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Standardized mouse phenotyping platform and integrative database system
S30
Abstracts / Neuroscience Research 58S (2007) S1–S244
S3P-J1 Impact of brain-behavior phenotypying of genetically
S3P-J4 The Genes to Cognition programme
engineered mice on research of neuropsychiatric disorders
Noboru H. Komiyama, Seth G.N. Grant Wellcome Trust Sanger Institute, UK
Tsuyoshi Miyakawa Frontier Technology Center, Kyoto, Japan Although all the genome sequences have been determined, we do not yet know the functions of most of these genes. To reveal the functions of genes expressed in the brain, and to accelerate the progress of neuroscience, we are proposing to develop a large-scale mouse phenotyping center and/or consortium in Japan (Aiba et al., 2007). In this project, phenotype data would be obtained systematically with reasonably standardized methods, and the data obtained in such projects should be included in a public database. So far, in collaboration with 50 laboratories, we have conducted comprehensive behavioral phenotyping of about 60 strains of genetically engineered mice. Surprisingly, more than 90% of those strains of mutant mice showed at least one behavioral phenotype, indicating that many of the genes expressed in the brain could have functions that affect behavior. The purpose and the current status of the project will be introduced and discussed, particularly focusing on its significance in understanding human neuropsychiatric disorders.
The Genes to Cognition programme (G2C) was established to provide a framework for studying genes, brain and behaviour in order to link basic molecular research from genomes and experimental genetic organisms with human clinical studies of cognition. The G2C programme will collect and integrate data in the areas of psychiatry, human and mouse psychology, cellular neurophysiology, proteomics and biochemistry, molecular biology, human and mouse genetics and genomics. Bioinformatics will be used to create new tools for presenting this diverse data in novel relationships that will facilitate mining of information across existing boundaries in neuroscience. G2C will create, and make publicly available, novel databases for basic and clinical neuroscience; and establish a general model for tackling diseases and brain function. G2C supports a consortium of scientists and offers research, training and collaborative opportunities and aims to become a network for neuroscientists. For more detailed information on G2C is available at the web site: http://www.genes2cognition.org/.
Research funds: KAKENHI (16680015, 18016012, 18023022, 18053015), JST BIRD, NIBIO
Research fund: Functional Genomics Development Initiative of the Wellcome Trust
S3P-J2 Systematic generation of conditional knockout mice
S3P-J5
using C57BL/6 ES cell RENKA
signaling
Kenji Sakimura, Manabu Abe, Maya Yamazaki, Hiroshi Usui Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Japan
Atsu Aiba Division of Molecular Genetics, Kobe University of Graduate School of Medicine, Kobe, Japan
Generation of conditional targeting mice has greatly contributed to the understanding of brain functions, although problems of their genetic background and time- and cost-consuming work remain to be solved. To make up for the problems in 129 strain mice, for which they are not suitable for behavioral analysis of brain higher functions, we established a new ES cell line RENKA derived from C57BL/6N mouse blastocyst. We then improved the culture condition of RENKA, chimera formation parameters, etc. to establish our original protocol, where germline transmission was as efficient as in the 129 strains. We also developed universal vectors and a unique recombination system using BAC clones to shorten the time for targeting vector construction. With the additional efforts made to increase homologous recombination ratio in RENKA leading to reduced screening colony number and the cost, we are entering a new era of systematic generation of the conditional targeting mouse having pure C57BL/6 genetic background.
We previously generated mGluR1 KO and mGluR1a- and mGluR1b-rescue mice to study the role of mGluR1 in cerebellar Purkinje cells (PCs). Using these mice, we showed that mGluR1 in PCs is required for motor coordination, motor learning, long-term depression and regression of multiple climbing fiber innervation. We recently established mGluR1 conditional KO (cKO) mice by using the tetracycline-controlled gene expression system. The mGluR1 cKO mice express mGluR1 only in PCs and its expression can be turned off with doxycycline administration. We found that suppression of the mGluR1 expression in the adult mGluR1 cKO mice led to impaired motor coordination, indicating that mGluR1 is essential for cerebellar function not only during postnatal development but also in adulthood. To identify mGluR1-signaling molecules responsible for these phenotypes, we analyzed mGluR1-complexes isolated from wild type, KO, mGluR1a- and mGluR1b-rescue cerebella by mass spectrometry. We found mGluR1 complexes include more than 100 proteins and some proteins interact with specific mGluR1 splicing variants, suggesting divergent signaling of mGluR1 variants in PCs.
Research funds: SORST JST, KAKENHI 17300099
Genetic and proteomic approaches to mGluR1-
Research fund: KAKENHI (17024038)
S3P-J3 Standardized mouse phenotyping platform and integrative database system
S3P-J6 The organizing principles of real-time memory encoding: Neural clique assemblies and universal neural codes
Hiroshi Masuya Functional Genomics Research Group, RIKEN GSC, Tsukuba, Japan
Joe Z. Tsien Center for Systems Neurobiology Departments of Pharmacology and Biomedical Engineering, Boston University, Boston, USA
Following whole mouse genome sequencing, collaborative programs of knockout (KO) mouse production of every mouse genes, such as KOMP and EuCOMM, have just started in USA and Europe. Beside, production of phenotype data of the expanded mouse mutants by a comprehensive phenotyping platform and standard operation procedures (SOPs) has also launched in Europe. In RIKEN, we established a comprehensive platform consisting of 14 packages of phenotyping assays, which was developed in a large-scale ENU mutagenesis program. Subsequently, we developed a new website, ‘PhenoSITE’ (http://www.gsc.riken.go.jp/Mouse/), which includes databases of terminology, operation procedures and baseline data. Furthermore, now we propose a new concept of format to describe procedures, which provides framework of procedure descriptions and enables direct comparison of parameters of the procedures. This is referred as ‘Standardized Description of Operating Procedures (SDOP)’. It will assist on-line publication and better sharing of not only the standardized procedures but also the local procedures used in multiple laboratories.
Recent identification of network-level coding units, termed neural cliques, in the hippocampus has allowed real-time patterns of memory traces to be mathematically described, directly visualized, and dynamically deciphered. Those neural clique assemblies are invariantly organized in a hierarchical and categorical manner, forming various ‘general-tospecific feature-encoding pyramids’. The hierarchical organization of memory coding units allows not only for encoding nearly infinite amount of episodic events, but also for converting daily experiences into generalized knowledge and concepts in the brain. This general-to-specific encoding mechanism is further supported by the recent discovery of hippocampal cells which appear to be involved in the encoding of abstract concepts from episodic experiences. Moreover, mathematical conversion of activation patterns of neural cliques into binary codes provides a potential framework for universal categorizations of various experiences across individuals and animal species.
Research fund: National Bioresources Project
Research funds: Keck Foundation, NIMH, and NIA
Abstracts / Neuroscience Research 58S (2007) S1–S244
S3P-J1 Impact of brain-behavior phenotypying of genetically
S3P-J4 The Genes to Cognition programme
engineered mice on research of neuropsychiatric disorders
Noboru H. Komiyama, Seth G.N. Grant Wellcome Trust Sanger Institute, UK
Tsuyoshi Miyakawa Frontier Technology Center, Kyoto, Japan Although all the genome sequences have been determined, we do not yet know the functions of most of these genes. To reveal the functions of genes expressed in the brain, and to accelerate the progress of neuroscience, we are proposing to develop a large-scale mouse phenotyping center and/or consortium in Japan (Aiba et al., 2007). In this project, phenotype data would be obtained systematically with reasonably standardized methods, and the data obtained in such projects should be included in a public database. So far, in collaboration with 50 laboratories, we have conducted comprehensive behavioral phenotyping of about 60 strains of genetically engineered mice. Surprisingly, more than 90% of those strains of mutant mice showed at least one behavioral phenotype, indicating that many of the genes expressed in the brain could have functions that affect behavior. The purpose and the current status of the project will be introduced and discussed, particularly focusing on its significance in understanding human neuropsychiatric disorders.
The Genes to Cognition programme (G2C) was established to provide a framework for studying genes, brain and behaviour in order to link basic molecular research from genomes and experimental genetic organisms with human clinical studies of cognition. The G2C programme will collect and integrate data in the areas of psychiatry, human and mouse psychology, cellular neurophysiology, proteomics and biochemistry, molecular biology, human and mouse genetics and genomics. Bioinformatics will be used to create new tools for presenting this diverse data in novel relationships that will facilitate mining of information across existing boundaries in neuroscience. G2C will create, and make publicly available, novel databases for basic and clinical neuroscience; and establish a general model for tackling diseases and brain function. G2C supports a consortium of scientists and offers research, training and collaborative opportunities and aims to become a network for neuroscientists. For more detailed information on G2C is available at the web site: http://www.genes2cognition.org/.
Research funds: KAKENHI (16680015, 18016012, 18023022, 18053015), JST BIRD, NIBIO
Research fund: Functional Genomics Development Initiative of the Wellcome Trust
S3P-J2 Systematic generation of conditional knockout mice
S3P-J5
using C57BL/6 ES cell RENKA
signaling
Kenji Sakimura, Manabu Abe, Maya Yamazaki, Hiroshi Usui Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Japan
Atsu Aiba Division of Molecular Genetics, Kobe University of Graduate School of Medicine, Kobe, Japan
Generation of conditional targeting mice has greatly contributed to the understanding of brain functions, although problems of their genetic background and time- and cost-consuming work remain to be solved. To make up for the problems in 129 strain mice, for which they are not suitable for behavioral analysis of brain higher functions, we established a new ES cell line RENKA derived from C57BL/6N mouse blastocyst. We then improved the culture condition of RENKA, chimera formation parameters, etc. to establish our original protocol, where germline transmission was as efficient as in the 129 strains. We also developed universal vectors and a unique recombination system using BAC clones to shorten the time for targeting vector construction. With the additional efforts made to increase homologous recombination ratio in RENKA leading to reduced screening colony number and the cost, we are entering a new era of systematic generation of the conditional targeting mouse having pure C57BL/6 genetic background.
We previously generated mGluR1 KO and mGluR1a- and mGluR1b-rescue mice to study the role of mGluR1 in cerebellar Purkinje cells (PCs). Using these mice, we showed that mGluR1 in PCs is required for motor coordination, motor learning, long-term depression and regression of multiple climbing fiber innervation. We recently established mGluR1 conditional KO (cKO) mice by using the tetracycline-controlled gene expression system. The mGluR1 cKO mice express mGluR1 only in PCs and its expression can be turned off with doxycycline administration. We found that suppression of the mGluR1 expression in the adult mGluR1 cKO mice led to impaired motor coordination, indicating that mGluR1 is essential for cerebellar function not only during postnatal development but also in adulthood. To identify mGluR1-signaling molecules responsible for these phenotypes, we analyzed mGluR1-complexes isolated from wild type, KO, mGluR1a- and mGluR1b-rescue cerebella by mass spectrometry. We found mGluR1 complexes include more than 100 proteins and some proteins interact with specific mGluR1 splicing variants, suggesting divergent signaling of mGluR1 variants in PCs.
Research funds: SORST JST, KAKENHI 17300099
Genetic and proteomic approaches to mGluR1-
Research fund: KAKENHI (17024038)
S3P-J3 Standardized mouse phenotyping platform and integrative database system
S3P-J6 The organizing principles of real-time memory encoding: Neural clique assemblies and universal neural codes
Hiroshi Masuya Functional Genomics Research Group, RIKEN GSC, Tsukuba, Japan
Joe Z. Tsien Center for Systems Neurobiology Departments of Pharmacology and Biomedical Engineering, Boston University, Boston, USA
Following whole mouse genome sequencing, collaborative programs of knockout (KO) mouse production of every mouse genes, such as KOMP and EuCOMM, have just started in USA and Europe. Beside, production of phenotype data of the expanded mouse mutants by a comprehensive phenotyping platform and standard operation procedures (SOPs) has also launched in Europe. In RIKEN, we established a comprehensive platform consisting of 14 packages of phenotyping assays, which was developed in a large-scale ENU mutagenesis program. Subsequently, we developed a new website, ‘PhenoSITE’ (http://www.gsc.riken.go.jp/Mouse/), which includes databases of terminology, operation procedures and baseline data. Furthermore, now we propose a new concept of format to describe procedures, which provides framework of procedure descriptions and enables direct comparison of parameters of the procedures. This is referred as ‘Standardized Description of Operating Procedures (SDOP)’. It will assist on-line publication and better sharing of not only the standardized procedures but also the local procedures used in multiple laboratories.
Recent identification of network-level coding units, termed neural cliques, in the hippocampus has allowed real-time patterns of memory traces to be mathematically described, directly visualized, and dynamically deciphered. Those neural clique assemblies are invariantly organized in a hierarchical and categorical manner, forming various ‘general-tospecific feature-encoding pyramids’. The hierarchical organization of memory coding units allows not only for encoding nearly infinite amount of episodic events, but also for converting daily experiences into generalized knowledge and concepts in the brain. This general-to-specific encoding mechanism is further supported by the recent discovery of hippocampal cells which appear to be involved in the encoding of abstract concepts from episodic experiences. Moreover, mathematical conversion of activation patterns of neural cliques into binary codes provides a potential framework for universal categorizations of various experiences across individuals and animal species.
Research fund: National Bioresources Project
Research funds: Keck Foundation, NIMH, and NIA