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Norio Kaneda - One of the best experts on this subject based on the ideXlab platform.
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establishment and characterization of a noradrenergic Adrenal chromaffin Cell line tsam5ne immortalized with the temperature sensitive sv40 t antigen
Cell Biology International, 2011Co-Authors: Susumu Kohno, Tomiyasu Murata, Kiyomi Hikita, Naoshi Koide, Norio KanedaAbstract:We established a clonal Adrenal Medullary Cell line, named tsAM5NE, from transgenic mice harbouring the temperature-sensitive Simian virus 40 large T-antigen gene, under the control of the tyrosine hydroxylase promoter. tsAM5NE Cells conditionally grew at a permissive temperature of 33°C and exhibited the noradrenergic chromaffin Cell phenotype. To understand the characteristics of tsAM5NE Cells, we first examined the responsiveness of the Cells to ligands of the GDNF (glial Cell line-derived neurotrophic factor) family. tsAM5NE Cells proliferated at the permissive temperature of 33°C in response to either GDNF or neurturin, but not artemin or persephin. At the non-permissive temperature of 39°C, GDNF or neurturin caused tsAM5NE Cells to differentiate into neuron-like Cells; however, the differentiated Cells died in a time-dependent manner. Interestingly, LIF (leukaemia inhibitory factor) did not affect the GDNF-mediated Cell proliferation at 33°C, but promoted the survival and differentiation of GDNF-treated Cells at 39°C. In the presence of GDNF plus LIF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal markers, indicating that the Cells had indeed undergone neuronal differentiation. Thus, we demonstrated that tsAM5NE Cells had the capacity to terminally differentiate into neuron-like Cells in response to GDNF plus LIF when the oncogene was inactivated by the temperature shift. Thus, this Cell line provides a useful model system for studying the mechanisms regulating neuronal differentiation.
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temperature dependent neurotrophic factor elicited neuronal differentiation in Adrenal chromaffin Cell line immortalized with temperature sensitive sv40 t antigen
Journal of Neurochemistry, 2003Co-Authors: Tomiyasu Murata, Masaru Tsuboi, Kiyomi Hikita, Kentaro Niwa, Misao Suzuki, Norio KanedaAbstract:We established Adrenal Medullary Cell lines from transgenic mice expressing an oncogene, the temperature-sensitive simian virus 40 large T-antigen, under the control of the tyrosine hydroxylase promoter. A clonal Cell line, named tsAM5D, conditionally grew at a permissive temperature of 33°C and exhibited the dopaminergic chromaffin Cell phenotype as exemplified by the expression pattern of mRNA for catecholamine-synthesizing enzymes and secretory vesicle-associated proteins. tsAM5D Cells proliferated at the permissive temperature in response to basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). At a non-permissive temperature of 39°C, bFGF and CNTF acted synergistically to differentiate tsAM5D Cells into neuron-like Cells. In addition, tsAM5D Cells caused to differentiate by bFGF plus CNTF at 39°C became dependent solely on nerve growth factor for their survival and showed markedly enhanced neurite outgrowth. In the presence of bFGF and CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal marker genes including neuron-specific enolase, growth-associated protein-43, microtubule-associated protein 2, neurofilament, and p75 neurotrophin receptor, indicating that the Cells underwent neuronal differentiation. Thus, we demonstrated that tsAM5D Cells could proliferate at permissive 33°C, and also had the capacity to terminally differentiate into neuron-like Cells in response to bFGF and CNTF when the oncogene was inactivated by shifting the temperature to non-permissive 39°C. These results suggest that tsAM5D Cells should be a good tool to allow a detailed study of mechanisms regulating neuronal differentiation.
Tomiyasu Murata - One of the best experts on this subject based on the ideXlab platform.
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establishment and characterization of a noradrenergic Adrenal chromaffin Cell line tsam5ne immortalized with the temperature sensitive sv40 t antigen
Cell Biology International, 2011Co-Authors: Susumu Kohno, Tomiyasu Murata, Kiyomi Hikita, Naoshi Koide, Norio KanedaAbstract:We established a clonal Adrenal Medullary Cell line, named tsAM5NE, from transgenic mice harbouring the temperature-sensitive Simian virus 40 large T-antigen gene, under the control of the tyrosine hydroxylase promoter. tsAM5NE Cells conditionally grew at a permissive temperature of 33°C and exhibited the noradrenergic chromaffin Cell phenotype. To understand the characteristics of tsAM5NE Cells, we first examined the responsiveness of the Cells to ligands of the GDNF (glial Cell line-derived neurotrophic factor) family. tsAM5NE Cells proliferated at the permissive temperature of 33°C in response to either GDNF or neurturin, but not artemin or persephin. At the non-permissive temperature of 39°C, GDNF or neurturin caused tsAM5NE Cells to differentiate into neuron-like Cells; however, the differentiated Cells died in a time-dependent manner. Interestingly, LIF (leukaemia inhibitory factor) did not affect the GDNF-mediated Cell proliferation at 33°C, but promoted the survival and differentiation of GDNF-treated Cells at 39°C. In the presence of GDNF plus LIF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal markers, indicating that the Cells had indeed undergone neuronal differentiation. Thus, we demonstrated that tsAM5NE Cells had the capacity to terminally differentiate into neuron-like Cells in response to GDNF plus LIF when the oncogene was inactivated by the temperature shift. Thus, this Cell line provides a useful model system for studying the mechanisms regulating neuronal differentiation.
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neurotrophic factor responsiveness of Adrenal Medullary Cell line tsam5d immortalized with temperature sensitive sv40 t antigen
Yakugaku Zasshi-journal of The Pharmaceutical Society of Japan, 2006Co-Authors: Tomiyasu MurataAbstract:We established Adrenal Medullary Cell lines from transgenic mice expressing an oncogene, the temperature-sensitive simian virus 40 large T-antigen, under the control of the tyrosine hydroxylase promoter. A clonal Cell line, named tsAM5D, conditionally grew at a permissive temperature of 33 degrees C and exhibited the dopaminergic chromaffin Cell phenotype as exemplified by the expression pattern of mRNA for catecholamine synthesizing-enzymes and secretory vesicle-associated proteins. tsAM5D Cells proliferated at the permissive temperature in response to glial Cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). At a nonpermissive temperature of 39 degrees C, GDNF and CNTF acted synergistically to differentiate tsAM5D Cells into neuron-like Cells. In addition, tsAM5D Cells caused to differentiate by GDNF plus CNTF at 39 degrees C became dependent solely on nerve growth factor for their survival and showed markedly enhanced neurite outgrowth. In the presence of GDNF plus CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal marker genes including microtubule-associated protein 2, neuron-specific enolase, neurofilament, and growth-associated protein-43, indicating that the Cells underwent neuronal differentiation. Thus, we demonstrated that tsAM5D Cells could proliferate at permissive 33 degrees C, and also had the capacity to terminally differentiate into neuron-like Cells in response to GDNF plus CNTF when the oncogene was inactivated by shifting the temperature to nonpermissive 39 degrees C. These results suggest that tsAM5D Cells should be a good tool to allow a detailed study of mechanisms regulating neuronal differentiation.
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temperature dependent neurotrophic factor elicited neuronal differentiation in Adrenal chromaffin Cell line immortalized with temperature sensitive sv40 t antigen
Journal of Neurochemistry, 2003Co-Authors: Tomiyasu Murata, Masaru Tsuboi, Kiyomi Hikita, Kentaro Niwa, Misao Suzuki, Norio KanedaAbstract:We established Adrenal Medullary Cell lines from transgenic mice expressing an oncogene, the temperature-sensitive simian virus 40 large T-antigen, under the control of the tyrosine hydroxylase promoter. A clonal Cell line, named tsAM5D, conditionally grew at a permissive temperature of 33°C and exhibited the dopaminergic chromaffin Cell phenotype as exemplified by the expression pattern of mRNA for catecholamine-synthesizing enzymes and secretory vesicle-associated proteins. tsAM5D Cells proliferated at the permissive temperature in response to basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). At a non-permissive temperature of 39°C, bFGF and CNTF acted synergistically to differentiate tsAM5D Cells into neuron-like Cells. In addition, tsAM5D Cells caused to differentiate by bFGF plus CNTF at 39°C became dependent solely on nerve growth factor for their survival and showed markedly enhanced neurite outgrowth. In the presence of bFGF and CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal marker genes including neuron-specific enolase, growth-associated protein-43, microtubule-associated protein 2, neurofilament, and p75 neurotrophin receptor, indicating that the Cells underwent neuronal differentiation. Thus, we demonstrated that tsAM5D Cells could proliferate at permissive 33°C, and also had the capacity to terminally differentiate into neuron-like Cells in response to bFGF and CNTF when the oncogene was inactivated by shifting the temperature to non-permissive 39°C. These results suggest that tsAM5D Cells should be a good tool to allow a detailed study of mechanisms regulating neuronal differentiation.
Kyoji Morita - One of the best experts on this subject based on the ideXlab platform.
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enhancement by f actin of mgatp dependent dopamine uptake into isolated chromaffin granules
Iubmb Life, 1996Co-Authors: Kyoji Morita, Stuart M. Tomares, Harvey B PollardAbstract:Interaction of chromaffin granules with the microfilament network in the cytoplasmic space of Adrenal Medullary Cells has been proposed, but the influence of chromaffin granule-microfilament interaction on granule function has not yet been studied. In the present study, the effect of F-actin on dopamine uptake into isolated chromaffin granules was examined. The MgATP-dependent component of dopamine uptake was significantly enhanced by F-actin. In contrast, catecholamine efflux from the granules was not significantly altered by F-actin under the conditions in which the enhancement of MgATP-dependent dopamine uptake was observed. These findings suggest that the interaction of chromaffin granules with the microfilament network results in the enhancement of MgATP-dependent dopamine uptake into the granules, thus modulating the accumulation and storage of catecholamines in the Adrenal Medullary Cell.
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ionic factors affecting the association of tyrosine hydroxylase with chromaffin granules in the Adrenal Medullary Cell
Neurochemistry International, 1994Co-Authors: Kyoji Morita, Shuichi Hamano, Motoo OkaAbstract:Chromaffin Cells were treated with digitonin in medium containing various ions and the efflux of tyrosine hydroxylase from these permeabilized Cells was then determined to elucidate a possible influence of cytoplasmic ionic environment on the association of this enzyme with the chromaffin granule. The enzyme efflux was observed with a distinct lag during exposure to low concentrations of digitonin in the medium containing isotonic sucrose. In contrast, a larger extent of the enzyme efflux was observed without any notable delay in the presence of isotonic NaCl. The results were thought to indicate the possibility that the dissociation of soluble enzyme from the granule surface within the permeabilized Cells might occur in the presence of NaCl. Furthermore, the interaction between tyrosine hydroxylase and isolated chromaffin granule membranes was directly examined, and this interaction was shown to be inhibited by NaCl. However, the enzyme-granule membrane interaction was also inhibited by KCl and choline chloride. It therefore seems possible to consider that the inhibitory action of NaCl on the association of soluble enzyme with the granule may not be due to the specific action of Na+, but presumably due to the non-specific chaotropic effect of Cl-. On the other hand, the enzyme efflux was markedly reduced by the presence of Ca2+ in the permeabilizing medium, but the enzyme-granule membrane interaction was not affected by Ca2+ at the same concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)
Don M Gash - One of the best experts on this subject based on the ideXlab platform.
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effect of mptp on primate chromaffin Cells in vitro relevance for Adrenal Medullary Cell transplantation
Restorative Neurology and Neuroscience, 1991Co-Authors: Mary F D Notter, Marcello Kaniuki, Suzanne Y Felten, John T Hansen, Don M GashAbstract:Primate Adrenal Medullary Cells were exposed to l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP) in vitro to examine the effect of this neurotoxic agent on chromaffin Cells. Chromaffin Cells from monkey and humans were cultured in the presence of 100 ng/ml nerve growth factor for 1 week and then exposed to 150 μM MPTP or its active metabolite methylpyridinium ion (MPP+) for an additional week. Cells which had extended neurites in the presence of NGF showed no morphological effect in response to MPTP or MPP+ at the light microscopic level. However, there was a significant loss in catecholamines as seen by histofluorescence and high performance liquid chromotography (HPLC). Electron microscopy revealed a depletion in dense-core vesicles in chromaffin Cells after chronic exposure to MPTP while the mitochondria appeared similar to those observed in control Cells. Replacement of MPTP medium with standard medium stimulated restoration of catecholamine histofluorescence after 7 days. An acute 15 min pretreatment of chromaffin Cells with MPTP or MPP+ induced secretion of catecholamines over a 1 h pulse, with MPP+ producing the maximum and more rapid secretion as determined by HPLC. These data indicate that MPTP induces a dramatic loss in catecholamines in primate chromaffin Cells in vitro after both acute and chronic exposures; however, removal of the toxic agent permits restoration of catecholamines without permanent effect on the integrity of these Cells.
Motoo Oka - One of the best experts on this subject based on the ideXlab platform.
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ionic factors affecting the association of tyrosine hydroxylase with chromaffin granules in the Adrenal Medullary Cell
Neurochemistry International, 1994Co-Authors: Kyoji Morita, Shuichi Hamano, Motoo OkaAbstract:Chromaffin Cells were treated with digitonin in medium containing various ions and the efflux of tyrosine hydroxylase from these permeabilized Cells was then determined to elucidate a possible influence of cytoplasmic ionic environment on the association of this enzyme with the chromaffin granule. The enzyme efflux was observed with a distinct lag during exposure to low concentrations of digitonin in the medium containing isotonic sucrose. In contrast, a larger extent of the enzyme efflux was observed without any notable delay in the presence of isotonic NaCl. The results were thought to indicate the possibility that the dissociation of soluble enzyme from the granule surface within the permeabilized Cells might occur in the presence of NaCl. Furthermore, the interaction between tyrosine hydroxylase and isolated chromaffin granule membranes was directly examined, and this interaction was shown to be inhibited by NaCl. However, the enzyme-granule membrane interaction was also inhibited by KCl and choline chloride. It therefore seems possible to consider that the inhibitory action of NaCl on the association of soluble enzyme with the granule may not be due to the specific action of Na+, but presumably due to the non-specific chaotropic effect of Cl-. On the other hand, the enzyme efflux was markedly reduced by the presence of Ca2+ in the permeabilizing medium, but the enzyme-granule membrane interaction was not affected by Ca2+ at the same concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)