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Intrathecal infusion of growth factors stimulates NG2-expressing progenitors in mutant SOD1-transgenic rats
S84
Abstracts / Neuroscience Research 58S (2007) S1–S244
P1-c36 Microarray analysis of neurospheres treated with mood stabilizers
P1-c42 Intrathecal infusion of growth factors stimulates NG2expressing progenitors in mutant SOD1-transgenic rats
Yuichi Mikito Seiji Kazuhiro Ikenaka 1,2 1 The Graduate University for Advanced Studies, Aichi, Japan; 2 NIPS, Japan
Hitoshi Warita, Masashi Aoki, Hideki Mizuno, Yasuto Itoyama Division of Neurology, Tohoku University, Sendai, Japan
Matsushita 1,2 ,
Higashi 2 ,
Hitoshi 2 ,
Neural stem cells (NSCs) are present in the subventricular zone in the adult brain and provide neurons in the olfactory bulb through the rostral migratory stream. We have shown that mood-stabilizing drugs enhance the self-renewal of NSCs and expand the size of NSC pool in the adult brain. In this study, we performed microarray analysis using neurospheres which were cultured in the presence of mood stabilizers. Several genes were isolated, which were upregulated or downregulated by the drug treatment and we focused a gene that is downregulated more than twofold by both lithium and valproic acid treatment. The gene contains a WW domain and seems to act as a transcriptional coactivator, but its function in the nervous system is not known. By in situ hybridization analysis, the expression of the gene is detected in the tissue surrounding the lateral ventricle of the embryonic and adult brain, where NSCs exist. We are investigating the function of the gene in the NSCs.
P1-c38 Decreased expression of Olig2 in response to amyloid accelerates the death of neural stem/progenitor cells
Yoko Uchida, Fujiya Gomi Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan Despite increased neurogenic differentiation markers in hippocampal CA1 in AD, neurons are not replaced in CA1 and neocortex in the disease. A might contribute to deteriorate brain microenvironment for neurogenesis and survival of immature neurons. We examined whether A alters the expression of cell fate determinants in cerebral cortical cultures and in an AD mouse model (PrP-APPSW). The up-regulation of Mash1 and down-regulation of Olig2 were found in cerebral cortical cultures treated with A1–42. A increased in the proportion of Olig2negative/Mash1-expressing cells, which are not labeled with cell-specific markers. The decrease in Olig2+ cells was also observed in cerebral cortex of adult PrP-APPSW mice. However, Mash1+ cells were not detected in either wild type or PrP-APPSW mice. Cotransfection experiments with Mash1 cDNA and Olig2 siRNA revealed that Olig2 may decide the fate of Mash1-overexpressing cells to differentiation or death. We conclude that down-regulation of Olig2 by A in Mash1-overexpressing cells switch the cell fate to death.
P1-c39 Expression pattern of Olig2 in the rat olfactory bulb following olfactory epithelium lesion Kazuho Hirata 1 , Takaaki Kanemaru 2 , Motozumi Minohara 3 , Jun-ichi Kira 3 1 Department of Anatomy & Cell Biology, Kyushu University, Fukuoka, Japan; 2 Morphology Core Unit, Japan; 3 Department of Neurology, Japan Our previous study showed that nestin positive reactive astrocytes appeared in glomeruli of the olfactory bulb at 3–7 days after the olfactory epithelium lesion. To clarify the detail of astroglial response in the olfactory bulb, immunohistochemistry for the transcription factor Olig2 was carried out. At 3–7 days after the lesion, glial progenitor cells with nuclear Olig2 expression significantly increased in the glomerular layer compared to those of the control animals that were sparsely distributed. Nestin positive reactive astrocytes showed the translocation of Olig2 expression into the cytoplasm. Thus the reactive astrocytes are thought to be derived from glial progenitor cells. Swift conversion from progenitors to reactive astrocytes following the lesion may play a crucial role on reinnervation in the olfactory bulb. Health and Labour Sciences research grants (H16-kokoro-026) from the Ministry of Health, Labour and Welfare.
NG2-expressing (NG2+ ) cells, previously known as oligodendrocyte progenitor cells, constitute the largest proliferating cell population in the adult central nervous system. Recent reports have shown that the NG2+ cells are intrinsically multipotent and have the ability to generate neurons. In the present study, we examined the NG2+ cells in the spinal cord of mutant SOD1-transgenic (Tg) rats characterized by adult-onset selective motoneuron degeneration. In addition, we studied whether exogenous growth factors could promote the NG2+ cell proliferation in the degenerating spinal cord. In parallel with the motoneuron loss, the NG2+ cells proliferated significantly as compared with wild-type rats. However, the number of newly generated mature oligodendrocytes was not significantly different between Tg and wild-type rats. Moreover, intrathecal infusion of EGF and FGF-2 in Tg rats significantly promoted the proliferation of NG2+ cells. Therefore, the NG2+ cells may be a potential target to develop a restoration therapy for neurodegenerative diseases. Research fund: KAKENHI (18790586)
P1-c43 Enhanced expression of TGF1 in cell cycle blockerinduced neuronal differentiation of neurosphere Sachiyo Misumi, Hideki Hida, Hye-Jung Kim, Hitoo Nishino Department. Neurophysiology & Brain Science, Nagoya City University, Nagoya, Japan We have shown that pre-treatment with deferoxamine (DFO: cell cycle blocker in G1/S phase) increase the number of neurons from neural stem/progenitor cells (NPCs) with relation to prolonged enhancement of p27kip1 mRNA expression. To investigate the molecular mechanism of neuronal differentiation by cell cycle blocker in detail, we focused on transforming growth factor beta (TGF). As TGF1 increases transcription of CKIs, such as p21cip1 and p27kip1 and inhibits cell proliferation, we investigated the expression of TGF mRNAs at the end point of DFO treatment and at 24 hours after differentiation when p27kip1 mRNA elevation has started. Expression of TGF1 mRNA was significantly elevated after the differentiation, although it was not altered during DFO treatment. On the other hand, different TGF isoforms expression did not change. Data suggest that enhancement of TGF1 expression along with p27kip1 elevation would be included in the mechanism of neuronal differentiation followed by cell cycle arrest.
P1-c44 Astrocyte-specific labeling with a recombinant lentiviral vector carrying DsRed protein driven by a human glial fibrillary acidic protein promoter Kaoru Sato 1 , Rachel E. Ventura 2 , James E. Goldman 2 , Ken Nakazawa 1 1 Division of Pharmacology, National Institute of Health Science, Tokyo, Japan; 2 Department of Pathology, Columbia University, NY, USA We have constructed a recombinant lentiviral vector carrying DsRed protein driven by a hGFAP promoter. We checked expression patterns of cell type-specific markers in DsRed-positive cells 1 day after infection of primary cultured cortical cells with this lentiviral vector. In DsRed-positive cells, the expression pattern was as follows: 100 ± 0.32% for GFAP (+), 98.1 ± 1.15% for S100 (+), 100 ± 2.61% for GLAST (+), 81.6% ± 3.31% for Nestin (+), 0.08 ± 0.43% for Tuj1 (+), and 0.00 ± 0.00% for O4 (+), indicating that this lentiviral vector achieved astrocyte-specific gene targeting. Currently we are investigating the in vivo expression pattern of these markers in DsRed-positive cells after direct injection of this lentiviral vector into the subventricular zone and white matter. This technique will be a strong tool for long term monitoring of morphology and differentiation of astrocytes and/or radial glia. Research funds: KAKENHI 18700373 (K.S.), NIH grants F30 NS-051997 (R.E.V.) and NS-017125 (J.E.G.)
Abstracts / Neuroscience Research 58S (2007) S1–S244
P1-c36 Microarray analysis of neurospheres treated with mood stabilizers
P1-c42 Intrathecal infusion of growth factors stimulates NG2expressing progenitors in mutant SOD1-transgenic rats
Yuichi Mikito Seiji Kazuhiro Ikenaka 1,2 1 The Graduate University for Advanced Studies, Aichi, Japan; 2 NIPS, Japan
Hitoshi Warita, Masashi Aoki, Hideki Mizuno, Yasuto Itoyama Division of Neurology, Tohoku University, Sendai, Japan
Matsushita 1,2 ,
Higashi 2 ,
Hitoshi 2 ,
Neural stem cells (NSCs) are present in the subventricular zone in the adult brain and provide neurons in the olfactory bulb through the rostral migratory stream. We have shown that mood-stabilizing drugs enhance the self-renewal of NSCs and expand the size of NSC pool in the adult brain. In this study, we performed microarray analysis using neurospheres which were cultured in the presence of mood stabilizers. Several genes were isolated, which were upregulated or downregulated by the drug treatment and we focused a gene that is downregulated more than twofold by both lithium and valproic acid treatment. The gene contains a WW domain and seems to act as a transcriptional coactivator, but its function in the nervous system is not known. By in situ hybridization analysis, the expression of the gene is detected in the tissue surrounding the lateral ventricle of the embryonic and adult brain, where NSCs exist. We are investigating the function of the gene in the NSCs.
P1-c38 Decreased expression of Olig2 in response to amyloid accelerates the death of neural stem/progenitor cells
Yoko Uchida, Fujiya Gomi Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan Despite increased neurogenic differentiation markers in hippocampal CA1 in AD, neurons are not replaced in CA1 and neocortex in the disease. A might contribute to deteriorate brain microenvironment for neurogenesis and survival of immature neurons. We examined whether A alters the expression of cell fate determinants in cerebral cortical cultures and in an AD mouse model (PrP-APPSW). The up-regulation of Mash1 and down-regulation of Olig2 were found in cerebral cortical cultures treated with A1–42. A increased in the proportion of Olig2negative/Mash1-expressing cells, which are not labeled with cell-specific markers. The decrease in Olig2+ cells was also observed in cerebral cortex of adult PrP-APPSW mice. However, Mash1+ cells were not detected in either wild type or PrP-APPSW mice. Cotransfection experiments with Mash1 cDNA and Olig2 siRNA revealed that Olig2 may decide the fate of Mash1-overexpressing cells to differentiation or death. We conclude that down-regulation of Olig2 by A in Mash1-overexpressing cells switch the cell fate to death.
P1-c39 Expression pattern of Olig2 in the rat olfactory bulb following olfactory epithelium lesion Kazuho Hirata 1 , Takaaki Kanemaru 2 , Motozumi Minohara 3 , Jun-ichi Kira 3 1 Department of Anatomy & Cell Biology, Kyushu University, Fukuoka, Japan; 2 Morphology Core Unit, Japan; 3 Department of Neurology, Japan Our previous study showed that nestin positive reactive astrocytes appeared in glomeruli of the olfactory bulb at 3–7 days after the olfactory epithelium lesion. To clarify the detail of astroglial response in the olfactory bulb, immunohistochemistry for the transcription factor Olig2 was carried out. At 3–7 days after the lesion, glial progenitor cells with nuclear Olig2 expression significantly increased in the glomerular layer compared to those of the control animals that were sparsely distributed. Nestin positive reactive astrocytes showed the translocation of Olig2 expression into the cytoplasm. Thus the reactive astrocytes are thought to be derived from glial progenitor cells. Swift conversion from progenitors to reactive astrocytes following the lesion may play a crucial role on reinnervation in the olfactory bulb. Health and Labour Sciences research grants (H16-kokoro-026) from the Ministry of Health, Labour and Welfare.
NG2-expressing (NG2+ ) cells, previously known as oligodendrocyte progenitor cells, constitute the largest proliferating cell population in the adult central nervous system. Recent reports have shown that the NG2+ cells are intrinsically multipotent and have the ability to generate neurons. In the present study, we examined the NG2+ cells in the spinal cord of mutant SOD1-transgenic (Tg) rats characterized by adult-onset selective motoneuron degeneration. In addition, we studied whether exogenous growth factors could promote the NG2+ cell proliferation in the degenerating spinal cord. In parallel with the motoneuron loss, the NG2+ cells proliferated significantly as compared with wild-type rats. However, the number of newly generated mature oligodendrocytes was not significantly different between Tg and wild-type rats. Moreover, intrathecal infusion of EGF and FGF-2 in Tg rats significantly promoted the proliferation of NG2+ cells. Therefore, the NG2+ cells may be a potential target to develop a restoration therapy for neurodegenerative diseases. Research fund: KAKENHI (18790586)
P1-c43 Enhanced expression of TGF1 in cell cycle blockerinduced neuronal differentiation of neurosphere Sachiyo Misumi, Hideki Hida, Hye-Jung Kim, Hitoo Nishino Department. Neurophysiology & Brain Science, Nagoya City University, Nagoya, Japan We have shown that pre-treatment with deferoxamine (DFO: cell cycle blocker in G1/S phase) increase the number of neurons from neural stem/progenitor cells (NPCs) with relation to prolonged enhancement of p27kip1 mRNA expression. To investigate the molecular mechanism of neuronal differentiation by cell cycle blocker in detail, we focused on transforming growth factor beta (TGF). As TGF1 increases transcription of CKIs, such as p21cip1 and p27kip1 and inhibits cell proliferation, we investigated the expression of TGF mRNAs at the end point of DFO treatment and at 24 hours after differentiation when p27kip1 mRNA elevation has started. Expression of TGF1 mRNA was significantly elevated after the differentiation, although it was not altered during DFO treatment. On the other hand, different TGF isoforms expression did not change. Data suggest that enhancement of TGF1 expression along with p27kip1 elevation would be included in the mechanism of neuronal differentiation followed by cell cycle arrest.
P1-c44 Astrocyte-specific labeling with a recombinant lentiviral vector carrying DsRed protein driven by a human glial fibrillary acidic protein promoter Kaoru Sato 1 , Rachel E. Ventura 2 , James E. Goldman 2 , Ken Nakazawa 1 1 Division of Pharmacology, National Institute of Health Science, Tokyo, Japan; 2 Department of Pathology, Columbia University, NY, USA We have constructed a recombinant lentiviral vector carrying DsRed protein driven by a hGFAP promoter. We checked expression patterns of cell type-specific markers in DsRed-positive cells 1 day after infection of primary cultured cortical cells with this lentiviral vector. In DsRed-positive cells, the expression pattern was as follows: 100 ± 0.32% for GFAP (+), 98.1 ± 1.15% for S100 (+), 100 ± 2.61% for GLAST (+), 81.6% ± 3.31% for Nestin (+), 0.08 ± 0.43% for Tuj1 (+), and 0.00 ± 0.00% for O4 (+), indicating that this lentiviral vector achieved astrocyte-specific gene targeting. Currently we are investigating the in vivo expression pattern of these markers in DsRed-positive cells after direct injection of this lentiviral vector into the subventricular zone and white matter. This technique will be a strong tool for long term monitoring of morphology and differentiation of astrocytes and/or radial glia. Research funds: KAKENHI 18700373 (K.S.), NIH grants F30 NS-051997 (R.E.V.) and NS-017125 (J.E.G.)