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Identification of genes specific to glutamatergic-neuron precursors in the developing mouse neocortex
S92
Abstracts
P1-d21 Proper segregation and connection between the CNS and PNS require SDF-1/CXCR4/CXCR7 signaling trio Yan Zhu 1 , Tomoko Matsumoto 1 , Sakae Mikami 2 , Takashi Sierro 3 , Fabienne Mackay 3 , Fujio Nagasawa 2 , Frederic Murakami 1 1
Osaka University, Japan; 2 Inst. for Frontier Medical Sciences, Kyoto, Japan; 3 Darlinghurst, Australia
Anatomical segregation of the functionally distinct central and peripheral nervous system (CNS and PNS) and the establishment of precise connectivity between them are fundamental in the development of the nervous system. Yet, the mechanisms underlying these processes are poorly understood. We provide evidence here that the signalling between the chemokine SDF-1 and its receptor CXCR4 contributes to the segregation and connection between the CNS and PNS in mice. SDF-1 or CXCR4 knockout mice show mis-positioned PNS boundary cap cells inside the spinal cord, disrupted glial limiting membrane and misrouted peripheral axons. Analysis of knockout mice of the second SDF-1 receptor CXCR7 showed phenotypes that are suggestive of CXCR7 being an integral component of SDF-1 signalling that controls the normal development of the boundary cap cells and the radial glial fibres. doi:10.1016/j.neures.2009.09.380
P1-d22 The expression and roles of Runx1 in the development of mouse embryo brainstem Ryota Ito, Mizuki Hirabayashi, Masaaki Yoshikawa, Satoru Takahashi, Kouji Senzaki, Takashi Shiga University of Tsukuba Graduate School of Comprehensive Human Science, Tsukuba, Japan Runx1 and Runx3, runt-related transcription factors, play important roles in the cell specification and axonal projections of the nociceptive and proprioceptive dorsal root ganglion neurons, respectively. Although Runx1 is expressed in the central nervous system as well as the peripheral nervous system, little is known about the roles in the brain development. In the present study, we first examined in detail the expression of the Runx1 in the brain of mouse embryos by immunohistochemistry. We found that Runx1 was expressed in selected nuclei in the brainstem including the trigeminal motor nucleus, mesencehalic trigeminal nucleus, ambiguus nucleus, nucleus of the solitary tract and hypoglossal nucleus. The expression in these nuclei was detected as early as embryonic day 13.5. Now we are analyzing roles of Runx in the development of these brainstem nuclei using Runx1-deficient mouse. doi:10.1016/j.neures.2009.09.381
P1-d23 Identification of genes specific to glutamatergic-neuron precursors in the developing mouse neocortex Keisuke Watanabe, Hirohide Takebayashi, Shigeyuki Esumi, Nobuaki Tamamaki Dept. Morphol. Neural. Sci., Kumamoto Univ., Japan Glutamatergic (Glu) and GABAergic neurons are characterized by distinct functions, cell morphologies and origins. We speculated that these distinct characters of the neurons are based on expression of distinct molecules. To address the issue, we compared gene-expression profiles between the two neuronal lineages in the developing neocortex. We identified newly born Glu-neuron precursors by GFP expression in the NEX/Math2-positive cell lineage and GABAergic neuronal lineages by GFP expression in the GAD67-GFP knock-in mice, respectively. Genetically labeled single cells were picked up and subjected to single cell microarray analysis. By cluster analysis using our database and reported one, we could reveal distinct gene expression profiles between the two populations. Furthermore, we could find several genes that were expressed specifically and/or strongly in the Glu neuronal lineages. Here, we report possible functions of a distinct gene in the neocortical Glu neurons. doi:10.1016/j.neures.2009.09.382
P1-d24 Gs-linked receptor GPR3 modulates proliferation and differentiation of cerebellar granule cell precursors Shigeru Tanaka 1,2 , Imran M. Shaikh 2 , E. Antonio chiocca 2 , Norio Sakai 1 , Yoshinaga Saeki 2 1
Hiroshima Univ. Grad. Sch. Biomed. Sci., Japan; The Ohio State Univ., Columbus OH, USA
2
Dept. Neurol. Surg.,
G-protein coupled receptor (GPR) 3, GPR6, and GPR12 are predominantly expressed in the mammalian central nervous system and constitutively activate adenylate cyclases. GPR3 is highly expressed in the internal granule cell layer (IGL) during
cerebellar development. Granule neuron precursors (GNPs) proliferate in the external germinal cell layer (EGL) and migrate past Purkinje neurons to form the IGL, where they mature into granule neurons. Sonic Hedgehog (Shh) from Purkinje neurons is one of the important mitogens for GNPs, and increased cAMP levels and PKA activation have been reported to antagonize Shh signaling. In this report, we show that the expression of GPR3 inhibits Shh-mediated proliferation of GNPs in vitro and in vivo. Upregulated GPR3 expression also stimulated p27 expression in GNPs, thereby inhibiting their cell cycle progression. These results indicated that GPR3 can be one of the cAMP generator that is responsible for the Shh inhibition and terminal differentiation of GNPs. doi:10.1016/j.neures.2009.09.383
P1-e01 Combinatorial role of the EGFR signaling pathway and a RUNX family transcription factor Lozenge in the subtype specification of the Drosophila olfactory sensory neurons Keita Endo 1 , Shohei Kuwa 2 , Kei Ito 1 1
IMCB, University of Tokyo, Japan; of Tokyo, Tokyo, Japan
2
Grad. Sch. of Frontier Sci., University
In the Drosophila olfactory system, olfactory sensory neurons (OSNs) are diversified into ∼50 classes, each of which expresses specific odorant receptors. We searched for molecular mechanisms that lead to such diverse neuron subtypes, and found that the EGFR signaling pathway is required for the differentiation of a specific subset of the OSN classes. Moreover, we found that both the EGFR-dependent and -independent OSN classes are further diversified into several subtypes through a dose-dependent regulation by a RUNX family transcription factor, Lozenge. These results suggest that the combinatorial regulation by these two mechanisms should be the key to generate the diverse OSN classes. We also found that the Lozenge function is required not for the post-mitotic OSNs but for their precursors. Thus, it is likely that Lozenge is expressed differentially among the OSN precursors to diversify their fate, which in turn determines the OSN classes they generate. doi:10.1016/j.neures.2009.09.384
P1-e02 The transcriptional repressor RP58 is crucial for celldivision patterning and neuronal survival in the developing cerebral cortex Haruo Okado 1 , Chiaki Ohtaka-Maruyama 1 , Yoshinobu Sugitani 2 , YuKo Fukuda 3 , Reiko Ishida 3 , Shinobu Hirai 1 , Akiko Miwa 1 , Akiyo Takahashi 1 , Katsunori Aoki 4 , Keiji Mochida 5 , Osamu Suzuki 3 , Takao Honda 6 , Kazunori Nakajima 6 , Masaharu Ogawa 2 , Toshio Terashima 7 , Junichiro Matsuda 3 , Hitoshi Kawano 1 , Masataka Kasai 3 1
Tokyo Metropolitan Institute for Neuroscience, Japan; 2 Brain Science Institute, Riken, Japan; 3 National Institute of Infectious Diseases, Japan; 4 The University of Tokyo, Japan; 5 Bioresource Center, RIKEN, Japan; 6 Keio University School of Medicine, Japan; 7 Graduate School of Medicine Kobe, Japan We show that, in the developing cerebral cortex, the RP58 transcription repressor protein was expressed both in postmitotic glutamatergic projection neurons and in their progenitor cells, but not in GABAergic interneurons. Targeted deletion of the RP58 gene led to dysplasia of the cerebral neocortex and of the hippocampus, reduction of the number of mature cortical neurons, and defects of laminar organization. We demonstrate an impairment of the cell-division patterning during the late embryonic stage and an enhancement of apoptosis of the postmitotic neurons in the RP58-deficient cortex. doi:10.1016/j.neures.2009.09.385
P1-e03 RP58 represses Id3 gene promoter activity Shinobu Hirai 1,2 , Chiaki Ohtaka-Maruyama 1 , Akiko Miwa 1 , Akiyo Takahashi 1 , Masataka Kasai 3 , Haruo Okado 1 1
Tokyo Metropolitan Inst for Neurosci, Tokyo, Japan; 2 Grad Sch of Tokyo Med and Dental Univ, Tokyo, Japan; 3 National Inst of Infectious Diseases, Tokyo, Japan Transcriptional repressor protein RP58 is strongly expressed in the developing mouse brain. RP58 is specifically required for maturation of the excitatory neurons of the cerebral cortex and hippocampus. However the mechanism by which RP58 contributes to neuronal maturation remains elusive. To identify downstream effectors of RP58, we performed the micro-array analysis using RNA isolated from WT and RP58 mutant cortex of E16. One of candidates was Id3, a member of Id fam-
Abstracts
P1-d21 Proper segregation and connection between the CNS and PNS require SDF-1/CXCR4/CXCR7 signaling trio Yan Zhu 1 , Tomoko Matsumoto 1 , Sakae Mikami 2 , Takashi Sierro 3 , Fabienne Mackay 3 , Fujio Nagasawa 2 , Frederic Murakami 1 1
Osaka University, Japan; 2 Inst. for Frontier Medical Sciences, Kyoto, Japan; 3 Darlinghurst, Australia
Anatomical segregation of the functionally distinct central and peripheral nervous system (CNS and PNS) and the establishment of precise connectivity between them are fundamental in the development of the nervous system. Yet, the mechanisms underlying these processes are poorly understood. We provide evidence here that the signalling between the chemokine SDF-1 and its receptor CXCR4 contributes to the segregation and connection between the CNS and PNS in mice. SDF-1 or CXCR4 knockout mice show mis-positioned PNS boundary cap cells inside the spinal cord, disrupted glial limiting membrane and misrouted peripheral axons. Analysis of knockout mice of the second SDF-1 receptor CXCR7 showed phenotypes that are suggestive of CXCR7 being an integral component of SDF-1 signalling that controls the normal development of the boundary cap cells and the radial glial fibres. doi:10.1016/j.neures.2009.09.380
P1-d22 The expression and roles of Runx1 in the development of mouse embryo brainstem Ryota Ito, Mizuki Hirabayashi, Masaaki Yoshikawa, Satoru Takahashi, Kouji Senzaki, Takashi Shiga University of Tsukuba Graduate School of Comprehensive Human Science, Tsukuba, Japan Runx1 and Runx3, runt-related transcription factors, play important roles in the cell specification and axonal projections of the nociceptive and proprioceptive dorsal root ganglion neurons, respectively. Although Runx1 is expressed in the central nervous system as well as the peripheral nervous system, little is known about the roles in the brain development. In the present study, we first examined in detail the expression of the Runx1 in the brain of mouse embryos by immunohistochemistry. We found that Runx1 was expressed in selected nuclei in the brainstem including the trigeminal motor nucleus, mesencehalic trigeminal nucleus, ambiguus nucleus, nucleus of the solitary tract and hypoglossal nucleus. The expression in these nuclei was detected as early as embryonic day 13.5. Now we are analyzing roles of Runx in the development of these brainstem nuclei using Runx1-deficient mouse. doi:10.1016/j.neures.2009.09.381
P1-d23 Identification of genes specific to glutamatergic-neuron precursors in the developing mouse neocortex Keisuke Watanabe, Hirohide Takebayashi, Shigeyuki Esumi, Nobuaki Tamamaki Dept. Morphol. Neural. Sci., Kumamoto Univ., Japan Glutamatergic (Glu) and GABAergic neurons are characterized by distinct functions, cell morphologies and origins. We speculated that these distinct characters of the neurons are based on expression of distinct molecules. To address the issue, we compared gene-expression profiles between the two neuronal lineages in the developing neocortex. We identified newly born Glu-neuron precursors by GFP expression in the NEX/Math2-positive cell lineage and GABAergic neuronal lineages by GFP expression in the GAD67-GFP knock-in mice, respectively. Genetically labeled single cells were picked up and subjected to single cell microarray analysis. By cluster analysis using our database and reported one, we could reveal distinct gene expression profiles between the two populations. Furthermore, we could find several genes that were expressed specifically and/or strongly in the Glu neuronal lineages. Here, we report possible functions of a distinct gene in the neocortical Glu neurons. doi:10.1016/j.neures.2009.09.382
P1-d24 Gs-linked receptor GPR3 modulates proliferation and differentiation of cerebellar granule cell precursors Shigeru Tanaka 1,2 , Imran M. Shaikh 2 , E. Antonio chiocca 2 , Norio Sakai 1 , Yoshinaga Saeki 2 1
Hiroshima Univ. Grad. Sch. Biomed. Sci., Japan; The Ohio State Univ., Columbus OH, USA
2
Dept. Neurol. Surg.,
G-protein coupled receptor (GPR) 3, GPR6, and GPR12 are predominantly expressed in the mammalian central nervous system and constitutively activate adenylate cyclases. GPR3 is highly expressed in the internal granule cell layer (IGL) during
cerebellar development. Granule neuron precursors (GNPs) proliferate in the external germinal cell layer (EGL) and migrate past Purkinje neurons to form the IGL, where they mature into granule neurons. Sonic Hedgehog (Shh) from Purkinje neurons is one of the important mitogens for GNPs, and increased cAMP levels and PKA activation have been reported to antagonize Shh signaling. In this report, we show that the expression of GPR3 inhibits Shh-mediated proliferation of GNPs in vitro and in vivo. Upregulated GPR3 expression also stimulated p27 expression in GNPs, thereby inhibiting their cell cycle progression. These results indicated that GPR3 can be one of the cAMP generator that is responsible for the Shh inhibition and terminal differentiation of GNPs. doi:10.1016/j.neures.2009.09.383
P1-e01 Combinatorial role of the EGFR signaling pathway and a RUNX family transcription factor Lozenge in the subtype specification of the Drosophila olfactory sensory neurons Keita Endo 1 , Shohei Kuwa 2 , Kei Ito 1 1
IMCB, University of Tokyo, Japan; of Tokyo, Tokyo, Japan
2
Grad. Sch. of Frontier Sci., University
In the Drosophila olfactory system, olfactory sensory neurons (OSNs) are diversified into ∼50 classes, each of which expresses specific odorant receptors. We searched for molecular mechanisms that lead to such diverse neuron subtypes, and found that the EGFR signaling pathway is required for the differentiation of a specific subset of the OSN classes. Moreover, we found that both the EGFR-dependent and -independent OSN classes are further diversified into several subtypes through a dose-dependent regulation by a RUNX family transcription factor, Lozenge. These results suggest that the combinatorial regulation by these two mechanisms should be the key to generate the diverse OSN classes. We also found that the Lozenge function is required not for the post-mitotic OSNs but for their precursors. Thus, it is likely that Lozenge is expressed differentially among the OSN precursors to diversify their fate, which in turn determines the OSN classes they generate. doi:10.1016/j.neures.2009.09.384
P1-e02 The transcriptional repressor RP58 is crucial for celldivision patterning and neuronal survival in the developing cerebral cortex Haruo Okado 1 , Chiaki Ohtaka-Maruyama 1 , Yoshinobu Sugitani 2 , YuKo Fukuda 3 , Reiko Ishida 3 , Shinobu Hirai 1 , Akiko Miwa 1 , Akiyo Takahashi 1 , Katsunori Aoki 4 , Keiji Mochida 5 , Osamu Suzuki 3 , Takao Honda 6 , Kazunori Nakajima 6 , Masaharu Ogawa 2 , Toshio Terashima 7 , Junichiro Matsuda 3 , Hitoshi Kawano 1 , Masataka Kasai 3 1
Tokyo Metropolitan Institute for Neuroscience, Japan; 2 Brain Science Institute, Riken, Japan; 3 National Institute of Infectious Diseases, Japan; 4 The University of Tokyo, Japan; 5 Bioresource Center, RIKEN, Japan; 6 Keio University School of Medicine, Japan; 7 Graduate School of Medicine Kobe, Japan We show that, in the developing cerebral cortex, the RP58 transcription repressor protein was expressed both in postmitotic glutamatergic projection neurons and in their progenitor cells, but not in GABAergic interneurons. Targeted deletion of the RP58 gene led to dysplasia of the cerebral neocortex and of the hippocampus, reduction of the number of mature cortical neurons, and defects of laminar organization. We demonstrate an impairment of the cell-division patterning during the late embryonic stage and an enhancement of apoptosis of the postmitotic neurons in the RP58-deficient cortex. doi:10.1016/j.neures.2009.09.385
P1-e03 RP58 represses Id3 gene promoter activity Shinobu Hirai 1,2 , Chiaki Ohtaka-Maruyama 1 , Akiko Miwa 1 , Akiyo Takahashi 1 , Masataka Kasai 3 , Haruo Okado 1 1
Tokyo Metropolitan Inst for Neurosci, Tokyo, Japan; 2 Grad Sch of Tokyo Med and Dental Univ, Tokyo, Japan; 3 National Inst of Infectious Diseases, Tokyo, Japan Transcriptional repressor protein RP58 is strongly expressed in the developing mouse brain. RP58 is specifically required for maturation of the excitatory neurons of the cerebral cortex and hippocampus. However the mechanism by which RP58 contributes to neuronal maturation remains elusive. To identify downstream effectors of RP58, we performed the micro-array analysis using RNA isolated from WT and RP58 mutant cortex of E16. One of candidates was Id3, a member of Id fam-