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Netrin-1 regulates cell surface distribution of its receptor DCC in hamster primary cortical neurons
S148
Abstracts / Neuroscience Research 58S (2007) S1–S244
P2-d28 Netrin-1 regulates cell surface distribution of its recep-
P2-d32 Neonatal heregulin/neuregulin signaling promotes
tor DCC in hamster primary cortical neurons
development of cortical GABA neurons
Hideko Matsumoto, Masabumi Nagashima Department of Anatomy, School of Medicine, Saitama Medical University, Saitama, Japan
Yuichi Abe, Hisaaki Namba, Hiroyuki Nawa Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, Japan
Deleted in colorectal cancer (DCC), a receptor for an axon guidance cue netrin-1, plays an important role in target innervation during development of central nervous system. In rat spinal commissural axons it is reported that intracellular vesicular pool of DCC exists and plasma membrane insertion of DCC occurs in response to netrin-1/protein kinase A. In addition, rapid depletion of cell surface DCC after netrin-1 treatment is reported in retinal growth cones of Xenopus laevis, which is dependent on endocytosis. In hamster cortical neurons netrin-1 is known to promote axon branching. In order to clarify whether translocation of DCC to, and/or endocytic removal of DCC from the plasma membrane occur by netrin-1 stimulation, we examined the changes in the level and distribution of DCC at the cell surface of hamster cortical neurons, employing total internal reflection fluorescence microscopy as well as cell-surface labeling techniques. A change in the distribution of cell surface DCC was observed in response to netrin-1 within 5 min.
Heregulin (HRG) is identical to neuregulin-1 and known to play crucial roles in the development of the central nervous system, such as glial differentiation, regulation of glutamate receptor expression, and neural migration. HRG receptor, ErbB4, is preferentially expressed in parvalbumin (PV)-positive GABAergic neurons. Here, we investigated the effect of HRG1 on the development of cortical GABAergic neuron and inhibitory synaptic transmission. In dissociated cortical culture, chronic application of the HRG1-EGF domain (25 ng/ml/day for 7days) enhances the expression of PV, whereas it has no effect on GAD67 levels. Brain slices were prepared from the prefrontal cortex of the postnatal mice receiving the HRG1-EGF domain and patch clump recording was performed in these slices. HRG1 administration enhances both frequency and amplitudes of spontaneous IPSCs in pyramidal neurons. These results suggest that ErbB4 stimulation enhances the development of PV-positive GABAergic neurons and/or termini.
P2-d29 Identification of a novel zebrafish semaphorin
P2-d33 Hypoxia inducible factor-1␣ participate in dopaminer-
Masahiko Taniguchi 1 , Yoshinori Mikami 2 , Tomoyuki Masuda 3 , Masafumi Kimura 4 , Tomoyuki Yoshida 2 , Masayoshi Mishina 2 , Takao Shimizu 4 1 Department of Biochemistry, Cancer Research Institute, Sapporo Medical University, Sapporo, Japan; 2 Department of Neurobiology and Pharmacology, University of Tokyo, Tokyo, Japan; 3 Department of Anatomy, Fukushima Medical University, Fukushima, Japan; 4 Department of Biochemistry and Molecular Biology, University of Tokyo, Tokyo, Japan Semaphorin gene family contains a large number of secreted or transmembrane proteins, and some of them function as the repulsive and attractive cues of axon guidance during development. Here we report a novel zebrafish semaphorin, semaphorin 6E (Sema6E). The ORF of Sema6E is 3192 bp (1064 aa). Zebrafish Sema6E protein showed a 45.1% identity with zebrafish Sema6D protein. To clarify the expression pattern of Sema6E, in situ hybridization was performed. In the embryos, Sema6E is expressed in the lens and hindbrain. We also found that Sema6E induces the repulsion of the axons. These results indicate that Sema6E repels specific types of the axons and functions in the nervous system development.
gic differentiation from ES-derived neural stem cells Tae-Sun Kim, Hideki Hida, Tadashi Masuda, Cha-Gyun Jung, Hitoo Nishino Department of Neuro-physiology & Brain Science, Nagoya City University, Nagoya, Japan To investigate how to induce DAergic differentiation from ES-derived neural stem cells (NSCs), low (3.5%) O2 or cytokine cocktail (IL-1, IL-11, LIF, GDNF) was treated to NSCs. Treatment with cytokine cocktail or low O2 increased the number of tyrosine hydroxylase (TH)-positive(+) cells as compared to controls (2.20-fold or 1.83-fold, respectively). Although major effect of cytokine cocktail was induced by LIF application alone, combination of low O2 with cytokine cocktail did not show any additive effect. To investigate the mechanism of DAergic differentiation by cytokine cocktail, we focused on hypoxia inducible factor (HIF)-1␣. As HIF-1␣ expression was enhanced by LIF treatment or low O2 , antisense ODNs to HIF-1␣ was treated to NSCs followed by TH immunnostaining, revealing that the number of TH (+) cells was significantly decreased in LIF-treated NSCs. Data suggest that enhanced differentiation to DAergic neurons from ES-derived NSCs by low O2 or cytokines was mediated by increased expression of HIF-1␣.
Research funds: KAKENHI on Priority Areas-Elucidation of neural network function in the brain- from the MEXT (18021031)
P2-d30
Mechanism of activation of transforming growth factor-beta in the brain by exercise
P2-d34 Neonatal EGF challenge attenuates GABAergic development in the neocortex and hippocampus
Teppei Fujikawa, Hiroyuki Yamada, Yoko Iwaki, Shigenobu Matsumura, Kazuo Inoue, Tohru Fushiki Division of Food Science and Biotechnology, Kyoto University, Kyoto, Japan
Hisaaki Namba, Yuichi Abe, Hiroyuki Nawa Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, Japan
Transforming growth factor-beta (TGF-beta) is a well-known pleiotropic cytokine. We previously reported the increase of active TGF-beta in the cerebrospinal fluid (CSF) after exercise in rats. TGF-beta is secreted as a latent form which consists of latency associated protein (LAP) and mature TGF-beta itself. To reveal its activity, latent form should be activated by cleavage or removal of LAP. In this study we investigated the mechanism of activation of TGF-beta in the brain during exercise. Among the factors that increase during exercise in the blood, intraperitonial (i.p.) injection of lactate and beta-hydroxybutylic acid could reproduce the increase in active TGF-beta in the CSF. Matrix metalloproteinases and plasmin which degrade LAP, and thrombospondin-1 (TSP-1) which causes structural change to LAP, are reported to active TGF-beta. We found that intracisternal administration of LSKL peptide that inhibits the interaction between LAP and TSP-1 could only block the activation of TGF-beta by i.p. administration of lactate.
We found that administration of epidermal growth factor (EGF) to neonatal mice down-regulates expression of AMPA-type glutamate receptors in developing GABAergic neurons. In parallel, the strength of recurrent inhibitory activities is also decreased in the dentate gyrus (Neurosci. Res. 52 (Suppl.) (2005) S133). Here, we examined phenotypic expressions of GABAergic markers in the frontal cortex and hippocampus of EGFtreated postnatal mice. EGF administration suppressed protein-levels of parvalbumin (PV) in the brain regions. The expression of GABA transporter, GAT-1 was also decreased in the frontal cortex. Slice patch-clamp recordings from the dentate gyrus of the EGF-administered mice revealed that the frequency of spontaneous inhibitory postsynaptic currents was decreased in the granule cells. Thus, during early postnatal development, activation of EGF receptors (ErbBs) negatively influences the phenotypic and functional maturation of PV-positive GABAergic neurons.
Abstracts / Neuroscience Research 58S (2007) S1–S244
P2-d28 Netrin-1 regulates cell surface distribution of its recep-
P2-d32 Neonatal heregulin/neuregulin signaling promotes
tor DCC in hamster primary cortical neurons
development of cortical GABA neurons
Hideko Matsumoto, Masabumi Nagashima Department of Anatomy, School of Medicine, Saitama Medical University, Saitama, Japan
Yuichi Abe, Hisaaki Namba, Hiroyuki Nawa Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, Japan
Deleted in colorectal cancer (DCC), a receptor for an axon guidance cue netrin-1, plays an important role in target innervation during development of central nervous system. In rat spinal commissural axons it is reported that intracellular vesicular pool of DCC exists and plasma membrane insertion of DCC occurs in response to netrin-1/protein kinase A. In addition, rapid depletion of cell surface DCC after netrin-1 treatment is reported in retinal growth cones of Xenopus laevis, which is dependent on endocytosis. In hamster cortical neurons netrin-1 is known to promote axon branching. In order to clarify whether translocation of DCC to, and/or endocytic removal of DCC from the plasma membrane occur by netrin-1 stimulation, we examined the changes in the level and distribution of DCC at the cell surface of hamster cortical neurons, employing total internal reflection fluorescence microscopy as well as cell-surface labeling techniques. A change in the distribution of cell surface DCC was observed in response to netrin-1 within 5 min.
Heregulin (HRG) is identical to neuregulin-1 and known to play crucial roles in the development of the central nervous system, such as glial differentiation, regulation of glutamate receptor expression, and neural migration. HRG receptor, ErbB4, is preferentially expressed in parvalbumin (PV)-positive GABAergic neurons. Here, we investigated the effect of HRG1 on the development of cortical GABAergic neuron and inhibitory synaptic transmission. In dissociated cortical culture, chronic application of the HRG1-EGF domain (25 ng/ml/day for 7days) enhances the expression of PV, whereas it has no effect on GAD67 levels. Brain slices were prepared from the prefrontal cortex of the postnatal mice receiving the HRG1-EGF domain and patch clump recording was performed in these slices. HRG1 administration enhances both frequency and amplitudes of spontaneous IPSCs in pyramidal neurons. These results suggest that ErbB4 stimulation enhances the development of PV-positive GABAergic neurons and/or termini.
P2-d29 Identification of a novel zebrafish semaphorin
P2-d33 Hypoxia inducible factor-1␣ participate in dopaminer-
Masahiko Taniguchi 1 , Yoshinori Mikami 2 , Tomoyuki Masuda 3 , Masafumi Kimura 4 , Tomoyuki Yoshida 2 , Masayoshi Mishina 2 , Takao Shimizu 4 1 Department of Biochemistry, Cancer Research Institute, Sapporo Medical University, Sapporo, Japan; 2 Department of Neurobiology and Pharmacology, University of Tokyo, Tokyo, Japan; 3 Department of Anatomy, Fukushima Medical University, Fukushima, Japan; 4 Department of Biochemistry and Molecular Biology, University of Tokyo, Tokyo, Japan Semaphorin gene family contains a large number of secreted or transmembrane proteins, and some of them function as the repulsive and attractive cues of axon guidance during development. Here we report a novel zebrafish semaphorin, semaphorin 6E (Sema6E). The ORF of Sema6E is 3192 bp (1064 aa). Zebrafish Sema6E protein showed a 45.1% identity with zebrafish Sema6D protein. To clarify the expression pattern of Sema6E, in situ hybridization was performed. In the embryos, Sema6E is expressed in the lens and hindbrain. We also found that Sema6E induces the repulsion of the axons. These results indicate that Sema6E repels specific types of the axons and functions in the nervous system development.
gic differentiation from ES-derived neural stem cells Tae-Sun Kim, Hideki Hida, Tadashi Masuda, Cha-Gyun Jung, Hitoo Nishino Department of Neuro-physiology & Brain Science, Nagoya City University, Nagoya, Japan To investigate how to induce DAergic differentiation from ES-derived neural stem cells (NSCs), low (3.5%) O2 or cytokine cocktail (IL-1, IL-11, LIF, GDNF) was treated to NSCs. Treatment with cytokine cocktail or low O2 increased the number of tyrosine hydroxylase (TH)-positive(+) cells as compared to controls (2.20-fold or 1.83-fold, respectively). Although major effect of cytokine cocktail was induced by LIF application alone, combination of low O2 with cytokine cocktail did not show any additive effect. To investigate the mechanism of DAergic differentiation by cytokine cocktail, we focused on hypoxia inducible factor (HIF)-1␣. As HIF-1␣ expression was enhanced by LIF treatment or low O2 , antisense ODNs to HIF-1␣ was treated to NSCs followed by TH immunnostaining, revealing that the number of TH (+) cells was significantly decreased in LIF-treated NSCs. Data suggest that enhanced differentiation to DAergic neurons from ES-derived NSCs by low O2 or cytokines was mediated by increased expression of HIF-1␣.
Research funds: KAKENHI on Priority Areas-Elucidation of neural network function in the brain- from the MEXT (18021031)
P2-d30
Mechanism of activation of transforming growth factor-beta in the brain by exercise
P2-d34 Neonatal EGF challenge attenuates GABAergic development in the neocortex and hippocampus
Teppei Fujikawa, Hiroyuki Yamada, Yoko Iwaki, Shigenobu Matsumura, Kazuo Inoue, Tohru Fushiki Division of Food Science and Biotechnology, Kyoto University, Kyoto, Japan
Hisaaki Namba, Yuichi Abe, Hiroyuki Nawa Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, Japan
Transforming growth factor-beta (TGF-beta) is a well-known pleiotropic cytokine. We previously reported the increase of active TGF-beta in the cerebrospinal fluid (CSF) after exercise in rats. TGF-beta is secreted as a latent form which consists of latency associated protein (LAP) and mature TGF-beta itself. To reveal its activity, latent form should be activated by cleavage or removal of LAP. In this study we investigated the mechanism of activation of TGF-beta in the brain during exercise. Among the factors that increase during exercise in the blood, intraperitonial (i.p.) injection of lactate and beta-hydroxybutylic acid could reproduce the increase in active TGF-beta in the CSF. Matrix metalloproteinases and plasmin which degrade LAP, and thrombospondin-1 (TSP-1) which causes structural change to LAP, are reported to active TGF-beta. We found that intracisternal administration of LSKL peptide that inhibits the interaction between LAP and TSP-1 could only block the activation of TGF-beta by i.p. administration of lactate.
We found that administration of epidermal growth factor (EGF) to neonatal mice down-regulates expression of AMPA-type glutamate receptors in developing GABAergic neurons. In parallel, the strength of recurrent inhibitory activities is also decreased in the dentate gyrus (Neurosci. Res. 52 (Suppl.) (2005) S133). Here, we examined phenotypic expressions of GABAergic markers in the frontal cortex and hippocampus of EGFtreated postnatal mice. EGF administration suppressed protein-levels of parvalbumin (PV) in the brain regions. The expression of GABA transporter, GAT-1 was also decreased in the frontal cortex. Slice patch-clamp recordings from the dentate gyrus of the EGF-administered mice revealed that the frequency of spontaneous inhibitory postsynaptic currents was decreased in the granule cells. Thus, during early postnatal development, activation of EGF receptors (ErbBs) negatively influences the phenotypic and functional maturation of PV-positive GABAergic neurons.