- Email: [email protected]
progenitor cells
Abstracts as NSCs cells surface molecules, were recognized by E-PHA. However, lectin blot analysis revealed that E-PHA bound to several molecules other than CD133 and EGF-R. These results suggest that the other molecules which could be participated in cell attachment could be existed on NSCs cells surface. doi:10.1016/j.neures.2009.09.373
P1-d18 Regulation of photoreceptor precursor differentiation in retinal development Kimiko Katoh 1,2 , Yoshihiro Omori 2 , Sigeru Sato 2 , Mineo Kondo 3 , Takahisa Furukawa 2 1 3
P1-d15 Molecular function of Tsukushi on neuronal stem cells Kunimasa Ohta 1 , Ayako Ito 1,2 , Sei kuriyama 3 , Giuseppe Lupo 4 , Rika Nakayama 5 , Naoko Ohshima 5 , Mitsuko Kosaka 5 , Shin-ichi Ohnuma 3 , Shinichi Nakagawa 6 , Hideaki Tanaka 1,2 1
Kumamoto Univ., Grad. Sch. of Med Sci., Japan; 2 Global COE, Kumamoto Univ, Japan; 3 UCL, Japan; 4 Univ of Cambridge, USA; 5 RIKEN CDB, Japan; 6 RIKEN FRS, Japan Multipotent stem/progenitor cells have been isolated from the ciliary body (CB) of the mouse retina, and shown to possess several universal characteristics of stem cells. Several lines of evidence suggest that Wnt signaling promotes proliferation of retinal stem/progenitor cells. Nonetheless, little is known about the molecular mechanisms that control stem cell self-renewal and restrict tissue-specific stem cells to specific compartments. We generated knock-out mice lacking TSK function and found that TSK inactivation results in expansion of the CB and the increased proliferation of the retinal stem/progenitor cells in vitro. Biochemical assays and overexpression experiments suggest that these effects are dependent on the ability of TSK to modulate Wnt signaling by binding to Frizzled. We propose that TSK is an important component of the stem/progenitor cell niche in the mammalian CB, controlling stem/progenitor cell proliferation by modulation of Wnt signaling. doi:10.1016/j.neures.2009.09.374
P1-d16 Macrophage migration inhibitory factor (MIF) promotes proliferation and cell survival of neural stem/progenitor cells Shigeki Ohta 1 , Yutaka Kawakami 1 , Hideyuki Okano 2 , Takeshi Kawase 3 , Masahiro Toda 3 1
Ctr. for Advanced Med. Res., Keio University, Tokyo, Japan; 2 Department of Physiol., Keio University, Tokyo, Japan; 3 Department of Neurosurgery, Keio University, Tokyo, Japan In previous studies, we demonstrated that mouse dendritic cells (DCs) can increase the number of neural stem/progenitor cells (NSPCs) in vitro and in vivo. In this study, we identified macrophage migration inhibitory factor (MIF) that is secreted from DCs and NSPCs as a novel factor that can support the proliferation and/or survival of NSPCs in vitro. To date the function of MIF in the normal brain remains largely unknown. FACS analyses showed that murine neurospheres express CD74 and CD44 which are known as MIF receptors. MIF increased the number of primary and secondary neurospheres. In contrast, MIF blocker and MIF shRNAi suppressed the neurosphere formations. Moreover, we found that MIF increases the phosphorylation of Akt and Erk in neurospheres that may support the cells? survival and proliferation. Taken together, MIF is a newly identified factor that can promote the survival and/or proliferation of NSPCs. doi:10.1016/j.neures.2009.09.375
P1-d17 Inhibitory effects of rotenone on migration of neural stem cells in neurosphere assay in vitro Masami Ishido, Junko Suzuki Natl. Inst. for Environmental Studies, Japan Parkinson’s disease is primarily a sporadic disorder and its specific etiology is incompletely understood. According to the working hypothesis of DOHaD (Developmental origins of health and disease), environmental origins of human sporadic Parkinson, s disease might be early in life. For the first step to substantiate this, in this study we examined the effect of the dopaminergic toxin, rotenone on neural stem cells in vitro, comparing with those of PC12 cells. Neurosphere was isolated from rat E15–16 mesencephalone in the presence of bFGF and EGF. To facilitate the stem cells to migrate from neurosphere in vitro, several neurospheres were cultured on a test well. Exposure of neurospheres to rotenone prevented the cell from migrating with the decrement of Ki67-positive cells; TUNEL-positive cells were increased. Thus, the data obtained would suggest the possibility that exposure to rotenone in utero might disturb the developmental programming, resulting in corresponding disease in later life. doi:10.1016/j.neures.2009.09.376
S91
Dept. Biostudies, Univ. of Kyoto, Kyoto, Japan; 2 Dev., OBI, Osaka, Japan; Dept. Ophthalmol., Univ. of Nagoya, Nagoya, Japan
The vertebrate retina consists of six types of neurons and one type of glial cells, which are all derived from multipotent progenitors. There is little information on the molecular mechanisms regulating the specification of cell type identity from multipotent progenitors in the developing retina. We have previously demonstrated that Otx2 acts as a master regulator of photoreceptor cell fate determination using photoreceptor-specific Otx2 conditional knockout (CKO) mice. Here, we performed microarray analysis comparing the retinal gene expression profiles of wild-type and Otx2 CKO mice. We have identified Blimp1 as one of the most reduced genes in the Otx2 CKO mice. In the Blimp1 CKO mice, we found that the number of photoreceptors was significantly reduced. Electroretinograms analysis also showed that the response of photoreceptor cells was impaired in adult Blimp1 CKO mice. These results suggest that Blimp1 plays an important role in the development of photoreceptors. doi:10.1016/j.neures.2009.09.377
P1-d19 The role of Ubc9, a sumo-conjugating enzyme, in retinal stem/progenitor proliferation Koji Terada, Takahisa Furukawa Depts. of Developmental Biology, Osaka Bio. Inst., Japan The mechanisms which underlie the regulation of stem/progenitor cell proliferation in the neural tissue during early development have been poorly understood. We previously found that xhmgb3 is expressed in the ciliary marginal zone where retinal stem cell exist in the Xenopus retina and that overexpression of xhmgb3 induced overproliferation of retinal progenitors. In this study, we performed yeast two-hybrid screening to identify the factors that act with xhmgb3, and we identified Ubc9. Ubc9 is involved in various cellular functions, including the control of a subcellular localization and the activity of target proteins by sumoylation. We performed overexpression experiments of xhmgb3 and Ubc9 in Xenopus embryos. As a result, we observed an eye enlargement more than two-fold in the frequency compared with that in only xhmgb3-injected embryos, suggesting that xhmgb3 and Ubc9 act cooperatively in eye development. Our results suggest that Ubc9 controls progenitor cell proliferation in the Xenopus retina. We are further investigating the mechanisms of progenitor cell proliferation regulated by xhmgb3 and Ubc9. doi:10.1016/j.neures.2009.09.378
P1-d20 Roles of Meis2 in the generation of medium spiny neurons in the developing striatum Saki Takami, Fujio Murakami, Yasuto Tanabe Grad. Sch. Front. Biosci., Osaka University, Suita, Japan Meis2 belongs to the family of homeodomain-containing TALE transcription factors that were proposed to be involved in the control of cell proliferation, differentiation and specification in both vertebrates and invertebrates. In the mammalian telencephalon, Meis2 is most strongly expressed in the developing striatum, however, its functions remained elusive. Our analyses showed that Meis2 is expressed in virtually all of the ventricular/subventricular zone progenitors and differentiating medium spiny neurons (MSNs), raising the possibility that Meis2 might be involved in the control of MSN generation by progenitors throughout the entire development of striatum. To test this possibility, we devised a gene-transfer method where the generation of patch and matrix MSNs can be monitored selectively. We found that perturbation of Meis2-mediated transcription severely disrupted neuronal differentiation, as demonstrated by retention of cells with radial glial morphology, in both patch and matrix MSN progenitors, indicating Meis2 as a critical regulator for striatal MSN generation. doi:10.1016/j.neures.2009.09.379
P1-d18 Regulation of photoreceptor precursor differentiation in retinal development Kimiko Katoh 1,2 , Yoshihiro Omori 2 , Sigeru Sato 2 , Mineo Kondo 3 , Takahisa Furukawa 2 1 3
P1-d15 Molecular function of Tsukushi on neuronal stem cells Kunimasa Ohta 1 , Ayako Ito 1,2 , Sei kuriyama 3 , Giuseppe Lupo 4 , Rika Nakayama 5 , Naoko Ohshima 5 , Mitsuko Kosaka 5 , Shin-ichi Ohnuma 3 , Shinichi Nakagawa 6 , Hideaki Tanaka 1,2 1
Kumamoto Univ., Grad. Sch. of Med Sci., Japan; 2 Global COE, Kumamoto Univ, Japan; 3 UCL, Japan; 4 Univ of Cambridge, USA; 5 RIKEN CDB, Japan; 6 RIKEN FRS, Japan Multipotent stem/progenitor cells have been isolated from the ciliary body (CB) of the mouse retina, and shown to possess several universal characteristics of stem cells. Several lines of evidence suggest that Wnt signaling promotes proliferation of retinal stem/progenitor cells. Nonetheless, little is known about the molecular mechanisms that control stem cell self-renewal and restrict tissue-specific stem cells to specific compartments. We generated knock-out mice lacking TSK function and found that TSK inactivation results in expansion of the CB and the increased proliferation of the retinal stem/progenitor cells in vitro. Biochemical assays and overexpression experiments suggest that these effects are dependent on the ability of TSK to modulate Wnt signaling by binding to Frizzled. We propose that TSK is an important component of the stem/progenitor cell niche in the mammalian CB, controlling stem/progenitor cell proliferation by modulation of Wnt signaling. doi:10.1016/j.neures.2009.09.374
P1-d16 Macrophage migration inhibitory factor (MIF) promotes proliferation and cell survival of neural stem/progenitor cells Shigeki Ohta 1 , Yutaka Kawakami 1 , Hideyuki Okano 2 , Takeshi Kawase 3 , Masahiro Toda 3 1
Ctr. for Advanced Med. Res., Keio University, Tokyo, Japan; 2 Department of Physiol., Keio University, Tokyo, Japan; 3 Department of Neurosurgery, Keio University, Tokyo, Japan In previous studies, we demonstrated that mouse dendritic cells (DCs) can increase the number of neural stem/progenitor cells (NSPCs) in vitro and in vivo. In this study, we identified macrophage migration inhibitory factor (MIF) that is secreted from DCs and NSPCs as a novel factor that can support the proliferation and/or survival of NSPCs in vitro. To date the function of MIF in the normal brain remains largely unknown. FACS analyses showed that murine neurospheres express CD74 and CD44 which are known as MIF receptors. MIF increased the number of primary and secondary neurospheres. In contrast, MIF blocker and MIF shRNAi suppressed the neurosphere formations. Moreover, we found that MIF increases the phosphorylation of Akt and Erk in neurospheres that may support the cells? survival and proliferation. Taken together, MIF is a newly identified factor that can promote the survival and/or proliferation of NSPCs. doi:10.1016/j.neures.2009.09.375
P1-d17 Inhibitory effects of rotenone on migration of neural stem cells in neurosphere assay in vitro Masami Ishido, Junko Suzuki Natl. Inst. for Environmental Studies, Japan Parkinson’s disease is primarily a sporadic disorder and its specific etiology is incompletely understood. According to the working hypothesis of DOHaD (Developmental origins of health and disease), environmental origins of human sporadic Parkinson, s disease might be early in life. For the first step to substantiate this, in this study we examined the effect of the dopaminergic toxin, rotenone on neural stem cells in vitro, comparing with those of PC12 cells. Neurosphere was isolated from rat E15–16 mesencephalone in the presence of bFGF and EGF. To facilitate the stem cells to migrate from neurosphere in vitro, several neurospheres were cultured on a test well. Exposure of neurospheres to rotenone prevented the cell from migrating with the decrement of Ki67-positive cells; TUNEL-positive cells were increased. Thus, the data obtained would suggest the possibility that exposure to rotenone in utero might disturb the developmental programming, resulting in corresponding disease in later life. doi:10.1016/j.neures.2009.09.376
S91
Dept. Biostudies, Univ. of Kyoto, Kyoto, Japan; 2 Dev., OBI, Osaka, Japan; Dept. Ophthalmol., Univ. of Nagoya, Nagoya, Japan
The vertebrate retina consists of six types of neurons and one type of glial cells, which are all derived from multipotent progenitors. There is little information on the molecular mechanisms regulating the specification of cell type identity from multipotent progenitors in the developing retina. We have previously demonstrated that Otx2 acts as a master regulator of photoreceptor cell fate determination using photoreceptor-specific Otx2 conditional knockout (CKO) mice. Here, we performed microarray analysis comparing the retinal gene expression profiles of wild-type and Otx2 CKO mice. We have identified Blimp1 as one of the most reduced genes in the Otx2 CKO mice. In the Blimp1 CKO mice, we found that the number of photoreceptors was significantly reduced. Electroretinograms analysis also showed that the response of photoreceptor cells was impaired in adult Blimp1 CKO mice. These results suggest that Blimp1 plays an important role in the development of photoreceptors. doi:10.1016/j.neures.2009.09.377
P1-d19 The role of Ubc9, a sumo-conjugating enzyme, in retinal stem/progenitor proliferation Koji Terada, Takahisa Furukawa Depts. of Developmental Biology, Osaka Bio. Inst., Japan The mechanisms which underlie the regulation of stem/progenitor cell proliferation in the neural tissue during early development have been poorly understood. We previously found that xhmgb3 is expressed in the ciliary marginal zone where retinal stem cell exist in the Xenopus retina and that overexpression of xhmgb3 induced overproliferation of retinal progenitors. In this study, we performed yeast two-hybrid screening to identify the factors that act with xhmgb3, and we identified Ubc9. Ubc9 is involved in various cellular functions, including the control of a subcellular localization and the activity of target proteins by sumoylation. We performed overexpression experiments of xhmgb3 and Ubc9 in Xenopus embryos. As a result, we observed an eye enlargement more than two-fold in the frequency compared with that in only xhmgb3-injected embryos, suggesting that xhmgb3 and Ubc9 act cooperatively in eye development. Our results suggest that Ubc9 controls progenitor cell proliferation in the Xenopus retina. We are further investigating the mechanisms of progenitor cell proliferation regulated by xhmgb3 and Ubc9. doi:10.1016/j.neures.2009.09.378
P1-d20 Roles of Meis2 in the generation of medium spiny neurons in the developing striatum Saki Takami, Fujio Murakami, Yasuto Tanabe Grad. Sch. Front. Biosci., Osaka University, Suita, Japan Meis2 belongs to the family of homeodomain-containing TALE transcription factors that were proposed to be involved in the control of cell proliferation, differentiation and specification in both vertebrates and invertebrates. In the mammalian telencephalon, Meis2 is most strongly expressed in the developing striatum, however, its functions remained elusive. Our analyses showed that Meis2 is expressed in virtually all of the ventricular/subventricular zone progenitors and differentiating medium spiny neurons (MSNs), raising the possibility that Meis2 might be involved in the control of MSN generation by progenitors throughout the entire development of striatum. To test this possibility, we devised a gene-transfer method where the generation of patch and matrix MSNs can be monitored selectively. We found that perturbation of Meis2-mediated transcription severely disrupted neuronal differentiation, as demonstrated by retention of cells with radial glial morphology, in both patch and matrix MSN progenitors, indicating Meis2 as a critical regulator for striatal MSN generation. doi:10.1016/j.neures.2009.09.379