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T Neuman

Publications and source records attributed to T Neuman.

At least 55 records · Page 3Linked to original sources
PubMed1993

Cell type-specific negative regulatory element in low-affinity nerve growth factor receptor gene.

Developmental changes in the expression pattern of the low-affinity nerve growth factor receptor (LNGFR) gene suggest a complex mechanism of gene regulation. We demonstrate the presence of a negative regulatory element (NRE) localized to a 40 base pair (bp) segment, -1731 to -1690 bp upstream from the translation start site in the LNGFR gene. The NRE possesses two tandemly arranged sequences with similarity to immunoglobulin gene enhancer E-boxes. The NRE is active in neurons and neuronal cell lines but not in astrocytes. Electrophoretic mobility shift analysis (EMSA) demonstrates changing expression pattern of proteins binding to the NRE in developing nervous system. Since the specific binding of the proteins to the NRE is competed with oligonucleotides containing E-box sequences we suggest that factor(s) responsible for down regulation of LNGFR gene include members of the helix-loop-helix class of transcription factors.

Animals↗
PubMed1993

Episodic angioedema with eosinophilia: a case associated with T cell activation and cytokine production.

Episodic angioedema with eosinophilia is a rare syndrome, characterized by periodic attacks of fever, myalgia, angioedema, and oliguria, associated with an increased eosinophil count. The pathogenesis of this disorder is incompletely understood. We describe an additional patient with episodic angioedema and eosinophilia, and review the literature regarding the clinical features and treatment of this disease. Previous hypotheses regarding disease mechanisms are discussed. In our patient, we found high basal levels of interleukin-1 (IL-1) and soluble interleukin-2 receptor (sIL-2R), with further increases in levels of both mediators during attacks. We suggest that IL-1 and T cell activation may play a role in the pathophysiology of episodic angioedema with eosinophilia.

Adult↗
PubMed1991

Freeze-fracture studies on vestibular secretory cells and melanocytes.

The fine structure of dark cells and melanocytes of the crista ampullaris was studied by using the freeze-fracture technique. For these studies we used normally pigmented guinea pigs, as well as those with experimentally induced endolymphatic hydrops. The results suggest the vestibular secretory region to be an area of unusually high dynamics. In the experimental animals the cell processes of the melanocytes gain closer contact with the dark cells as well as with the subepithelial capillaries. There is considerable evidence of secretory transfer of different substances, especially ions.

Animals↗
PubMed1990

Oct-6: a POU transcription factor expressed in embryonal stem cells and in the developing brain.

A family of octamer binding proteins is expressed during mouse development. Oct-4 and Oct-6 have been identified as two octamer binding proteins present in embryonal stem cells. Here we report the complementary DNA cloning and characterization of the mouse Oct-6 gene. The protein of 448 amino acids contains a glycine/alanine-rich amino terminal region, a histidine-rich sequence with homology to a region of kininogen associated with clotting, a POU domain and a short proline/histidine-rich carboxy terminal region. Expression of Oct-6 in HeLa cells is sufficient for transcriptional activation from the octamer motif, identifying Oct-6 as a transcription factor. The Oct-6 expression is downregulated upon embryonic stem cell differentiation increasing again during brain development. Expression in brain is present in certain areas of telencephalon, mesencephalon and brain stem with abundant expression in the cortex anlagen and in the developing colliculi. Thus Oct-6 is a new octamer binding transcription factor specifically regulated during mouse development.

Amino Acid Sequence↗
PubMed1990

Nerve growth factor-induced rapid reorganization of microfilaments in PC12 cells: possible roles of different second messenger systems.

Nerve growth factor (NGF) induces in 2 to 10 min the redistribution of F-actin in rat pheochromocytoma PC12 cells. The NGF specificity of this phenomenon was shown by blocking it with anti-NGF antibodies. We used the rapid F-actin redistribution as an assay to study NGF second messenger systems and their inhibition or activation by specific agents. The results show that the NGF-induced effect on the microfilament system of PC12 cells can be specifically inhibited by lithium chloride and neomycin, inhibitors of the phosphoinositol system, but cannot be mimicked by TPA and acetylcholine, the activators of the phosphoinositol system. An increase in the intracellular concentration of cyclic AMP by addition of dBcAMP (but not dBcGMP) caused rapid F-actin redistribution that nonetheless differed from the NGF-induced effect. Changes in the intracellular calcium level did not have any influence on the microfilament system of PC12 cells. The specificity of the inhibition of NGF-induced effects by methylase inhibitors was questionable, since MTA- or SAH-treated PC12 cells acquired an altered morphology even in the absence of NGF or dBcAMP. Using the microfilament- and microtubule-disrupting drugs cytochalasin B and colchicine, we showed that the microtubule system in PC12 cells is required for the initiation of neurite outgrowth and that microfilament-associated filopodial activity does not appear to be necessary.

Actin Cytoskeleton↗
PubMed1990

Structure and expression of the mouse Oct2a and Oct2b, two differentially spliced products of the same gene.

A large family of tissue-specific nuclear proteins interact with the octamer motif ATTTGCAT, a transcriptional regulatory element found in the promoter and enhancer sequences of many genes. As a step towards elucidating the mechanism of this regulation, cDNA clones of the mouse Oct2 protein were isolated. One, called here Oct2b, encodes a larger variant of the previously described Oct2a proteins. The Oct2b cDNA has an insertion of 74 bp close to the 3' end which creates an open reading frame distinct from Oct2a. As a result, the Oct2b protein has a carboxy end which is similar to that of the ubiquitous octamer-binding protein Oct1. Analysis of the Oct2 gene shows that Oct2a and Oct2b are differentially spliced products of the same gene. The insertion in the Oct2b cDNA results from the inclusion of an additional exon in the mRNA which would otherwise reside in an intron sequence of the Oct2a transcript. RNA analysis demonstrates that both Oct2a and 2b mRNAs are most abundant in B-cells but they are also expressed in a variety of tissues including brain, intestine, testis, kidney, as well as in embryos. Interestingly, the ratio of Oct2a and 2b varies among tissues. In situ hybridization studies during mouse embryogenesis show that the Oct2 gene is widely expressed in the developing nervous system. In contrast, expression in the adult brain is confined to very specific areas which include the suprachiasmatic and medial mammillary nuclei, hippocampus, olfactory tract and the olfactory bulb. Oct2 proteins are present in both neuronal and oligodendroglial cells, although they are more abundant in glial cells.

Amino Acid Sequence↗
PubMed1989

Sensitive time-resolved fluoroimmunoassay of nerve growth factor and the disappearance of nerve growth factor from rat pheochromocytoma PC12 cell culture medium.

A sensitive time-resolved fluoroimmunoassay of nerve growth factor (NGF) has been developed. The method is based on the unique property of the lanthanides for delayed fluorescence, which reduces substantially the endogenous fluorescence of biological substances, because the excitation of the sample and detection of the fluorescence signal are separated in time and in wavelength. Using the europium-conjugated antibodies to the NGF from Vipera lebetina (snake) venom and to the beta NGF from mouse submandibular gland in a solid-phase quantitative two-site fluoroimmunoassay, we obtained a maximal sensitivity of 10 pg/ml (0.38 pM)for mouse NGF and 40 pg/ml (1.2 pM) for snake NGF. Using this method, we investigated the disappearance of NGF from rat pheochromocytoma PC12 cell culture medium. Mouse beta NGF (5-10 ng/ml) disappeared completely after 12 h of incubation, whereas snake NGF was not substantially internalized even after 48 h.

Animals↗
PubMed1989

Induction of morphological differentiation of human neuroblastoma cells is accompanied by induction of tissue-type plasminogen activator.

Human SH-SY5Y neuroblastoma cells treated with retinoic acid, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or nerve growth factor differentiated morphologically to neuronlike cells with increased amounts of neurofilament protein and mRNA. All three effectors induced an increase in the amount of relative molecular weight (Mr) 70,000 tissue-type plasminogen activator (t-PA) and its mRNA, as determined by immunocapture, enzyme activity, and Northern blotting analyses. About 90% of the t-PA activity was secreted to the culture medium. In contrast, of the three effectors studied, only TPA induced transcription of the proto-oncogene c-fos, studied as a control gene responsive to various stimuli, and induced a rapid increase in urokinase-type PA (u-PA). Most of the u-PA activity induced by TPA remained cell-associated. Because induction of differentiation correlated closely with induction of t-PA, and not u-PA, the authors propose that t-PA may have a functional role in the morphological differentiation of neuronal cells.

Cell Differentiation↗
PubMed1988

Nerve growth factor increases the cyclic GMP level and activates the cyclic GMP phosphodiesterase in PC12 cells.

Nerve growth factor (NGF) rapidly increases the cyclic GMP (cGMP) level about 2-3-fold and enhances the cGMP phosphodiesterase (PDE) activity about 2-fold in rat pheochromocytoma PC12 cells. No changes in the level of cyclic AMP (cAMP) and in the activity of cAMP PDE were found. GTP and a nonhydrolysable analog of GTP, GMP-PCP, at 100 microM, were able to mimic the effect of NGF on the cGMP PDE activity. These results suggest that the cGMP system may be one of the second messengers of NGF action in PC12 cells.

1-Methyl-3-isobutylxanthine↗
PubMed1988

Nerve growth factor induces rapid redistribution of F-actin in PC12 cells.

Nerve growth factor (NGF) induces the redistribution of F-actin in rat pheochromocytoma PC12 cells within 2-10 min, whereas epidermal growth factor (EGF) has no effect on microfilament organization. This redistribution of F-actin in PC12 cells is not protein synthesis dependent, but can be blocked by methyltransferase inhibitors.

Actins↗
PubMed1987

Monoclonal antibodies against Vipera lebetina venom nerve growth factor cross-react with other snake venom nerve growth factors.

Nerve growth factor (NGF) was isolated from the venom of Vipera lebetina and was purified to homogeneity as judged by SDS gel electrophoresis. The biologically active NGF was used to immunize BALB/c mouse, and the spleen cells from immunized mouse were fused with mouse PAI myeloma cells. Forty-seven hybrid cell lines, secreting monoclonal antibodies to V. lebetina NGF, were isolated and nine of them purified from ascitic fluids. The isolated antibodies define two partially overlapping epitopes of the V. lebetina NGF which are not involved in the biological activity of the molecule. Both epitopes are also present on the beta-NGF from the mouse salivary gland and on the NGFs from the following snake venoms: V. lebetina, V, ursini, V, berus berus, Echis carinatus, Bungarus caeruleus, Agkistrodon halys, Naja naja oxiana, Naja naja atra and Naja naja, but not on the bovine seminal plasma NGF. The mol. wts of the NGFs in these snake venoms were determined by Western immunoblot with monoclonal antibodies. The mol. wts of the NGFs from V. ursini (37,000), E. carinatus (36,000, 44,000) and A. halys (29,000) were determined for the first time.

Animals↗
PubMed1987

Monoclonal antibody immunoaffinity chromatography of the nerve growth factor from snake venoms.

1. Pure monoclonal antibodies to Vipera lebetina venom nerve growth factor have been isolated by affinity chromatography using CNBr-agarose bound antigen. 2. Nerve growth factors from ten snake venoms (Vipera lebetina, Vipera russellii, Vipera berus berus, Vipera ursini, Echis carinatus, Agkistrodon halys, Bungarus caeruleus, Naja naja oxiana, Naja naja, Naja naja atra) were purified using monoclonal antibodies against NGF linked to BrCN-activated agarose.

Animals↗
PubMed1987

The muscarinic receptor-mediated action of acetylcholine in the gastrulating chick embryo.

Some of the muscarinic receptor-mediated effects of acetylcholine in early chick embryo cells at stages three-four by Hamburger and Hamilton were studied. Acetylcholine increased the intracellular level of cGMP about two-fold. Acetylcholine raised the intracellular level of free calcium from the basal level of 120 nM to 140 nM. Atropine, a muscarinic antagonist, blocked both the above-mentioned responses.

Acetylcholine↗
PubMed1986

Nerve growth factor induces changes in (2'-5')oligo(A) synthetase and 2'-phosphodiesterase activities during differentiation of PC12 pheochromocytoma cells.

Treatment of rat pheochromocytoma cell line PC12 with Vipera lebetina (snake) nerve growth factor (NGF) induces a rapid increase (from 5 to 25-fold) in the level of (2'-5')oligo(A) synthetase activity and a simultaneous decrease (from 2 to 5-fold) in the activity of 2'-5' A degrading enzymes--2'-phosphodiesterases (2'-PDE). These changes in the enzyme activities led to the significant increase in the intracellular concentration of 2'-5' A. We have found that the serum starvation of PC12 cells causes a 1.5 to 2.0-fold increase in the level of 2'-5' A-synthetase activity, but the activities of 2'-PDE and the intracellular concentration of 2'-5' A remain unaltered. These results show that NGF modulates the activity of (2'-5')oligo(A) enzymes and intracellular concentration of 2'-5' A during the neural differentiation of PC12 cells.

2',5'-Oligoadenylate Synthetase↗
PubMed1986

Isolation and characterization of nerve growth factor from Vipera berus berus (common viper) venom.

Nerve growth factor from Vipera berus berus venom was purified by gel filtration on Sephadex G-100 (superfine), ion-exchange-chromatography on DEAE-Sephadex A-50 and chromatofocusing on PBE 118. The Vipera berus berus venom NGF consists of multiple molecular forms with pls in the interval 9.1-9.7. All isoforms have identical mol. wts approximately 35,000 +/- 3000 (in gel filtration) and 17,000 +/- 2000, 15,000 +/- 2000 (by SDS electrophoresis with beta-mercaptethanol). V. berus berus venom NGF reacted with monoclonal antibodies against Viper lebetina NGF and caused differentiation of pheochromocytoma PC12 cells.

Animals↗