21 results on '"Takashi Kuramochi"'
Search Results
2. Establishment and characterization of CAG/EGFP transgenic rabbit line
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Takashi Kuramochi, Masatsugu Ueda, Ri Ichi Takahashi, Noriyuki Kasai, Yoji Hakamata, Kazuki Aoyagi, Ichiro Miyoshi, Shu Hashimoto, and Eiji Kobayashi
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Male ,Transgene ,Genetic Vectors ,Green Fluorescent Proteins ,Gene Expression ,Breeding ,Biology ,Regenerative medicine ,Fluorescence ,Green fluorescent protein ,Animals, Genetically Modified ,Gene expression ,Genetics ,medicine ,Animals ,Tissue Distribution ,Enhancer ,In Situ Hybridization, Fluorescence ,Expression vector ,medicine.diagnostic_test ,Molecular biology ,Recombinant Proteins ,Transgenesis ,Female ,Animal Science and Zoology ,Rabbits ,Genetic Engineering ,Agronomy and Crop Science ,Biotechnology ,Fluorescence in situ hybridization - Abstract
Cell marking is a very important procedure for identifying donor cells after cell and/or organ transplantation in vivo. Transgenic animals expressing marker proteins such as enhanced green fluorescent protein (EGFP) in their tissues are a powerful tool for research in fields of tissue engineering and regenerative medicine. The purpose of this study was to establish transgenic rabbit lines that ubiquitously express EGFP under the control of the cytomegalovirus immediate early enhancer/beta-actin promoter (CAG) to provide a fluorescent transgenic animal as a bioresource. We microinjected the EGFP expression vector into 945 rabbit eggs and 4 independent transgenic candidate pups were obtained. Two of them died before sexual maturation and one was infertile. One transgenic male candidate founder rabbit was obtained and could be bred by artificial insemination. The rabbit transmitted the transgene in a Mendelian manner. Using fluorescence in situ hybridization analysis, we detected the transgene at 7q11 on chromosome 7 as a large centromeric region in two F1 offspring (one female and one male). Eventually, one transgenic line was established. Ubiquitous EGFP fluorescence was confirmed in all examined organs. There were no gender-related differences in fluorescence. The established CAG/EGFP transgenic rabbit will be an important bioresource and a useful tool for various studies in tissue engineering and regenerative medicine.
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- 2006
3. A Testicular Germ Cell-Associated Serine-Threonine Kinase, MAK, Is Dispensable for Sperm Formation
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Tomoya Kato, Takashi Kuramochi, Mikiko Fukuda, Emiko Chiba, Yoshihiko Araki, Hideo Satoh, Yoichi Shinkai, and Naoki Takeda
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Male ,Protein Serine-Threonine Kinases ,Biology ,Mice ,Testis ,Gene expression ,Mammalian Genetic Models with Minimal or Complex Phenotypes ,Animals ,Spermatogenesis ,Protein kinase A ,Molecular Biology ,Crosses, Genetic ,Sperm motility ,Mice, Knockout ,Serine/threonine-specific protein kinase ,Genetics ,Cell Biology ,Spermatozoa ,Sperm ,In vitro ,Cell biology ,Fertility ,Sperm Motility ,Female ,Protein Kinases ,Function (biology) - Abstract
A member of the mitogen-activated protein kinase superfamily, MAK, has been proposed to have an important role in spermatogenesis, since Mak gene expression is highly restricted to testicular germ cells. To assess the biological function of MAK, we have established MAK-deficient (Mak(-/-)) mice. Mak(-/-) mice developed normally, and no gross abnormalities were observed. Spermatogenesis of the Mak(-/-) mice was also intact, and most of the mice were fertile. However, Mak(-/-) male-derived litter sizes and their sperm motility in vitro were mildly reduced. These data show that function of MAK is not essential for spermatogenesis and male fertility.
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- 2002
4. Prophylactic Effect of FK463, a Novel Antifungal Lipopeptide, against Pneumocystis carinii Infection in Mice
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Fumiaki Ikeda, Ryoko Nozu, Kyoji Hioki, Shuzo Suzuki, Mamoru Ito, Takashi Kuramochi, Toshio Itoh, and Natsuko Eguchi
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Pathology ,medicine.medical_specialty ,Antifungal Agents ,Lipoproteins ,Mice, SCID ,Biology ,Peptides, Cyclic ,Polymerase Chain Reaction ,Microbiology ,Echinocandins ,Immunocompromised Host ,Lipopeptides ,Mice ,chemistry.chemical_compound ,parasitic diseases ,medicine ,Pneumocystosis ,Animals ,Experimental Therapeutics ,Pharmacology (medical) ,Pharmacology ,Lung ,Pneumocystis ,Pneumonia, Pneumocystis ,Respiratory disease ,Lipopeptide ,Histology ,Antibiotic Prophylaxis ,medicine.disease ,respiratory tract diseases ,Disease Models, Animal ,Infectious Diseases ,medicine.anatomical_structure ,chemistry ,Pneumocystis carinii ,Micafungin ,Female ,Nasal administration ,Pentamidine ,medicine.drug - Abstract
The prophylactic effect of FK463, a new water-soluble echinocandin-like lipopeptide with inhibitory activity against 1,3-β- d -glucan synthase, against Pneumocystis carinii infection was investigated with the severe combined immunodeficient (SCID) mouse model. Treatment with FK463, pentamidine, and saline only was performed for 6 weeks from the day after the SCID mice were inoculated intranasally with infected lung homogenates. FK463 at 0.2 or 1.0 mg/kg of body weight, pentamidine at 4 mg/kg, or saline was subcutaneously administered daily into the backs of the SCID mice. The effects of the drugs were evaluated by detection of P. carinii cysts in mouse lung homogenates by toluidine blue O staining, lung histology, and PCR amplification of a P. carinii -specific DNA fragment from the lungs. P. carinii cysts were detected in the lungs of all mice administered saline. In contrast, no cysts were detected in mice administered both doses of FK463 and pentamidine. A specific DNA fragment was amplified from all mice administered saline and at least half or more of the mice administered FK463 and pentamidine. These results indicate that FK463 acts on cyst wall formation but not on trophozoite proliferation and is extremely effective in preventing P. carinii -associated pneumonia. These results suggest that FK463 is potentially useful as a prophylactic agent against P. carinii infection.
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- 2000
5. CD4+Cells Are Indispensable for Ulcer Development in Murine Cutaneous Leishmaniasis
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Takashi Kuramochi, Chizu Sanjoba, Takashi Onodera, Kwang Poo Chang, Masaki Terabe, Yoshitsugu Matsumoto, Mamoru Ito, and Toshimitsu Hatabu
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CD4-Positive T-Lymphocytes ,Male ,Pathology ,medicine.medical_specialty ,Immunology ,Population ,Leishmaniasis, Cutaneous ,Spleen ,Mice, SCID ,CD8-Positive T-Lymphocytes ,Biology ,Microbiology ,Mice ,Cutaneous leishmaniasis ,Skin Ulcer ,medicine ,Splenocyte ,Animals ,education ,Skin ,Mice, Inbred BALB C ,Severe combined immunodeficiency ,education.field_of_study ,Leishmaniasis ,medicine.disease ,Leishmania ,biology.organism_classification ,Leukocyte Transfusion ,Infectious Diseases ,medicine.anatomical_structure ,Parasitology ,Fungal and Parasitic Infections ,CD8 - Abstract
One of the most characteristic clinical features in cutaneous leishmaniasis is the development of nodules followed by ulcerations at the site of infection.Leishmania amazonensis-infected mice show similar ulcerative lesions.Leishmania-infected severe combined immunodeficiency (SCID) mice, however, have been shown to develop nonulcerative nodules. In the present study, the roles of T cells in ulceration were examined using SCID mice in cell reconstitution experiments. After development of nonulcerative nodules, SCID mice were inoculated with splenocytes from eitherLeishmania-infected or naive immunocompetent mice, resulting in ulceration in all mice. When naive splenocytes were depleted of CD4+, CD8+, or B220+cell populations and the remaining cells were injected intoLeishmania-infected SCID mice after the development of nodules, only SCID mice inoculated with splenocytes depleted of CD4+cells did not show ulceration. The evidence obtained in this study clearly shows that the CD4+cell population is indispensable for ulceration in leishmaniasis lesions of SCID mice.
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- 2000
6. Cytokine dependent growth of human TF-1 leukemic cell line in human GM-CSF and IL-3 producing transgenic SCID mice
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Kimio Kobayashi, Tatsuji Nomura, Takashi Kuramochi, Norikazu Tamaoki, Yumi Fukuchi, Mamoru Ito, Kazuo Shimamura, Yoshitaka Miyakawa, and Yoshito Ueyama
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Male ,Genetically modified mouse ,Cancer Research ,medicine.medical_treatment ,Transgene ,Transplantation, Heterologous ,Mice, Transgenic ,Mice, SCID ,Biology ,Mice ,In vivo ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Growth Substances ,Interleukin 3 ,Granulocyte-Macrophage Colony-Stimulating Factor ,Hematology ,Molecular biology ,Disease Models, Animal ,Haematopoiesis ,Cytokine ,Granulocyte macrophage colony-stimulating factor ,Oncology ,Cell culture ,Immunology ,Female ,Interleukin-3 ,Leukemia, Erythroblastic, Acute ,Neoplasm Transplantation ,medicine.drug - Abstract
Although severe combined immunodeficient (SCID) mice are considered useful as an animal model for human hematopoietic diseases, the complete reconstruction of human hematopoietic cells can not be established even in these mice. This appears to be because human cytokines, adhesion molecules and extracellular matrices which support differentiation and growth of human hematopoietic cells differ from those in animals. To improve this animal model, we attempted to produce transgenic (Tg) mice producing human interleukin 3 (hIL-3) and human granulocyte macrophage colony stimulating factor (hGM-CSF) with the homozygote of the scid gene. We established two Tg mouse lines, one releasing both 0.5-1 ng/ml of hIL-3 and 0.05-0.2 ng/ml of hGM-CSF in their sera and another releasing only high (2-10 ng/ml) levels of hGM-CSF. When human cytokine-dependent myeloid cell line, TF-1, was subcutaneously transplanted into these two Tg-SCID mouse lines, TF-1 could be successfully engrafted and grew in all lines of Tg-SCID mice but not in control mice. We also observed that TF-1 grows in GM-CSF Tg-SCID mice in a dose dependent manner in vivo and IL-3 shows an additive effect on its growth. These results indicated that these Tg-SCID mice were an useful in vivo model for investigating human leukemogenesis, especially the role of IL-3 and GM-CSF in leukemogenesis.
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- 1998
7. [Establishment and characterization of transgenic mice expressing human platelet glycoprotein Ib alpha]
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Tatsuji Nomura, Takashi Kuramochi, Tetsuya Kitaguchi, Yoshiki Hiraoka, Yoshitaka Miyakawa, Kimio Kobayashi, Mamoru Ito, Yoshito Ueyama, Yasuo Ikeda, Sadakazu Aiso, Makoto Handa, Mitsuru Murata, and Ken Hikichi
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Genetically modified mouse ,Receptor complex ,Platelet Aggregation ,Transgene ,Blotting, Western ,Molecular Sequence Data ,Biophysics ,Gene Expression ,Mice, Transgenic ,Biology ,Platelet membrane glycoprotein ,Polymerase Chain Reaction ,Biochemistry ,Mice ,chemistry.chemical_compound ,von Willebrand Factor ,Animals ,Humans ,Platelet ,Ristocetin ,Molecular Biology ,Base Sequence ,Platelet Glycoprotein GPIb-IX Complex ,Cell Biology ,Flow Cytometry ,Molecular biology ,Blotting, Southern ,chemistry ,Platelet aggregation inhibitor ,Megakaryocytes ,Platelet Aggregation Inhibitors - Abstract
The platelet glycoprotein (GP) Ib/IX/V is a hetero-oligomeric receptor complex for von Willebrand factor (vWF) and mediates platelet adhesion and aggregation under high shear stress conditions. It is composed of alpha and beta chain of GP Ib, GP IX, AND and GP V. To establish transgenic mice carrying human GP Ib alpha, we injected into mouse zygotes a 6 kb DNA fragment containing human GP Ib alpha gene that included entire coding sequence and putative promoter region. One hundred and thirteen offsprings were screened, and only one was found to express human GP Ib alpha protein and has passed the human GP Ib alpha gene as well as the expression of the gene to next generation. The expression of human GP Ib alpha in transgenic mice was limited to platelets and megakaryocytes. Glycocalicin, a proteolytic fragment of human GP Ib alpha found in normal human plasma, was not detected in transgenic mouse plasma. Human vWF in the presence of ristocetin supported agglutination of transgenic mouse platelets, but not of control mouse platelets.
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- 1997
8. Pneumocystis carinii Cysts are Susceptible to Inactivation by Chemical Disinfectants
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Mamoru Ito, Takashi Kuramochi, and Kyoji Hioki
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Hypochlorous acid ,Disinfectant ,Sodium chlorite ,Mice, SCID ,Biology ,Polymerase Chain Reaction ,General Biochemistry, Genetics and Molecular Biology ,Microbiology ,Mice ,chemistry.chemical_compound ,parasitic diseases ,Animals ,DNA, Fungal ,Hydrogen peroxide ,Lung ,DNA Primers ,General Veterinary ,Pneumocystis ,Pneumonia, Pneumocystis ,General Medicine ,Ribosomal RNA ,Iodoform ,respiratory tract diseases ,Staining ,Disinfection ,chemistry ,Pneumocystis carinii ,Female ,Severe Combined Immunodeficiency ,Animal Science and Zoology ,Disinfectants - Abstract
The inactivation efficacy of eight disinfectants commonly used in laboratories and animal rooms to inactivate Pneumocystis carinii cysts was estimated by experimental infection in C.B-17-scid mice. The disinfectants examined in this study were 70% ethyl alcohol, 10% iodoform, 0.5% hypochlorous acid, two 1% quanternary ammonium salts, 3% hydrogen peroxide, sodium chlorite and 1% cresol soap. The lung homogenates from P. carinii infected C.B-17-scid mice were treated with each disinfectant for 15 min at room temperature, washed with saline, and inoculated into C.B-17-scid mice. Eight weeks after inoculation, lungs from these mice were examined by staining with toluidine blue O to detect P. carinii cysts. PCR amplifying 346 bp of P. carinii specific mitochondrial ribosomal RNA large segments was also performed using DNA extracted from the lungs of the mice. As a result, seven disinfectants, excepting for 0.5% hypochlorous acid, were effective in the inactivation of P. carinii cysts. These results suggest that P. carinii cysts were sensitive to chemical disinfectants even though they have been commonly considered as insensitive.
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- 1997
9. Refined Porcine Follicle Stimulating Hormone Promotes the Responsiveness of Rabbits to Multiple-Ovulation Treatment
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Mikako Kamei, Kazuo Hirasawa, Kazuki Aoyagi, Kensaku Kitada, Masatsugu Ueda, Takashi Kuramochi, Masao Hirao, Shu Hashimoto, and Ri-ichi Takahashi
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Ovulation ,medicine.medical_specialty ,General Veterinary ,media_common.quotation_subject ,Superovulation ,General Medicine ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Human chorionic gonadotropin ,Follicle-stimulating hormone ,Endocrinology ,Pregnancy ,Internal medicine ,medicine ,Animals ,Female ,Animal Science and Zoology ,Rabbits ,Follicle Stimulating Hormone ,media_common - Abstract
We investigated whether refined follicle stimulating hormone (FSH) with only a little contaminating LH can promote the responsiveness of rabbits to multiple-ovulation treatment. One group of female rabbits was stimulated with refined porcine FSH (pFSH), an FSH source with low LH activity, and another group was treated with pFSH. The mean number of eggs recovered from donors stimulated with refined pFSH (27 +/- 3) was significantly greater (P0.05) than that with pFSH (20 +/- 2). Furthermore, the mean number of remaining follicles of donors stimulated with refined pFSH (19 +/- 4) was significantly greater (P0.05) than that with pFSH (12 +/- 1). To decrease the number of remaining follicles in donors treated with refined pFSH, the dose of human chorionic gonadotropin (hCG) was increased from 75 to 150. However, there were no differences in the numbers of eggs and remaining follicles. The results of the present study suggest that refined pFSH with little contaminating LH promotes the responsiveness of rabbits to multiple-ovulation treatment compared with pFSH.
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- 2004
10. Responsiveness of rabbits to superovulation treatment by a single injection of follicle-stimulating hormone with aluminum hydroxide gel
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Koji Kimura, Makoto Hirako, Kazuo Hirasawa, Masatsugu Ueda, Kazuki Aoyagi, Kensaku Kitada, Takashi Kuramochi, Hisataka Iwata, Mamoru Kawaguchi, Masao Hirao, and Shu Hashimoto
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endocrine system ,medicine.medical_specialty ,medicine.medical_treatment ,Aluminum Hydroxide ,Superovulation ,Biology ,Animals, Genetically Modified ,Follicle-stimulating hormone ,Ovulation Induction ,Internal medicine ,Genetics ,medicine ,Animals ,Saline ,Embryo ,Cell Biology ,Single injection ,Endocrinology ,Aluminum hydroxide gel ,Female ,Rabbits ,Follicle Stimulating Hormone ,Adjuvant ,Blood stream ,Gels ,hormones, hormone substitutes, and hormone antagonists ,Developmental Biology ,Hormone - Abstract
Aluminum hydroxide gel (Al-gel), which is used as an adjuvant, can absorb macromolecules. We investigated the applicability of Al-gel to the sustained release of follicle-stimulating hormone (FSH) as a simplified method of superovulation (SOV) in rabbits. The responsiveness of rabbits to SOV by a single injection of FSH dissolved in Al-gel suspension (3.2 mg Al/ml) and in 10% (w/v) polyvinylpyrrolidone (PVP), and by multiple injections of FSH in saline was examined. The numbers of total and fertilized eggs recovered from rabbits treated with FSH in Al-gel (40.5 and 26.3, respectively) were similar to multiple injections (47.4 and 28.6, respectively) and were significantly greater (P
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- 2007
11. Small eye phenotypes observed in a human tau gene transgenic rat
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Masahiko Yasuda, Takashi Kuramochi, Noriyuki Azuma, Mamoru Ito, Ayako Sugawara, Toshio Itoh, and Kazuo Goto
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Male ,genetic structures ,Transgene ,tau Proteins ,In situ hybridization ,Biology ,Extraocular muscles ,Microphthalmia ,Polymerase Chain Reaction ,Animals, Genetically Modified ,Rats, Sprague-Dawley ,Cellular and Molecular Neuroscience ,Ciliary body ,Complementary DNA ,Lens, Crystalline ,medicine ,Animals ,Microphthalmos ,Transgenes ,Microinjection ,In Situ Hybridization, Fluorescence ,Chromosome Mapping ,medicine.disease ,Molecular biology ,Chromosomes, Mammalian ,eye diseases ,Sensory Systems ,Epithelium ,Rats ,Ophthalmology ,medicine.anatomical_structure ,Phenotype ,Female ,sense organs - Abstract
We developed a rat line showing small eye from transgenic rats that were obtained by microinjection of a DNA segment containing the human (h)tau cDNA (GenBank: BC000558: 31-677,774-1180) expressed under control of CAG promoter, which is related to Alzheimer disease, into the pronuclei rat embryos. The rat line was established by selective brother-sister mating of rats showing small eyes. Of 11 offspring in the 11th generation, there were eight animals with microphthalmia and the transgene. The remaining three rats without transgene did not show the small eyes phenotype. The globes of affected rats were 1.2 mm in length compared with normal globes (3.5 mm), and all other ocular structures were normal. The expression of hTau protein was evident immunohistochemically in the ciliary body, extraocular muscle, lens epithelium, and pigment epithelium. Cytogenetic analysis suggested that the chromosome location of the transgene was chromosome 1 (1p12). This region may include genes related to lens development, such as Cat5.
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- 2006
12. Serological Studies on Porcine Pneumocystis carinii Pneumonia: Kinetics of the Antibody Titers in Swine Herds and the Association of Porcine Reproductive and Respiratory Syndrome Virus Infection
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Masaji Taguchi, Takashi Kuramochi, Hiroshi Kondo, and Mamoru Ito
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Aging ,Swine ,animal diseases ,Porcine Reproductive and Respiratory Syndrome ,Antibodies, Viral ,Serology ,Orthomyxoviridae Infections ,medicine ,Animals ,Porcine respiratory and reproductive syndrome virus ,Fluorescent Antibody Technique, Indirect ,Antibodies, Fungal ,Swine Diseases ,Pseudorabies ,General Veterinary ,biology ,Pneumocystis ,Pneumonia, Pneumocystis ,Antibody titer ,Porcine reproductive and respiratory syndrome virus ,biology.organism_classification ,medicine.disease ,Herpesvirus 1, Suid ,Virology ,Titer ,Pneumonia ,Pneumocystis carinii ,Influenza A virus ,Immunology ,biology.protein ,Herd ,Female ,Antibody - Abstract
Serological titers to Pneumocystis carinii (Pc) and porcine reproductive and respiratory syndrome virus (PRRSV) were measured on a herd with epidemic Pc pneumonia (case herd) and two comparison herds, by an indirect fluorescent-antibody technique. In the case herd, the geometric mean titer (GMT) for Pc were 1:80 in pigs 1 week old, 1:10 in pigs 5 weeks old, and 1:80 to 1:190 in pigs over 6 weeks old. GMTs for PRRSV were >1:145 in most of age groups over 7 weeks old. In comparison herds, Pc and PRRSV antibody titers were low in weanling pigs. The results clarified the kinetics of antibodies to Pc and concurrent infection of PRRSV in the case herd.
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- 1997
13. Catalase in manipulation buffer enhances the developmental competence of DNA-injected embryos
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Masatsugu Ueda, Kazuki Aoyagi, Shu Hashimoto, Takashi Kuramochi, and Ri-ichi Takahashi
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Male ,Genotype ,Ontogeny ,Transgene ,Green Fluorescent Proteins ,Green fluorescent protein ,Embryo Culture Techniques ,Mice ,Animals ,Transgenes ,Microinjection ,chemistry.chemical_classification ,Cell Nucleus ,Reactive oxygen species ,biology ,Gene Transfer Techniques ,Embryo ,DNA ,Catalase ,Embryo, Mammalian ,Molecular biology ,Spermatozoa ,Transgenesis ,Mice, Inbred C57BL ,Blastocyst ,chemistry ,Genetic Techniques ,embryonic structures ,biology.protein ,Animal Science and Zoology ,Female ,Reactive Oxygen Species - Abstract
To improve the efficiency of transgenesis, we investigated the effects of a radical scavenger during microinjection on the development to blastocysts or pups of mouse pronuclear embryos, microinjected with the enhanced green fluorescent protein (EGFP) transgene. When embryos were microinjected in medium containing 0-1,000 units/ml catalase, the developmental rate to blastocysts was significantly higher (P
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- 2005
14. Effect of a null mutation of the oviduct-specific glycoprotein gene on mouse fertilization
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Yutaka Sendai, Takashi Kuramochi, Mamoru Ito, Hiromi Yoshida-Komiya, Hiroyoshi Hoshi, Yoshihiko Araki, Yoichi Shinkai, and Makoto Nohara
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Male ,Fertilization in Vitro ,Biology ,Transfection ,Biochemistry ,Mice ,Human fertilization ,In vivo ,OVGP1 ,medicine ,Animals ,Gene Silencing ,Molecular Biology ,Glycoproteins ,chemistry.chemical_classification ,Genetics ,Mice, Knockout ,Spermatozoon ,Stem Cells ,Cell Biology ,Null allele ,In vitro ,Cell biology ,medicine.anatomical_structure ,chemistry ,Fertilization ,Mutation ,Oviduct ,Female ,Glycoprotein ,Research Article - Abstract
The mammalian fertilization process takes place in a complex microenvironment within the female genital tract. A member of the chitinase protein family, oviduct-specific glycoprotein (OGP), has been identified in oviductal fluid from various mammalian species, including humans. Although OGP is widely believed to be involved in the process of mammalian fertilization, including spermatozoon function and gamete interactions, based on experimental results obtained in vitro, its physiological significance remains controversial. The present study established OGP gene-null (ogp−/−) mice, and primarily characterized their reproductive properties to study the physiological function(s) of OGP. Results obtained from studies using an in vivo or in vitro system showed that the fertility of ogp−/− females was within normal limits. These results indicate that OGP is not essential for the process of in vivo fertilization, at least in mice.
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- 2003
15. Successful cryopreservation of mouse ovaries by vitrification
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Fujio, Migishima, Rika, Suzuki-Migishima, Si-Young, Song, Takashi, Kuramochi, Sadahiro, Azuma, Masahiro, Nishijima, and Minesuke, Yokoyama
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Cryopreservation ,Male ,Mice, Inbred ICR ,Histocytochemistry ,Green Fluorescent Proteins ,Ovary ,Mice, Transgenic ,Fertilization in Vitro ,Embryo Transfer ,Specific Pathogen-Free Organisms ,Mice, Inbred C57BL ,Luminescent Proteins ,Mice ,Random Allocation ,Cryoprotective Agents ,Animals, Newborn ,Microscopy, Fluorescence ,Pregnancy ,Propylene Glycols ,Acetamides ,Animals ,Dimethyl Sulfoxide ,Female - Abstract
We developed a new method of cryopreservation of whole ovaries by vitrification using DAP213 (2 M dimethyl sulfoxide, 1 M acetamide, and M propylene glycol) as a cryoprotectant. Four-week-old C57BL/6 mice that underwent partial ovariectomy were orthotopically transplanted with cryopreserved or fresh ovaries (experimental or control group) isolated from 10-day-old green fluorescent protein (GFP)-transgenic mice (+/+). GFP-positive pups were similarly obtained from both groups by natural mating or in vitro fertilization (IVF) followed by embryo transfer, indicating that the cryopreserved ovaries by vitrification retain their fecundity. However, a statistically significant difference (P0.05) was found between both groups with respect to the following parameters: the number of GFP-positive pups born by natural mating/grafted ovary (0.8 +/- 0.3 for the experimental group versus 2.0 +/- 0.7 for the control group, mean +/- SEM), the number of collected oocytes by superovulation per mouse (7.0 +/- 1.7 for the experimental group versus 22.7 +/- 3.2 for the control group), the percentage of two-cell embryos obtained from GFP-positive oocytes by IVF (38.5% for the experimental group versus 90.0% for the control group). Histologically, normal development of follicles and formation of corpora lutea were observed in frozen-thawed grafts. However, estimated number of follicles decreased in frozen-thawed ovaries compared with fresh ovaries. Taken together, cryopreservation of the ovary by vitrification seems a promising method to preserve ovarian function, but further studies are required to overcome the possible inhibitory effects of this method on the growth of the ovarian graft.
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- 2003
16. Effects of cryopreservation of mouse embryos and in vitro fertilization on genotypic frequencies in colonies
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Kazuo Goto, Takashi Kuramochi, Toshio Itoh, Nobuhiro Shimozawa, Kaori Muguruma, Kyoji Hioki, and Michi Ebukuro
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Genetic Markers ,Male ,Genotype ,Ratón ,medicine.medical_treatment ,Fertilization in Vitro ,Biology ,Cryopreservation ,Mice ,Genetic drift ,Gene Frequency ,Pregnancy ,Genetics ,medicine ,Animals ,Allele ,Mice, Inbred ICR ,In vitro fertilisation ,Chi-Square Distribution ,Polymorphism, Genetic ,Embryo ,Cell Biology ,Microsatellite ,Female ,Developmental Biology ,Microsatellite Repeats - Abstract
To evaluate the effects of cryopreservation and in vitro fertilization (IVF) on genotypic frequencies in mouse colonies, genotypic frequencies at 15 biochemical, 4 immunological and 20 microsatellite loci were examined in three colonies of MCH (ICR) mice derived from noncryopreserved embryos obtained by natural mating without the induction of superovulation, cryopreserved embryos obtained by natural mating with the induction of superovulation, and cryopreserved embryos obtained by the induction of superovulation and IVF. Three (Pgm-1, Ldr-1 and Hbb) out of the 15 biochemical loci, two (Thy-1 and H2K) out of four immunological loci and five (D5Mit18, D6Mit15, D12Mit5, D13Mit26, and D14Mit7) out of 20 microsatellite loci that showed polymorphisms in every colony were used for detection of genotypic frequencies. The genotypic frequencies of the loci in the three colonies did not differ from the predicted genotypic frequencies (P > 0.05). The results suggested that genetic drift does not occur among colonies established from treated and untreated embryos, and it was clear that the embryo banking by cryopreservation is suitable for preservation of outbred stock without genetic drift.
- Published
- 2002
17. Lack of B cell leakiness in BALB/cA-nu, scid double mutant mice
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Yoshito Ueyama, Takashi Kuramochi, Sachio Endoh, Ehji Terada, Mamoru Ito, and Kyoji Hioki
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Lymphoma ,Lymphocyte ,medicine.medical_treatment ,T-Lymphocytes ,Longevity ,Immunoglobulins ,Mice, Nude ,Enzyme-Linked Immunosorbent Assay ,Mice, SCID ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Flow cytometry ,Interferon-gamma ,Mice ,medicine ,Animals ,Lymphocyte Count ,RNA, Messenger ,B cell ,Thymic Lymphoma ,B-Lymphocytes ,Mice, Inbred BALB C ,General Veterinary ,medicine.diagnostic_test ,Interleukin-6 ,Reverse Transcriptase Polymerase Chain Reaction ,RNA ,General Medicine ,Thymus Neoplasms ,medicine.disease ,Flow Cytometry ,Virology ,Molecular biology ,Phenotype ,medicine.anatomical_structure ,Cytokine ,Interleukin-2 ,Animal Science and Zoology ,Interleukin-4 - Abstract
BALB/cA mice homozygous for both nu and scid mutations (BALB/cA-nu/nu, scid/scid) were developed by mating between BALB/cA-scid and BALB/cA-nu. These mice have greater longevity than C.B-17-scid because no thymic lymphoma occurs in them unlike in the latter. C.B-17-scid is known to show the leaky phenomenon in which a few clones of functional T and B cells develop in aged C.B-17-scid. Unexpectedly, the leaky B cells and T cells were absent or suppressed in BALB/cA-nu, scid mice when cytokine expressions were determined by RT-PCR, lymphocyte phenotypes by flow cytometry and serum immunoglobulin levels by ELISA. These results indicate that B cell leakiness may be induced by leaked T cells. BALB/cA-nu, scid mice may be useful as a recipient in allo- and xeno-transplantation experiments because of the absence of both thymic lymphomas and leakiness, in addition to lack of hair.
- Published
- 2001
18. Non-ulcerative cutaneous lesion in immunodeficient mice with Leishmania amazonensis infection
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Ken Katakura, Shin-ichiro Kawazu, Takashi Onodera, Masaki Terabe, Yoshihito Ueyama, Takashi Kuramochi, Yoshitsugu Matsumoto, Mamoru Ito, and Toshimitsu Hatabu
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Pathology ,medicine.medical_specialty ,Leishmaniasis, Cutaneous ,Mice, SCID ,Mice ,Immune system ,Cutaneous leishmaniasis ,Skin Ulcer ,medicine ,Animals ,Skin ,Leishmania ,Mice, Knockout ,Leishmania amazonensis ,Mice, Inbred BALB C ,biology ,Inoculation ,Cutaneous lesion ,Leishmaniasis ,Skin ulcer ,biology.organism_classification ,medicine.disease ,DNA-Binding Proteins ,Disease Models, Animal ,Infectious Diseases ,Immunology ,Parasitology ,medicine.symptom - Abstract
Cutaneous leishmaniasis begins as papules or nodules at the site of promastigote inoculation. The next key pathogenic event in this disease is the formation of an ulcer at this site. Leishmania infection in immunodeficient mice, however, showed non-ulcerative cutaneous lesions suggesting the involvement of the immune system in ulcer formation. Severe combined immunodeficient (SCID), recombination-activating gene 2 knockout (RAG-2-/-), and immunocompetent mice were inoculated subcutaneously with cultured L. amazonensis promastigotes. Macroscopic nodules appeared at the inoculation site within 2 weeks of infection in all the mice and gradually extended to the surrounding skin tissue. Although nodules of immunocompetent mice ulcerated within 6 weeks, immunodeficient mice did not form ulcers even after 25 weeks of inoculation. These results strongly suggest the importance of functional T and B cells in ulcer formation of cutaneous leishmaniasis and are consistent with clinical features of non-ulcerative cutaneous leishmaniasis in some AIDS patients. The present study also indicates that the L. amazonensis-infected immunodeficient mouse model might be suitable for studying the mechanisms of ulcer formation in cutaneous leishmaniasis.
- Published
- 2001
19. Human acute myeloblastic leukemia-ascites model using the human GM-CSF- and IL-3-releasing transgenic SCID mice
- Author
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Mamoru Ito, Yumi Fukuchi, Takashi Kuramochi, Norikazu Tamaoki, Jun-ichi Hata, Kimio Kobayashi, Masahiro Kizaki, Tatsuji Nomura, Kazuo Shimamura, Akihiro Umezawa, Yasuo Ikeda, Yoshitaka Miyakawa, and Yoshito Ueyama
- Subjects
Pathology ,medicine.medical_specialty ,Myeloid ,Acute myeloblastic leukemia ,Mice, Transgenic ,Mice, SCID ,Biology ,Mice ,medicine ,Tumor Cells, Cultured ,Animals ,Humans ,Interleukin 3 ,Ascites ,Granulocyte-Macrophage Colony-Stimulating Factor ,Hematology ,General Medicine ,Aminolevulinic Acid ,medicine.disease ,Molecular biology ,Haematopoiesis ,Leukemia ,Disease Models, Animal ,Leukemia, Myeloid, Acute ,Granulocyte macrophage colony-stimulating factor ,medicine.anatomical_structure ,Cell culture ,Interleukin-3 ,medicine.drug ,Differentiation Inducer - Abstract
To generate an appropriate model for human acute myeloblastic leukemia (AML), we have successfully established a human hematopoietic growth factor-dependent AML cell line (TF-1 and UT-7/GM)-ascites model using human granulocyte-macrophage colony-stimulating factor (hGM-CSF)- and human interleukin 3 (hIL-3)-releasing transgenic (Tg)-SCID mice. When 1 x 10(7) cells of TF-1, a human erythroleukemia cell line, were transplanted into the peritoneum of irradiated Tg-SCID mice (TF-1 ip/Tg-SCID mice), TF-1 cells grew in both the single cell suspension form (asTF-1) and solid form in ascites and invaded various tissues: lungs, liver, pancreas, and genitals, 3-6 weeks following transplantation. Subsequently, 0.5-1 x 10(7) cells of UT-7/GM, a subline of the UT-7 human megakaryoblastic leukemia cell line, grown in the back of hGM-CSF Tg-SCID mice after subcutaneous inoculation, were transplanted into the peritoneum of other irradiated hGM-CSF Tg-SCID mice. After 4 weeks, UT-7/GM cells (asUT-7/GM) also grew in the same manner as TF-1 cells in hGM-CSF Tg-SCID mice. Analysis of the cells from the peritoneum and tissues by PCR amplifying ALU and human GM-CSF receptor beta sequences and by immunohistochemical staining using anti-human CD45 revealed that they possessed the original characteristics of the parental cells. To confirm the usefulness of this human AML-ascites model, experimental treatment of AML cells grown in these mice was carried out with a differentiation inducer, delta-aminolevulinic acid (deltaALA), which induces hemoglobin synthesis for TF-1 in vitro and is thus regarded as an anti-leukemia drug candidate. Unexpectedly, growth promotion of TF-1 cells was observed in the treated TF-1 ip/hIL-3 Tg-SCID mice without differentiation to erythroid cells after treatment with delta-ALA (5 mM) for 7 days. These results indicate that Tg-SCID mice can support the growth of human hematopoietic growth factor-dependent AML cell lines which are usually rejected by SCID mice, without modification of the parental cell characteristics. In addition, this Tg-SCID leukemia-ascites model may become a useful preclinical tool for estimation of drug efficacy in vivo, since the drug candidate which was promising in vitro did not act in the same manner in vivo.
- Published
- 1999
20. Mutant presenilin 2 transgenic mouse: effect on an age-dependent increase of amyloid beta-protein 42 in the brain
- Author
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Jochen Walter, Fumitaka Oyama, Maho Morishima-Kawashima, Kimio Kobayashi, Naoya Sawamura, Anja Capell, Takaomi C. Saido, Christian Haass, Yasuo Ihara, Taisuke Tomita, Mamoru Ito, Jürgen Grünberg, Takeshi Iwatsubo, Yoshito Ueyama, Takashi Kuramochi, and Kei Maruyama
- Subjects
Genetically modified mouse ,Aging ,Amyloid ,Transgene ,Mutant ,Gene Expression ,Mice, Transgenic ,Biology ,medicine.disease_cause ,Cell Fractionation ,Biochemistry ,Presenilin ,Cellular and Molecular Neuroscience ,Mice ,Alzheimer Disease ,Gene expression ,Presenilin-2 ,medicine ,Missense mutation ,Animals ,Humans ,RNA, Messenger ,skin and connective tissue diseases ,Aged ,Brain Chemistry ,Mutation ,Amyloid beta-Peptides ,Membrane Proteins ,Molecular biology ,Peptide Fragments ,Immunology ,hormones, hormone substitutes, and hormone antagonists - Abstract
The N141I missense mutation in presenilin (PS) 2 is tightly linked with a form of autosomal dominant familial Alzheimer's disease (AD) in the Volga German families. We have generated transgenic mouse lines overexpressing human wild-type or mutant PS2 under transcriptional control of the chicken beta-actin promoter. In the brains of transgenic mice, the levels of human PS2 mRNA were found to be five- to 15-fold higher than that of endogenous mouse PS2 mRNA. The amyloid beta-protein (Abeta) 42 levels in the brains of mutant PS2 transgenic mice were higher than those in wild-type PS2 transgenic mice at the age of 2, 5, or 8 months. In addition, the Abeta42 levels appeared to increase steadily in the mutant PS2 transgenic mouse brains from 2 to 8 months of age, whereas there was only a small increase in wild-type transgenic mice between the ages of 5 and 8 months. There was no definite difference in the levels of N-terminal and C-terminal fragments between wild-type and mutant PS2 transgenic mice at the age of 2, 5, or 8 months. These data show a definite effect of the PS2 mutation on an age-dependent increase of Abeta42 content in the brain.
- Published
- 1998
21. Establishment of human granulocyte-macrophage colony stimulating factor producing transgenic SCID mice
- Author
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Yoshitaka Miyakawa, Tatsutoshi Nakahata, Kimio Kobayashi, Takashi Kuramochi, Yoshito Ueyama, Toshiyuki Tanaka, Yasuo Ikeda, Yumi Fukuchi, Yutaka Takebe, Mamoru Ito, Kazuo Shimamura, Tatsuji Nomura, Norikazu Tamaoki, and Masayuki Miyasaka
- Subjects
Pathology ,medicine.medical_specialty ,Transgene ,Spleen ,Enzyme-Linked Immunosorbent Assay ,Mice, SCID ,Biology ,Kidney ,Mice ,Bone Marrow ,medicine ,Animals ,Peripheral blood cell ,Lung ,Leukemia, Experimental ,Myocardium ,Granulocyte-Macrophage Colony-Stimulating Factor ,Hematology ,medicine.disease ,Colony-stimulating factor ,Molecular biology ,Transplantation ,Leukemia ,Disease Models, Animal ,medicine.anatomical_structure ,Granulocyte macrophage colony-stimulating factor ,Liver ,Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ,Bone marrow ,Neoplasm Transplantation ,medicine.drug - Abstract
Previous work has shown the usefulness of severe combined immunodeficient (SCID) mice as in vivo models for the growth of normal human haemopoietic cells and leukaemic cells. Many approaches have been made to improve the engraftment of human haemopoietic cells in SCID mice. We established transgenic mice producing human granulocyte-macrophage colony stimulating factor (hGM-CSF) with the homozygote of the scid gene. Endogenous serum hGM-CSF levels were detected by ELISA [mean 9585 pg/ml (line A, n = 4); mean 1610 pg/ml (line B, n = 4)]. Expression of hGM-CSF was observed in all organs tested including the heart, lung, liver, kidney, spleen, thymus, bone marrow and brain of hGM-CSF transgenic (hGMTg) mice. Morphological analysis of organs and peripheral blood cell counts showed no differences between hGMTg mice and their littermates. Murine Ba/F3 cells expressing functional hGM-CSF alpha beta receptor (BAF/alpha beta cells) could be successfully engrafted in hGMTg SCID mice. The cells invaded multiple organs and caused death within a few weeks of transplantation, although they infiltrated only the spleen of their littermates. These results showed that these hGM-CSF-producing SCID mice are useful as an in vivo assay system for investigating leukaemogenesis.
- Published
- 1996
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