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B Bloch

Publications and source records attributed to B Bloch.

At least 127 records · Page 7Linked to original sources
PubMed1992

Phenotypical characterization of the rat striatal neurons expressing muscarinic receptor genes.

Neurons expressing the m1, m2, and m4 muscarinic receptor genes in the adult rat striatum were identified and characterized by using several in situ hybridization and immunohistochemical procedures. Combined in situ hybridization for the simultaneous detection of two mRNAs in the same section or in adjacent sections as well as in situ hybridization and immunohistochemistry on adjacent sections permitted us to identify the neurons containing m1, m2, or m4 receptor mRNA. Our observations demonstrate that m1, m2, and m4 receptor genes are expressed in one or several phenotypically distinct neuronal populations. The m1 receptor gene was the most widely expressed (85% of the striatal neurons). Most cholinergic neurons (80% or more) contain m1, m2, and m4 receptor mRNAs. Almost all the substance P neurons contain m1 and m4 receptor mRNA. All enkephalinergic neurons contained m1 receptor mRNA, but only 39% contained m4 receptor mRNA. Most somatostatin and neurotensin neurons expressed the m1 receptor gene, but only a few (15% and 9%, respectively) contained m4 receptor mRNA. The present study offers anatomical evidence that ACh may act directly in complex ways on the main neuronal populations of the striatum through muscarinic receptors. The m1, m2, and m4 receptors may act as autoreceptors to control ACh release and possibly other parameters of ACh neurons. On the other hand, the m1 and m4 receptors may act as heteroreceptors in cholinoceptive efferent neurons (enkephalin and substance P neurons) and other neurons (somatostatin/neuropeptide Y and neurotensin neurons). The presence of m4 receptor mRNA in only parts of the enkephalin, somatostatin, and neurotensin neuronal populations indicates that muscarinic receptor gene expression contributes to the functional and anatomical heterogeneity of the striatum that may relate to higher order of organization, including patch-matrix compartmentalization. The wide expression of m1 and m4 receptor genes in the striatum suggests that ACh may directly influence neurotransmitter release and synthesis in striatal efferent and intrinsic neurons. Our results imply that the specific pattern of expression of the muscarinic receptor genes mediates direct effects of ACh on activities and functions of chemically and topologically defined striatal neuronal populations. Since the expression of muscarinic receptors occurred in the three main neuronal populations of the striatum, namely ACh, enkephalins, and substance P neurons that also express dopamine receptors, it is highly probable that ACh and dopamine may act together at the single-cell level to influence striatal functions.

Animals↗
PubMed1992

The influence of initial residual disease on the outcome of second-look laparotomy in patients with carcinoma of the ovary.

Sixty four patients with epithelial ovarian cancer underwent second-look laparotomy. Influencing factors on the outcome of the second-look laparotomy were analysed. Both grade and stage appeared to have an effect on the outcome of the procedure. However, this effect appeared to be dependent on the amount of residual disease after primary surgery. The amount of residual disease appeared to be independently associated with the outcome of second-look laparotomy.

Adult↗
PubMed1991

[Tumor necrosis factor in graft rejection. In situ hybridization study].

The development of necrosis and macrophage infiltration increases the risk of renal graft rejection. But the macrophages secrete the alpha form of the tumour necrosing factor (TNF) which is also involved in several immunologic and inflammatory phenomena. We therefore studied the expression of the gene TNF alpha by in situ hybridization during advanced stage rejection after renal transplantation: the grafts were infiltrated with macrophage-like cells expressing the mRNA of the TNF alpha gene, particularly deep in the cortex and in the medulla. These cells then secrete the TNF alpha molecule since they are recognized by anti-TNF alpha antibodies. These antibodies also recognize certain other glomerular endothelial and tubular epithelial cells which do not express the TNF alpha gene: these cells are undoubtedly the TNF target cells. These findings confirm the synthesis of TNF alpha in advanced stage renal graft rejection.

Colorectal Neoplasms↗
PubMed1991

Repeat laparotomy in ovarian carcinoma after primary surgery.

Thirty-two patients with malignant ovarian disease were referred after primary surgery to the Gynaecological Oncology Unit of Groote Schuur Hospital, Cape Town. All 32 patients underwent a re-laparotomy with a view to accurate staging and possible cytoreductive surgery. On referral, 24 patients (75%) had stage I or II disease and the remaining 8 patients (25%) had stage III and IV disease. Twenty-seven patients (81%) had ovarian malignant disease of epithelial origin while the remaining 5 patients (19%) had ovarian disease of nonepithelial origin. Five (20.8%) of a total of 24 patients with stage I or II disease had their disease stage raised after repeat laparotomy. The overall success rate of cytoreductive surgery, i.e. less than 2 cm residual disease, was 58%.

Adult↗
PubMed1991

Aromatic L-amino-acid decarboxylase (DOPA decarboxylase) gene expression in dopaminergic and serotoninergic cells of the rat brainstem.

In situ hybridization was performed in the rat brain to detect aromatic L-amino acid decarboxylase (AADC) mRNA using 35S-labeled oligonucleotide probes derived from rat kidney AADC cDNA. Results demonstrated AADC mRNA in areas containing dopaminergic and serotoninergic cell bodies. Combined immunohistochemistry for tyrosine- or tryptophan hydroxylase and in situ hybridization for AADC mRNA demonstrated the dopaminergic or serotoninergic nature of cells containing AADC mRNA. Tyrosine hydroxylase-positive mesencephalic neurons containing a very low or no AADC mRNA signal were also observed.

5-Hydroxytryptophan↗
PubMed1991

D2 dopamine receptor gene expression in the rat striatum during ontogeny: an in situ hybridization study.

D2 dopamine receptor (D2R) gene expression in the rat striatum was studied by in situ hybridization throughout the pre- and the postnatal period from gestational day 12 to postnatal day 8. D2R mRNA was detected with 35S-labelled oligonucleotide probes, one that hybridized equally to the two isoforms of the D2R mRNA (D2(415) and D2(444)) and the other that hybridized specifically to the large isoform (D2(444)). D2R mRNA was first detected in the striatal primordium at day 14 of gestation with the probe that recognizes indifferently the two isoforms and with the probe specific for the D2(444) mRNA. At day 16, D2R mRNA was present in the lateral part of the striatum and in the germinal ventricular zone lining the lateral ventricle. At day 18, D2R mRNA was found in neurons of the caudate-putamen, the nucleus accumbens, the olfactory tubercle and the subependymal zone lining the lateral ventricle. The microautoradiographic analysis demonstrated that the labelled cells have a neuroblastic and immature aspect before birth. After birth the topography and aspect of labelled cells was similar to the one observed in the adult animals. D2R mRNA was present in neurons of the caudate-putamen, the nucleus accumbens and the olfactory tubercle. In the caudate-putamen there was a latero-medial gradient of labelling. From postnatal day 2 onward the D2R gene was expressed in two striatal cell types, small neurons probably enkephalinergic, and large-sized neurons with prominent cytoplasm, most probably cholinergic.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Phenotypical characterization of the rat striatal neurons expressing the D1 dopamine receptor gene.

In situ hybridization experiments were performed in rat brain sections from normal and 6-hydroxydopamine-treated rats in order to map and identify the neurons expressing the D1 receptor gene in the striatum and the substantia nigra. Procedures of combined in situ hybridization, allowing the simultaneous detection of two mRNAs in the same section or in adjacent sections, were used to characterize the phenotypes of the neurons expressing the D1 receptor gene. D1 receptor mRNA was found in neurons all over the caudate-putamen, the accumbens nucleus, and the olfactory tubercle but not in the substantia nigra. In the caudate-putamen and accumbens nucleus, most of the neurons containing D1 receptor mRNA were characterized as medium-sized substance P neurons and distinct from those containing D2 receptor mRNA. Nevertheless, 15-20% of the substance P neurons did not contain D1 receptor mRNA. The neurons containing preproenkephalin A mRNA did not contain D1 receptor mRNA but contained D2 receptor mRNA. A small number of cholinergic and somatostatinergic neurons exhibited a weak reaction for D1 receptor mRNA. These results demonstrate that dopamine acts on efferent striatal neurons through expression of distinct receptors--namely, D1 and D2 in separate cell populations (substance P and preproenkephalin A neurons, respectively)--and can also act on nonprojecting neurons through D1 receptor expression.

Animals↗
PubMed1991

Rat striatal and mesencephalic neurons contain the long isoform of the D2 dopamine receptor mRNA.

Cloning of the D2 dopamine receptor gene has demonstrated that two D2 mRNA isoforms are generated through alternative splicing: a short mRNA (D2(415)) and a long one (D2(444)) that differs by an additional exon transcript. D2 receptor gene expression was demonstrated in striatal dopamine terminal fields as well as in the mesencephalon by in situ hybridization using a probe that does not differentiate the two isoforms or a probe specific for the D2(444)) mRNA. We report here that the long D2(444)) mRNA isoform is present in all the neurons expressing the D2 receptor gene in the striatum and the mesencephalon. In the striatum, medium-sized neurons (previously identified as enkephalinergic) and large-sized neurons (previously identified as cholinergic) contain the D2(444)) mRNA. Our results with the D2(444)) probe confirm that 20% of the cholinergic neurons in the striatum do not express the D2 gene. The present results also demonstrate that the neurons expressing the D2 receptor gene in the substantia nigra are dopamine neurons and that D2 mRNA can be present in proximal dendrites.

Animals↗
PubMed1991

Striatal neurons express increased level of dopamine D2 receptor mRNA in response to haloperidol treatment: a quantitative in situ hybridization study.

In the present study, quantitative in situ hybridization was used to analyse the effect of haloperidol treatment on D2 dopamine receptor gene expression in the rat caudate-putamen nucleus. Variations of D2 receptor mRNA level were studied and measured at the macroscopic level of densitometric analysis of X-ray film and at the microscopic level by counting of autoradiographic silver grains in striatal cells. Macroscopic analysis demonstrated that haloperidol treatment two times 1 mg/kg per day during seven, 14 and 21 days increased D2 receptor mRNA level in the caudate-putamen. Detailed microscopic analysis demonstrated a significant increase in D2 receptor mRNA in the two neuronal populations known to express the D2 receptor gene: medium-sized neurons previously identified as enkephalinergic neurons, and large-sized neurons previously identified as cholinergic neurons. The increase was more important in cholinergic neurons (+119%) than in enkephalinergic neurons (+54%). Haloperidol treatment did not modify the number of medium-sized enkephalinergic neurons expressing the D2 receptor mRNA. In contrast, it significantly increased the percentage of large-sized neurons containing D2 receptor mRNA (from 80 to 94%). These results demonstrate that haloperidol treatment acts at the gene level to modulate D2 receptor content in striatal dopaminoceptive neurons, and that the D2 receptor mRNA increase in postsynaptic neurons contributes to dopamine supersensitivity induced by neuroleptics in the rat. This suggests that dopamine acts trans-synaptically to control D2 receptor gene expression in target striatal neurons. These results suggest that modifications of D2 receptor gene expression may be part of the biological events that lead to the movement disorders induced by neuroleptic drugs or Parkinson's disease.

Animals↗
PubMed1991

Simultaneous detection of two messenger RNAs in the central nervous system: a simple two-step in situ hybridization procedure using a combination of radioactive and non-radioactive probes.

We present here a method enabling the simultaneous detection of two messenger RNAs in tissue sections by use of a two-step in situ hybridization procedure. Tissue sections were hybridized with a radioactive probe and coated with emulsion. The emulsion was processed for development, fixed, and a second hybridization was performed through the emulsion with a biotinylated probe subsequently revealed with streptavidin-alkaline phosphatase. This procedure allows the detection of two mRNAs without loss of signal, removal of the emulsion, or spurious reaction. The simultaneous detection of oxytocin and vasopressin mRNAs in the hypothalamus, and of dopamine receptor and neuropeptide mRNAs in the striatum, demonstrated the efficiency of the procedure. Such a two-step procedure provides a simple and flexible way to make possible comparative analysis of the localization of two mRNAs within the same tissue section.

Animals↗
PubMed1991

The value of magnetic resonance imaging in patients with carcinoma of the cervix (a pilot study).

Magnetic resonance imaging (MRI) was performed in 11 patients with untreated squamous carcinoma of the cervix. The MRI stage and clinical stage were both compared with the surgical stage. MRI stage was only accurate in three of the 11 patients (27.3%) while the clinical stage was correct in eight of the 11 cases (72.7%). MRI assessment of parametrial and lymph-node involvement was difficult to interpret. These preliminary findings suggest that MRI is not, at present, a useful adjunct to staging in the evaluation of patients with carcinoma of the cervix.

Adult↗
PubMed1990

D2 dopamine receptor gene expression by cholinergic neurons in the rat striatum.

In situ hybridization with D2 receptor probe and immunohistochemistry with choline acetyltransferase (ChAT) antibody performed on adjacent sections demonstrate dopamine D2 receptor gene expression in cholinergic neurons of the rat caudate-putamen and nucleus accumbens. Eighty per cent of cholinergic neurons in the striatum contain detectable D2 receptor mRNA. The other neurons without detectable D2 mRNA do not display specific localization or aspect in the caudate-putamen and nucleus accumbens as compared to the other cholinergic neurons. The absence of detectable D2 mRNA in certain cholinergic neurons can be due to the limited sensitivity of the procedure that would not detect low mRNA levels, or alternatively can reflect the existence of two cholinergic cell populations in the striatum, one of which would not express the D2 receptor gene. The other forebrain cholinergic neurons do not contain D2 mRNA.

Animals↗
PubMed1990

Anatomical distribution of LHRH-immunoreactive neurons in the human infant hypothalamus and extrahypothalamic regions.

The morphological features and distribution of luteinizing hormone-releasing hormone (LHRH)-immunoreactive cell bodies and fibers of the hypothalamic and the neighboring mesencephalic regions were studied in the normal newborn infant by immunohistochemistry. Within the hypothalamus, numerous LHRH-immunoreactive like (IL) cell bodies were found mainly in the ventral portion of the infundibular nucleus close to the median eminence and at a lower extent in the medial preoptic area. In addition, sparse immunoreactive cell bodies were displayed in the paraventricular and medial mammillary nuclei. The mesencephalon also exhibited rare immunoreactive cell bodies in the periaqueductal gray. LHRH-IL fibers, predominantly varicose, formed a continuum from the septo-preoptico level to the mesencephalon. In the hypothalamus, the median eminence exhibited the highest LHRH innervation. LHRH-IL fibers are also observed in the lamina terminalis, the medial preoptic area, the suprachiasmatic, the supraoptic, the peri- and the paraventricular nuclei. In the last two nuclei, some fibers projected to the dorsomedial and ventromedial nuclei whereas others were in close relation with the ependyma. The mesencephalon displayed low LHRH-IL fibers, present essentially in the raphe and interpeduncular nuclei and around the ependyma. When compared with data obtained in other mammals, the present findings agree well with the general distribution and morphological features of LHRH-IL neuronal structures reported elsewhere.

Brain↗
PubMed1990

Presence of neuropeptide messenger RNAs in neuronal processes.

The messenger RNAs coding for vasopressin, oxytocin, luteinizing hormone releasing-hormone and somatostatin have been detected in tissue sections of the rat brain, especially in the hypothalamus with radioactive and biotinylated oligonucleotide probes. The results demonstrate that neuropeptide mRNAs are present in the cytoplasm of cell bodies, in processes and in punctate structures in the vicinity of the cell bodies. These results demonstrate that neuropeptide mRNAs can be transported outside the cell body most probably in proximal dendrites but also in some of their branching, and possibly at synaptic contacts. These data suggest that neuropeptide mRNA could undergo a specific compartmentation that could contribute to the targetting of the corresponding peptide inside neurons.

Animals↗
PubMed1990

Histochemical detection of the messenger RNAs coding for calcitonin and calcitonin gene-related peptide in medullary thyroid carcinomas with radioactive and biotinylated oligonucleotide probes.

The present study has been undertaken to investigate the efficiency of biotinylated synthetic oligonucleotide probes in detection by in situ hybridization of the mRNAs coding for calcitonin (CT) or calcitonin gene-related peptide (CGRP) in human medullary thyroid carcinomas (MTCs). Tissue sections fixed with formaldehyde were hybridized with 45-base long oligonucleotides, specific for CT or CGRP mRNA. Recombinant DNA probe or synthetic oligonucleotides radioactively labelled with 32P or 35S were used as controls to detect by autoradiography the corresponding mRNAs in the tumour cells. Oligonucleotide probes labelled by fixation of one biotin molecule at their 5'-end, or by incorporation of a tail of biotin-11-dUTP at their 3'-end, were used and were revealed by incubation with streptavidin-alkaline phosphatase associated with the corresponding substrate. Each biotinylated probe stained exclusively the cytoplasm of the tumour cells, the CT probe giving a much higher level of staining than the CGRP probe. The same cells were found to contain CT and CGRP mRNAs. Controls performed with either radioactive or biotinylated probes confirmed the specificity of the staining. These results demonstrate that biotinylated synthetic oligonucleotides can be used as efficient tools to investigate gene expression in tissue sections, thus avoiding the various inconveniences connected with the use of radioactive probes, especially bio-hazards, the use of autoradiography, the limited histological resolution, and the delay in obtaining results.

Autoradiography↗
PubMed1990

Topography and ontogeny of the neurons expressing vasopressin, oxytocin, and somatostatin genes in the rat brain: an analysis using radioactive and biotinylated oligonucleotides.

1. The use of radioactive and biotinylated oligonucleotide probes has been optimized to detect and analyze by in situ hybridization, neurons expressing neuropeptide genes (vasopressin, oxytocin, somatostatin). 2. In situ hybridization was performed on cryostat-cut sections obtained from tissues perfused with 1% formaldehyde. Radioactive probes were labeled by tailing with 35S-dATP and revealed with autoradiography. Biotinylated probes were obtained either by the incorporation of 11-biotin dUTP or by the addition of biotinylated nucleotides to the oligonucleotide during its synthesis. Biotin was revealed with streptavidin alkaline phosphatase and the appropriate substrate. 3. In the adult rat brain, radioactive and biotinylated probes revealed peptidergic neurons. The biotinylated probes provided an optimal cellular and subcellular resolution with a sensitivity similar to that observed with radioactive probes. Staining was selectively restricted to the cytoplasm and to the proximal part of processes. 4. Biotinylated vasopressin probes with 10 biotins added demonstrated magnocellular neurons and parvocellular neurons in the suprachiasmatic nucleus and the bed nucleus stria terminalis. 5. Vasopressin gene expression was studied during ontogeny in the rat fetus and neonate. Vasopressin mRNA was first detectable at gestational day 16 in the supraoptic nucleus in neurons of neuroblastic appearance. An aspect similar to the one present in adult was found at gestational day 19 in magnocellular neurons and at day 3 postnatal in parvocellular neurons. 6. The results confirm that radioactive oligonucleotide probes are efficient tools to investigate neuropeptide gene expression by in situ hybridization and demonstrate that biotinylated oligonucleotides are very efficient and provide a much higher resolution than radioactive probes with a reasonable sensitivity.

Animals↗
PubMed1990

Dopamine receptor gene expression by enkephalin neurons in rat forebrain.

In situ hybridization experiments were performed with brain sections from normal, control and haloperidol-treated rats to identify and map the cells expressing the D2 dopamine receptor gene. D2 receptor mRNA was detected with radioactive or biotinylated oligonucleotide probes. D2 receptor mRNA was present in glandular cells of the pituitary intermediate lobe and in neurons of the substantia nigra, ventral tegmental area, and forebrain, especially in caudate putamen, nucleus accumbens, olfactory tubercle, and piriform cortex. Hybridization with D2 and preproenkephalin A probes in adjacent sections, as well as combined hybridization with the two probes in the same sections, demonstrated that all detectable enkephalin neurons in the striatum contained the D2 receptor mRNA. Large neurons in caudate putamen, which were unlabeled with the preproenkephalin A probe and which may have been cholinergic, also expressed the D2 receptor gene. Haloperidol treatment (14 or 21 days) provoked an increase in mRNA content for D2 receptor and preproenkephalin A in the striatum. This suggests that the increase in D2 receptor number observed after haloperidol treatment is due to increased activity of the D2 gene. These results indicate that in the striatum, the enkephalin neurons are direct targets for dopamine liberated from mesostriatal neurons.

Animals↗