The Experts below are selected from a list of 72 Experts worldwide ranked by ideXlab platform

Yu Zheng - One of the best experts on this subject based on the ideXlab platform.

  • construction of fusion expression vector of human derived Neurotrophin 6 gene encoding mature peptide and purification of its expressed product
    Journal of Biomedical Engineering, 2005
    Co-Authors: Chengwu Zhang, Chaoyang Zhai, Zicheng Li, Yu Zheng
    Abstract:

    : To get the mature peptide of human-derived Neurotrophin-6 (NT-6), NT-6 gene encoding mature peptide was amplified by PCR, using the NT-6 cDNA that had been cloned as templet. The gene encoding mature peptide of NT-6 gene was cloned into pGEX1-lambdaT plasmid to construct the fusion expression vector. Expression of fusion protein in Escherichia coli was defected after induction by isopropyl beta-D-thiogalactoside(IPTG). The mature peptide of NT-6 was collected with GST fusion protein purifying kit. It was shown that a fragment of 460bp was gained by PCR. With the techniques of double-cleave and electrophoresis, the recombinant vector was identified as pGEX1-NT-6. The recombinant vector pGEX1-NT-6 transformed Escherichia coli expressed fusion protein of 41KD after induction by IPTG. Cleaved by thrombin, the mature peptide of NT-6 was obtained; its molecular weight was about 15KD. The cloning and expression of human-derived NT-6 gene encoding mature protein has provided a basis for further studies on the function and clinical application of NT-6.

  • protecting effect of human derived Neurotrophin 6 on retrogradely degenerated motoneurons of facial nucleus of rats
    Chinese journal of applied physiology, 2005
    Co-Authors: Chengwu Zhang, Zicheng Li, Li Li, Yu Zheng
    Abstract:

    AIM: The protecting effect of human-derived Neurotrophin-6 (NT-6) on injured neurons was investigated to test the neurobiological characteristics of human-derived NT-6. METHODS: Adult SD rats were used and divided into two groups, normal control group and experiment group. The right facial nerve of the experimental rats was sectioned. The experimental animals were subdivided into three groups, blank control group, NT-6 group and saline control group. The rats were raised for two weeks. Brain stem of the rats was removed and transversely sliced. Nissle stain and ChAT immunohistochemical stain of the slices were carried out to observe the role of NT-6 in protecting facial motoneurons. RESULTS: The number of the ChAT positive neurons and the intensity of Nissle stain in the facial nucleus of NT-6 group were significantly augmented compared with that of the blank and saline control groups. CONCLUSION: The human-derived NT-6 could partially protect the facial motoneurons from retrograde degeneration induced by their axon damage.

  • cloning of human brain derived Neurotrophin 6 gene and its expression in procaryotic cell
    Chinese journal of medical genetics, 2002
    Co-Authors: Chengwu Zhang, Xu Zhuang, Chaoyang Zhai, Yu Zheng
    Abstract:

    OBJECTIVE: To clone human brain-derived Neurotrophin-6(NT-6) gene and to observe its expression in the procaryotic cell. METHODS: Total RNA was extracted from aborted antenatal cerebral cortex, and cDNA fragment of NT-6 was amplified through reverse transcript-polymerase chain reaction. After being incised and recovered, the NT-6 gene was cloned into pBK-CMV plasmid to construct a NT-6 gene expression vector. Expression of NT-6 gene in Escherichia coli was studied after being induced by isopropyl beta-D-thiogalactoside(IPTG). RESULTS: The NT-6 gene expression vector was constructed and Escherichia coli with recombinant vector expressed specific protein after induction by IPTG. CONCLUSION: The cloning of human brain-derived NT-6 gene provides a basis for further studying the structure, function and clinical application of NT-6.

Rudolf Gotz - One of the best experts on this subject based on the ideXlab platform.

  • recombinant fish Neurotrophin 6 is a heparin binding glycoprotein implications for a role in axonal guidance
    Biochemical Journal, 1997
    Co-Authors: Xiaoling Li, Rudolf Gotz, Friedrich Lottspeich, Juliane Franz
    Abstract:

    Neurotrophin-6 (NT-6) was identified in the teleost fish Xiphophorus as a new member of the Neurotrophin gene family. NT-6 binds specifically the glycosaminoglycan heparin. In this study NT-6 was expressed in a stably transfected mammalian cell line, and in insect cells via a recombinant baculovirus. It was purified to homogeneity and characterized by MS and N-terminal sequencing. NT-6 from both expression systems was proteolytically processed at one of two protease cleavage motifs and was found to be glycosylated. It supported the survival of embryonic chick sensory neurons; half-maximal survival was observed at 100 ng/ml. Furthermore, NT-6 elicited neurite outgrowth in explanted embryonic dorsal root ganglia. Addition of heparin into the medium did not potentiate the activity of NT-6 in survival assays. However, when a sensory ganglion explant was cultured in a collagen gel matrix assay adjacent to a heparin bead coated with NT-6, neurite outgrowth directed towards the bead was observed. This indicated that NT-6 was slowly released from the heparin bead generating a concentration gradient of NT-6 instrumental for axonal guidance in vitro. Thus the interaction of NT-6 with heparin might not be required for the activation of the cellular receptor for NT-6 on responsive cells but rather may serve to control, in vivo, the distribution of NT-6.

  • Neurotrophin 6 is a new member of the nerve growth factor family
    Nature, 1994
    Co-Authors: Rudolf Gotz, Christoph Winkler, Franz Raulf, Manfred Schartl, Friedrich Lottspeich, Reinhard W Koster, Hans Thoenen
    Abstract:

    DURING vertebrate development, many neurons depend for survival and differentiation on their target cells1–3. The best documented mediator of such a retrograde trophic action is the Neurotrophin nerve growth factor (NGF)1. NGF and the other known members of the Neurotrophin family, brain-derived neurotrophic factor (BDNF), Neurotrophin-3 (NT-3) and Neurotrophin-4/5 (NT-4/5) are conserved as distinct genes over large evolutionary distances4–6. Here we report the cloning of Neurotrophin-6 (NT-6), a new mem-ber of this family from the teleost fish Xiphophorus. NT-6 dis-tinguishes itself from the other known Neurotrophins in that it is not found as a soluble protein in the medium of producing cells. The addition of heparin (but not chondroitin) effects the release of NT-6 from cell surface and extracellular matrix molecules. Recombinant purified NT-6 has a spectrum of actions similar to NGF on chick sympathetic and sensory neurons, albeit with a lower potency. NT-6 is expressed in the embryonic valvulla cerebelli; expression persists in some adult tissues. The interaction of NT-6 with heparin-binding molecules may modulate its action in the nervous system.

Chengwu Zhang - One of the best experts on this subject based on the ideXlab platform.

  • construction of fusion expression vector of human derived Neurotrophin 6 gene encoding mature peptide and purification of its expressed product
    Journal of Biomedical Engineering, 2005
    Co-Authors: Chengwu Zhang, Chaoyang Zhai, Zicheng Li, Yu Zheng
    Abstract:

    : To get the mature peptide of human-derived Neurotrophin-6 (NT-6), NT-6 gene encoding mature peptide was amplified by PCR, using the NT-6 cDNA that had been cloned as templet. The gene encoding mature peptide of NT-6 gene was cloned into pGEX1-lambdaT plasmid to construct the fusion expression vector. Expression of fusion protein in Escherichia coli was defected after induction by isopropyl beta-D-thiogalactoside(IPTG). The mature peptide of NT-6 was collected with GST fusion protein purifying kit. It was shown that a fragment of 460bp was gained by PCR. With the techniques of double-cleave and electrophoresis, the recombinant vector was identified as pGEX1-NT-6. The recombinant vector pGEX1-NT-6 transformed Escherichia coli expressed fusion protein of 41KD after induction by IPTG. Cleaved by thrombin, the mature peptide of NT-6 was obtained; its molecular weight was about 15KD. The cloning and expression of human-derived NT-6 gene encoding mature protein has provided a basis for further studies on the function and clinical application of NT-6.

  • protecting effect of human derived Neurotrophin 6 on retrogradely degenerated motoneurons of facial nucleus of rats
    Chinese journal of applied physiology, 2005
    Co-Authors: Chengwu Zhang, Zicheng Li, Li Li, Yu Zheng
    Abstract:

    AIM: The protecting effect of human-derived Neurotrophin-6 (NT-6) on injured neurons was investigated to test the neurobiological characteristics of human-derived NT-6. METHODS: Adult SD rats were used and divided into two groups, normal control group and experiment group. The right facial nerve of the experimental rats was sectioned. The experimental animals were subdivided into three groups, blank control group, NT-6 group and saline control group. The rats were raised for two weeks. Brain stem of the rats was removed and transversely sliced. Nissle stain and ChAT immunohistochemical stain of the slices were carried out to observe the role of NT-6 in protecting facial motoneurons. RESULTS: The number of the ChAT positive neurons and the intensity of Nissle stain in the facial nucleus of NT-6 group were significantly augmented compared with that of the blank and saline control groups. CONCLUSION: The human-derived NT-6 could partially protect the facial motoneurons from retrograde degeneration induced by their axon damage.

  • cloning of human brain derived Neurotrophin 6 gene and its expression in procaryotic cell
    Chinese journal of medical genetics, 2002
    Co-Authors: Chengwu Zhang, Xu Zhuang, Chaoyang Zhai, Yu Zheng
    Abstract:

    OBJECTIVE: To clone human brain-derived Neurotrophin-6(NT-6) gene and to observe its expression in the procaryotic cell. METHODS: Total RNA was extracted from aborted antenatal cerebral cortex, and cDNA fragment of NT-6 was amplified through reverse transcript-polymerase chain reaction. After being incised and recovered, the NT-6 gene was cloned into pBK-CMV plasmid to construct a NT-6 gene expression vector. Expression of NT-6 gene in Escherichia coli was studied after being induced by isopropyl beta-D-thiogalactoside(IPTG). RESULTS: The NT-6 gene expression vector was constructed and Escherichia coli with recombinant vector expressed specific protein after induction by IPTG. CONCLUSION: The cloning of human brain-derived NT-6 gene provides a basis for further studying the structure, function and clinical application of NT-6.

Hans Thoenen - One of the best experts on this subject based on the ideXlab platform.

  • ultrastructural identification of storage compartments and localization of activity dependent secretion of Neurotrophin 6 in hippocampal neurons
    Molecular and Cellular Neuroscience, 2000
    Co-Authors: Annette Gartner, Yuri Shostak, Natalie Hackel, Iryna M Ethell, Hans Thoenen
    Abstract:

    A modulatory role of Neurotrophins (NTs) in activity-dependent neuronal plasticity by pre- and postsynaptic mechanisms is now well established. In this context, it is important to identify the storage compartments and to localize the precise site(s) and mechanism of NT secretion in order to deduce the spatial and temporal availability of NTs. We approached these questions at the ultrastructural level, exploiting the unique property of NT6 to bind tightly to heparan sulfate proteoglycans at the neuronal surface (R. Gotz et al., 1994, Nature372, 266–269), permitting the localization of secretion sites excluding diffusion artifacts. The myc tagging of NT6 permitted glutaraldehyde fixation and hence good preservation of the membrane structure, permitting immunogold labeling of NT6myc at the neuronal surface. NT6myc is preferentially secreted from neurites compared to neuronal cell bodies. In agreement with light-microscopic observations, the ultrastructural localization of NT6myc by postembedding procedures showed a predominant localization in ER-like membrane-confined compartments, partially associated with microtubules.

  • Neurotrophin 6 is a new member of the nerve growth factor family
    Nature, 1994
    Co-Authors: Rudolf Gotz, Christoph Winkler, Franz Raulf, Manfred Schartl, Friedrich Lottspeich, Reinhard W Koster, Hans Thoenen
    Abstract:

    DURING vertebrate development, many neurons depend for survival and differentiation on their target cells1–3. The best documented mediator of such a retrograde trophic action is the Neurotrophin nerve growth factor (NGF)1. NGF and the other known members of the Neurotrophin family, brain-derived neurotrophic factor (BDNF), Neurotrophin-3 (NT-3) and Neurotrophin-4/5 (NT-4/5) are conserved as distinct genes over large evolutionary distances4–6. Here we report the cloning of Neurotrophin-6 (NT-6), a new mem-ber of this family from the teleost fish Xiphophorus. NT-6 dis-tinguishes itself from the other known Neurotrophins in that it is not found as a soluble protein in the medium of producing cells. The addition of heparin (but not chondroitin) effects the release of NT-6 from cell surface and extracellular matrix molecules. Recombinant purified NT-6 has a spectrum of actions similar to NGF on chick sympathetic and sensory neurons, albeit with a lower potency. NT-6 is expressed in the embryonic valvulla cerebelli; expression persists in some adult tissues. The interaction of NT-6 with heparin-binding molecules may modulate its action in the nervous system.

Friedrich Lottspeich - One of the best experts on this subject based on the ideXlab platform.

  • recombinant fish Neurotrophin 6 is a heparin binding glycoprotein implications for a role in axonal guidance
    Biochemical Journal, 1997
    Co-Authors: Xiaoling Li, Rudolf Gotz, Friedrich Lottspeich, Juliane Franz
    Abstract:

    Neurotrophin-6 (NT-6) was identified in the teleost fish Xiphophorus as a new member of the Neurotrophin gene family. NT-6 binds specifically the glycosaminoglycan heparin. In this study NT-6 was expressed in a stably transfected mammalian cell line, and in insect cells via a recombinant baculovirus. It was purified to homogeneity and characterized by MS and N-terminal sequencing. NT-6 from both expression systems was proteolytically processed at one of two protease cleavage motifs and was found to be glycosylated. It supported the survival of embryonic chick sensory neurons; half-maximal survival was observed at 100 ng/ml. Furthermore, NT-6 elicited neurite outgrowth in explanted embryonic dorsal root ganglia. Addition of heparin into the medium did not potentiate the activity of NT-6 in survival assays. However, when a sensory ganglion explant was cultured in a collagen gel matrix assay adjacent to a heparin bead coated with NT-6, neurite outgrowth directed towards the bead was observed. This indicated that NT-6 was slowly released from the heparin bead generating a concentration gradient of NT-6 instrumental for axonal guidance in vitro. Thus the interaction of NT-6 with heparin might not be required for the activation of the cellular receptor for NT-6 on responsive cells but rather may serve to control, in vivo, the distribution of NT-6.

  • Neurotrophin 6 is a new member of the nerve growth factor family
    Nature, 1994
    Co-Authors: Rudolf Gotz, Christoph Winkler, Franz Raulf, Manfred Schartl, Friedrich Lottspeich, Reinhard W Koster, Hans Thoenen
    Abstract:

    DURING vertebrate development, many neurons depend for survival and differentiation on their target cells1–3. The best documented mediator of such a retrograde trophic action is the Neurotrophin nerve growth factor (NGF)1. NGF and the other known members of the Neurotrophin family, brain-derived neurotrophic factor (BDNF), Neurotrophin-3 (NT-3) and Neurotrophin-4/5 (NT-4/5) are conserved as distinct genes over large evolutionary distances4–6. Here we report the cloning of Neurotrophin-6 (NT-6), a new mem-ber of this family from the teleost fish Xiphophorus. NT-6 dis-tinguishes itself from the other known Neurotrophins in that it is not found as a soluble protein in the medium of producing cells. The addition of heparin (but not chondroitin) effects the release of NT-6 from cell surface and extracellular matrix molecules. Recombinant purified NT-6 has a spectrum of actions similar to NGF on chick sympathetic and sensory neurons, albeit with a lower potency. NT-6 is expressed in the embryonic valvulla cerebelli; expression persists in some adult tissues. The interaction of NT-6 with heparin-binding molecules may modulate its action in the nervous system.