 grains), B (11-20 grains), C (21-30 grains), and D (> 30 grains). The number of neurons belonging to each of these groups was then evaluated for their occurrence in each cortical layer. Immunocytochemistry with subunit-specific antibodies showed that 1) GluR1-immunoreactive neurons were mostly layers V and VI nonpyramidal neurons; 2) GluR2/3-immunoreactive neurons were more numerous in layers II-III and V-VI, and most of them were pyramidal; and 3) GluR4-positive cells were the least numerous, and they were either neurons (pyramidal and nonpyramidal) or astrocytes. These observations indicate that cortical neurons exhibit a remarkable degree of heterogeneity with regard to both the composition and the number of AMPA receptors and suggest that this diversity might be correlated with the functional attributes of neurons receiving glutamatergic afferents and with the physiological features of corticifugal neurons.

Animals↗
PubMed1994

Cellular localization and laminar distribution of NMDAR1 mRNA in the rat cerebral cortex.

N-methyl-D-aspartate (NMDA) receptors, which play a critical role in many cortical functions, are composed of a heteromeric assembly of different subunits: of these, the NMDA receptor subunit 1 (NMDAR1) is a constant component of, and thus an excellent marker for, NMDA receptors. In this study, we have investigated the cellular localization and laminar distribution of NMDAR1 mRNA in the cerebral cortex of adult rats by in situ hybridization histochemistry with a 35S-labeled cRNA probe. Specificity and background levels were determined in adjacent sections incubated with a 35S-labeled sense RNA. In sections incubated with the antisense RNA probe, specific hybridization signal was observed in a large number of cells. Some cells, however, did not appear to contain NMDAR1 mRNA. The vast majority of these unlabeled cells were small, suggesting that they are astrocytes or other small nonneuronal cells. Double-labeling studies with in situ hybridization histochemistry and immunocytochemistry with antibodies to glial fibrillary acidic protein (GFAP) showed that about 95.7% of the GFAP-positive cells did not express NMDAR1 mRNA, indicating that virtually all astrocytes do not contain this transcript. A semiquantitative evaluation of cortical neurons, defined as those cells larger than the GFAP-positive astrocytes, revealed that about 80% were associated with silver grains. The number of silver grains associated with every neuron was determined from sections exposed for 15 days, the background level was subtracted, and all labeled neurons were grouped into five groups: A (< or = 10 grains), B (11-20 grains), C (21-30 grains), D (31-40 grains), and E (> 40 grains). The number of neurons belonging to each group was then evaluated according to their occurrence in each cortical layer. In layer I all labeled neurons were in group A, whereas in layers II-III and V-VI positive neurons were in group A-E. In layer IV most neurons were in groups A and B, whereas only a few were in group E. These observations indicate that 1) virtually all cortical cells containing NMDAR1 mRNA in adult rats are neurons; 2) about 80% of all cortical neurons express NMDAR1 mRNA; and 3) labeled neurons can be divided into several groups on the basis of NMDAR1 mRNA levels expressed, which presumably reflect the number of NMDA receptors. The existence of neurons with a different number of receptors may be a critical factor for determining the physiological effect of NMDA receptor activation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Topographical relations between ipsilateral cortical afferents and callosal neurons in the second somatic sensory area of cats.

Experiments were carried out on the second somatic sensory area (SII) of cats to study 1) the laminar distribution of axon terminals from the ipsilateral first somatic sensory cortex (SI); and 2) the topographical relations between their terminal field and the callosal neurons projecting to the contralateral homotopic cortex. To label simultaneously in SII both ipsilateral cortical afferents and callosal cells, cats were given iontophoretic injections of Phaseolus vulgaris-leucoagglutinin (PHA-L) in the forepaw zone of ipsilateral SI, and pressure injections of horseradish peroxidase (HRP) in the same zone of contralateral SII. The possibility that ipsilateral cortical axon terminals synapse callosal neurons was investigated with the electron microscope by combining lesion-induced degeneration with retrograde HRP labelling. Fibers and terminations immunolabelled with PHA-L from ipsilateral SI were distributed in SII in a typical patchy pattern and were mostly concentrated in supragranular layers. Labelled fibers formed a very dense plexus in layer III and ramified densely also in layers I and II. Labelled axon terminals were both en passant and single-stalked boutons. Counts of 8,303 PHA-L-labelled terminals of either type showed that 82.40% were in supragranular layers. The highest concentration was in layer III (43.99%), followed by layers II (30.32%) and I (8.09%). The remaining terminals were distributed among layers IV (6.96%), V (4.93%), and VI (5.68%). The same region of SII containing anterogradely labelled axons and terminals also contained numerous neurons retrogradely labelled with HRP from contralateral SII. Callosal projection neurons were pyramidal, dwelt mainly in layer III, and were distributed tangentially in periodic patches. Patches of anterograde and retrograde labelling either interdigitated or overlapped both areally and laminarly. In the zones of overlap, numerous PHA-L-labelled axon terminals were seen in close apposition to HRP-labelled pyramidal cell dendrites. Combined HRP-electron microscopic degeneration experiments showed that in SII axon terminals from ipsilateral SI form asymmetric synapses with HRP-labelled dendrites and dendritic spines pertaining to callosal projection neurons. These results are discussed in relation to the layering and function of the SI to SII projection, and to the evidence that SII neurons projecting to the homotopic area of the contralateral hemisphere have direct access to the sensory information transmitted from ipsilateral SI.

Animals↗
PubMed1994

Commissural connections of the cat periaqueductal gray matter studied with anterograde and retrograde tract-tracing techniques.

The commissural connections of the periaqueductal gray matter were investigated by light and electron microscopy by using the anterograde tracer Phaseolus vulgaris leucoagglutinin and the retrograde tracer horseradish peroxidase. In the first group of seven animals (1-7), single injections of Phaseolus vulgaris leucoagglutinin were performed iontophoretically (4.5 microA for 30 min) into various subdivisions of the periaqueductal gray matter. On light microscopic examination, injection sites were characterized by several immunolabeled neurons of different sizes and morphology, with the cytoplasm, nucleus and neuronal processes intensely stained. Many labeled fibers turned from injection sites toward all contralateral periaqueductal gray matter subdivisions, but anterograde labeling was densest in the regions homotopic to those injected. Commissural fibers bore along their course many en passant boutons of different sizes and morphology, and gave off spine-like processes, at the end of which one terminal bouton was observed. Labeled fibers branched into numerous collaterals which ended in a terminal array of 10-20 en passant and en grappe boutons. At the electron microscopic level, commissural axons were observed in close proximity to the cytoplasmic membranes of cells. Axon terminals formed symmetric or asymmetric synapses mainly on dendritic shafts of neurons and rarely on vesicle-containing profiles. Horseradish peroxidase experiments were carried out in four cats (1-4). The tracer was injected iontophoretically into different regions of the periaqueductal gray matter of three cats (1-3). Retrogradely labeled neurons giving rise to commissural connections had a morphology similar to that of polygonal, triangular and fusiform cells described in previous Golgi studies. The perikaryal cross-sectional area of commissural neurons was smaller than that of neurons projecting outside the periaqueductal gray matter (mean value of commissural neurons 149.77 microns 2 vs 261.19 microns 2 for projecting neurons), which were retrogradely labeled by pressure-injecting horseradish peroxidase into several targets of periaqueductal gray matter (4). Moreover, since the distribution of sizes of the two populations of the periaqueductal gray matter overlapped in the range of 90-300 microns 2, a considerable number of projecting neurons were as small as commissural neurons. The present results suggest that commissural fibers could reciprocally connect zones of the periaqueductal gray matter with similar functions, and originate from small and medium-sized neurons, some of which are also projecting neurons.

Animals↗
PubMed1994

Glutamate immunoreactivity in rat cerebral cortex is reversibly abolished by 6-diazo-5-oxo-L-norleucine (DON), an inhibitor of phosphate-activated glutaminase.

Glutamate (Glu) immunocytochemistry has been widely used to identify presumed gluergic neurons and synapses, but several problems related to the fact that Glu is both a synaptic transmitter and a compound used for metabolic purposes are still unsolved. One of these concerns the intense perikaryal staining observed in perfusion-fixed tissue. Phosphate-activated glutaminase, a key enzyme for the synthesis of releasable glutamate, is inhibited by the diazoketone 6-diazo-5-oxo-L-norleucine (DON), which greatly reduces glutamate release. In the present experiments, DON was either injected intraparenchymally or applied epipially to the sensorimotor cortex of adult Sprague-Dawley rats at concentrations of 0.25-1 mM. Both intraparenchymal and epipial applications of the chemical abolished Glu immunoreactivity in neuron perikarya. Adjacent sections processed for cytochrome oxidase histochemistry, for aspartate immunoreactivity, or stained with thionine showed no changes. The effects of DON application are reversible, as shown in a second series of experiments in which, after 30 min of DON application, animals were allowed to survive for 5-10 days. In these cases, Glu immunoreactivity in cortical neurons was identical to that observed in normal untreated animals. The results reported here suggest that Glu immunoreactivity demonstrated by the present procedure in neuron perikarya is mainly due to Glu produced via phosphate-activated glutaminase.

Animals↗
PubMed1993

D8S7 is consistently deleted in inverted duplications of the short arm of chromosome 8 (inv dup 8p).

Ten patients with inverted duplication of 8p (inv dup 8p) were studied with cytogenetic, biochemical and molecular techniques. The duplication for the region 8p12-p22 was always associated with a deletion of the locus D8S7 (mapped in 8p23.1) as demonstrated with the probe pSW50 by both in situ hybridization and Southern blot. Restriction fragment length polymorphisms detected by probes pSW50 (1 case) and by pG2LPL35 (locus LPL) (two cases) were informative as to a maternal origin of the anomaly. The activity of glutathione reductase, whose gene maps in the duplicated region at 8p21.1, was increased in all patients. The recognizable phenotype of inv dup 8p includes neonatal hypotonia, prominent forehead, large mouth with everted lower lip, abnormally shaped large ears, brain malformations and severe mental retardation. Our findings indicate that the chromosome rearrangement is homogeneous at least for the presence of the deletion and support the hypothesis of a common mechanism of origin.

Abnormalities, Multiple↗
PubMed1993

Effects of adenine nucleotides on low Km 5' nucleotidase from human seminal plasma.

1. Low Km 5' nucleotidase purified from human seminal plasma has been used in this study to investigate the response of the enzyme ot adenine nucleoside di- and triphosphates in the presence of AMP and IMP as substrates. 2. In the presence of AMP, the addition of 0.5 mM ATP to the enzyme Mg-free results into the highest Vmax/Km ratio value and other experimental combinations of effectors tested cause variation of the kinetic parameters of the enzyme, indicating a control of AMP dephosphorylation by adenine nucleotides. 3. In the presence of IMP, ATP and ADP activate the enzyme but the response to various experimental combinations of effectors shows no significant difference in the kinetic properties of the enzyme, indicating a different control of the dephosphorylation of IMP.

5'-Nucleotidase↗
PubMed1992

Numerous SP-positive pyramidal neurons in cat neocortex are glutamate-positive.

An immunocytochemical technique that allows visualization of two antigens in the same neuron was used to verify the possibility that some neocortical pyramidal neurons contain both glutamate (Glu) and substance P (SP) immunoreactivity. The results show that a large fraction of SP-positive pyramidal neurons are also Glu-positive, and indicate that in a small population of cortical neurons a fast excitatory synaptic transmitter and a slow peptidic modulator coexist.

Animals↗
PubMed1992

Prenatal diagnosis of metabolic diseases on chorionic villi obtained before the ninth week of pregnancy.

Nine pregnancies at risk for various metabolic disorders were monitored by prenatal diagnosis on chorionic villi obtained between the sixth and ninth weeks of pregnancy. A diagnosis of an affected fetus was made in five cases (Sandhoff, Tay-Sachs (2), Pompe's, GM1), while metachromatic leukodystrophy, GM1 (2), and Pompe's were excluded in four cases. It is concluded that chorionic villi are a reliable tissue for prenatal diagnosis of metabolic disorders also when obtained before the ninth week.

Cerebroside-Sulfatase↗
PubMed1992

A homozygous missense arginine to histidine substitution at position 482 of the beta-galactosidase in an Italian infantile GM1-gangliosidosis patient.

We have studied, by the polymerase chain reaction, the beta-galactosidase cDNA from several Italian patients with infantile GM1-gangliosidosis. One homozygote for a previously undiscovered G > A mutation at position 1479, causing an arginine to histidine change, was detected. The same mutation, in heterozygosis, was identified in 6 unrelated patients, but not in 100 normal chromosomes.

Arginine↗
PubMed1992

Metachromatic leukodystrophy: on an atypical case.

We describe an atypical case of juvenile metachromatic leukodystrophy. Motor conduction velocity was still within the normal range 3 years after clinical onset, in contrast to what is commonly found in this disease. Another unusual feature is the normal level of CSF protein. These data are discussed in the light of the sural nerve biopsy findings, which revealed only slight impairment.

Child↗
PubMed1992

Thalamic connections of the second somatic sensory area in cats studied with anterograde and retrograde tract-tracing techniques.

The thalamic connections of the second somatosensory area in the anterior ectosylvian gyrus of cats have been investigated using the retrograde tracer horseradish peroxidase and the anterograde tracer Phaseolus vulgaris leucoagglutinin. Horseradish peroxidase was injected iontophoretically in several somatotopic zones of the second somatosensory area map of six cats. Sites of horseradish peroxidase delivery were identified preliminarily by recording with microelectrodes the responses of neurons to skin stimulation. Phaseolus vulgaris leucoagglutinin was iontophoretically injected within the ventrobasal complex (one cat) or in the posterior complex (one cat). Horseradish peroxidase injections into cytoarchitectonic area SII retrogradely labeled neurons in the ipsilateral ventrobasal complex and in the posterior complex. Counts of labeled neurons from the ipsilateral thalamus showed that the overwhelming majority of horseradish peroxidase-labeled neurons were in the ventrobasal complex (96.3-96.9%) and few were in the posterior complex (3.1-3.7%). Neurons labeled in the ventrobasal complex were observed throughout the anteroposterior extent of the nucleus, while their mediolateral distribution varied with the site of horseradish peroxidase delivery in the body map of the second somatosensory area, which indicates that the projections from the ventrobasal complex to the second somatosensory area are somatotopically organized. In the cat in which the horseradish peroxidase injection involved both the second somatosensory area proper and the second somatosensory area medial, which lies in the lower bank of suprasylvian sulcus, labeled neurons were almost as numerous in the ventrobasal complex as in the posterior complex. Phaseolus vulgaris leucoagglutinin injected in the ventrobasal complex anterogradely labeled thalamocortical fibers in the ipsilateral anterior ectosylvian gyrus. In this case, patches of labeled fibers and terminals were distributed exclusively within the cytoarchitectonic borders of the second somatosensory area proper. Labeled terminals were numerous in layer IV and lower layer III, but terminal boutons and fibers with axonal swellings, probably forming synapses en passant, were frequently observed also in layers VI and I. Injection of Phaseolus vulgaris leucoagglutinin in the posterior complex labeled thalamocortical fibers in two distinct regions in the ipsilateral anterior ectosylvian gyrus, one lying laterally and the other medially, which correspond, respectively, to the fourth somatosensory area and the second somatosensory area medial. In both areas the densest plexus of labeled fibers and axon terminals was in layer IV and lower layer III, but numerous labeled fibers and terminals were also observed in layer I. In this case, only rare fragments of labeled fibers were present in second somatosensory area proper, but no labeled terminals could be observed.

Animals↗
PubMed1991

Purification and partial characterization of the soluble low Km 5'-nucleotidase from human seminal plasma.

Soluble low Km 5'-nucleotidase from human seminal plasma has been purified to homogeneity by one affinity and two gel-filtration chromatographic steps. The pure enzyme had a specific activity of 2000 nmol min-1 mg-1. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of purified low Km 5'-nucleotidase revealed a single polypeptide band of 40 +/- 7 kDa and a tetrameric structure of 160 +/- 10 kDa has been proposed for the native enzyme. The kinetic properties of low Km 5'-nucleotidase have been determined and rather unique characteristics have been found for this soluble low Km 5'-nucleotidase: the substrate efficiency was slightly higher for IMP with an optimum pH at 7.5; the enzyme showed an absolute dependence on Mg2+ ions. Ca2+ could replace Mg2+ ions for activity while other divalent cations could not substitute for Mg2+; the enzymes were equally activated by ATP and ADP up to 0.1 mM concentrations. At higher concentrations up to 1 mM, ADP was still an activator while ATP caused a gradual decrease of activation to the native activity. This effect could not be related to the Mg-ATP = complexes since the enzymic preparation Mg(2+)-free still showed the same biphasic pattern of activation.

5'-Nucleotidase↗
PubMed1991

Self-similarity in biological classifications.

Size distributions of supraspecific taxa, e.g. genera, measured as the number of included subtaxa, e.g. species, are found to follow a power law. This behaviour has been verified for a number of taxa of different size and taxonomic rank, thus suggesting a fractal structure of biological classifications. This is regarded as probably dependent on evolutionary processes shaping the phylogenetic tree, especially speciation and extinction, as well as on the topological properties of developmental constraints and/or of the ecospace(s) with which the group has been confronted during its history. The role of taxonomic bias is deemphasized.

Animals↗
PubMed1990

Gene dosage effect in acquired monosomy 7: distinct behaviour of beta-glucuronidase and phosphoserine phosphatase.

Evidence for gene dosage effect for beta-glucuronidase (GUSB) and phosphoserine phosphatase (PSP), whose genes are mapped on chromosome 7, was searched in a group of 13 patients with myeloproliferative disorders and acquired monosomy 7. The monosomy 7 was the sole anomaly in nine patients and was associated with other chromosome changes in four. A group of 19 patients with similar diseases but with normal karyotype or with anomalies not involving chromosome 7 served as control. beta-galactosidase and arylsulphatase A, whose genes are not on chromosome 7, were tested as control enzymes. We obtained evidence for a gene dosage effect for GUSB, but not for PSP. When all cases with monosomy 7 were compared with controls, no dosage effect was observed for PSP, but when this group was split into two, according to the presence of anomalies additional to the monosomy 7, the values of activity in the group with additional anomalies were significantly lower than in the controls. Thus, in the case of PSP, the loss of one allele is not followed immediately by reduction in activity, and this could be due to the specific importance of PSP in nucleic acid metabolism. We postulate that some regulatory mechanisms are able to keep normal levels of PSP even in the presence of only one allele, and that they are overwhelmed only when additional chromosome changes are present. These changes tend to involve chromosomes carrying genes for enzymes involved in a metabolic pathway closely related to PSP functions, and only then is a gene dosage effect for PSP detectable.

Alleles↗
PubMed1990

Involvement of the region 13q14 in a patient with adamantinoma of the long bones.

A cytogenetic analysis of a lung metastasis of an adamantinoma of the tibia, a rare tumor of the long bones, revealed a karyotype 52, XY, t(7;13)(q32;q14), +7, +12, +13, +19, +der(7) t(7;13), +der(13) t(7;13). The t(7;13) was found to be constitutional in the patient and was also present in peripheral blood lymphocytes of his unaffected father. However, both subjects displayed normal levels of esterase D enzyme activity.

Ameloblastoma↗