研究者総覧

國田 智 (クニタ サトシ)

  • 実験医学センター 教授
Last Updated :2021/11/23

研究者情報

学位

  • 獣医学博士

ホームページURL

J-Global ID

研究キーワード

  • 人獣共通感染症   実験動物倫理   疾患モデル   感染症   

研究分野

  • ライフサイエンス / 実験動物学

経歴

  • 2011年 - 現在  自治医科大学実験医学センター教授
  • 2002年 - 2011年  筑波大学 人間総合科学研究科 人間総合科学研究科 生命システム医学専攻講師

所属学協会

  • 日本獣医学会   日本実験動物学会   Japanese Society of Veterinary Science   Japanese Association for Laboratory Animal Science   

研究活動情報

論文

  • Shinji Kawaguchi, Yusuke Soma, Kazuaki Nakajima, Hideaki Kanazawa, Shugo Tohyama, Ryota Tabei, Akinori Hirano, Noriko Handa, Yoshitake Yamada, Shigeo Okuda, Shuji Hishikawa, Takumi Teratani, Satoshi Kunita, Yoshikazu Kishino, Marina Okada, Sho Tanosaki, Shota Someya, Yuika Morita, Hidenori Tani, Yujiro Kawai, Masataka Yamazaki, Akira Ito, Rei Shibata, Toyoaki Murohara, Yasuhiko Tabata, Eiji Kobayashi, Hideyuki Shimizu, Keiichi Fukuda, Jun Fujita
    JACC. Basic to translational science 6 3 239 - 254 2021年03月 
    The severe shortage of donor hearts hampered the cardiac transplantation to patients with advanced heart failure. Therefore, cardiac regenerative therapies are eagerly awaited as a substitution. Human induced pluripotent stem cells (hiPSCs) are realistic cell source for regenerative cardiomyocytes. The hiPSC-derived cardiomyocytes are highly expected to help the recovery of heart. Avoidance of teratoma formation and large-scale culture of cardiomyocytes are definitely necessary for clinical setting. The combination of pure cardiac spheroids and gelatin hydrogel succeeded to recover reduced ejection fraction. The feasible transplantation strategy including transplantation device for regenerative cardiomyocytes are established in this study.
  • 原 弘真, 伊藤 拓哉, 菱川 修司, 中野 和明, 阿部 朋行, 柴田 宏昭, 魚崎 英毅, 渡邊 將人, 國田 智, 長嶋 比呂志, 花園 豊
    Organ Biology 26 3 89 - 89 (一社)日本臓器保存生物医学会 2019年10月
  • 原 弘真, 菱川 修司, 伊藤 拓哉, 阿部 朋行, 柴田 宏昭, 魚崎 英毅, 國田 智, 花園 豊
    Organ Biology 26 3 121 - 121 (一社)日本臓器保存生物医学会 2019年10月
  • 和食の健康維持・増進効果の機序解明を目指した研究
    原 弘真, 永山 学, 須田 亙, 柴田 宏昭, 菱川 修司, 國田 智, 阿部 朋行, 魚崎 英毅, 新 幸二, 本田 賢也, 花園 豊
    自治医科大学紀要 41 92 - 92 (学)自治医科大学 2019年03月
  • Ryota Tabei, Shinji Kawaguchi, Hideaki Kanazawa, Shugo Tohyama, Akinori Hirano, Noriko Handa, Shuji Hishikawa, Takumi Teratani, Satoshi Kunita, Junichi Fukuda, Yoshihiro Mugishima, Tsuneyoshi Suzuki, Kazuaki Nakajima, Tomohisa Seki, Yoshikazu Kishino, Marina Okada, Masataka Yamazaki, Kazuma Okamoto, Hideyuki Shimizu, Eiji Kobayashi, Yasuhiko Tabata, Jun Fujita, Keiichi Fukuda
    The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation 38 2 203 - 214 2019年02月 [査読有り][通常論文]
     
    BACKGROUND: Induced pluripotent stem cell (iPSC)‒based regenerative therapy is a promising strategy for cardiovascular disease treatment; however, the method is limited by the myocardial retention of grafted iPSCs. Thus, an injection protocol that efficiently introduces and retains human iPSC-derived cardiomyocytes (hiPSC-CMs) within the myocardium is urgently needed. The objective of the present study was to develop a method to improve the retention of hiPSCs in the myocardium for cardiac therapy. METHODS: We efficiently produced hiPSC-CM spheroids in 3-dimensional (3D) culture using microwell plates, and developed an injection device for optimal 3D distribution of the spheroids in the myocardial layer. Device biocompatibility was assessed with purified hiPSC-CM spheroids. Device effectiveness was evaluated in 10- to 15-month-old farm pigs (n = 15) and 5- to 24-month-old micro-minipigs (n = 20). The pigs were euthanized after injection, and tissues were harvested for retention and histologic analysis. RESULTS: We demonstrated an injection device for direct intramyocardial transplantation of hiPSC-CM spheroids from large-scale culture. The device had no detrimental effects on cell viability, spheroid shape, or size. Direct epicardial injection of spheroids mixed with gelatin hydrogel into beating porcine hearts using this device resulted in better distribution and retention of transplanted spheroids in a layer within the myocardium than did conventional needle injection procedures. CONCLUSIONS: The combination of the newly developed transplant device and spheroid formation promotes the retention of transplanted CMs. These findings support the clinical application of hiPSC-CM spheroid‒based cardiac regenerative therapy in patients with heart failure.
  • Hiromasa Hara, Hiroaki Shibata, Kazuaki Nakano, Tomoyuki Abe, Hideki Uosaki, Takahiro Ohnuki, Shuji Hishikawa, Satoshi Kunita, Masahito Watanabe, Osamu Nureki, Hiroshi Nagashima, Yutaka Hanazono
    Experimental animals 67 2 139 - 146 2018年05月 [査読有り][通常論文]
     
    Pigs with X-linked severe combined immunodeficiency (X-SCID) caused by a mutation of the interleukin-2 receptor gamma chain gene (IL2RG) are of value for a wide range of studies. However, they do not survive longer than 8 weeks because of their susceptibility to infections. To allow longer survival of X-SCID pigs, the animals must be born and reared under germ-free conditions. Here, we established an efficient system for piglet derivation by hysterectomy and used it to obtain and maintain a germ-free X-SCID pig. In four trials using pregnant wild-type pigs, 66% of piglets after hysterectomy started spontaneous breathing (range of 20-100% per litter). The resuscitation rate was found to negatively correlate with elapsed time from the uterus excision to piglet derivation (r=-0.97, P<0.05). Therefore, it is critical to deliver piglets within 5 min to achieve a high resuscitation rate (82% estimated from regression analysis). In a fifth trial with an IL2RG+/- pig, four piglets were delivered within 4.2 min of uterus excision and three were alive (75%). One of the live born piglets was genotypically and phenotypically determined to be X-SCID and was reared for 12 weeks. The X-SCID piglet was free from both bacteria and fungi at all time points tested by microbial culture and grew without any abnormal signs or symptoms. This study showed successful production and rearing of germ-free pigs, enabling experiments involving long-term follow-up of X-SCID pigs.
  • Tanggis, Tominari Kobayashi, Masaharu Takahashi, Suljid Jirintai, Tsutomu Nishizawa, Shigeo Nagashima, Takashi Nishiyama, Satoshi Kunita, Emiko Hayama, Takeshi Tanaka, Mulyanto, Hiroaki Okamoto
    Virus research 249 16 - 30 2018年04月 [査読有り][通常論文]
     
    Rat hepatitis E virus (ratHEV) genome has four open reading frames (ORFs: ORF1, ORF2, ORF3 and ORF4). The functions of ORF3 and ORF4 are unknown. An infectious cDNA clone (pUC-ratELOMB-131L_wt, wt) and its derivatives including ORF3-defective (ΔORF3) and ORF4-defective (ΔORF4) mutants, were constructed and their full-length RNA transcripts transfected into PLC/PRF/5 cells. ΔORF3 replicated as efficiently as wt in cells. However, ≤1/1000 of the number of progenies were detectable in the culture supernatant of ΔORF3-infected cells compared with wt-infected cells. ORF4 protein was not detectable in ratHEV-infected cells or in the liver tissues of ratHEV-infected rats. No marked differences were noted between wt and ΔORF4 regarding the viral replication and protein expression. ORF3 mutants with proline-to-leucine mutations at amino acids (aa) 93, 96 and/or 98 in ORF3 were constructed and transfected into PLC/PRF/5 cells. Wt and an ORF3 mutant with leucine at aa 98 (ORF3-L98) replicated efficiently (density 1.15-1.16 g/cm3), while ORF3-L93 + L96 exhibited a decreased viral release and banded at 1.26-1.27 g/cm3, similar to ΔORF3. In conclusion, the ORF3 protein, especially its proline residues at aa 93 and 96, is essential for the release of membrane-associated ratHEV particles, and ORF4 is unnecessary for the replication of ratHEV.
  • Eiji Kobayashi, Yutaka Hanazono, Satoshi Kunita
    Experimental animals 67 1 7 - 13 2018年02月 [査読有り][通常論文]
     
    Center for Development of Advanced Medical Technology (CDAMTec) in Jichi Medical University was established in 2009. It is the first educational research facility specialized for medical research and training using swine in Japan. Preclinical studies on large animals are essential prior to clinical trials to develop regenerative medical products and medical equipment. We have continued comprehensively considering using miniature swine for experiments to develop advanced medical technologies and train physicians with advanced clinical abilities, while paying attention to animal welfare. The center plays a pioneering role in this field by accumulating know-how such as (1) Construction and effective utilization of research facilities, (2) Procurement of quality animal resources, (3) Education and training of technical staff, (4) Establishment of support system for physicians and researchers. We now open up widely these expertise and foundation for medical research and training not only within our university but also outside the university, so as to move faster to practical use of advanced medical technology and contribute to human health and welfare.
  • Takuya Okada, Kazuko Keino-Masu, Satoshi Nagamine, Fuyuki Kametani, Tatsuyuki Ohto, Masato Hasegawa, Toin H van Kuppevelt, Satoshi Kunita, Satoru Takahashi, Masayuki Masu
    Scientific reports 7 1 13847 - 13847 2017年10月 [査読有り][通常論文]
     
    Heparan sulfate (HS) has been implicated in a wide range of cell signaling. Here we report a novel mechanism in which extracellular removal of 6-O-sulfate groups from HS by the endosulfatases, Sulf1 and Sulf2, is essential for axon guidance during development. In Sulf1/2 double knockout (DKO) mice, the corticospinal tract (CST) was dorsally displaced on the midbrain surface. In utero electroporation of Sulf1/2 into radial glial cells along the third ventricle, where Sulf1/2 mRNAs are normally expressed, rescued the CST defects in the DKO mice. Proteomic analysis and functional testing identified Slit2 as the key molecule associated with the DKO phenotype. In the DKO brain, 6-O-sulfated HS was increased, leading to abnormal accumulation of Slit2 protein on the pial surface of the cerebral peduncle and hypothalamus, which caused dorsal repulsion of CST axons. Our findings indicate that postbiosynthetic desulfation of HS by Sulfs controls CST axon guidance through fine-tuning of Slit2 presentation.
  • Putu Prathiwi Primadharsini, Masao Miyake, Satoshi Kunita, Tsutomu Nishizawa, Masaharu Takahashi, Shigeo Nagashima, Tanggis, Hiroshi Ohnishi, Tominari Kobayashi, Takashi Nishiyama, Suljid Jirintai, Hiroaki Okamoto
    Virus research 240 147 - 153 2017年08月 [査読有り][通常論文]
     
    Hepatitis E virus (HEV) causes acute or chronic hepatitis in humans and can be transmitted via the fecal-oral route. Pigs are one of the main reservoirs for this infection. Sixty pigs, 4-5 months of age, on a swine herd in Japan had detectable anti-HEV IgG antibodies, and five (8.3%) of them had ongoing infection of genotype 3 HEV. Five HEV strains obtained from the viremic pigs shared 98.8-100% nucleotide identity, and one representative strain (swHE1606845), whose entire genomic sequence was determined in this study, differed by 14.1-19.6% from the reported HEV strains of subtypes 3a-3k and by 14.7-19.1% from other genotype 3 HEV strains whose subtypes have not yet been assigned. swHE1606845 showed a higher nucleotide p-distance value of ≥0.143 with the genotype 3 HEV strains of subtypes 3a-3k and ≥0.152 with other genotype 3 strains of unassigned subtypes. A SimPlot analysis revealed a lack of recombination events. These results indicate that swHE1606845 is a candidate member of a novel subtype of genotype 3. Further efforts to identify the swHE1606845-like novel strain are warranted to clarify the origin of this strain and to determine the complete nucleotide sequences of two additional swHE1606845-like strains for assigning a new subtype.
  • Masaharu Takahashi, Tominari Kobayashi, Tanggis, Suljid Jirintai, Mulyanto, Shigeo Nagashima, Tsutomu Nishizawa, Satoshi Kunita, Hiroaki Okamoto
    Archives of virology 161 12 3391 - 3404 2016年12月 [査読有り][通常論文]
     
    Eight murine monoclonal antibodies (MAbs) against a synthetic peptide corresponding to the C-terminal 15-amino-acid portion of the ORF3 protein of rat hepatitis E virus (ratHEV) were produced and characterized. Immunofluorescence assays using the anti-ratHEV ORF3 MAbs revealed the accumulation of ORF3 protein in the cytoplasm of PLC/PRF/5 cells transfected with ORF3-expressing plasmids or inoculated with cell-culture-generated ratHEV strains. Anti-ORF3 MAbs could capture ratHEV particles in culture supernatant and serum following treatment with 0.5 % deoxycholate, but not those without prior detergent treatment or fecal ratHEV particles. Following treatment with 0.5 % deoxycholate and 0.5 % trypsin, the buoyant density of ratHEV particles in culture supernatant with ORF3 protein on the surface shifted from 1.15 g/cm3 to 1.26 g/cm3 in a sucrose gradient; the resulting particles were capturable by an anti-ORF2 MAb but not by an anti-ORF3 MAb. This indicates that the ORF3 protein (at least its C-terminal portion) is incorporated into the enveloped ratHEV virions released from infected cells but that it is not found in the virions in the feces, supporting the hypothesis that the ratHEV ORF3 protein is associated with the egress of virions from infected cells, similar to human HEV, despite the fact that the ratHEV ORF3 protein lacks a PSAP amino acid motif.
  • Tomoyuki Abe, Shota Kono, Takahiro Ohnuki, Shuji Hishikawa, Satoshi Kunita, Yutaka Hanazono
    Experimental animals 65 4 345 - 351 2016年11月 [査読有り][通常論文]
     
    Animal models of thrombocytopenia are indispensable for evaluating the in vivo efficacy of hemostatic agents, cryopreserved platelets, and artificial platelets, but no large animal models are available. In this study, we generated a swine model of acute thrombocytopenia with prolonged bleeding times by administering the chemotherapeutic drug busulfan. First, we tested multiple doses of busulfan (4, 6, and 8 mg/kg) in pigs, and found that 6 mg/kg of busulfan is an optimal dose for producing a safe and moderate thrombocytopenia, with a platelet count of less than 30,000/µl. The pigs administered 6 mg/kg of busulfan (n=8) reached half their initial counts at day 7, counts below 30,000/µl at day 12, and their nadirs at day 15 (on average). The minimal platelet count was 14,000/µl. With this dose of busulfan (6 mg/kg), bleeding times were significantly prolonged in addition to the decrease in platelet counts (r=-0.63, P<0.01), while there were no cases of apparent hemorrhage. White blood cell counts were maintained at over 5,000/µl, and there were no infections or other adverse events including anemia or appetite or body weight loss. All pigs were sacrificed on day 16, with subsequent examination showing a significant reduction in cellularity and colony-forming units in the bone marrow, indicating that thrombocytopenia was the result of myelosuppression. In summary, administration with 6 mg/kg of busulfan induces safe and moderate thrombocytopenia with a prolonged bleeding time in swine.
  • Seiya Mizuno, Dinh T H Tra, Atsushi Mizobuchi, Hiroyoshi Iseki, Saori Mizuno-Iijima, Jun-Dal Kim, Junji Ishida, Yoichi Matsuda, Satoshi Kunita, Akiyoshi Fukamizu, Fumihiro Sugiyama, Ken-ichi Yagami
    Laboratory investigation; a journal of technical methods and pathology 94 3 321 - 30 2014年03月 [査読有り][通常論文]
     
    Agenesis of the corpus callosum (ACC) is a congenital abnormality of the brain structure. More than 60 genes are known to be involved in corpus callosum development. However, the molecular mechanisms underlying ACC are not fully understood. Previously, we produced a novel transgenic mouse strain, TAS, carrying genes of the tetracycline-inducible expression system that are not involved in brain development, and inherited ACC was observed in the brains of all homozygous TAS mice. Although ACC was probably induced by transgene insertion mutation, the causative gene and the molecular mechanism of its pathogenesis remain unclear. Here, we first performed interphase three-color fluorescence in situ hybridization (FISH) analysis to determine the genomic insertion site. Transgenes were inserted into chromosome 18 ∼12.0 Mb from the centromere. Gene expression analysis and genomic PCR walking showed that the genomic region containing exon 4 of Cables1 was deleted by transgene insertion and the other exons of Cables1 were intact. The mutant allele was designated as Cables1(TAS). Interestingly, Cables1(TAS) mRNA consisted of exons 1-3 of Cables1 and part of the transgene that encoded a novel truncated Cables1 protein. Homozygous TAS mice exhibited mRNA expression of Cables1(TAS) in the fetal cerebrum, but not that of wild-type Cables1. To investigate whether a dominant negative effect of Cables1(TAS) or complete loss of function of Cables1 gives rise to ACC, we produced Cables1-null mutant mice. ACC was not observed in Cables1-null mutant mice, suggesting that a dominant negative effect of Cables1(TAS) impairs callosal formation. Moreover, ACC frequency in Cables1(+/TAS) mice was significantly lower than that in Cables1(-/TAS) mice, indicating that wild-type Cables1 interfered with the dominant negative effect of Cables1(TAS). This study indicated that truncated Cables1 causes ACC and wild-type Cables1 contributes to callosal formation.
  • Tomohiro Ishimaru, Junji Ishida, Shoko Nakamura, Misuzu Hashimoto, Tanomu Matsukura, Ayumi Nakamura, Satoshi Kunita, Fumihiro Sugiyama, Ken-Ichi Yagami, Akiyoshi Fukamizu
    Molecular medicine reports 6 1 28 - 32 2012年07月 [査読有り][通常論文]
     
    Pregnancy-induced hypertension or pre-eclampsia is a major disorder that may result in serious complications for the mother and fetus. It is characterized from maternal hypertension in late pregnancy and peripheral tissue damage, including kidney, heart and placenta, and the fetus suffers from intrauterine growth retardation (IUGR) and high perinatal mortality. Recently, it has been postulated that angiotensin II (Ang II), a potent vasoconstrictor in the renin-angiotensin system (RAS), plays a pivotal role in the pathogenesis of pre-eclampsia; however, the beneficial effect of the suppression of RAS has not yet been fully elucidated. Previously, we generated a transgenic mouse model that developed pregnancy-associated hypertension (PAH) by the overproduction of Ang II in maternal circulation during late pregnancy. In addition, mice with PAH exhibited maternal and fetal abnormalities, such as proteinuria, cardiac hypertrophy, placental morphological changes and IUGR. In this study, in order to attenuate the activity of redundant RAS during the advanced stages of PAH, we administered olmesartan (Olm), an angiotensin receptor blocker, and captopril (Cp), an angiotensin converting enzyme inhibitor, from E17 to E19 days of gestation, and evaluated its effect on cardiac and placental abnormalities and fetal growth. Olm and Cp administration significantly lowered the blood pressure of mice with PAH, and placental histological change and severe IUGR were markedly ameliorated in both groups. On the contrary, Olm or Cp treatment had little effect on cardiac remodeling during the advanced stages of PAH. These findings highlight a variety of therapeutic actions of RAS repression on the progressive pathology of PAH in mice.
  • Satoshi Nagamine, Michiko Tamba, Hisako Ishimine, Kota Araki, Kensuke Shiomi, Takuya Okada, Tatsuyuki Ohto, Satoshi Kunita, Satoru Takahashi, Ronnie G P Wismans, Toin H van Kuppevelt, Masayuki Masu, Kazuko Keino-Masu
    The Journal of biological chemistry 287 12 9579 - 90 2012年03月 [査読有り][通常論文]
     
    Heparan sulfate endosulfatases Sulf1 and Sulf2 hydrolyze 6-O-sulfate in heparan sulfate, thereby regulating cellular signaling. Previous studies have revealed that Sulfs act predominantly on UA2S-GlcNS6S disaccharides and weakly on UA-GlcNS6S disaccharides. However, the specificity of Sulfs and their role in sulfation patterning of heparan sulfate in vivo remained unknown. Here, we performed disaccharide analysis of heparan sulfate in Sulf1 and Sulf2 knock-out mice. Significant increases in ΔUA2S-GlcNS6S were observed in the brain, small intestine, lung, spleen, testis, and skeletal muscle of adult Sulf1(-/-) mice and in the brain, liver, kidney, spleen, and testis of adult Sulf2(-/-) mice. In addition, increases in ΔUA-GlcNS6S were seen in the Sulf1(-/-) lung and small intestine. In contrast, the disaccharide compositions of chondroitin sulfate were not primarily altered, indicating specificity of Sulfs for heparan sulfate. For Sulf1, but not for Sulf2, mRNA expression levels in eight organs of wild-type mice were highly correlated with increases in ΔUA2S-GlcNS6S in the corresponding organs of knock-out mice. Moreover, overall changes in heparan sulfate compositions were greater in Sulf1(-/-) mice than in Sulf2(-/-) mice despite lower levels of Sulf1 mRNA expression, suggesting predominant roles of Sulf1 in heparan sulfate desulfation and distinct regulation of Sulf activities in vivo. Sulf1 and Sulf2 mRNAs were differentially expressed in restricted types of cells in organs, and consequently, the sulfation patterns of heparan sulfate were locally and distinctly altered in Sulf1 and Sulf2 knock-out mice. These findings indicate that Sulf1 and Sulf2 differentially contribute to the generation of organ-specific sulfation patterns of heparan sulfate.
  • Junji Ishida, Toshiki Matsuoka, Tomoko Saito-Fujita, Saki Inaba, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami, Akiyoshi Fukamizu
    Journal of biochemistry 150 1 5 - 14 2011年07月 [査読有り][通常論文]
     
    Physiological alterations occur in many organ systems during pregnancy. These changes are necessary for the adaptation to pregnancy-specific physiological processes in mother and fetus, and the placenta plays a critical role in the maintenance of homeostasis in pregnancy. Dysregulation of these functional feto-maternal interactions leads to severe complications. There have been many attempts to create animal models that mimic the hypertensive disorders of pregnancy, especially pre-eclampsia. In this review, we summarize the physiology of pregnancy and placental function, and discuss the placental gene expression in normal pregnancy. In addition, we assess a number of established animal models focusing on a specific pathogenic mechanism of pre-eclampsia, including genetically modified mouse models involving the renin-angiotensin system. Validation of these animal models would contribute significantly to understanding the basic principles of pregnancy-associated homeostasis and the pathogenesis of pre-eclampsia.
  • Satoshi Kunita, Kanako Kato, Miyuki Ishida, Kozue Hagiwara, Shuko Kameda, Tomoko Ishida, Akira Takakura, Kazuo Goto, Fumihiro Sugiyama, Ken-Ichi Yagami
    Clinical and vaccine immunology : CVI 18 5 758 - 66 2011年05月 [査読有り][通常論文]
     
    We describe a new microsphere-based multiplex fluorescent immunoassay (MFI) using recombinant mouse hepatitis virus (MHV) proteins to detect antibodies to coronaviruses in mouse and rat sera. All the recombinant proteins, including nucleocapsid (N) and 3 subunits of spike protein, S1, S2, and Smid, showed positive reactivity in MFI with mouse antisera to 4 MHV strains (MHV-S, -A59, -JHM, and -Nu67) and rat antiserum to a strain of sialodacryoadenitis virus (SDAV-681). The MFI was evaluated for its diagnostic power, with panels of mouse sera classified as positive or negative for anti-MHV antibodies by enzyme-linked immunosorbent assay (ELISA) using MHV virion antigen and indirect fluorescent antibody assay. The reactivities of 236 naturally infected mouse sera were examined; 227 samples were positive by MFI using S2 antigen (96% sensitivity), and 208 samples were positive using N antigen (88% sensitivity). Based on the assessment by MFI using the S2 and N antigens, only 3 serum samples showed double-negative results, indicating a false-negative rate of 1.3%. In 126 uninfected mouse sera, including 34 ELISA false-positive sera, only 7 samples showed false-positive results by MFI using either the S2 or N antigen (94% specificity). Similarly, the S2 and N antigen-based MFI was 98% sensitive and 100% specific in detecting anticoronavirus antibodies in rat sera. Thus, this MFI-based serologic assay using the S2 and N antigens promises to be a reliable diagnostic method, representing a highly sensitive and specific alternative to traditional ELISA for detection of coronavirus infections in laboratory mouse and rat colonies.
  • Seiya Mizuno, Saori Iijima, Tomoko Okano, Noriko Kajiwara, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami
    Experimental animals 60 2 161 - 7 2011年 [査読有り][通常論文]
     
    We found 6 spontaneous mutant mice with long pelage hair in our ICR breeding colony. The abnormal trait was restricted to long hair in these mice, which we named moja. They were fertile and showed the same growth and behavior as wild-type mice. To investigate the manner of the genetic inheritance of the moja allele, offspring were bred by mating the moja mice; all offspring had long pelage hair. Furthermore, we performed a reciprocal cross between moja mice and wild-type ICR mice with normal hair. All offspring exhibited normal hair suggesting an autosomal recessive inheritance of the trait. The moja/moja hair phenotype was maintained in skin grafted onto nude mice, suggesting that circulating or diffusible humoral factors regulating the hair cycle are not involved in the abnormal trait. The phenotype of moja/moja mice is similar to that of Fgf5-deficient mice. Therefore, we examined the expression of Fgf5 by RT-PCR in moja/moja mice. As expected, no Fgf5 expression was found in moja/moja mouse skin. PCR and DNA sequence analyses were performed to investigate the structure of the Fgf5 gene. We found a deletion of a 9.3-kb region in the Fgf5 gene including exon 3 and its 5' and 3' flanking sequences. Interestingly, the genomic deletion site showed insertion of a 498-bp early transposon element long terminal repeat. Taken together, these results suggest that the long hair mutation of moja/moja mice is caused by disruption of Fgf5 mediated by insertion of a retrotransposon.
  • Seiya Mizuno, Atsushi Mizobuchi, Hiroyoshi Iseki, Saori Iijima, Yoichi Matsuda, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami
    Mammalian genome : official journal of the International Mammalian Genome Society 21 11-12 525 - 33 2010年12月 [査読有り][通常論文]
     
    Agenesis of the corpus callosum (ACC) is a congenital abnormality of the brain structure. We have produced transgenic mice expressing both reverse tetracycline-controlled transactivator (rtTA) and transcriptional silencer (tTS) ubiquitously. Although the transgene products do not affect development of the mouse brain, one of the founder lines, TAS, showed ACC, suggesting transgenic disruption of endogenous gene(s). To identify the causative gene and its role in ACC, we performed pathological investigations of the brain and chromosomal mapping of foreign genes in TAS mice. Sixty-two percent of the heterozygous TAS mice showed ACC accompanied with formation of Probst bundles, as seen in human. Complete penetrance of ACC was observed in homozygous TAS mice. Furthermore, homozygous TAS fetuses revealed that ACC is a congenital anomaly. Moreover, axons of the corpus callosum were not repelled by the midline glial structures in TAS mice. These findings suggested that the causative gene for ACC is involved in critical steps in corpus callosum development. Multiple FISH analyses were performed to determine the site of transgene insertion. On 1-color FISH analyses, rtTA and tTS were detected on the A/B region of chromosome 18, suggesting cointegration of the transgenes. On 2-color FISH analyses, tTS signal was observed in a region from 9.3 to 16.9 Mb on chromosome 18. The TAS mice may serve as a useful model to identify a novel gene regulating corpus callosum development and to gain a new insight into molecular genetics of ACC.
  • Saori Iijima, Yoko Tanimoto, Seiya Mizuno, Yoko Daitoku, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami
    Cellular reprogramming 12 6 679 - 88 2010年12月 [査読有り][通常論文]
     
    As the phenotype of a given single-gene mutation in mice is modulated by the genetic background of the inbred strain, embryonic stem (ES) cells derived from various inbred mouse strains are required to produce gene-targeted mice without the need for backcrossing and for detailed analysis of gene function in vivo. Here, we performed a comparative investigation of the effects of three culture conditions, LIF + KSR/ES medium described previously, High LIF + KSR/ES medium and iSTEM + LIF medium containing three inhibitors of glycogen synthase kinase 3, mitogen-activated protein kinase kinase, and fibroblast growth factor receptor signaling (3i), on the establishment of germline-competent ES cells derived from strains BALB/c and NZB mice. The results indicated that LIF + KSR/ES medium was permissive for the derivation of ES cells from NZB mice, which contribute to the somatic lineage in vivo, but not to the germline lineage. In contrast, ES cells that contribute to the makeup of chimeric mice were not propagated from blastocysts of BALB/c mice. Both germline and somatic competency were improved by increased LIF concentration in cultures of BALB/c ES cells, although we failed to establish germline-competent NZB ES cells using the same concentration of LIF. Unexpectedly, iSTEM + LIF medium containing 3i showed a negative effect on the derivation of NZB ES cells with normal chromosome numbers, but not on the maintenance of previously established ES cells. Our findings suggest that the stability of pluripotency in the inner cell mass isolated from blastocyst embryos may differ according to the genetic background of inbred mouse strains, and that although the concentration of LIF is a determinant for authentic pluripotency, including germline and somatic competency in BALB/c ES cells, additional factor(s) are required for commitment to germline lineage independent of somatic lineage in NZB ES cells.
  • Iijima,S, Tanimoto,Y, Mizuno,S, Daitoku,Y, Kunita,S, Sugiyama,F, Yagami,K
    Cell Reprogram 12 6 679 - 688 2010年12月 [査読有り][通常論文]
     
    As the phenotype of a given single-gene mutation in mice is modulated by the genetic background of the inbred strain, embryonic stem (ES) cells derived from various inbred mouse strains are required to produce gene-targeted mice without the need for backcrossing and for detailed analysis of gene function in vivo. Here, we performed a comparative investigation of the effects of three culture conditions, LIF + KSR/ES medium described previously, High LIF + KSR/ES medium and iSTEM + LIF medium containing three inhibitors of glycogen synthase kinase 3, mitogen-activated protein kinase kinase, and fibroblast growth factor receptor signaling (3i), on the establishment of germline-competent ES cells derived from strains BALB/c and NZB mice. The results indicated that LIF + KSR/ES medium was permissive for the derivation of ES cells from NZB mice, which contribute to the somatic lineage in vivo, but not to the germline lineage. In contrast, ES cells that contribute to the makeup of chimeric mice were not propagated from blastocysts of BALB/c mice. Both germline and somatic competency were improved by increased LIF concentration in cultures of BALB/c ES cells, although we fa
  • A novel mouse model for agenesis of corpus callosum
    Mizuno, S, Ishige, T, Mizobuchi, A, Kunita, S, Sugiyama, F, Yagami, K
    60th AALAS National Meeting 2009年11月 [査読有り][通常論文]
  • Okada, Takuya, Keino-Masu, Kazuko, Nagamine, Satoshi, Kunita, Satoshi, Takahashi, Satoru, Masu, Masayuki
    NEUROSCIENCE RESEARCH 65 Suppl. 1 0 - 0 ELSEVIER IRELAND LTD 2009年01月 [査読有り][通常論文]
  • Restoration of axon guidance defects in SulfFP-deficient mice by introducing exogenous SulfFPs
    Okada, Takuya, Keino-Masu, Kazuko, Nagamine, Satoshi, Kunita, Satoshi, Takahashi, Satoru, Masu, Masayuki
    NEUROSCIENCE RESEARCH 61 Suppl. 1 0 - 0 ELSEVIER IRELAND LTD 2008年01月 [査読有り][通常論文]
  • Motor learning in the mice lacking a heparan sulfate 6-O-endosulfatase SulfFP1
    Keino-Masu, Kazuko, Okamoto, Takehito, Shutoh, Fumihiro, Yamasaki, Nobuyuki, Miyakawa, Tsuyoshi, Ohto, Tatsuyuki, Kunita, Satoshi, Takahashi, Satoru, Nagao, Soichi, Masu, Masayuki
    NEUROSCIENCE RESEARCH 55 Suppl. 1 0 - 0 ELSEVIER IRELAND LTD 2006年01月 [査読有り][通常論文]
  • Heparan sulfate 6-O-endosulfatases are required for normal brain development
    Okada, Takuya, Nagamine, Satoshi, Keino-Masu, Kazuko, Ohto, Tatsuyuki, Kunita, Satoshi, Takahashi, Satoru, Masu, Masayuki
    NEUROSCIENCE RESEARCH 55 Suppl. 1 0 - 0 ELSEVIER IRELAND LTD 2006年01月 [査読有り][通常論文]
  • Cerebellar development in the mice lacking a heparan sulfate 6-O-endosulfatase SulfFP1
    Ishibashi, Noriyo, Keino-Masu, Kazuko, Ohto, Tatsuyuki, Kunita, Satoshi, Takahashi, Satoru, Masu, Masayuki
    NEUROSCIENCE RESEARCH 55 Suppl. 1 0 - 0 ELSEVIER IRELAND LTD 2006年01月 [査読有り][通常論文]
  • Takeshi Sakurai, Ruby Nagata, Akihiro Yamanaka, Hiroko Kawamura, Natsuko Tsujino, Yo Muraki, Haruaki Kageyama, Satoshi Kunita, Satoru Takahashi, Katsutoshi Goto, Yoshimasa Koyama, Seiji Shioda, Masashi Yanagisawa
    Neuron 46 5 837  2005年06月 [査読有り][通常論文]
  • Sakurai, T, Nagata, R, Yamanaka, A, Kawamura, H, Tsujino, N, Muraki, Y, Kageyama, H, Kunita, S, Takahashi, S, Goto, K, Koyama, Y, Shioda, S, Yanagisawa, M
    NEURON 46 5 837 - 837 CELL PRESS 2005年06月 [査読有り][通常論文]
  • Kazuo Goto, Ryoko Nozu, Toshio Itoh, Satoshi Kunita, Eiji Terada
    Experimental Animals 44 2 159 - 161 1995年 [査読有り][通常論文]
     
    Restriction endonuclease analysis of amplified nucleocapsid protein genes from mouse hepatitis virus (MHV) was used to differentiate 12 strains isolated from mouse liver or transplantable tumors from five facilities, and the restriction patterns of the isolates were compared with those of five well-defined MHV strains, A59, JHM, 2, S and Nu-67. The patterns of 10 isolates from three facilities were the same as that of Nu-67. The remaining two isolates revealed different patterns from the five reference strains. This study showed that reverse transcription and the polymerase chain reaction assay based restriction analysis are feasible for the detection and genotyping of MHV, and the Nu-67 related strain was the most prevalent type found in the clinical samples. © 1995, Japanese Association for Laboratory Animal Science. All rights reserved.

書籍

  • National Research Council (U.S.). Committee for the Update of the Guide for the Care and Use of Laboratory Animals, 日本実験動物学会, 鍵山, 直子, 大和田, 一雄, 國田, 智, 久原, 孝俊, 黒澤, 努 ()
    アドスリー,丸善出版 (発売) 2011年06月 ISBN: 9784904419236 xxiii, 212p
  • 國田, 智 ()
    [國田智] 2007年05月 43枚
  • マウスにおけるパルボウイルス感染
    ()
    アニテックス 2007年
  • 血液脳関門とRAS
    ()
    Journal of Angiotensin Research 2004年
  • 実験動物の技術と応用 実践編
    ()
    アドスリー 2004年
  • 実験動物施設 -施設の実例:筑波大学生命科学動物資源センター
    ()
    これからの大学等研究施設「生命科学編」(社)文教施設協会 2003年
  • トランスジェニック動物由来細胞の品質・安全性の確保
    ()
    ㈱エル・アイ・シー 「バイオ医薬品の品質・安全性評価」(共著) 2001年

作品等

  • マウス肝炎ウイルス感染症診断に用いる組換え抗原の開発
    2005年

MISC

  • 原弘真, 永山学, 永山学, 永山学, 須田亙, 柴田宏昭, 柴田宏昭, 大貫貴広, 菱川修司, 國田智, 阿部朋行, 阿部朋行, 魚崎英毅, 魚崎英毅, 新幸二, 本田賢也, 本田賢也, 花園豊, 花園豊 日本実験動物学会総会講演要旨集 65th 157 2018年04月 [査読無し][通常論文]
  • 原弘真, 柴田宏昭, 中野和明, 阿部朋行, 阿部朋行, 魚崎英毅, 大貫貴広, スブド ビャンバー, ナーウィン ジャントラー, 菱川修司, 國田智, 長嶋比呂志, 濡木理, 花園豊, 花園豊 日本実験動物学会総会講演要旨集 64th 171 2017年04月 [査読無し][通常論文]
  • 河野正太, 菱川修司, 阿部朋行, 長田直希, 國田智, 花園豊 日本獣医学会学術集会講演要旨集 158th 441 -441 2015年08月 [査読無し][通常論文]
  • Masayuki Masu, Takuya Okada, Satoshi Nagamine, Tatsuyuki Ohto, Fuyuki Kametani, Masato Hasegawa, Satoshi Kunita, Satoru Takahashi, Kazuko Keino-Masu NEUROSCIENCE RESEARCH 71 E10 -E10 2011年 [査読無し][通常論文]
  • Takuya Okada, Kazuko Keino-Masu, Satoshi Nagamine, Satoshi Kunita, Satoru Takahashi, Arie Oosterhof, Toin van Kuppevelt, Masayuki Masu NEUROSCIENCE RESEARCH 68 E247 -E247 2010年 [査読無し][通常論文]
  • A. Fuke, S. Kunita, K. Goto, F. Sugiyama, K. Yagami JOURNAL OF THE AMERICAN ASSOCIATION FOR LABORATORY ANIMAL SCIENCE 48 (5) 634 -635 2009年09月 [査読無し][通常論文]
  • Hiraku Sasaki, Eiichi Kawamoto, Yoshikazu Tanaka, Takuo Sawada, Satoshi Kunita, Ken-ichi Yagami JOURNAL OF BACTERIOLOGY 191 (11) 3698 -3705 2009年06月 [査読無し][通常論文]
     
    Pasteurella pneumotropica is an opportunistic pathogen that causes lethal pneumonia in immunodeficient rodents. The virulence factors of this bacterium remain unknown. In this study, we identified the genes encoding two RTX toxins, designated as pnxI and pnxII, from the genomic DNA of P. pneumotropica ATCC 35149 and characterized with respect to hemolysis. The pnxI operon was organized according to the manner in which the genes encoded the structural RTX toxin (pnxIA), the type I secretion systems (pnxIB and pnxID), and the unknown orf. The pnxII gene was involved only with the pnxIIA that coded for a structural RTX toxin. Both the structural RTX toxins of deduced PnxIA and PnxIIA were involved in seven of the RTX repeat and repeat-like sequences. By quantitative PCR analysis of the structural RTX toxin-encoding genes in P. pneumotropica ATCC 35149, the gene expression of pnxIA was found to have increased from the early log phase, while that of pnxIIA increased from the late log to the early stationary phase. As expressed in Escherichia coli, both the recombinant proteins of PnxIA and PnxIIA showed weak hemolytic activity in both sheep and murine erythrocytes. On the basis of the results of the Southern blotting analysis, the pnxIA gene was detected in 82% of the isolates, while the pnxIIA gene was detected in 39%. These results indicate that the products of both pnxIA and pnxIIA were putative associations of virulence factors in the rodent pathogen P. pneumotropica.
  • Hiraku Sasaki, Eiichi Kawamoto, Yoshikazu Tanaka, Takuo Sawada, Satoshi Kunita, Ken-ichi Yagami Journal of bacteriology 191 (11) 3698 -705 2009年06月 [査読有り][通常論文]
     
    Pasteurella pneumotropica is an opportunistic pathogen that causes lethal pneumonia in immunodeficient rodents. The virulence factors of this bacterium remain unknown. In this study, we identified the genes encoding two RTX toxins, designated as pnxI and pnxII, from the genomic DNA of P. pneumotropica ATCC 35149 and characterized with respect to hemolysis. The pnxI operon was organized according to the manner in which the genes encoded the structural RTX toxin (pnxIA), the type I secretion systems (pnxIB and pnxID), and the unknown orf. The pnxII gene was involved only with the pnxIIA that coded for a structural RTX toxin. Both the structural RTX toxins of deduced PnxIA and PnxIIA were involved in seven of the RTX repeat and repeat-like sequences. By quantitative PCR analysis of the structural RTX toxin-encoding genes in P. pneumotropica ATCC 35149, the gene expression of pnxIA was found to have increased from the early log phase, while that of pnxIIA increased from the late log to the early stationary phase. As expressed in Escherichia coli, both the recombinant proteins of PnxIA and PnxIIA showed weak hemolytic activity in both sheep and murine erythrocytes. On the basis of the results of the Southern blotting analysis, the pnxIA gene was detected in 82% of the isolates, while the pnxIIA gene was detected in 39%. These results indicate that the products of both pnxIA and pnxIIA were putative associations of virulence factors in the rodent pathogen P. pneumotropica.
  • Hiraku Sasaki, Eiichi Kawamoto, Yoshikazu Tanaka, Takuo Sawada, Satoshi Kunita, Ken-ichi Yagami Antonie van Leeuwenhoek 95 (4) 311 -7 2009年05月 [査読有り][通常論文]
     
    Selected biochemical and genetic characteristics of the wild-type strains of Pasteurella pneumotropica isolated from mice and rats were investigated and compared in order to determine the significant differences among the isolates. The isolates were divided into six groups on the basis of the patterns of carbon source utilization in the host rodents. The genome sizes were determined by electrophoretic analysis, and the mean genome size of the isolates from mice was larger than that of the isolates from rats (P < 0.05). Cluster analysis of the rpoB sequences discriminated five clusters; the differences might have correlated with the host associations. Principal component analysis (PCA) based on both the biochemical and genetic characteristics revealed total 44 strains discriminated into three groups comprising the host-dependent and host-independent groups. Although the P. pneumotropica isolates were mainly classified on the basis of the host rodents by the examinations, the existence of isolates that could not be discriminated on the basis of the host rodents alone was confirmed by the PCA. These results indicated that the P. pneumotropica isolates could be further classified by taxonomic analysis and also suggested the existence of a host-independent group in addition to the host-dependent groups.
  • Hiraku Sasaki, Eiichi Kawamoto, Yoshikazu Tanaka, Takuo Sawada, Satoshi Kunita, Ken-ichi Yagami ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY 95 (4) 311 -317 2009年05月 [査読無し][通常論文]
     
    Selected biochemical and genetic characteristics of the wild-type strains of Pasteurella pneumotropica isolated from mice and rats were investigated and compared in order to determine the significant differences among the isolates. The isolates were divided into six groups on the basis of the patterns of carbon source utilization in the host rodents. The genome sizes were determined by electrophoretic analysis, and the mean genome size of the isolates from mice was larger than that of the isolates from rats (P < 0.05). Cluster analysis of the rpoB sequences discriminated five clusters; the differences might have correlated with the host associations. Principal component analysis (PCA) based on both the biochemical and genetic characteristics revealed total 44 strains discriminated into three groups comprising the host-dependent and host-independent groups. Although the P. pneumotropica isolates were mainly classified on the basis of the host rodents by the examinations, the existence of isolates that could not be discriminated on the basis of the host rodents alone was confirmed by the PCA. These results indicated that the P. pneumotropica isolates could be further classified by taxonomic analysis and also suggested the existence of a host-independent group in addition to the host-dependent groups.
  • Taizo Matsuki, Mika Nomiyama, Hitomi Takahira, Noriko Hirashima, Satoshi Kunita, Satoru Takahashi, Ken-ichi Yagami, Thomas S. Kilduff, Bernhard Bettler, Masashi Yanagisawa, Takeshi Sakurai PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 106 (21) 8790 -8790 2009年05月 [査読無し][通常論文]
  • 今野裕土, 伊関大敬, 秋野雅一, 國田智, 杉山文博, 奥泉久人, 八神健一 日本実験動物学会総会講演要旨集 56th 212 2009年04月 [査読無し][通常論文]
  • Taizo Matsuki, Mika Nomiyama, Hitomi Takahira, Noriko Hirashima, Satoshi Kunita, Satoru Takahashi, Ken-ichi Yagami, Thomas S Kilduff, Bernhard Bettler, Masashi Yanagisawa, Takeshi Sakurai Proceedings of the National Academy of Sciences of the United States of America 106 (11) 4459 -64 2009年03月 [査読有り][通常論文]
     
    Hypothalamic neurons that contain the neuropeptide orexin (hypocretin) play important roles in the regulation of sleep/wake. Here we analyze the in vivo and in vitro phenotype of mice lacking the GABA(B1) gene specifically in orexin neurons (oxGKO mice) and demonstrate that GABA(B) receptors on orexin neurons are essential in stabilizing and consolidating sleep/wake states. In oxGKO brain slices, we show that the absence of GABA(B) receptors decreases the sensitivity of orexin neurons to both excitatory and inhibitory inputs because of augmented GABA(A)-mediated inhibition that increases the membrane conductance and shunts postsynaptic currents in these neurons. This increase in GABA(A)-mediated inhibitory tone is apparently the result of an orexin receptor type 1-mediated activation of local GABAergic interneurons that project back onto orexin neurons. oxGKO mice exhibit severe fragmentation of sleep/wake states during both the light and dark periods, without showing an abnormality in total sleep time or signs of cataplexy. Thus, GABA(B) receptors on orexin neurons are crucial in the appropriate control of the orexinergic tone through sleep/wake states, thereby stabilizing the state switching mechanisms.
  • Taizo Matsuki, Mika Nomiyama, Hitomi Takahira, Noriko Hirashima, Satoshi Kunita, Satoru Takahashi, Ken-ichi Yagami, Thomas S. Kilduff, Bernhard Bettler, Masashi Yanagisawa, Takeshi Sakurai PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 106 (11) 4459 -4464 2009年03月 [査読無し][通常論文]
     
    Hypothalamic neurons that contain the neuropeptide orexin (hypocretin) play important roles in the regulation of sleep/wake. Here we analyze the in vivo and in vitro phenotype of mice lacking the GABA(B1) gene specifically in orexin neurons (oxGKO mice) and demonstrate that GABA(B) receptors on orexin neurons are essential in stabilizing and consolidating sleep/wake states. In oxGKO brain slices, we show that the absence of GABAB receptors decreases the sensitivity of orexin neurons to both excitatory and inhibitory inputs because of augmented GABA(A)-mediated inhibition that increases the membrane conductance and shunts postsynaptic currents in these neurons. This increase in GABA(A)-mediated inhibitory tone is apparently the result of an orexin receptor type 1-mediated activation of local GABAergic interneurons that project back onto orexin neurons. oxGKO mice exhibit severe fragmentation of sleep/wake states during both the light and dark periods, without showing an abnormality in total sleep time or signs of cataplexy. Thus, GABAB receptors on orexin neurons are crucial in the appropriate control of the orexinergic tone through sleep/wake states, thereby stabilizing the state switching mechanisms.
  • Yoko Tanimoto, Saori Iijima, Yoshikazu Hasegawa, Yuko Suzuki, Yoko Daitoku, Seiya Mizuno, Taichiro Ishige, Takashi Kudo, Satoru Takahashi, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami Comparative medicine 58 (4) 347 -52 2008年08月 [査読有り][通常論文]
     
    Mouse embryonic stem (ES) cells with the C57BL/6 genetic background allow the generation of knockout mice without the need to backcross to C57BL/6. However, C57BL/6 ES cells whose pluripotency after homologous recombination has been confirmed are not yet available from public cell banks. To facilitate the use of ES cells derived from C57BL/6 sublines in both biologic and medical research, we demonstrated that the use of knockout serum replacement as a medium supplement and 8-cell blastomeres as recipient embryos allowed establishment of ES cells and production of germline chimeric mice, respectively. Under effective conditions, a large number of ES cell lines were established from C57BL/6J and C57BL/6N blastocysts. The majority of ES cells in many cell lines obtained from both strains showed a normal chromosome number. Germline chimeric mice were generated from C57BL/6J and C57BL/6N ES cells. Finally, the ES cell line B6J-S1UTR, derived from C57BL/6J, was used for successful production of gene knockout mice. C57BL/6J ES (B6J-S1UTR and B6J-23UTR) and C57BL/6N ES (B6N-22UTR) cells are available from the cell bank of the BioResource Center at RIKEN Tsukuba Institute (http://www.brc.riken.jp/lab/cell/english/).
  • Yoko Tanimoto, Saori Iijima, Yoshikazu Hasegawa, Yuko Suzuki, Yoko Daitoku, Seiya Mizuno, Taichiro Ishige, Takashi Kudo, Satoru Takahashi, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami COMPARATIVE MEDICINE 58 (4) 347 -352 2008年08月 [査読無し][通常論文]
     
    Mouse embryonic stem (ES) cells with the C57BL/6 genetic background allow the generation of knockout mice without the need to backcross to C57BL/6. However, C57BL/6 ES cells whose pluripotency after homologous recombination has been confirmed are not yet available from public cell banks. To facilitate the use of ES cells derived from C57BL/6 sublines in both biologic and medical research, we demonstrated that the use of knockout serum replacement as a medium supplement and 8-cell blastomeres 44 as recipient embryos allowed establishment of ES cells and production of germline chimeric mice, respectively. Under effective conditions, a large number of ES cell lines were established from C57BL/6J and C57BL/6N blastocysts. The majority of ES cells in many cell lines obtained from both strains showed a normal chromosome number. Germline chimeric mice were generated from C57BL/6J and C57BL/6N ES cells. Finally, the ES cell line B6J-S1(UTR), derived from C57BL/6J, was used for successful production of gene knockout mice. C57BL/6J ES (B6J-S1(UTR) and B6J-23(UTR)) and C57BL/6N ES (B6N-22(UTR)) cells are available from the cell bank of the BioResource Center at RIKEN Tsukuba Institute (http://www.brc.riken.jp/lab/cell/english/).
  • Yoko Shigematsu, Naoki Yoshida, Yoshihiro Miwa, Atsushi Mizobuti, Yuko Suzuki, Yoko Tanimoto, Satoru Takahashi, Satoshi Kunita, Fumihiro Sugiyama, Ken-Ichi Yagami INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 20 (4) 439 -444 2007年10月 [査読無し][通常論文]
     
    Kaede protein is a photoconvertible tracer that emits green fluorescence after synthesis, which changes to stable red fluorescence upon irradiation with violet or UV illumination. This color-change characteristic is a very effective means of optically marking living cells of interest. We established novel embryonic stem (ES) cell lines, B6KED-1 and -2, from C57BL/6J transgenic mouse blastocysts ubiquitously expressing tandem dimeric Kaede (tdKaede) protein. Undifferentiated B6KED-1 and -2 cells showed bright green fluorescence and mRNAs of pluripotent marker genes. Photoconversion of tdKaede protein in undifferentiated and differentiated B6KED cells in vitro occurred upon short-term UV irradiation. B6KED cells completely generated ES cell-derived females on transfer into tetraploid blastomeres. All organs showed strong green emission in the females derived completely from B6KED cells. These novel ES cell lines ubiquitously expressing photoconvertible Kaede protein, B6KED-1 and -2, are useful for basic research in developmental biology and regenerative medicine.
  • Yoko Shigematsu, Naoki Yoshida, Yoshihiro Miwa, Atsushi Mizobuti, Yuko Suzuki, Yoko Tanimoto, Satoru Takahashi, Satoshi Kunita, Fumihiro Sugiyama, Ken-Ichi Yagami INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 20 (4) 439 -444 2007年10月 [査読無し][通常論文]
     
    Kaede protein is a photoconvertible tracer that emits green fluorescence after synthesis, which changes to stable red fluorescence upon irradiation with violet or UV illumination. This color-change characteristic is a very effective means of optically marking living cells of interest. We established novel embryonic stem (ES) cell lines, B6KED-1 and -2, from C57BL/6J transgenic mouse blastocysts ubiquitously expressing tandem dimeric Kaede (tdKaede) protein. Undifferentiated B6KED-1 and -2 cells showed bright green fluorescence and mRNAs of pluripotent marker genes. Photoconversion of tdKaede protein in undifferentiated and differentiated B6KED cells in vitro occurred upon short-term UV irradiation. B6KED cells completely generated ES cell-derived females on transfer into tetraploid blastomeres. All organs showed strong green emission in the females derived completely from B6KED cells. These novel ES cell lines ubiquitously expressing photoconvertible Kaede protein, B6KED-1 and -2, are useful for basic research in developmental biology and regenerative medicine.
  • Hiraku Sasaki, Eiichi Kawamoto, Satoshi Kunita, Ken-Ichi Yagami Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 19 (5) 557 -60 2007年09月 [査読有り][通常論文]
     
    The objectives of this study were to determine and compare the in vitro enrofloxacin susceptibility of 94 Pseudomonas aeruginosa isolates obtained from enrofloxacin-treated and untreated mice and that of 40 Pasteurella pneumotropica strains and also to assess the efficacy and effects of enrofloxacin treatment of laboratory mice. The minimum inhibitory concentrations (MICs) of enrofloxacin against all the Ps. aeruginosa isolates were in the range of 1 to 4 microg/ml, whereas those against all the P. pneumotropica strains were less than 0.5 microg/ml. The mutation frequency in 54% of the Ps. aeruginosa isolates on treatment with enrofloxacin ranged from 10(-6) to 10(-8); however, none of the P. pneumotropica strains could grow on medium containing more than 3 microg/ml enrofloxacin. Comparison of in vitro enrofloxacin susceptibilities suggested that enrofloxacin was effective for eliminating P. pneumotropica but not for eliminating Ps. aeruginosa for which the MIC of enrofloxacin was more than 1 microg/ml. These results indicated that the enrofloxacin susceptibility of P. pneumotropica was higher than that of Ps. aeruginosa, and that the enrofloxacin treatment might not affect the susceptibility of Ps. aeruginosa.
  • Hiraku Sasaki, Eiichi Kawamoto, Satoshi Kunita, Ken-ichi Yagami JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION 19 (5) 557 -560 2007年09月 [査読無し][通常論文]
     
    The objectives of this study were to determine and compare the in vitro enrofloxacin susceptibility of 94 Pseudomonas aeruginosa isolates obtained from enrofloxacin-treated and untreated mice and that of 40 Pasteurella pneumotropica strains and also to assess the efficacy and effects of enrofloxacin treatment of laboratory mice. The minimum inhibitory concentrations (MICs) of enrofloxacin against all the Ps. aeruginosa isolates were in the range of 1 to 4 mu g/ml, whereas those against all the P. pneumotropica strains were less than 0.5 mu g/ml. The mutation frequency in 54% of the Ps. aeruginosa isolates on treatment with enrofloxacin ranged from 10(-6) to 10(-8); however, none of the P. pneumotropica strains could grow on medium containing more than 3 mu g/ml enrofloxacin. Comparison of in vitro enrofloxacin susceptibilities suggested that enrofloxacin was effective for eliminating P. pneumotropica but not for eliminating Ps. aeruginosa for which the MIC of enrofloxacin was more than 1 mu g/ml. These results indicated that the enrofloxacin susceptibility of P. pneumotropica was higher than that of Ps. aeruginosa, and that the enrofloxacin treatment might not affect the susceptibility of Ps. aeruginosa.
  • Eri Nishihara, Shirng-Wern Tsaih, Chieko Tsukahara, Sarah Langley, Susan Sheehan, Keith DiPetrillo, Satoshi Kunita, Ken-ichi Yagami, Gary A. Churchill, Beverly Paigen, Fumihiro Sugiyama MAMMALIAN GENOME 18 (8) 573 -583 2007年08月 [査読無し][通常論文]
     
    In a previous study in 15 inbred mouse strains, we found highest and lowest systolic blood pressures in NZO/HILtJ mice (metabolic syndrome) and C3H/HeJ mice (common lean strain), respectively. To identify the loci involved in hypertension in metabolic syndrome, we performed quantitative trait locus (QTL) analysis for blood pressure with direction of cross as a covariate in segregating F-2 males derived from NZO/HILtJ and C3H/HeJ mice. We detected three suggestive main-effect QTLs affecting systolic and diastolic blood pressures (SBP and DBP). We analyzed the first principle component (PC1) generated from SBP and DBP to investigate blood pressure. In addition to all the suggestive QTLs (Chrs 1, 3, and 8) in SBP and DBP, one suggestive QTL on Chr 4 was found in PC1 in the main scan. Simultaneous search identified two significant epistatic locus pairs (Chrs 1 and 4, Chrs 4 and 8) for PC1. Multiple regression analysis revealed three blood pressure QTLs (Bpq10, 100 cM on Chr 1; Bpq11, 6 cM on Chr 4; Bpq12, 29 cM on Chr 8) accounting for 29.4% of blood pressure variance. These were epistatic interaction QTLs constructing a small network centered on Chr 4, suggesting the importance of genetic interaction for development of hypertension. The blood pressure QTLs on Chrs 1, 4, and 8 were detected repeatedly in multiple studies using common inbred nonobese mouse strains, implying substantial QTL independent of development of obesity and insulin resistance. These results enhance our understanding of complicated genetic factors of hypertension in metabolic diseases.
  • Eri Nishihara, Shirng-Wern Tsaih, Chieko Tsukahara, Sarah Langley, Susan Sheehan, Keith DiPetrillo, Satoshi Kunita, Ken-ichi Yagami, Gary A. Churchill, Beverly Paigen, Fumihiro Sugiyama MAMMALIAN GENOME 18 (8) 573 -583 2007年08月 [査読無し][通常論文]
     
    In a previous study in 15 inbred mouse strains, we found highest and lowest systolic blood pressures in NZO/HILtJ mice (metabolic syndrome) and C3H/HeJ mice (common lean strain), respectively. To identify the loci involved in hypertension in metabolic syndrome, we performed quantitative trait locus (QTL) analysis for blood pressure with direction of cross as a covariate in segregating F-2 males derived from NZO/HILtJ and C3H/HeJ mice. We detected three suggestive main-effect QTLs affecting systolic and diastolic blood pressures (SBP and DBP). We analyzed the first principle component (PC1) generated from SBP and DBP to investigate blood pressure. In addition to all the suggestive QTLs (Chrs 1, 3, and 8) in SBP and DBP, one suggestive QTL on Chr 4 was found in PC1 in the main scan. Simultaneous search identified two significant epistatic locus pairs (Chrs 1 and 4, Chrs 4 and 8) for PC1. Multiple regression analysis revealed three blood pressure QTLs (Bpq10, 100 cM on Chr 1; Bpq11, 6 cM on Chr 4; Bpq12, 29 cM on Chr 8) accounting for 29.4% of blood pressure variance. These were epistatic interaction QTLs constructing a small network centered on Chr 4, suggesting the importance of genetic interaction for development of hypertension. The blood pressure QTLs on Chrs 1, 4, and 8 were detected repeatedly in multiple studies using common inbred nonobese mouse strains, implying substantial QTL independent of development of obesity and insulin resistance. These results enhance our understanding of complicated genetic factors of hypertension in metabolic diseases.
  • エンロフロキサシンに対する緑膿菌と肺パスツレラ菌の感受性の比較
    佐々木 啓, 川本 英一, 國田 智, 八神 健一 日本獣医学会学術集会講演要旨集 143回 226 -226 2007年03月 [査読無し][通常論文]
  • マウスにおけるパルボウイルス感染
    研成社アニテックス 19 (3) 53 -55 2007年 [査読無し][通常論文]
  • Satoshi Kunita, Miyuki Chaya, Kozue Hagiwara, Tomoko Ishida, Akira Takakura, Tatsuya Sugimoto, Hiroyoshi Iseki, Kumiko Fuke, Fumihiro Sugiyama, Ken-ichi Yagami Experimental animals 55 (2) 117 -24 2006年04月 [査読有り][通常論文]
     
    Nucleotide sequences of mouse parvovirus (MPV) isolate, named MPV/UT, and mouse minute virus (MMV) were analyzed and used for expressing recombinant proteins in E. coli. ELISA tests using recombinant major capsid protein (rVP2) and recombinant major non-structural protein (rNS1) as antigens were developed and their performance in serologic detection of rodent parvovirus infection was assessed. MPV-rVP2 and MMV-rVP2 ELISAs reacted specifically with anti-MPV and anti-MMV mouse sera, respectively. MMV-rNS1 antigen had a wide reaction range with antisera to rodent parvoviruses including MPV, MMV, Kilham rat virus (KRV) and H-1 virus. All mice oronasally infected with MPV were seropositive at 4 weeks post-infection in screening by ELISAs using MPV-rVP2 and MMV-rNS1 antigens, but were negative by conventional ELISA using whole MMV antigen. A contact transmission experiment revealed that transmission of MPV occurred up to 4 weeks post-infection, and all cage mates were seropositive in screening with MPV-rVP2 and MMV-rNS1 ELISAs. These results indicate that MPV-rVP2 and MMV-rVP2 are specific ELISA antigens which distinguish between MPV and MVM infection, while MMV-rNS1 antigen can be used in generic ELISA for a variety of rodent parvoviruses. The higher sensitivity of MPV-rVP2 ELISA than conventional ELISA for detecting seroconversion to MPV in oronasally infected mice as well as in cage mates suggests the usefulness of MPV-rVP2 ELISA in quarantine and microbiological monitoring of MPV infection in laboratory mice.
  • S Kunita, M Chaya, K Hagiwara, T Ishida, A Takakura, T Sugimoto, H Iseki, K Fuke, F Sugiyama, K Yagami EXPERIMENTAL ANIMALS 55 (2) 117 -124 2006年04月 [査読無し][通常論文]
     
    Nucleotide sequcences of mouse parvovirus (MPV) isolate, named MPV/UT, and mouse minute virus (MMV) were analyzed and used for expressing recombinant proteins in E. coli. ELISA tests using recombinant major capsid protein (rVP2) and recombinant major non-structural protein (rNS1) as antigens were developed and their performance in serologic detection of rodent parvovirus infection was assessed. MPV-rVP2 and MMV-rVP2 ELISAs reacted specifically with anti-MPV and anti-MMV mouse sera, respectively. MMV-rNS1 antigen had a wide reaction range with antisera to rodent parvoviruses including MPV, MMV, Kilham rat virus (KRV) and H-1 virus. All mice oronasally infected with MPV were seropositive at 4 weeks post-infection in screening by ELISAs using MPV-rVP2 and MMV-rNS1 antigens, but were negative by conventional ELISA using whole MMV antigen. A contact transmission experiment revealed that transmission of MPV occurred up to 4 weeks post-infection, and all cage mates were seropositive in screening with MPV-rVP2 and MMV-rNS1 ELISAs. These results indicate that MPV-rVP2 and MMV-rVP2 are specific ELISA antigens which distinguish between MPV and MVM infection, while MMV-rNS1 antigen can be used in generic ELISA for a variety of rodent parvoviruses. The higher sensitivity of MPV-rVP2 ELISA than conventional ELISA for detecting seroconversion to MPV in oronasally infected mice as well as in cage mates suggests the usefulness of MPV-rVP2 ELISA in quarantine and microbiological monitoring of MPV infection in laboratory mice.
  • Y Shimizu, N Motohashi, H Iseki, S Kunita, F Sugiyama, KI Yagami INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 17 (1) 21 -28 2006年01月 [査読無し][通常論文]
     
    We characterized murine spermatogonial stem cells (SSCs) using a multi-parameter selection strategy, combining Oct4 expression determined by monitoring green fluorescent protein (GFP) expression, and the testicular side population (SP) showing weak fluorescence on Hoechst 33342 dye staining, as markers of stem cell purification. Testicular cells were collected from Oct4/GFP transgenic mice and analyzed using a fluorescence-activated cell sorter (FACS). SP was detected in testicular cell suspensions at an average rate of 0.10%. Multicolor analysis indicated that 96% of SP cells were negative for Oct4. The cells did not express SSC marker genes, but expressed Bcrp1. While the main population was 93% positive for pyronin Y staining, this was limited to 51% in SP. We found a novel subpopulation with reduced RNA content lacking Oct4 expression in testicular SP. These results suggest that the cells isolated by FACS represent a novel population of SSCs in the G0 quiescent state.
  • Motoko Yanagita, Tomohiko Okuda, Shuichiro Endo, Mari Tanaka, Katsu Takahashi, Fumihiro Sugiyama, Satoshi Kunita, Satoru Takahashi, Atsushi Fukatsu, Masashi Yanagisawa, Toru Kita, Takeshi Sakurai The Journal of clinical investigation 116 (1) 70 -9 2006年01月 [査読有り][通常論文]
     
    Dialysis dependency is one of the leading causes of morbidity and mortality in the world, and once end-stage renal disease develops, it cannot be reversed by currently available therapy. Although administration of large doses of bone morphogenetic protein-7 (BMP-7) has been shown to repair established renal injury and improve renal function, the pathophysiological role of endogenous BMP-7 and regulatory mechanism of its activities remain elusive. Here we show that the product of uterine sensitization-associated gene-1 (USAG1), a novel BMP antagonist abundantly expressed in the kidney, is the central negative regulator of BMP function in the kidney and that mice lacking USAG-1 (USAG1 mice) are resistant to renal injury. USAG1 mice exhibited prolonged survival and preserved renal function in acute and chronic renal injury models. Renal BMP signaling, assessed by phosphorylation of Smad proteins, was significantly enhanced in USAG1 mice with renal injury, indicating that the preservation of renal function is attributable to enhancement of endogenous BMP signaling. Furthermore, the administration of neutralizing antibody against BMP-7 abolished renoprotection in USAG1 mice, indicating that USAG-1 plays a critical role in the modulation of renoprotective action of BMP and that inhibition of USAG-1 is a promising means of development of novel treatment for renal diseases.
  • Yuko Shimizu, Noriko Motohashi, Hiroyoshi Iseki, Satoshi Kunita, Fumihiro Sugiyama, Ken-Ichi Yagami International journal of molecular medicine 17 (1) 21 -8 2006年01月 [査読有り][通常論文]
     
    We characterized murine spermatogonial stem cells (SSCs) using a multi-parameter selection strategy, combining Oct4 expression determined by monitoring green fluorescent protein (GFP) expression, and the testicular side population (SP) showing weak fluorescence on Hoechst 33342 dye staining, as markers of stem cell purification. Testicular cells were collected from Oct4/GFP transgenic mice and analyzed using a fluorescence-activated cell sorter (FACS). SP was detected in testicular cell suspensions at an average rate of 0.10%. Multicolor analysis indicated that 96% of SP cells were negative for Oct4. The cells did not express SSC marker genes, but expressed Bcrp1. While the main population was 93% positive for pyronin Y staining, this was limited to 51% in SP. We found a novel subpopulation with reduced RNA content lacking Oct4 expression in testicular SP. These results suggest that the cells isolated by FACS represent a novel population of SSCs in the G0 quiescent state.
  • M Yanagita, T Okuda, S Endo, M Tanaka, K Takahashi, F Sugiyama, S Kunita, S Takahashi, A Fukatsu, M Yanagisawa, T Kita, T Sakurai JOURNAL OF CLINICAL INVESTIGATION 116 (1) 70 -79 2006年01月 [査読無し][通常論文]
     
    Dialysis dependency is one of the leading causes of morbidity and mortality in the world, and once end-stage renal disease develops, it cannot be reversed by currently available therapy. Although administration of large doses of bone morphogenetic protein-7 (BMP-7) has been shown to repair established renal injury and improve renal function, the pathophysiological role of endogenous BMP-7 and regulatory mechanism of its activities remain elusive. Here we show that the product of uterine sensitization-associated gene-1 (USAG1), a novel BMP antagonist abundantly expressed in the kidney, is the central negative regulator of BMP function in the kidney and that mice lacking USAG-1 (USAG1(-/-) mice) are resistant to renal injury. USAG1(-/-) mice exhibited prolonged survival and preserved renal function in acute and chronic renal injury models. Renal BMP signaling, assessed by phosphorylation of Smad proteins, was significantly enhanced in USAG1(-/-) mice with renal injury, indicating that the preservation of renal function is attributable to enhancement of endogenous BMP signaling. Furthermore, the administration of neutralizing antibody against BMP-7 abolished renoprotection in USAG1(-/-) mice, indicating that USAG-1 plays a critical role in the modulation of renoprotective action of BMP and that inhibition of USAG-1 is a promising means of development of novel treatment for renal diseases.
  • H Iseki, R Shimizukawa, F Sugiyama, S Kunita, A Iwama, M Onodera, H Nakauchi, K Yagami JOURNAL OF VIROLOGY 79 (14) 8886 -8893 2005年07月 [査読無し][通常論文]
     
    Several malignant tumor cells become apoptotic and revert to the benign phenotype upon parvovirus infection. Recently, we demonstrated that the rat parvovirus RPV/UT also induces apoptosis in the rat thymic lymphoma cell line C58(NT)D. However, a minority of cells that escaped apoptosis showed properties different from the parental cells, such as resistance to apoptosis, enhanced cell adherence, and suppressed tumorigenicity. The present study was performed to determine the molecular mechanism of parvovirus-induced phenotypic modification, including oncosuppression. We demonstrated that the nonstructural (NS) proteins of RPV/UT induced apoptosis in C58(NT)D cells and suppressed tumor growth in vivo. Interestingly, NS proteins induced the expression of ciliary neurotrophic factor receptor alpha, which is up-regulated in revertant cell clones, and enhanced histone acetylation of its gene. These results indicate that parvoviral NS regulate host gene expression through histone acetylation, suggesting a possible mechanism of oncosuppression.
  • Hiroyoshi Iseki, Rie Shimizukawa, Fumihiro Sugiyama, Satoshi Kunita, Atsushi Iwama, Masafumi Onodera, Hiromitsu Nakauchi, Ken-ichi Yagami Journal of virology 79 (14) 8886 -93 2005年07月 [査読有り][通常論文]
     
    Several malignant tumor cells become apoptotic and revert to the benign phenotype upon parvovirus infection. Recently, we demonstrated that the rat parvovirus RPV/UT also induces apoptosis in the rat thymic lymphoma cell line C58(NT)D. However, a minority of cells that escaped apoptosis showed properties different from the parental cells, such as resistance to apoptosis, enhanced cell adherence, and suppressed tumorigenicity. The present study was performed to determine the molecular mechanism of parvovirus-induced phenotypic modification, including oncosuppression. We demonstrated that the nonstructural (NS) proteins of RPV/UT induced apoptosis in C58(NT)D cells and suppressed tumor growth in vivo. Interestingly, NS proteins induced the expression of ciliary neurotrophic factor receptor alpha, which is up-regulated in revertant cell clones, and enhanced histone acetylation of its gene. These results indicate that parvoviral NS regulate host gene expression through histone acetylation, suggesting a possible mechanism of oncosuppression.
  • Rie Shimizukawa, Aya Sakata, Michiko Hirose, Akio Takahashi, Hiroyoshi Iseki, Ying Liu, Satoshi Kunita, Fumihiro Sugiyama, Ken-ichi Yagami Genesis (New York, N.Y. : 2000) 42 (1) 47 -52 2005年05月 [査読有り][通常論文]
     
    Transgenic mice ubiquitously expressing enhanced green fluorescent protein (EGFP) are useful as marker lines in chimera experiments. We established a new embryonic stem (ES) cell line (named B6G-2) from a C57BL/6 blastocyst showing ubiquitous EGFP expression. Undifferentiated B6G-2 cells showed strong green fluorescence and mRNAs of pluripotent marker genes. B6G-2 cells were transferred into a C57BL/6 blastocyst to generate a germline chimera, the progeny of which inherited ubiquitous EGFP expression. Mice derived completely from B6G-2 cells were also developed from the ES cells; these were tetraploid chimeras. The established B6G-2 cells were shown to be pluripotent and to be capable of differentiating into cells of all lineages. Thus, the new ES cell line expressing EGFP ubiquitously is useful for basic research in the field of regenerative medicine. The B6G-2 cell line is freely available from the BioResource Center, RIKEN Tsukuba Institute (http://www.brc.riken.jp/lab/cell/english/).
  • R Shimizukawa, A Sakata, M Hirose, A Takahashi, H Iseki, Y Liu, S Kunita, F Sugiyama, K Yagami GENESIS 42 (1) 47 -52 2005年05月 [査読無し][通常論文]
     
    Transgenic mice ubiquitously expressing enhanced green fluorescent protein (EGFP) are useful as marker lines in chimera experiments. We established a new embryonic stem (ES) cell line (named B6G-2) from a C57BL/6 blastocyst showing ubiquitous EGFP expression. Undifferentiated B6G-2 cells showed strong green fluorescence and mRNAs of pluripotent marker genes. B6G-2 cells were transferred into a C57BL/6 blastocyst to generate a germline chimera, the progeny of which inherited ubiquitous EGFP expression. Mice derived completely from B6G-2 cells were also developed from the ES cells; these were tetraploid chimeras. The established B6G-2 cells were shown to be pluripotent and to be capable of differentiating into cells of all lineages. Thus, the new ES cell line expressing EGFP ubiquitously is useful for basic research in the field of regenerative medicine. The B6G-2 cell line is freely available from the BioResource Center, RIKEN Tsukuba Institute (http:// www.brc.riken.jp/lab/cell/english/). (c) 2005 Wiley-Liss, Inc.
  • Takeshi Sakurai, Ruby Nagata, Akihiro Yamanaka, Hiroko Kawamura, Natsuko Tsujino, Yo Muraki, Haruaki Kageyama, Satoshi Kunita, Satoru Takahashi, Katsutoshi Goto, Yoshimasa Koyama, Seiji Shioda, Masashi Yanagisawa Neuron 46 (2) 297 -308 2005年04月 [査読有り][通常論文]
     
    The finding of orexin/hypocretin deficiency in narcolepsy patients suggests that this hypothalamic neuropeptide plays a crucial role in regulating sleep/wakefulness states. However, very little is known about the synaptic input of orexin/hypocretin-producing neurons (orexin neurons). We applied a transgenic method to map upstream neuronal populations that have synaptic connections to orexin neurons and revealed that orexin neurons receive input from several brain areas. These include the amygdala, basal forebrain cholinergic neurons, GABAergic neurons in the preoptic area, and serotonergic neurons in the median/paramedian raphe nuclei. Monoamine-containing groups that are innervated by orexin neurons do not receive reciprocal connections, while cholinergic neurons in the basal forebrain have reciprocal connections, which might be important for consolidating wakefulness. Electrophysiological study showed that carbachol excites almost one-third of orexin neurons and inhibits a small population of orexin neurons. These neuroanatomical findings provide important insights into the neural pathways that regulate sleep/wakefulness states.
  • T Sakurai, R Nagata, A Yamanaka, H Kawamura, N Tsujino, Y Muraki, H Kageyama, S Kunita, S Takahashi, K Goto, Y Koyama, S Shioda, M Yanagisawa NEURON 46 (2) 297 -308 2005年04月 [査読無し][通常論文]
     
    The finding of orexin/hypocretin deficiency in narcolepsy patients suggests that this hypothalamic neuropeptide plays a crucial role in regulating sleep/wakefulness states. However, very little is known about the synaptic input of orexin/hypocretin-producing neurons (orexin neurons). We applied a transgenic method to map upstream neuronal populations that have synaptic connections to orexin neurons and revealed that orexin neurons receive input from several brain areas. These include the amygdala, basal forebrain cholinergic neurons, GABAergic neurons in the preoptic area, and serotonergic neurons in the median/paramedian raphe nuclei. Monoamine-containing groups that are innervated by orexin neurons do not receive reciprocal connections, while cholinergic neurons in the basal forebrain have reciprocal connections, which might be important for consolidating wakefulness. Electrophysiological study showed that carbachol excites almost one-third of orexin neurons and inhibits a small population of orexin neurons. These neuroanatomical findings provide important insights into the neural pathways that regulate sleep/ wakefulness states.
  • Chieko Tsukahara, Fumihiro Sugiyama, Beverly Paigen, Satoshi Kunita, Ken-Ichi Yagami Mammalian genome : official journal of the International Mammalian Genome Society 15 (12) 943 -50 2004年12月 [査読有り][通常論文]
     
    We characterized the systolic and diastolic blood pressures of 10-week-old males from 15 inbred mouse strains and found that blood pressures among strains were continuously distributed and that strain C3H/HeJ had the lowest mean systolic and diastolic pressure (100.5 +/- 3.2 and 66.8 +/- 3.5 mmHg), and a strain with obesity and diabetes, NZO/HILtJ, had the highest (132.4 +/- 3.1 and 86.6 +/- 6.9 mmHg). To understand the relationship of blood pressure with insulin resistance and obesity, we produced F1 and F2 progeny from reciprocal crosses of NZO, the strain with obesity, diabetes, and high blood pressure, and the strain with the lowest blood pressures, C3H/HeJ. Mean systolic pressures of 10-week-old (NZO x C3H)F1 and (C3H x NZO)F1 males were similar to each other (114.9 +/- 3.8 and 117.2 +/- 5.0 mmHg) and were intermediate to those of the parental strains. Systolic pressure of F2 males (n = 223) was distributed normally about the mean, suggesting that blood pressure is a polygenic trait. The body mass index (BMI) and plasma insulin levels of F2 progeny correlated significantly and positively with plasma leptin levels, suggesting that obesity is associated with insulin resistance. In contrast, systolic pressure did not correlate with BMI, plasma leptin levels, and plasma insulin levels, suggesting that genes underlying the development of hypertension in this intercross are not associated with the development of obesity and insulin resistance. Our results demonstrate that the progeny of NZO and C3H intercrosses are a practical and powerful tool for identifying blood pressure genes and for understanding human polygenic hypertension.
  • C Tsukahara, F Sugiyama, B Paigen, S Kunita, K Yagami MAMMALIAN GENOME 15 (12) 943 -950 2004年12月 [査読無し][通常論文]
     
    We characterized the systolic and diastolic blood pressures of 10-week-old males from 15 inbred mouse strains and found that blood pressures among strains were continuously distributed and that strain C3H/HeJ had the lowest mean systolic and diastolic pressure (100.5 +/- 3.2 and 66.8 +/- 3.5 mmHg), and a strain with obesity and diabetes, NZO/HILtJ, had the highest (132.4 +/- 3.1 and 86.6 +/- 6.9 mmHg). To understand the relationship of blood pressure with insulin resistance and obesity, we produced F-1 and F-2 progeny from reciprocal crosses of NZO, the strain with obesity, diabetes, and high blood pressure, and the strain with the lowest blood pressures, C3H/HeJ. Mean systolic pressures of 10-week-old (NZO x C3H)F-1 and (C3H x NZO)F-1 males were similar to each other (114.9 +/- 3.8 and 117.2 +/- 5.0 mmHg) and were intermediate to those of the parental strains. Systolic pressure of F-2 males n = 223 was distributed normally about the mean, suggesting that blood pressure is a polygenic trait. The body mass index (BMI) and plasma insulin levels of F-2 progeny correlated significantly and positively with plasma leptin levels, suggesting that obesity is associated with insulin resistance. In contrast, systolic pressure did not correlate with BMI, plasma leptin levels, and plasma insulin levels, suggesting that genes underlying the development of hypertension in this intercross are not associated with the development of obesity and insulin resistance. Our results demonstrate that the progeny of NZO and C3H intercrosses are a practical and powerful tool for identifying blood pressure genes and for understanding human polygenic hypertension.
  • 血液脳関門とRAS -その生理と病態-
    血液脳関門とRAS -その生理と病態- 1 (1) 35 41 2004年 [査読無し][通常論文]
  • 遺伝子改変マウスの位置づけ
    日本実験動物学会シンポジウム「医学・医療における実験動物の位置付けとその役割」 2004年 [査読無し][通常論文]
  • 伊関大敬, 国田智, 杉山文博, 八神健一 日本分子生物学会年会プログラム・講演要旨集 26th 573 2003年11月 [査読無し][通常論文]
  • M Masuzawa, T Fujimura, M Tsubokawa, S Nishiyama, K Katsuoka, E Terada, S Kunita, Y Sakurai, H Kato JOURNAL OF DERMATOLOGICAL SCIENCE 16 (2) 91 -98 1998年01月 [査読無し][通常論文]
     
    A cell line, designated ISOS-1, was established from a tumor formed by transplantation of a human angiosarcoma into mice with severe combined immunodeficiency (SCID). The cells showed endothelial properties, based on the uptake of Dil-Ac-LDL and binding of UEA-I/GSA-I lectins, but were negative for CD11b and Pan Cytokeratin. However, the cells lost differentiated characteristics such as expression of von Willebrand factor, contact inhibition growth and tube formation activity. These findings indicate that ISOS-1 is a poorly-differentiated endothelial cell line. At the 81st, passage, all of the cells were positive for H-2D(d) in various intensity, but not HLA-ABC. The metaphase chromosomes consistently showed a characteristic mouse, but not human, telocentric form. Furthermore, this cell line produced fatal tumor growth in SCID mice and also in BALB/c mice. These results suggest that ISOS-1 is a murine-phenotypic angiosarcoma cell line. (C) 1998 Elsevier Science Ireland Ltd.
  • M Masuzawa, T Fujimura, M Tsubokawa, S Nishiyama, K Katsuoka, E Terada, S Kunita, Y Sakurai, H Kato JOURNAL OF DERMATOLOGICAL SCIENCE 16 (2) 91 -98 1998年01月 [査読無し][通常論文]
     
    A cell line, designated ISOS-1, was established from a tumor formed by transplantation of a human angiosarcoma into mice with severe combined immunodeficiency (SCID). The cells showed endothelial properties, based on the uptake of Dil-Ac-LDL and binding of UEA-I/GSA-I lectins, but were negative for CD11b and Pan Cytokeratin. However, the cells lost differentiated characteristics such as expression of von Willebrand factor, contact inhibition growth and tube formation activity. These findings indicate that ISOS-1 is a poorly-differentiated endothelial cell line. At the 81st, passage, all of the cells were positive for H-2D(d) in various intensity, but not HLA-ABC. The metaphase chromosomes consistently showed a characteristic mouse, but not human, telocentric form. Furthermore, this cell line produced fatal tumor growth in SCID mice and also in BALB/c mice. These results suggest that ISOS-1 is a murine-phenotypic angiosarcoma cell line. (C) 1998 Elsevier Science Ireland Ltd.
  • K GOTO, R NOZU, S KUNITA, E TERADA, T ITOH EXPERIMENTAL ANIMALS 44 (2) 159 -161 1995年04月 [査読無し][通常論文]
     
    Restriction endonuclease analysis of amplified nucleocapsid protein genes from mouse hepatitis virus (MHV) was used to differentiate 12 strains isolated from mouse liver or transplantable tumors from five facilities, and the restriction patterns of the isolates were compared with those of five well-defined MHV strains, A59, JHM, 2, S and Nu-67. The patterns of 10 isolates from three facilities were the same as that of Nu-67. The remaining two isolates revealed different patterns from the five reference strains. This study showed that reverse transcription and the polymerase chain reaction assay based restriction analysis are feasible for the detection and genotyping of MHV, and the Nu-67 related strain was the most prevalent type found in the clinical samples.
  • K GOTO, R NOZU, S KUNITA, E TERADA, T ITOH EXPERIMENTAL ANIMALS 44 (2) 159 -161 1995年04月 [査読無し][通常論文]
     
    Restriction endonuclease analysis of amplified nucleocapsid protein genes from mouse hepatitis virus (MHV) was used to differentiate 12 strains isolated from mouse liver or transplantable tumors from five facilities, and the restriction patterns of the isolates were compared with those of five well-defined MHV strains, A59, JHM, 2, S and Nu-67. The patterns of 10 isolates from three facilities were the same as that of Nu-67. The remaining two isolates revealed different patterns from the five reference strains. This study showed that reverse transcription and the polymerase chain reaction assay based restriction analysis are feasible for the detection and genotyping of MHV, and the Nu-67 related strain was the most prevalent type found in the clinical samples.
  • S KUNITA, LN ZHANG, FR HOMBERGER, COMPTON, SR VIRUS RESEARCH 35 (3) 277 -289 1995年03月 [査読無し][通常論文]
     
    Enterotropic strains of murine coronaviruses (MHV-Y and MHV-RI) differ extensively in their pathogenesis from the prototypic respiratory strains of murine coronaviruses. In an effort to determine which viral proteins might be determinants of enterotropism, immunoblots of MHV-Y and MHV-RI virions using anti-S, -N and -M protein-specific antisera were performed. The uncleaved MHV-Y and MHV-RI S proteins migrated slightly faster than the MHV-A59 S protein. The MHV-Y S protein was inefficiently cleaved. The MHV-Y, MHV-RI and MHV-A59 N and M proteins showed only minor differences in their migration. The S genes of MHV-Y and MI-IV-RI were cloned, sequenced and found to encode 1361 and 1376 amino acid long proteins, respectively. The presence of several amino acids changes upstream from the predicted cleavage site of the MHV-Y S protein may contribute its inefficient cleavage. A high degree of homology was found between the MHV-RI and MHV-4 S proteins, whereas the homology between the MHV-Y S protein and the S proteins of other MHV strains was much lower. These results indicate that the enterotropism of MHV-RI and MHV-Y may be determined by different amino acid changes in the S protein and/or by changes in other viral proteins.
  • S KUNITA, LN ZHANG, FR HOMBERGER, COMPTON, SR VIRUS RESEARCH 35 (3) 277 -289 1995年03月 [査読無し][通常論文]
     
    Enterotropic strains of murine coronaviruses (MHV-Y and MHV-RI) differ extensively in their pathogenesis from the prototypic respiratory strains of murine coronaviruses. In an effort to determine which viral proteins might be determinants of enterotropism, immunoblots of MHV-Y and MHV-RI virions using anti-S, -N and -M protein-specific antisera were performed. The uncleaved MHV-Y and MHV-RI S proteins migrated slightly faster than the MHV-A59 S protein. The MHV-Y S protein was inefficiently cleaved. The MHV-Y, MHV-RI and MHV-A59 N and M proteins showed only minor differences in their migration. The S genes of MHV-Y and MI-IV-RI were cloned, sequenced and found to encode 1361 and 1376 amino acid long proteins, respectively. The presence of several amino acids changes upstream from the predicted cleavage site of the MHV-Y S protein may contribute its inefficient cleavage. A high degree of homology was found between the MHV-RI and MHV-4 S proteins, whereas the homology between the MHV-Y S protein and the S proteins of other MHV strains was much lower. These results indicate that the enterotropism of MHV-RI and MHV-Y may be determined by different amino acid changes in the S protein and/or by changes in other viral proteins.
  • S KUNITA, LN ZHANG, FR HOMBERGER, COMPTON, SR VIRUS RESEARCH 35 (3) 277 -289 1995年03月 [査読無し][通常論文]
     
    Enterotropic strains of murine coronaviruses (MHV-Y and MHV-RI) differ extensively in their pathogenesis from the prototypic respiratory strains of murine coronaviruses. In an effort to determine which viral proteins might be determinants of enterotropism, immunoblots of MHV-Y and MHV-RI virions using anti-S, -N and -M protein-specific antisera were performed. The uncleaved MHV-Y and MHV-RI S proteins migrated slightly faster than the MHV-A59 S protein. The MHV-Y S protein was inefficiently cleaved. The MHV-Y, MHV-RI and MHV-A59 N and M proteins showed only minor differences in their migration. The S genes of MHV-Y and MI-IV-RI were cloned, sequenced and found to encode 1361 and 1376 amino acid long proteins, respectively. The presence of several amino acids changes upstream from the predicted cleavage site of the MHV-Y S protein may contribute its inefficient cleavage. A high degree of homology was found between the MHV-RI and MHV-4 S proteins, whereas the homology between the MHV-Y S protein and the S proteins of other MHV strains was much lower. These results indicate that the enterotropism of MHV-RI and MHV-Y may be determined by different amino acid changes in the S protein and/or by changes in other viral proteins.
  • S KUNITA, LN ZHANG, FR HOMBERGER, COMPTON, SR VIRUS RESEARCH 35 (3) 277 -289 1995年03月 [査読無し][通常論文]
     
    Enterotropic strains of murine coronaviruses (MHV-Y and MHV-RI) differ extensively in their pathogenesis from the prototypic respiratory strains of murine coronaviruses. In an effort to determine which viral proteins might be determinants of enterotropism, immunoblots of MHV-Y and MHV-RI virions using anti-S, -N and -M protein-specific antisera were performed. The uncleaved MHV-Y and MHV-RI S proteins migrated slightly faster than the MHV-A59 S protein. The MHV-Y S protein was inefficiently cleaved. The MHV-Y, MHV-RI and MHV-A59 N and M proteins showed only minor differences in their migration. The S genes of MHV-Y and MI-IV-RI were cloned, sequenced and found to encode 1361 and 1376 amino acid long proteins, respectively. The presence of several amino acids changes upstream from the predicted cleavage site of the MHV-Y S protein may contribute its inefficient cleavage. A high degree of homology was found between the MHV-RI and MHV-4 S proteins, whereas the homology between the MHV-Y S protein and the S proteins of other MHV strains was much lower. These results indicate that the enterotropism of MHV-RI and MHV-Y may be determined by different amino acid changes in the S protein and/or by changes in other viral proteins.
  • マウス肝炎ウイルスの分子生物学
    アニテックス 7: 4-9 1995年 [査読無し][通常論文]
  • K GOTO, S KUNITA, E TERADA, T ITOH EXPERIMENTAL ANIMALS 43 (3) 413 -415 1994年07月 [査読無し][通常論文]
     
    The polymerase chain reaction (PCR) and culture methods were used to detect Mycoplasma pulmonis in nasal, tracheal and oral swab samples of rats derived from 5 mycoplasma-contaminated and 2 mycoplasma-free facilities, and the results of both methods were compared. Thirty-four/54 and 30/54 in nasal samples, 31/54 and 28/54 in tracheal samples, and 12/39 and 30/39 in oral samples were positive in PCR and cultures, respectively. Agreements in the results of both tests were 48/54 (88.9%) in nasal samples and 49/54 (90.7%) in tracheal samples. This indicates that nasal and tracheal samples are useful for detecting M. pulmonis by PCR. The detection number for M. pulmonis from the oral cavity by PCR was remarkably lower than those of nasal and tracheal sites. These results show that the PCR method has significant potential as a rapid and sensitive method for detecting M. pulmonis in clinical samples collected from the nasal cavity and trachea.
  • K GOTO, S KUNITA, E TERADA, T ITOH EXPERIMENTAL ANIMALS 43 (3) 413 -415 1994年07月 [査読無し][通常論文]
     
    The polymerase chain reaction (PCR) and culture methods were used to detect Mycoplasma pulmonis in nasal, tracheal and oral swab samples of rats derived from 5 mycoplasma-contaminated and 2 mycoplasma-free facilities, and the results of both methods were compared. Thirty-four/54 and 30/54 in nasal samples, 31/54 and 28/54 in tracheal samples, and 12/39 and 30/39 in oral samples were positive in PCR and cultures, respectively. Agreements in the results of both tests were 48/54 (88.9%) in nasal samples and 49/54 (90.7%) in tracheal samples. This indicates that nasal and tracheal samples are useful for detecting M. pulmonis by PCR. The detection number for M. pulmonis from the oral cavity by PCR was remarkably lower than those of nasal and tracheal sites. These results show that the PCR method has significant potential as a rapid and sensitive method for detecting M. pulmonis in clinical samples collected from the nasal cavity and trachea.
  • S KUNITA, M MORI, E TERADA VIROLOGY 193 (1) 520 -523 1993年03月 [査読無し][通常論文]
  • S KUNITA, M MORI, E TERADA VIROLOGY 193 (1) 520 -523 1993年03月 [査読無し][通常論文]
  • S KUNITA, E TERADA, K GOTO, N KAGIYAMA LABORATORY ANIMAL SCIENCE 42 (6) 593 -598 1992年12月 [査読無し][通常論文]
     
    Sequence analysis of the nucleocapsid protein genes of five strains of mouse hepatitis virus (MHV) disclosed that the 3' region of the nucleocapsid protein gene contains highly conserved sequences unique to MHV. We designed a pair of primers to amplify cDNA from such sequences of MHV by using the Polymerase chain reaction (PCR). Six isolates of wild-type MHV, as well as prototype viruses, were amplified successfully and detected in ethidium bromide-stained agarose gels. The sequence identity of PCR products was readily verified by confirming target size and a MflI site within the target. The sensitivity of our PCR assay was estimated to be sufficient to detect a single cell infected with MHV. This new approach may permit more sensitive and rapid detection of MHV in biologic materials than current methods such as virus isolation, the infant mouse bioassay, and the mouse antibody production test.
  • S KUNITA, E TERADA, K GOTO, N KAGIYAMA LABORATORY ANIMAL SCIENCE 42 (6) 593 -598 1992年12月 [査読無し][通常論文]
     
    Sequence analysis of the nucleocapsid protein genes of five strains of mouse hepatitis virus (MHV) disclosed that the 3' region of the nucleocapsid protein gene contains highly conserved sequences unique to MHV. We designed a pair of primers to amplify cDNA from such sequences of MHV by using the Polymerase chain reaction (PCR). Six isolates of wild-type MHV, as well as prototype viruses, were amplified successfully and detected in ethidium bromide-stained agarose gels. The sequence identity of PCR products was readily verified by confirming target size and a MflI site within the target. The sensitivity of our PCR assay was estimated to be sufficient to detect a single cell infected with MHV. This new approach may permit more sensitive and rapid detection of MHV in biologic materials than current methods such as virus isolation, the infant mouse bioassay, and the mouse antibody production test.
  • K GOTO, T ITOH, A TAKAKURA, S KUNITA, E TERADA, N KAGIYAMA EXPERIMENTAL ANIMALS 40 (2) 231 -233 1991年04月 [査読無し][通常論文]
     
    A total of 544 rabbit sera obtained from 6 commercial breeding facilities and 9 research institutions during 1985-1990 were tested for Bacillus piliformis antibody by an enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent antibody test (IFAT). The antibody was detected in 53 (14.2%) rabbits from 3 breeding facilities and 30 (17.4%) rabbits from 6 research institutions, indicating the prevalence of B. piliformis infection among laboratory rabbits in Japan. The overall agreement with ELISA for immune status was 96.9% (527/544) with IFAT. In tests of the ability of ELISA and IFAT to quantitate antibody, a correlation coefficient (r) of 0.86 (P < 0.01) was obtained by plotting the measured average log of the ELISA titer against the corresponding log of the IFAT titer.
  • K GOTO, T ITOH, A TAKAKURA, S KUNITA, E TERADA, N KAGIYAMA EXPERIMENTAL ANIMALS 40 (2) 231 -233 1991年04月 [査読無し][通常論文]
     
    A total of 544 rabbit sera obtained from 6 commercial breeding facilities and 9 research institutions during 1985-1990 were tested for Bacillus piliformis antibody by an enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent antibody test (IFAT). The antibody was detected in 53 (14.2%) rabbits from 3 breeding facilities and 30 (17.4%) rabbits from 6 research institutions, indicating the prevalence of B. piliformis infection among laboratory rabbits in Japan. The overall agreement with ELISA for immune status was 96.9% (527/544) with IFAT. In tests of the ability of ELISA and IFAT to quantitate antibody, a correlation coefficient (r) of 0.86 (P < 0.01) was obtained by plotting the measured average log of the ELISA titer against the corresponding log of the IFAT titer.
  • H KOYAMA, M KOZAKAI, S KUNITA, T HOHDATSU, T NASU, H SAITO JOURNAL OF VETERINARY MEDICINE SERIES B-ZENTRALBLATT FUR VETERINARMEDIZIN REIHE B-INFECTIOUS DISEASES AND VETERINARY PUBLIC HEALTH 37 (1) 9 -18 1990年02月 [査読無し][通常論文]
  • H KOYAMA, M KOZAKAI, S KUNITA, T HOHDATSU, T NASU, H SAITO JOURNAL OF VETERINARY MEDICINE SERIES B-ZENTRALBLATT FUR VETERINARMEDIZIN REIHE B-INFECTIOUS DISEASES AND VETERINARY PUBLIC HEALTH 37 (1) 9 -18 1990年02月 [査読無し][通常論文]
  • S KUNITA, E TERADA, K GOTO, N KAGIYAMA EXPERIMENTAL ANIMALS 39 (1) 103 -107 1990年01月 [査読無し][通常論文]
  • S KUNITA, E TERADA, K GOTO, N KAGIYAMA EXPERIMENTAL ANIMALS 39 (1) 103 -107 1990年01月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 201 -206 1989年07月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, Y SAKURAI, H SUZUKI, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 207 -213 1989年07月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, Y SAKURAI, H SUZUKI, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 215 -219 1989年07月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 201 -206 1989年07月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, Y SAKURAI, H SUZUKI, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 207 -213 1989年07月 [査読無し][通常論文]
  • S KUNITA, E TERADA, A GHODA, Y SAKURAI, H SUZUKI, A TAKAKURA, N KAGIYAMA EXPERIMENTAL ANIMALS 38 (3) 215 -219 1989年07月 [査読無し][通常論文]
  • N. Kobayasi, E. Terada, S. Kunita, Y. Sakurai, H. Suzuki, N. Kagiyama, A. Takakura, A. Ghoda Kitazato Archives of Experimental Medicine 62 (1) 45 -51 1989年 [査読無し][通常論文]
  • N. Kobayasi, E. Terada, S. Kunita, Y. Sakurai, H. Suzuki, N. Kagiyama, A. Takakura, A. Ghoda Kitazato Archives of Experimental Medicine 62 (1) 45 -51 1989年 [査読無し][通常論文]
  • Preparation and Characterization of Monoclonal Antibodies against Bovine B Lymphocyte Antigens
    Vet. Pathol. Immunopathol. 18: 201-212 1988年 [査読無し][通常論文]
  • Preparation and Characterization of Monoclonal Antibodies against Bovine B Lymphocyte Antigens
    Vet. Pathol. Immunopathol. 18: 201-212 1988年 [査読無し][通常論文]
  • H KOYAMA, S KUNITA, N UEHARA, H SAITO JAPANESE JOURNAL OF VETERINARY SCIENCE 49 (3) 433 -437 1987年06月 [査読無し][通常論文]
  • H KOYAMA, S KUNITA, N UEHARA, H SAITO JAPANESE JOURNAL OF VETERINARY SCIENCE 49 (3) 433 -437 1987年06月 [査読無し][通常論文]

共同研究・競争的資金等の研究課題

  • 実験動物の微生物学的品質管理法の研究
    遺伝子科学研究
    研究期間 : 2002年 -2006年
  • 感染症の疾患モデルマウスの開発・解析
    遺伝子科学研究
    研究期間 : 2002年 -2006年
  • Study on Microbiological Quality Control of Laboratory Animals
    Gene Science Research
    研究期間 : 2002年 -2006年
  • Study on Animal Models for Infectious Diseases
    Gene Science Research
    研究期間 : 2002年 -2006年

委員歴

  • 2012年 - 現在   日本獣医学会   評議員
  • 2010年 - 現在   日本実験動物学会   評議員   日本実験動物学会
  • 2014年 - 2019年   日本実験動物学会   理事


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