Updated on 2023/11/10

写真a

 
Asano Atsushi
 
Organization
Research Field in Veterinary Medicine, Agriculture, Fisheries and Veterinary Medicine Area Joint faculty of Veterinary Medicine Joint Department of Veterinary Medicine Professor
Title
Professor
Profile
Analysis of gene regulation in spermatogenesis Analysis of functions of interferon-inducible genes in host defense Development of serological diagnosis of viral and bacterial infection in laboratory animals
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Degree

  • Ph.D. ( 1999.3   Hokkaido University )

  • Doctor of Veterinary Medicine ( 1995.3   Hokkaido University )

Research Interests

  • spermatogenesis

  • host defense

  • interferon-inducible genes

Research Areas

  • Life Science / Veterinary medical science

Education

  • Hokkaido University   Graduate School of Veterinary Medicine

    1995.4 - 1999.3

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    Country: Japan

  • Hokkaido University   School of Veterinary Medicine

    1989.4 - 1995.3

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    Country: Japan

Research History

  • Kagoshima University   Research Field in Veterinary Medicine, Agriculture, Fisheries and Veterinary Medicine Area Joint faculty of Veterinary Medicine Department of Veterinary Medicine   Professor

    2015.9

  • Tottori University   Faculty of Agriculture, School of Veterinary Medicine   Associate Professor

    2007.4 - 2015.9

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    Country:Japan

  • Tottori University   Faculty of Agriculture, School of Veterinary Medicine   Associate Professor (as old post name)

    2006.10 - 2007.3

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    Country:Japan

  • Hokkaido University   Research Assistant

    1999.6 - 2006.9

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    Country:Japan

  • 日本学術振興会特別研究員

    1998 - 1999

Professional Memberships

  • 九州実験動物研究会

    2015.11

  • Japanese Association for Laboratory Animal Medicine

    2015.11

  • Japanese College of Laboratory Animal Medicine

    2015.11

  • 日本実験動物学会

    2015.10

  • 日本獣医学会

    2015.10

Committee Memberships

  • 日本実験動物医学会   理事  

    2023.10   

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    Committee type:Academic society

  • Japanese College of Laboratory Animal Medicine   President  

    2022.5   

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    Committee type:Academic society

  • 日本実験動物学会   評議委員  

    2022.4   

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    Committee type:Academic society

  • 日本実験動物医学専門医協会   理事  

    2021.10   

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    Committee type:Academic society

  • AAALAC International   ad hoc Specialist  

    2021.9   

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    Committee type:Other

  • 日本実験動物学会   動物実験外部検証事業 外部検証専門員  

    2021.4   

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    Committee type:Academic society

  • 九州実験動物研究会   理事  

    2019.12   

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    Committee type:Academic society

  • 九州実験動物研究会   評議員  

    2017.1   

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    Committee type:Academic society

  • 日本実験動物医学会   実験動物学教育委員  

    2016.9   

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    Committee type:Academic society

  • 日本実験動物学会   評議員  

    2016.6 - 2017.5   

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    Committee type:Other

  • 日本実験動物学会   編集委員  

    2010.6 - 2018.5   

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    Committee type:Other

  • 日本獣医学会   評議委員  

    2010.4   

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    Committee type:Other

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Qualification acquired

  • Diplomate of Japanese College of Laboratory Animal Medicine (DJCLAM)

  • Veterinarian

 

Papers

  • Uno Y, Morikuni S, Shiraishi M, Asano A, Murayama N, Yamazaki H .  Novel Cytochrome P450 2C94 Functionally Metabolizes Diclofenac and Omeprazole in Dogs. .  Drug metabolism and disposition: the biological fate of chemicals51 ( 5 ) 637 - 644   2023.5

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    Language:English   Publisher:Drug metabolism and disposition: the biological fate of chemicals  

    Cytochromes P450 (P450s or CYPs) are important drug-metabolizing enzymes. Because dogs are frequently used in drug metabolism studies, knowledge of dog CYP2C enzymes is essential because in humans these enzymes are abundant and play major roles in liver and intestine. The present study identified and characterized novel dog CYP2C94 along with previously identified dog CYP2C21 and CYP2C41. Dog CYP2C21, CYP2C41, and CYP2C94 cDNAs, respectively, contained open reading frames of 490, 489, and 496 amino acids and shared high-sequence identities (70%, 75%, and 58%) with human CYP2Cs. Dog CYP2C94 mRNA was preferentially expressed in liver, just as dog CYP2C21 and CYP2C41 mRNAs were. In dog liver, CYP2C21 mRNA was the most abundant, followed by CYP2C94 and CYP2C41 mRNAs. Moreover, the hepatic expressions of all three dog CYP2C mRNAs varied in four individual dogs, two of which did not express CYP2C41 mRNA. The three dog CYP2C genes had similar gene structures, and CYP2C94, although located on the same chromosome, was in a genomic region far from the gene cluster containing CYP2C21 and CYP2C41 Metabolic assays with recombinant proteins showed that dog CYP2C94, along with CYP2C21 and CYP2C41, efficiently catalyzed oxidations of diclofenac, warfarin, and/or omeprazole, indicating that dog CYP2C94 is a functional enzyme. Novel dog CYP2C94 is expressed abundantly in liver and encodes a functional enzyme that metabolizes human CYP2C substrates; it is, therefore, likely responsible for drug clearances in dogs. SIGNIFICANCE STATEMENT: Novel dog cytochrome P450 2C94 (CYP2C94) was identified and characterized along with dog CYP2C21 and CYP2C41. Dog CYP2C94, isolated from liver, had 58% sequence identity and a close phylogenetic relationship with its human homologs and was expressed in liver at the mRNA level. Dog CYP2C94 (and CYP2C21 and CYP2C41) catalyzed oxidations of diclofenac and omeprazole, human CYP2C9 and CYP2C19 substrates, respectively, but CYP2C41 also hydroxylated warfarin. CYP2C94 is therefore a functional drug-metabolizing enzyme likely responsible for drug clearances in dogs.

    DOI: 10.1124/dmd.122.001236

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  • A comprehensive analysis of six forms of cytochrome P450 2C (CYP2C) in pigs. .      1 - 10   2022.11A comprehensive analysis of six forms of cytochrome P450 2C (CYP2C) in pigs.Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1080/00498254.2022.2148139

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  • Uno Y, Uehara S, Ijiri M, Kawaguchi H, Asano A, Shiraishi M, Banju K, Murayama N, Yamazaki H .  Molecular and Functional Characterization of <i>N</i>-Acetyltransferases in Common Marmosets and Pigs. .    50 ( 11 ) 1429 - 1433   2022.6Reviewed

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    DOI: 10.1124/dmd.122.000919

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  • Uno Y, Murayama N, Ijiri M, Kawaguchi H, Yamato O, Shiraishi M, Asano A, Teraoka H, Mizukawa H, Nakayama SMM, Ikenaka Y, Ishizuka M, Yamazaki H .  Cytochrome P450 2J Genes Are Expressed in Dogs, Cats, and Pigs, and Encode Functional Drug-Metabolizing Enzymes. .  Drug metabolism and disposition50 ( 11 ) 1434 - 1441   2022.6Reviewed

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    DOI: 10.1124/dmd.122.000930

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  • Nakamura,Y., Asano, A.,, Hosaka, Y.,Takeuchi, T.,Iwanaga, T., Yamano, Y. .  Expression and intracellular localization of TBC1D9, a Rab GTPase-accelerating protein, in mouse testes. .  Exp. Anim.64   415 - 424   2015.4Expression and intracellular localization of TBC1D9, a Rab GTPase-accelerating protein, in mouse testes.Reviewed

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1538/expanim.15-0016

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  • Taiki Moriya, Rina Shibasaki, Tomohiko Kayano, Nami Takebuchi, Momoko Ichimura, Naoki Kitamura, Atsushi Asano, Yoshinao Z Hosaka, Oksana Forostyak, Alexei Verkhratsky, Govindan Dayanithi, Izumi Shibuya .  Full-length transient receptor potential vanilloid 1 channels mediate calcium signals and possibly contribute to osmoreception in vasopressin neurones in the rat supraoptic nucleus. .  Cell calcium57 ( 1 ) 25 - 37   2015.1Full-length transient receptor potential vanilloid 1 channels mediate calcium signals and possibly contribute to osmoreception in vasopressin neurones in the rat supraoptic nucleus.Reviewed International journal

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    Neurones in the supraoptic nucleus (SON) of the hypothalamus possess intrinsic osmosensing mechanisms, which are lost in transient receptor potential vanilloid 1 (Trpv1)-knock-out mice. The molecular nature of the osmosensory mechanism in SON neurones is believed to be associated with the N-terminal splice variant of Trpv1, although their entire molecular structures have not been hitherto identified. In this study, we sought for TRPV1-related molecules and their function in the rat SON. We performed RT-PCR and immunohistochemistry to detect TRPV1-related molecules in the SON, and patch-clamp and imaging of the cytosolic Ca(2+) concentration ([Ca(2+)]i) to measure responses to osmolality changes and TRPV-related drugs in acutely dissociated SON neurones of rats. RT-PCR analysis revealed full-length Trpv1 and a new N-terminal splice variant, Trpv1_SON (LC008303) in the SON. Positive immunostaining was observed using an antibody against the N-terminal portion of TRPV1 in arginine vasopressin (AVP)-immunoreactive neurones, but not in oxytocin (OT)-immunoreactive neurones. Approximately 20% of SON neurones responded to mannitol (50 mM) with increased action potential firing, inward currents, and [Ca(2+)]i mobilization. Mannitol-induced responses were observed in AVP neurones isolated from AVP-eGFP transgenic rats and identified by GFP fluorescence, but not in OT neurones isolated from OT-mRFP transgenic rats and identified by RFP fluorescence. The mannitol-induced [Ca(2+)]i responses were reversibly blocked by the non-selective TRPV antagonist, ruthenium red (10 μM) and the TRPV1 antagonists, capsazepine (10 μM) and BCTC (10 μM). Although the TRPV1 agonist, capsaicin (100 nM) evoked no response at room temperature, it triggered cationic currents and [Ca(2+)]i elevation when the temperature was increased to 36°C. These results suggest that AVP neurones in the rat SON possess functional full-length TRPV1. Moreover, differences between the responses to capsaicin or hyperosmolality obtained in rat SON neurones and those obtained from dorsal root ganglion neurones or TRPV1-expressing cells indicate that the osmoreceptor expressed in the SON may be a heteromultimer in which TRPV1 is co-assembled with some other, yet unidentified, molecules.

    DOI: 10.1016/j.ceca.2014.11.003

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  • Kasem Rattanapinyopituk, Akinori Shimada, Takehito Morita, Masashi Sakurai, Atsushi Asano, Tatsuya Hasegawa, Kenichiro Inoue, Hirohisa Takano .  Demonstration of the clathrin- and caveolin-mediated endocytosis at the maternal-fetal barrier in mouse placenta after intravenous administration of gold nanoparticles. .  The Journal of veterinary medical science76 ( 3 ) 377 - 87   2014.3Demonstration of the clathrin- and caveolin-mediated endocytosis at the maternal-fetal barrier in mouse placenta after intravenous administration of gold nanoparticles.Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC VET SCI  

    Exposure to nanoparticles during pregnancy is a public concern, because nanoparticles may pass from the mother to the fetus across the placenta. The purpose of this study was to determine the possible translocation pathway of gold nanoparticles across the maternal-fetal barrier as well as the toxicity of intravenously administered gold nanoparticles to the placenta and fetus. Pregnant ICR mice were intravenously injected with 0.01% of 20- and 50-nm gold nanoparticle solutions on the 16th and 17th days of gestation. There was no sign of toxic damage to the placentas as well as maternal and fetal organs of the mice treated with 20- and 50-nm gold nanoparticles. ICP-MS analysis demonstrated significant amounts of gold deposited in the maternal livers and placentas, but no detectable level of gold in the fetal organs. However, electron microscopy demonstrated an increase of endocytic vesicles in the cytoplasm of syncytiotrophoblasts and fetal endothelial cells in the maternal-fetal barrier of mice treated with gold nanoparticles. Clathrin immunohistochemistry and immunoblotting showed increased immunoreactivity of clathrin protein in the placental tissues of mice treated with 20- and 50-nm gold nanoparticles; clathrin immunopositivity was observed in syncytiotrophoblasts and fetal endothelial cells. In contrast, caveolin-1 immunopositivity was observed exclusively in the fetal endothelium. These findings suggested that intravenous administration of gold nanoparticles may upregulate clathrin- and caveolin-mediated endocytosis at the maternal-fetal barrier in mouse placenta.

    DOI: 10.1292/jvms.13-0512

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  • Atsushi Asano, Daisuke Torigoe, Nobuya Sasaki, Takashi Agui .  Development of an ELISA Using a Recombinant P46-Like Lipoprotein for Diagnosis of Mycoplasma pulmonis Infection in Rodents .  JOURNAL OF VETERINARY MEDICAL SCIENCE76 ( 2 ) 151 - 157   2014.2Development of an ELISA Using a Recombinant P46-Like Lipoprotein for Diagnosis of Mycoplasma pulmonis Infection in RodentsReviewed

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    Mycoplasma pulmonis is one of the most prevalent bacterial pathogens that infects laboratory mice and rats. To develop an M. pulmonis-specific antigen for serological diagnosis, we cloned the cDNA of P46-like lipoprotein (P46L), an M. pulmonis putative periplasmic protein. P46L is a homolog of P46, an M. hyopneumoniae antigen. We produced recombinant P46L fused to glutathione S-transferase (GST) in Escherichia coli. Immunoblot analysis revealed that sera from Mycoplasma-infected mice and rats contained anti-P46L antibodies. We developed an ELISA using the recombinant P46L-GST protein as an antigen. Thirteen of the 14 samples from rats naturally infected with M. pulmonis were determined to be positive according to the commercial ELISA (MONILISA Myco) and positive by our ELISA. Furthermore, 18/19 samples from mice experimentally infected with M. pulmonis were positive using our P46L-GST ELISA. In contrast, only 8/19 samples from infected mice were positive by the commercial ELISA. Our results indicate that P46L-GST was an appropriate antigen for developing a serological test to determine M. pulmonis infection in laboratory mice and rats.

    DOI: 10.1292/jvms.13-0308

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  • Tomonori Ichihashi, Atsushi Asano, Tatsufumi Usui, Takashi Takeuchi, Yasuko Watanabe, Yoshiaki Yamano .  Antiviral and antiproliferative effects of canine interferon-λ1. .  Veterinary immunology and immunopathology156 ( 1-2 ) 141 - 6   2013.11Antiviral and antiproliferative effects of canine interferon-λ1.Reviewed International journal

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Interferon (IFN)-λs, members of the type III IFN group, were recently identified in several vertebrates. Although IFN-λs have the potential to be utilized as antiviral and antitumor agents in veterinary medicine, the biological properties of IFN-λs have not yet been studied in companion animals. In this study, we analyzed the expression of canine IFN-λs and their receptors, produced a recombinant canine IFN-λ1 protein, and investigated its antiviral and antiproliferative activities using a canine kidney epithelial cell line, MDCK cells. MDCK cells were found to express type III IFN molecules, IFN-λ1 and IFN-λ3, and the receptors, IFNλR1 and IL10R2. IFN-λ1 was induced faster than IFN-λ3 by stimulation with poly (I:C). His-tagged IFN-λ1 protein expressed in Escherichia coli inhibited cytolytic plaque formation by influenza A virus infection, and induced the expression of interferon-stimulated genes, Mx1 and OAS1, in MDCK cells. In addition, recombinant IFN-λ1 inhibited the proliferation of MDCK cells slightly. These effects were observed in a dose-dependent manner. These results indicate that canine IFN-λ1 has antiviral effect, and suggest the potential applicability of canine IFN-λ1 as a therapeutic agent.

    DOI: 10.1016/j.vetimm.2013.09.013

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  • Ahmed Magzoub Khalid, Atsushi Asano, Yoshinao Z. Hosaka, Takashi Takeuchi, Yoshiaki Yamano .  Tumor Suppressor Candidate TUSC3 Expression during Rat Testis Maturation .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY77 ( 10 ) 2019 - 2024   2013.10Tumor Suppressor Candidate TUSC3 Expression during Rat Testis MaturationReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    Analysis of microarray data obtained by comparing gene expression between 2-week-old infant and 7-week-old mature SD rat testes revealed novel targets involved in tumor suppression. Reverse-transcription polymerase chain reaction and Northern blotting indicated that Tusc3 gene expression was upregulated in the normal maturing testis and prostate and other organs such as the cerebrum and ovary. Tumor suppressor candidate 3 protein expression was detected in these same organs at a size of about 40 kDa, in accord with the predicted molecular size. In situ hybridization and immunohistochemistry showed that mRNA and protein localization were prevalent in the testis spermatocytes and interstitial cells such as the Leydig cells, as well as prostate epithelial cells. These data suggest that TUSC3 is deeply involved in spermatogenesis in the testis, inducing sperm differentiation and maturation, and plays a role in normal prostate development and tumor suppression.

    DOI: 10.1271/bbb.130327

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  • Atsushi Asano, Daisuke Torigoe, Nobuya Sasaki, Takashi Agui .  Epitope Mapping of the Nucleocapsid Protein of Sendai Virus and Application of Antigenic Epitopes for the ELISA-Based Diagnosis of Sendai Virus Infection .  JOURNAL OF VETERINARY MEDICAL SCIENCE75 ( 7 ) 909 - 916   2013.7Epitope Mapping of the Nucleocapsid Protein of Sendai Virus and Application of Antigenic Epitopes for the ELISA-Based Diagnosis of Sendai Virus InfectionReviewed

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    Sendai virus (SeV) is one of the most prevalent viral pathogens infecting laboratory mice and rats. To date, mature SeV virions have been used as antigens for serological diagnosis. To develop antigens that are more specific and easier to prepare for diagnosis, we examined the antigenic sites in the nucleocapsid protein (NP) of SeV with antisera from experimentally SeV-infected mice and a peptide array membrane containing overlapping 10-mer peptides covering the entire NP. We found antigenic linear sequences in two regions, amino acids 120-160 and 420-500, of the SeV-NP. From these antigenic sequences, we applied two synthesized peptides, IVKTRDMEYERTTEWL and FVTLHGAERLEEETNDE, which correspond to positions 119-134 and 458-474 of the SeV-NP, respectively, as antigens in an enzyme-linked immunosorbent assay (ELISA). Evaluation of the ELISAs using these peptides revealed that they were specific to anti-SeV antisera. Furthermore, the ELISAs using these peptides were able to distinguish between SeV-positive and SeV-negative mouse sera to the same extent as a commercial ELISA kit. These results indicate that these peptides are useful for the serological diagnosis of SeV infection.

    DOI: 10.1292/jvms.12-0496

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  • Takashi Takeuchi, Yusuke Sugimoto, Atsushi Asano, Tetsuya Shimokawa, Motowo Nabeta, Tatsufumi Usui .  Gene Transcriptions of Toll-Like Receptors in the Mouse Uterus during Gestation .  JOURNAL OF VETERINARY MEDICAL SCIENCE75 ( 5 ) 547 - 551   2013.5Gene Transcriptions of Toll-Like Receptors in the Mouse Uterus during GestationReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC VET SCI  

    Toll-like receptors (TLRs) play critical roles in innate immunity by recognizing a broad range of microbial components as ligands. The activation of TLRs is an important step not only for the innate immune response, but also for the development of the subsequent antigen-specific adaptive immune response. However, little is known about TLR expression in the female genital mucosa during gestation. In the present study, gene transcriptions of TLRs 1 to 9 were investigated in both the mesometrial side and the anti-mesometrial side of the uterus during gestation in the mouse reproductive organ during the gestation period. In the mesometrial side, gene transcriptions of TLR 1, 3,4 and 9 were decreased in the late gestation period, whereas an increase of gene transcriptions of TLR 4 and 9 was seen in the early gestation period. In the anti-mesometrial side, gene transcriptions of TLR 1 and 9 were also decreased in the late gestation period, and TLR 9 gene transcription was increased in the early gestation period. On the other hand, gene transcriptions of TLR 3 and 4 were not changed in the late gestation period, but they were increased in the early gestation period. Gene transcriptions of TLR 2, 5, 6, 7 and 8 were not changed statistically in either side during the gestation period. These results suggest that the expressions of particular TLRs may be regulated in the uterus during the gestation period to maintain the pregnant state.

    DOI: 10.1292/jvms.12-0457

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  • Takashi Takeuchi, Masahiro Yoshida, Takuro Shimizu, Atsushi Asano, Tetsuya Shimokawa, Motowo Nabeta, Tatsufumi Usui .  Differential expressions of toll-like receptor genes in the vagina of pregnant mice .  Journal of Veterinary Medical Science75 ( 5 ) 561 - 565   2013Differential expressions of toll-like receptor genes in the vagina of pregnant miceReviewed

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    The mammalian immune system is classified into two categories, innate and adaptive immunity, and innate immunity is an immunological first line of defense for the mucosal immune system. Toll-like receptors (TLRs) play critical roles in innate immunity, as they recognize specific molecular patterns found in microbial pathogens, and the activation of TLRs is an important step not only for the innate immune response, but also for the development of the subsequent antigen-specific adaptive immune response. Despite the importance of TLRs in mucosal immunity, little is known about their expression in the female genital mucosa during gestation. In the present study, gene expressions of TLRs 1 to 9 were investigated together with NF-κB and FoxP3 gene expressions in the vaginae of pregnant mice to understand the immune response of the female genital mucosa during pregnancy. We found that mRNA expressions of TLR4, TLR5, TLR6, TLR7 and TLR9 were significantly decreased during the late gestation period, whereas temporary increases were seen in the middle gestation period. Gene transcriptions of TLR1, TLR2, TLR3 and TLR8 were not changed specifically during the gestation period. The mRNA expression of NF-κB was not changed at any time during the gestation period, while the FoxP3 mRNA expression was increased in the middle gestation period. These results suggest that expressions of particular TLRs would be down-regulated during gestation so as to maintain the pregnant state. © 2013 The Japanese Society of Veterinary Science.

    DOI: 10.1292/jvms.12-0458

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  • Yasumitsu Masuda, Akiko Matsuda, Tatsufumi Usui, Toru Sugai, Atsushi Asano, Yoshiaki Yamano .  Biological Effects of Chicken Type III Interferon on Expression of Interferon-Stimulated Genes in Chickens: Comparison with Type I and Type II Interferons .  JOURNAL OF VETERINARY MEDICAL SCIENCE74 ( 11 ) 1381 - 1386   2012.11Biological Effects of Chicken Type III Interferon on Expression of Interferon-Stimulated Genes in Chickens: Comparison with Type I and Type II InterferonsReviewed

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    Interferons (IFNs) are key mediators that activate host defense mechanisms against viruses. The recently identified mammalian Type III IFN has biological effects similar to type I IFN. However, the biological effects of type III IFN have not yet been characterized in birds. We compared the effects of chicken type III IFN (IFN-lambda) with type I (IFN-beta) and type II (IFN-gamma) IFNs on IFN-stimulated genes (ISGs) using recombinant proteins expressed in Escherichia coli. Recombinant chicken IFN-lambda inhibited influenza virus replication and induced the mRNA expression of the ISGs, Mx and OAS, in chicken embryonic fibroblasts (CEFs) in a dose-dependent manner. However, the effective dose of IFN-lambda was higher than that of IFN-beta and IFN-gamma. Furthermore, the effect of IFN-lambda on induction of Mx and OAS was lesser than that of IFN-beta, but comparable to that of IFN-gamma. These results indicate that chicken IFN-lambda has the potential to induce ISGs and inhibit viral replication in chicken cells.

    DOI: 10.1292/jvms.11-0517

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  • Taiki Moriya, Tomohiko Kayano, Naoki Kitamura, Yoshinao Z. Hosaka, Atsushi Asano, Oksana Forostyak, Alexei Verkhratsky, Cedric Viero, Govindan Dayanithi, Emil C. Toescu, Izumi Shibuya .  Vasopressin-induced intracellular Ca2+ concentration responses in non-neuronal cells of the rat dorsal root ganglion .  BRAIN RESEARCH1483   1 - 12   2012.11Vasopressin-induced intracellular Ca2+ concentration responses in non-neuronal cells of the rat dorsal root ganglionReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Arginine-vasopressin (AVP) is a nonapeptide of hypothalamic origin that has been shown to exert many important cognitive and physiological functions in neurons and terminals of both the central and peripheral nervous system (CNS and PNS). Here we report for the first time that AVP induced an increase in intracellular Ca2+ concentration ([Ca2+](i)) in non-neuronal cells isolated from the rat dorsal root ganglion (DRG) and cultured in vitro. The ratiometric [Ca2+](i) measurements showed that AVP evoked [Ca2+](i) responses in the non-neuronal cells and these concentration-dependent (100 pM to 1 mu M) responses increased with days in vitro in culture, reaching a maximum amplitude after 4-5 day. Immunostaining by anti-S-100 antibody revealed that more than 70% of S-100 positive cells were AVP-responsive, indicating that glial cells responded to AVP and increased their [Ca2+](i). The responses were inhibited by depletion of the intracellular Ca2+ stores or in the presence of inhibitors of phospholipase C, indicating a metabotropic response involving inositol trisphosphate, and were mediated by the Vi subclass of AVP receptors, as evidenced by the use of the specific blockers for V-1 and OT receptors, (d(CH2)(5)(1),Tyr(Me)(2),Arg(8))-Vasopressin and (d(CH2)(5)(1), Tyr(Me)(2),Thr(4),Orn(8),des-Gly-NH29)-Vasotocin, respectively. V-1a but not V-1b receptor mRNA was expressed sustainably through the culture period in cultured DRG cells. These results suggest that AVP modulates the activity of DRG glial cells via activation of V-1a receptor. (C) 2012 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.brainres.2012.08.028

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  • Ahmed Magzoub Khalid, Atsushi Asano, Yoshinao Z. Hosaka, Masanori Ohta, Kenji Ohyama, Yoshiaki Yamano .  Rat Stem-Cell Leukemia Gene Expression Increased during Testis Maturation .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY76 ( 11 ) 2118 - 2123   2012.11Rat Stem-Cell Leukemia Gene Expression Increased during Testis MaturationReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    We found that stem-cell leukemia (SCL), also known as T cell acute-lymphocytic leukemia (Tal-1) gene expression, was upregulated in the maturing rat testis. Strong expression of Tal-1 was detected in the normal maturing rat testis by Northern blotting. Western blotting revealed the protein size to be about 34 kDa. Protein expression was wide-spread in spermatocytes, spermtids and spermatogonia in accordance with the seminiferous epithelium cycle, as determined by an analysis of immunohistochemistry. Gene expression of Tal-1 regulatory gene, NKX3.1, was negatively correlated with Tal-1 expression. Human Tal-1 expression in the maturing testis as well as in bone marrow was observed, which suggests that the gene product is a novel cancer-testis antigen candidate. Taken together, TAL-1 may be involved in cell division, morphological changes, and the development of spermatogenic cells in the normal rat testis.

    DOI: 10.1271/bbb.120503

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  • Ahmed Magzoub Khalid, Atsushi Asano, Yoshinao Z. Hosaka, Kenji Ohyama, Masanori Ohta, Yoshiaki Yamano .  Expression of a Novel Sphingosine 1-Phosphate Receptor Motif Protein Gene in Maturing Rat Testes .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY76 ( 9 ) 1769 - 1773   2012.9Expression of a Novel Sphingosine 1-Phosphate Receptor Motif Protein Gene in Maturing Rat TestesReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    We screened a novel sphingosine 1-phosphate receptor type 5 motif-containing gene, LOC290876, from maturing rat testes by differential display. Gene expression was testis-specific, increased at week 7, and continued for 15 weeks. PCR analysis clarified two gene transcript isoforms, which were expressed at the same level in all samples detected in Northern blot. The deduced amino acid sequences of the two isoforms revealed differences in carboxyl terminal sequences. Gene and protein expression in the testes was dominant in the spermatocytes, and protein expression was localized to the nucleus. Taken together, these findings suggest that the LOC290876-encoded gene product is not involved in sphingosine signaling, but has distinct roles in the nucleus during the processes of spermatocyte maturation and meiosis producing spermatids.

    DOI: 10.1271/bbb.120379

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  • Masanori Ohta, Kenji Ohyama, Atsushi Asano, Shin-Ichi Yokota, Ahmed Magzoub Khalid, Yoshiaki Yamano .  Regulation of Rat Tetratricopeptide Repeat Domain 29 Gene Expression by Follicle-Stimulating Hormone .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY76 ( 8 ) 1540 - 1543   2012.8Regulation of Rat Tetratricopeptide Repeat Domain 29 Gene Expression by Follicle-Stimulating HormoneReviewed

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    We screened the gene that encodes tetratricopeptide repeat domain 29 (Ttc29) in the maturing rat testis. Gene expression was determined by Northern blotting of 7-week-old rat testes, and a strong signal was detected close to the 18S rRNA band in addition to two weak high-molecular-weight signals. In situ hybridization revealed that Ttc29 was expressed primarily in the spermatocytes. We evaluated the effect of gonadotropin on Ttc29 expression using hypophysectomized rats. The pituitary was removed from 3-week-old rats, gonadotropin was injected at 5 weeks, and Ttc29 expression was determined at 7 weeks. Although testicular development and hyperplasia of interstitial cells were observed following chorionic gonadotropin treatment after hypophysectomy, Ttc29 expression was upregulated by treatment with follicle-stimulating hormone. Ttc29 encodes axonemal dynein, a component of sperm flagella. Taken together, these data indicate that axonemal dynein expression starts in the spermatocytes and is regulated by follicle-stimulating hormone.

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  • Youhei Yasunaga, Takashi Takeuchi, Tetsuya Shimokawa, Motowo Nabeta, Aya Matsuu, Atsushi Asano, Yasuhiko Ohta .  Sugar Expressions on the Vaginal Epithelium in Pregnant Mice .  JOURNAL OF VETERINARY MEDICAL SCIENCE74 ( 6 ) 805 - 808   2012.6Sugar Expressions on the Vaginal Epithelium in Pregnant MiceReviewed

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    Sugar expressions were examined on the epithelium of both the middle portion of the vagina and the vaginal portion of the cervical canal (CC) in pregnant mice to understand the pathogenesis of bacterial infection in the female reproductive organ by using a panel of lectins. As a result, N-acetylglucosamine was positive before pregnant day (P) 7 but negative after P10 and at diestrus on both the vagina and the CC. In addition, some differences in sugar expressions were seen between them. These results suggest that sugar expressions on the mucosal surface would change not only site-specifically but also time-dependently, and these sugar differences indicate the possibility of the alteration of the settled bacterial species on the vaginal mucosa in pregnancy.

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  • Atsushi Asano, Daisuke Torigoe, Nobuya Sasaki, Takashi Agui .  Identification of antigenic peptides derived from B-cell epitopes of nucleocapsid protein of mouse hepatitis virus for serological diagnosis .  JOURNAL OF VIROLOGICAL METHODS177 ( 1 ) 107 - 111   2011.10Identification of antigenic peptides derived from B-cell epitopes of nucleocapsid protein of mouse hepatitis virus for serological diagnosisReviewed

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    Mouse hepatitis virus (MHV) infection is found commonly in laboratory mice and this virus has been known to cause various diseases such as subclinical infection, enteritis, hepatitis, and encephalitis. Serological tests are used commonly to diagnose MHV infection. Complete MHV virions have been used primarily as antigens for serological diagnosis to date. To develop an antigen that is more specific, more sensitive, and easier to prepare for serological diagnosis, the antigenic sites in the MHV-nucleocapsid (N) protein were screened in this study. Sixteen antigenic linear sequences in the N protein were found using antisera obtained from mice infected naturally with MHV and a peptide array containing overlapping 10-mer peptides covering the entire N protein. From these antigenic sequences, two synthesized peptides, ILKKTTWADQTERGL and RFDSTLPGFETIMKVL, which were consistent with positions 24-38 and 357-372 of the N protein respectively, were used as antigens in ELISA. Evaluation of ELISA with these peptides revealed that both peptides were specific to anti-MHV antisera. Furthermore, ELISA performed using these peptides was more sensitive than commercial ELISA used for a screening sera from mice infected accidentally to MHV maintained in cages, suggesting that these peptides are useful for serological diagnosis of MHV infection. (C) 2011 Elsevier B.V. All rights reserved.

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  • Yumi Kimoto, Akihiko Sugiyama, Masaaki Nishinohara, Atsushi Asano, Aino Masuda, Tairin Ochi, Takashi Takeuchi .  Expressions of Protein Oxidation Markers, Dityrosine and Advanced Oxidation Protein Products in Cisplatin-Induced Nephrotoxicity in Rats .  JOURNAL OF VETERINARY MEDICAL SCIENCE73 ( 3 ) 403 - 407   2011.3Expressions of Protein Oxidation Markers, Dityrosine and Advanced Oxidation Protein Products in Cisplatin-Induced Nephrotoxicity in RatsReviewed

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    The purpose of this study was to evaluate whether dityrosine and advanced oxidation protein products (AOPP) reflect the severity of cisplatin-induced nephrotoxicity. Immunoexpression of dityrosine in kidneys and plasma AOPP concentration were examined up to day 4 post-cisplatin injection in rats. Cisplatin injection induced tubular injury on days 2-4 after injection and increased scrum creatinine and BUN on days 3 and 4. On days 2-4, dityrosine was immunostained in the cytoplasm of damaged tubular cells, and their immunostaining intensity increased time-dependently. Plasma AOPP levels were significantly increased on days 3 and 4. These results suggest that expressions of dityrosine and AOPP were associated with the severity of renal injury and may be useful markers for the development of cisplatin-induced nephrotoxicity.

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  • Takashi Kamio, Atsushi Asano, Yoshinao Z. Hosaka, Ahmed Magzoub Khalid, Shin-ichi Yokota, Masanori Ohta, Kenji Ohyama, Yoshiaki Yamano .  Expression of the Centrosomal Colon Cancer Autoantigen Gene during Spermatogenesis in the Maturing Rat Testis .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY74 ( 7 ) 1466 - 1469   2010.7Expression of the Centrosomal Colon Cancer Autoantigen Gene during Spermatogenesis in the Maturing Rat TestisReviewed

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    We analyzed the gene and protein expression of serologically defined colon cancer antigen 8. Gene expression was upregulated in the maturing rat testis, and was localized to the spermatocytes. Protein was detected in the spermatids and at the sites of mRNA expression. Specific expression of colon cancer antigen 8 was observed in the maturing rat testis.

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  • Simon A. Y., Moritoh K., Torigoe D., Asano A., Sasaki N., Agui T. .  Multigenic control of resistance to Sendai virus infection in mice .  Infection, Genetics and Evolution9 ( 6 ) 1253 - 1259   2009.12

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  • Yoshiaki Yamano, Atsushi Asano, Masanori Ohta, Shuji Hirata, Tomoko Shoda, Kenji Ohyama .  Expression of Rat Sperm Flagellum-Movement Associated Protein Genes under 2,3,7,8-Tetrachlorodibenzo-p-dioxin Treatment .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY73 ( 4 ) 946 - 949   2009.4Expression of Rat Sperm Flagellum-Movement Associated Protein Genes under 2,3,7,8-Tetrachlorodibenzo-p-dioxin TreatmentReviewed

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    We analyzed the gene expression of sperm flagellum-movement associated proteins, namely, adenylate kinase domain containing protein (RGD1303144) and outer dense fiber protein 1. These gene expressions were upregulated in a maturing rat testis, and RGD1303144 gene was expressed dominantly in spermatocytes. Although 2,3,7,8-tetrachlorodibenzo-p-dioxin administration reduced sperm motility, these gene expressions were not affected.

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  • Yoshiaki Yamano, Atsushi Asano, Kenji Ohyama, Masanori Ohta, Ren Nishio, Isao Morishima .  Expression of the Ha-ras suppressor family member 5 gene in the maturing rat testis .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY72 ( 5 ) 1360 - 1363   2008.5Expression of the Ha-ras suppressor family member 5 gene in the maturing rat testisReviewed

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    We analyzed the gene expression of Ha-ras suppressor family member 5 (Hrasls5), which is considered to modulate the Ha-ras signaling cascade, from maturing rat testis. Expression was detected primarily in the spermatocytes in the maturing rat testis. The Hrasls5 gene product might function as a tumor suppressor as well as in spermatogenesis, as deduced from its amino acid sequence.

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  • Hiroaki Kariwa, Hiroshi Noda, Mina Nakauchi, Mariko Ishizuka, Kazuaki Hashiguchi, Shingo Hashimoto, Kentaro Yoshii, Atsushi Asano, Takashi Agui, Hiroyuki Kogaki, Yoshihiro Kurano, Yoshiaki Uchida, Nobuyuki Fuji, Masahisa Okada, Ikuo Takashima .  Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus .  JAPANESE JOURNAL OF VETERINARY RESEARCH55 ( 4 ) 115 - 127   2008.2Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirusReviewed

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    The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKW(250) on SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.

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  • Atsushi Asano, Kouta Tsubomatsu, Cha-Gyun Jung, Nobuya Sasaki, Takashi Agui .  A deletion mutation of the protein tyrosine phosphatase kappa (Ptprk) gene is responsible for T-helper immunodeficiency (thid) in the LEC rat .  MAMMALIAN GENOME18 ( 11 ) 779 - 786   2007.11A deletion mutation of the protein tyrosine phosphatase kappa (Ptprk) gene is responsible for T-helper immunodeficiency (thid) in the LEC ratReviewed

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    Bone marrow (BM)-derived T-cell progenitors differentiate into CD4 or CD8 single-positive (SP) cells in the thymus. We have previously reported that a single autosomal mutation, thid, causes a defect in the maturation of CD4 SP thymocytes and an abnormality of peripheral helper T cells in the LEC rat. In this study we attempted to identify a gene responsible for the thid mutation. We first performed genetic linkage analysis and mapped the thid locus between Myb and D1Rat392 on Chr 1. In this region we found an approximately 380-kb deletion from intron 3 of the Ptprk gene, which encodes a receptor-like protein tyrosine phosphatase type kappa (RPTP kappa) to intron 1 of the RGD1560849 predicted gene in the LEC rat genome. Reconstitution with syngenic BM cells transduced Ptprk but not the RGD1560849 predicted gene rescued development of CD4 SP cells in the LEC rat thymus. It is confirmed by this result that the Ptprk gene is responsible for the thid mutation in the LEC rat. Our results further suggest that RPTP kappa plays a critical role in the development of CD4 SP cells in the thymus.

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  • Nobuya Sasaki, Yayoi Hosoda, Aogu Nagata, Ming Ding, Ji-Ming Cheng, Tomomi Miyamoto, Shinya Okano, Atsushi Asano, Ichiro Miyoshi, Takashi Agui .  A mutation in Tpst2 encoding tyrosylprotein sulfotransferase causes dwarfism associated with hypothyroidism .  MOLECULAR ENDOCRINOLOGY21 ( 7 ) 1713 - 1721   2007.7A mutation in Tpst2 encoding tyrosylprotein sulfotransferase causes dwarfism associated with hypothyroidismReviewed

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    The growth-retarded (grt) mouse has an autosomal recessive, fetal-onset, severe thyroid hypoplasia related to TSH hyporesponsiveness. Through genetic mapping and complementation experiments, we show that grt is a missense mutation of a highly conserved region of the tyrosylprotein sulfotransferase 2 (Tpst2) gene, encoding one of the two Tpst genes implicated in posttranslational tyrosine Osulfation. We present evidence that the grt mutation leads to a loss of TPST2 activity, and TPST2 isoform has a high degree of substrate preference for TSH receptor (TSHR). The expression of TPST2 can restore TSH-TSHR-mediated cAMP production in fibroblasts derived from grt mice. Therefore, we propose that the tyrosine sulfation of TSHR by TPST2 is crucial for TSH signaling and resultant thyroid gland function.

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  • AR Cho, K Uchio-Yamada, T Torigai, T Miyamoto, Miyoshi, I, J Matsuda, T Kurosawa, Y Kon, A Asano, N Sasaki, T Agui .  Deficiency of the tensin2 gene in the ICGN mouse: an animal model for congenital nephrotic syndrome .  MAMMALIAN GENOME17 ( 5 ) 407 - 416   2006.5Deficiency of the tensin2 gene in the ICGN mouse: an animal model for congenital nephrotic syndromeReviewed

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    The ICGN mouse is a model for nephrotic syndrome (NS) which presents with proteinuria, hyperlipidemia, and edema. In this study we attempted to identify the gene(s) responsible for NS. By analyzing albuminuria in 160 (ICGN x MSM)F-1 x ICGN backcross progenies, we found that NS in the ICGN mouse is caused by more than one gene. We then performed a quantitative trait locus (QTL) analysis and detected a QTL with a very high LOD score peak in the telomeric region of Chr 15. By analyzing the nucleotide sequence of 22 genes located close to the QTL, we found that the tensin2 gene of the ICGN mouse possessed an 8-nucleotide deletion mutation in exon 18, leading to a frameshift and giving rise to a terminal codon at a premature position. Analyses of in situ hybridization and immunohistochemistry revealed that tensin2 was expressed in podocytes and tubular epithelial cells in normal mice but not in the ICGN mouse. These data raise the possibility that a mutation of the tensin2 gene is responsible for NS of the ICGN mouse and tensin2 is a prerequisite for the normal kidney function.

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  • T Nakamura, A Asano, S Okano, JH Ko, Y Kon, T Watanabe, T Agui .  Intracellular localization and antiviral property of canine Mx proteins .  JOURNAL OF INTERFERON AND CYTOKINE RESEARCH25 ( 3 ) 169 - 173   2005.3Intracellular localization and antiviral property of canine Mx proteinsReviewed

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    Mx is an interferon (IFN)-induced protein that shows antiviral activities against RNA viruses. We examined an expression of mRNA, an intracellular localization of protein, and an antiviral property of canine Mx1 and Mx2. Both Mx1 and Mx2 mRNAs were induced in a canine kidney cell line Madin-Darby canine kidney (MDCK), stimulated with an IFN-inducer, poly(I) (.) poly(C) for 12 h, suggesting the presence of regulatory mechanisms consistent with Mx genes in other species. By immunostaining BALB/3T3 fibroblasts transiently transfected FLAG epitope-tagged canine Mx1 and Mx2 cDNAs with an anti-FLAG tag, it was revealed that both Mx1 and Mx2 proteins are localized in cytoplasm. BALB/3T3 fibroblasts expressing stably Mx2 but not Mx1 had an antiviral activity against recombinant vesicular stomatitis virus (VSV). This is the first report demonstrating the functional analysis of canine Mx proteins.

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  • S Okano, A Asano, N Sasaki, Y Kon, T Watanabe, T Agui .  Examination of the Lunatic fringe and Uncx4.1 expression by whole-mount in situ hybridization in the embryo of the CKH-Jsr (jumbled spine and ribs) mouse .  JAPANESE JOURNAL OF VETERINARY RESEARCH52 ( 4 ) 145 - 149   2005.2Examination of the Lunatic fringe and Uncx4.1 expression by whole-mount in situ hybridization in the embryo of the CKH-Jsr (jumbled spine and ribs) mouseReviewed

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    The CKH-Jsr (jumbled spine and ribs) mouse was found as a spontaneous mutant with malformation of vertebrae, that is, a short trunk and kinky tail. We examined Lunatic Fringe (Lfng) and Uncx4.1 expression in the presomitic mesoderm (PSM) and somites of Jsr-mutant (CKH-Jsr/+) embryos to elucidate pathogenesis of the Jsr mutation. Expression pattern of Lfng in the PSM of Jsr-mutant embryos was similar to that of the normal (C57BL/6) embryos. However, expression pattern of Uncx4.1 in the somites of Jsr-mutant embryos was impaired to be irregular and mosaic, suggesting that the anterior-posterior (A-P) polarity is disordered in the Jsr mutant. These results indicate that the Jsr mutation disrupts the A-P polarity of somites during the somitogenesis without altering Lfng expression pattern in the PSM.

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  • Y Namiki, Y Kon, K Kazusa, A Asano, N Sasaki, T Agui .  Quantitative trait loci analysis of heat stress resistance of spermatocytes in the MRL/MpJ mouse .  MAMMALIAN GENOME16 ( 2 ) 96 - 102   2005.2Quantitative trait loci analysis of heat stress resistance of spermatocytes in the MRL/MpJ mouseReviewed

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    The MRL/MpJ mouse has previously been reported to possess an interesting phenotype in which spermatocytes are resistant to the abdominal temperature heat shock. In this study genetic analysis for it was performed. The phenotypes of F-2 progenies produced by mating MRL/MpJ and control strain C57BL/6 mice were not segregated into two types as parental phenotypes, suggesting that the phenotype is controlled by multiple genetic loci. Thus, quantitative trait loci (QTL) analysis was performed using 98 microsatellite markers. The weight ratio of the cryptorchid testis to the intact testis (testis weight ratio) and the Sertoli cell index were used for quantitative traits. QTL analysis revealed two significant QTLs located on Chrs 1 and 11 for testis weight ratio and one significant QTL located in the same region of Chr 1 for the Sertoli cell index. A microsatellite marker locus located in the peak of the QTL on Chr 1 did not recombine with the exonuclease 1 (Exo1) gene locus in 140 F-2 progenies. Mutation of the Exo1 gene was previously reported to be responsible for metaphase-specific apoptosis (MSA) of spermatocytes in the MRL/MpJ mouse. These results raise the possibility that mutation of the Exo1 gene is responsible for both MSA and heat stress resistance of spermatocytes in the MRL/MpJ mouse.

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  • A Asano, Y Kon, T Agui .  The mRNA regulation of porcine double-stranded RNA-activated protein kinase gene .  JOURNAL OF VETERINARY MEDICAL SCIENCE66 ( 12 ) 1523 - 1528   2004.12The mRNA regulation of porcine double-stranded RNA-activated protein kinase geneReviewed

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    The double-stranded RNA (dsRNA)-activated protein kinase (PKR), which is one of the products of interferon (IFN)-stimulated genes, participates in the biological actions of IFN such as antiviral effects and immune response. In the present study, we identified the primary structure of porcine PKR proteins by cDNA cloning. Porcine PKR protein consisted of 537 amino acids and had two dsRNA-binding domains similarly existing in PKR proteins of other species. The treatment with IFN-alpha induced the expression of PKR 3.9-fold in a porcine kidney cell line, LLC-PK1. The same results were obtained when the cells were treated with poly(I)-poly(C), but treatment with either IFN-gamma or LPS did not induce this gene in LLC-PK1 cells. These results suggest similarity of the regulatory mechanisms in the PKR gene among mammalian species.

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  • M Okumura, K Yoshimatsu, K Araki, BH Lee, A Asano, T Agui, J Arikawa .  Epitope analysis of monoclonal antibody E5/G6, which binds to a linear epitope in the nucleocapsid protein of hantaviruses .  ARCHIVES OF VIROLOGY149 ( 12 ) 2427 - 2434   2004.12Epitope analysis of monoclonal antibody E5/G6, which binds to a linear epitope in the nucleocapsid protein of hantavirusesReviewed

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    Monoclonal antibody E5/G6 recognized a linear epitope common to hantavirus nucleocapsid proteins. Using synthetic peptides, we identified epitope E5/G6 as the 9 merYEDVNGIRK (NP 165 - 173), in which D167, G170, I171, and R172 are indispensable. Furthermore, all the peptides synthesized using various hantavirus sequences bound MAb E5/G6 consistently, despite the existence of several amino acid variations in this region. These results indicate that MAb E5/G6 is a useful tool for detecting hantavirus antigen in rodent or patient tissues using Western blotting or other immunohistochemical assays.

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  • K Kazusa, Y Namiki, A Asano, Y Kon, D Endoh, T Agui .  Differences in spermatogenesis in cryptorchid testes among various strains of mice .  COMPARATIVE MEDICINE54 ( 2 ) 179 - 184   2004.4Differences in spermatogenesis in cryptorchid testes among various strains of miceReviewed

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    The purpose of the study reported here was to define strain differences in spermatogenesis in cryptorchid testes in mice. Mice of strains AM, BALB/c, CBA/N, C3H/He, C57BL/6 (136), ddY and ICR were found to be sensitive to heat stress attributable to experimentally induced cryptorchidism. In contrast, mice of strains AKR/N (AKR), MRL/MpJ-+/+ (M+) and MRL/MpJ-lpr/lpr (lpr) were resistant to heat stress. Relative increases of apoptotic cells were detected in the sensitive group, but not in the resistant group. A decrease of proliferating cell nuclear antigen-immunoreactive cells after experimentally induced cryptorchidism was observed only in the sensitive group. These results suggested that heat stress-resistant germ cells were present in MRL and AKR strains, possibly originating from the genetic background.

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  • Kazusa K, Namiki Y, Asano A, Kon Y, Endoh D, Agui T .  Differences in spermatogenesis in cryptorchid testes among various strains of mice. .  Comparative medicine54 ( 2 ) 179 - 84   2004.4Differences in spermatogenesis in cryptorchid testes among various strains of mice.

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  • JH Ko, A Asano, Y Kon, T Watanabe, T Agui .  Characterization of the chicken PKR: Polymorphism of the gene and antiviral activity against vesicular stomatitis virus .  JAPANESE JOURNAL OF VETERINARY RESEARCH51 ( 3-4 ) 123 - 133   2004.2Characterization of the chicken PKR: Polymorphism of the gene and antiviral activity against vesicular stomatitis virusReviewed

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    The double-stranded RNA-dependent protein kinase (PKR) is induced in mouse and human cells on treatment with interferon. In this study, we have cloned and determined the nucleotide sequences of chicken PKR cDNA in various chicken breeds. Chicken PKR was a 550 - amino-acid protein as deduced from the cDNA open reading frame (ORF), and there were specific domains (two double-stranded RNA binding domains (DRBDs) and numerous kinase subdomains) characterized in RNA binding proteins and kinase families. Furthermore, it was suggested that chicken PKR was polymorphic. Transfected cell clones expressing chicken PKR mRNA were demonstrated to confer antiviral responses to vesicular stomatitis virus, except for Koshamo type - 3 (KS - 3). KS - 3 PKR, which has an amino acid substitution at position 507 (Arg to Gln), showed amphibious antiviral responses. This specific amino acid substitution was considered to determine the antiviral function of chicken PKR in addition to essential domains as DRBDs and kinase subdomains.

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  • A Asano, HK Jin, T Watanabe .  Mouse Mx2 gene: organization, mRNA expression and the role of the interferon-response promoter in its regulation .  GENE306   105 - 113   2003.3Mouse Mx2 gene: organization, mRNA expression and the role of the interferon-response promoter in its regulationReviewed

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    Mx is an interferon (IFN)-inducible intracellular protein found in various vertebrates that mediates resistance against negative-strand RNA viruses. We have demonstrated previously that feral mouse strains are able to express a functional Mx2 mRNA, while that of the inbred laboratory strains was non-functional because of a single nucleotide insertion in the open reading frame, and under the detectable level. In the present study, we examined the regulation of Mx2 expression in vivo using a congenic mouse carrying Mx1 and Mx2 genes derived from feral strain SPR. Mx2 mRNA was induced strongly in the spleen, ovary and white adipose tissue after the treatment with IFNalpha/beta. Furthermore, we identified the structure of the Mx2 gene. It consists of 14 exons, greatly homologous to the Mx1 gene. The promoter region of Mx2 contained two putative IFN-stimulated response elements (ISREs). We found that the proximal ISRE site positioned between -69 and -55 was essential to IFNalpha/beta-induced transcription by transient transfection assay using reporter gene constructs with mutants of the Mx2 promoter. These results indicate the similarity of the mechanisms of mRNA induction among Mx genes. (C) 2003 Elsevier Science B.V. All rights reserved.

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  • A Asano, JH Ko, T Morozumi, N Hamashima, T Watanabe .  Polymorphisms and the antiviral property of porcine Mx1 protein .  JOURNAL OF VETERINARY MEDICAL SCIENCE64 ( 12 ) 1085 - 1089   2002.12Polymorphisms and the antiviral property of porcine Mx1 proteinReviewed

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    We determined the cDNA sequences of the type I interferon-inducible proteins, pig Mx1 from PK(15) and LLC-PK1 cells, and compared the antiviral activities of both Mx proteins, including Mx1 polymorphisms against vesicular stomatitis virus (VSV). Mv1 cDNA derived from PK( 15) cells had an 11 bp-deletion in the 3' end of the coding region, and was estimated to encode 8 amino acid substitutions and a 23 amino acid extension compared to that from LLC-PK1 cells. VSV replication was inhibited in the 3T3 cells expressing Mx1 mRNA after the cDNA was transfected. However, the efficiency of this inhibition was not different between the cells expressing Mx1 mPNA from both PK and LLC, These results indicate that pig Mx1 protein confers resistance to VSV.

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  • S Okano, A Asano, Y Kon, H Miyoshi, T Watanabe .  Genetic analysis of jumbled spine and ribs (Jsr) mutation affecting the vertebral development in mice .  BIOCHEMICAL GENETICS40 ( 9-10 ) 311 - 322   2002.10Genetic analysis of jumbled spine and ribs (Jsr) mutation affecting the vertebral development in miceReviewed

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    The jumbled spine and ribs (Jsr) mouse was derived from a spontaneous mutation. As the phenotype, a shortened trunk and kinky tail are characteristic Jsr traits. In this study, on high resolution mapping it was found that Lunatic fringe (Lfng) mapped at the same position as Jsr. Lfng was identified as the candidate gene for Jsr, but sequence analysis of this gene revealed no substitution in the coding region of cDNA. Therefore, we adopted the strategy of positional cloning for Jsr using a mouse bacterial artificial chromosome (BAC) library. A BAC contig was constructed from three BAC clones showing positive signals of Lfng and 11MMHAP75FRD8.seq near the Jsr locus on chromosome 5. Based on the genetic mapping of both T7 and sp6 ends of a clone of BAC382-O-7 (BAC382), the Jsr gene was considered to exist in BAC382 and to be positioned near the sp6 side.

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  • JH Ko, HK Jin, A Asano, A Takada, A Ninomiya, H Kida, H Hokiyama, M Ohara, M Tsuzuki, M Nishibori, M Mizutani, T Watanabe .  Polymorphisms and the differential antiviral activity of the chicken Mx gene .  GENOME RESEARCH12 ( 4 ) 595 - 601   2002.4Polymorphisms and the differential antiviral activity of the chicken Mx geneReviewed

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    The nucleotide sequence of chicken Mx cDNA was reported earlier using the White Leghorn breed in Germany, but it showed no enhanced resistance to viruses. In this study, the nucleotide sequences of chicken Mx cDNA were determined in many breeds. A total of 25 nucleotide substitutions, of which 14 were deduced to cause amino acid exchanges, were detected, Suggesting that the chicken Mx gene is very polymorphic. Transfected cell clones expressing chicken Mx mRNA were established after the Mx cDNA was constructed with an expression vector and introduced into mouse 3T3 cells, and the Mx genes from some breeds were demonstrated to confer positive antiviral responses to influenza virus and vesicular stomatitis virus. On the basis of the comparison among the antiviral activities associated with many Mx variations, a specific amino acid substitution at position 631 (Ser to Asn) was considered to determine the antivirally positive or negative Mx gene. Thus, a single amino acid Substitution influences the antiviral activity of Mx in domesticated chickens.

    DOI: 10.1101/gr.210702

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  • S El-Shazly, S Okano, A Asano, T Watanabe .  Developmental study of the different effects on the hybrid sterility of Kit(W) and Kit(W-v) alleles paired with Kit(S) from Mus spretus .  DEVELOPMENT GROWTH & DIFFERENTIATION43 ( 5 ) 611 - 617   2001.10Developmental study of the different effects on the hybrid sterility of Kit(W) and Kit(W-v) alleles paired with Kit(S) from Mus spretusReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    The combination of the Kit(W) or Kit(W-n) mutant alleles and Kits from Mus spretus results in male hybrid sterility with small testes. In the present study, reproduction of the combination between Kit(W-v) and Kit(s) alleles was examined. The Kit(W-v)/Kit(s) male was fertile and the histologic structure was normal; the seminiferous tubules showed all of the normal stages of spermatogenesis. The postnatal development of the testis at 8, 12, 16 and 20 days was also studied in the fertile +(Kit)/+(Kit) and Kit(W-v)/Kit(s) males and the sterile Kit(W)/Kit(s). The results showed that at 8 days there was no noticeable difference among the three genotype combinations, while from 12 to 20 days spermatogenesis in the Kit(w)/Kit(s) male nearly stopped before the meiosis stage. The expression of Kit receptor protein from the Kit(s) allele in the sterile testis of the Kit(w)/Kit(s) male was confirmed using western blot analysis. The Kit ligand derived from M. spretus showed two amino acid changes in the extracellular domain compared with that from C57BL and it appears that the ligand-receptor interaction between C57BL and SPR may influence the male hybrid sterility of Kit(w)/Kit(s).

    DOI: 10.1046/j.1440-169X.2001.00598.x

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  • J Ohuchi, T Arai, Y Kon, A Asano, H Yamauchi, T Watanabe .  Characterization of a novel gene, sperm-tail-associated protein (Stap), in mouse post-meiotic testicular germ cells .  MOLECULAR REPRODUCTION AND DEVELOPMENT59 ( 4 ) 350 - 358   2001.8Characterization of a novel gene, sperm-tail-associated protein (Stap), in mouse post-meiotic testicular germ cellsReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-LISS  

    During mammalian spermatogenesis, many specific molecules show the dynamics of expression and elimination, corresponding with the morphological differentiation of germ cells. We have isolated a novel cDNA designated F77 from mouse testis by cDNA subtractive hybridization between normal and sterile mice, using the C57BL/6 congenic strain for the hybrid sterility-3 (Hst-3) allele from Mus spretus. The full-length F77 mRNA was 3.4 kb and showed significant nonmatching with entries in the databases. F77 was mapped at a proximal position between D8Mit212 and D8Mit138 on mouse chromosome 8, in which no corresponding genes related to its nucleotide sequence were found. F77 mRNA was not detected in any other organs except the testis of adult fertile mice. F77 protein was only seen in normal adult testis and epididymis. in contrast to normal C57BL/6 mice, F77 mRNA and protein were not seen in germ cell-deficient Kit(W)/Kit(Wv) mice. By in situ hybridization, F77 mRNA was detected mainly at round spermatids in the sexually mature testis, and immunohistochemical analysis revealed that F77 protein was located at the tail of elongated spermatids. We are proposing the name, sperm-tail-associated protein (Stap), for the gene encoding F77 cDNA. (C) 2001 Wiley-Liss, Inc.

    DOI: 10.1002/mrd.1041

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  • T Morozumi, C Sumantri, E Nakajima, E Kobayashi, A Asano, T Oishi, T Mitsuhashi, T Watanabe, N Hamasima .  Three types of polymorphisms in exon 14 in porcine Mx1 gene .  BIOCHEMICAL GENETICS39 ( 7-8 ) 251 - 260   2001.8Three types of polymorphisms in exon 14 in porcine Mx1 geneReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:KLUWER ACADEMIC/PLENUM PUBL  

    Much is known about the antiviral activity of Mx proteins in species such as mouse and human. In the mouse, loss of resistibility to influenza virus has been shown to be due to specific polymorphisms in the Mx gene. This gene is therefore an interesting candidate gene for disease resistance in farm animals. The porcine Mx1 gene has already been identified and characterized based on its homology with mouse Mx1; however until now no evidence of polymorphisms in the porcine gene has been reported. In this study, we have found two new polymorphisms in exon 14 of porcine Mx I by DNA sequencing and confirmed their presence in different breeds, using polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) with NarI and NaeI restriction enzymes. On the basis of the deduced amino acid sequence, one allele contains a deletion that may result in a frameshift to yield several amino acid substitutions and extension of the carboxyl terminal region of Mx1 protein. The deletion allele, Mx1(c), was found to be segregating in Landrace, Berkshire, Duroc, Hampshire, and Yucatan miniature pig. A second point mutation, Mx1(b), was detected in Meishan and two Vietnamese native pig breeds. All other breeds tested were fixed for the Mx1(a) allele that is identical to the sequence reported previously. It will be interesting to determine if the Mx1(c) deletion is associated with variation in resistance to the myxovirus family in the pig.

    DOI: 10.1023/A:1010230715605

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  • SD El-Shazly, KW Seo, A El-Nahas, GP Ables, A Asano, T Watanabe .  Male hybrid sterility of mice with the genomic region of the Kit(W) mutation and the Kit(S) allele from Mus spretus .  BIOCHEMICAL GENETICS39 ( 3-4 ) 127 - 137   2001.4Male hybrid sterility of mice with the genomic region of the Kit(W) mutation and the Kit(S) allele from Mus spretusReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:KLUWER ACADEMIC/PLENUM PUBL  

    Two congenic strains, C57BL-Kit(w) and C57BL-Kit(s), were generated. The Kit(w) allele originated from strain WB-Kit(w) and the Kit(s) allele from Mus spretus. The Kit(w)/Kit(s) males showed hybrid sterility with small testes, but the females were fertile. The development of the seminiferous tubules of Kit(W)/Kit(s) males stopped before the spermatocyte stage and they were almost free of sperm. The Kit gene is located at position 42 on chromosome 5. We investigated in the C57BL-Kits congenic strain which part of the chromosomal region adjacent to the Kits allele is introduced from SPR into a C57BL background. The region between positions 42 and 44 was derived from SPR. Eleven amino acid substitutions of the Kit(s) cDNA were detected by comparison with the sequence data of the +(Kit) cDNA from C57BL; seven were in the extracellular domain, one in the transmembrane domain, two in the kinase I domain, and one in the carboxy-terminal tail. The Kit mRNA derived from both Kit(w) and Kit(s) alleles was expressed in the sterile testes of Kit(w)/Kit(s) males.

    DOI: 10.1023/A:1010217924328

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  • Y Nakamura, Nagase, I, A Asano, N Sasaki, T Yoshida, T Umekawa, N Sakane, M Saito .  beta 3-adrenergic agonist up-regulates uncoupling proteins 2 and 3 in skeletal muscle of the mouse .  JOURNAL OF VETERINARY MEDICAL SCIENCE63 ( 3 ) 309 - 314   2001.3beta 3-adrenergic agonist up-regulates uncoupling proteins 2 and 3 in skeletal muscle of the mouseReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC VET SCI  

    Chronic stimulation of the beta3-adrenergic receptor (AR) in obese animals resulted in a reduced adiposity associated with an increased expression of thermogenic uncoupling protein (UCP)1 in adipose tissues. In this study, the mRNA expression of newly cloned UCP isoforms (UCP2 and UCP3) were examined in obese yellow KK and C57BL control mice. UCP2 mRNA was found in all tissues examined, with higher levels in adipose tissues and skeletal muscle of the obese mice. UCP3 mRNA was expressed in skeletal muscle, heart and brown adipose tissue similarly in the two mouse strains. Daily injection of a selective beta3-adrenergic agonist, CL316,243 (0.1 mg/kg), for 10 days resulted in a marked reduction of white fat pad weight and 1.8 similar to4.8-fold increase in the mRNA levels of UCP2 and UCP3 in skeletal muscle of obese mice. No noticeable change in the UCP2 and 3 mRNA levels was found in brown and white adipose tissues. It was also found that CL316,243 injection produced a marked and sustained elevation of the plasma free fatty acid level. These results, together with our previous Endings of the fatty acid-induced UCP expression in a myocyte cell line in vitro, suggest that the beta3-AR agonist-induced UCP expression in skeletal muscle may be mediated through the elevated plasma free fatty acids. It was also suggested that anti-obesity effect of beta3-AR agonists is attributable to increased thermogenesis not only by UCP1 but also by UCP2 and UCP3.

    DOI: 10.1292/jvms.63.309

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  • HK Jin, K Yoshimatsu, A Takada, M Ogino, A Asano, J Arikawa, T Watanabe .  Mouse Mx2 protein inhibits hantavirus but not influenza virus replication .  ARCHIVES OF VIROLOGY146 ( 1 ) 41 - 49   2001Mouse Mx2 protein inhibits hantavirus but not influenza virus replicationReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER-VERLAG WIEN  

    The antiviral potential of Mx2 protein remains unknown, because the Mx2 gene in commonly used strains of laboratory mice is nonfunctional. Our previous study showed that functional Mx2 protein in some feral-origin strains was induced upon interferon treatment, was localized in the cytoplasm, and inhibited vesicular stomatitis virus replication. In the present study, we have demonstrated chat the embryonic fibroblastic cells from a feral-origin strain (SPR) expressed 74 kDa Mx2 protein, which prevented the accumulation of viral transcripts and proteins of hantaviruses when the Mx2 gene was constitutively expressed in transfected Vero cells. Furthermore, the cells showed significantly lower titers of the virus than control cells. In contrast, influenza virus replication was not affected by the expression of Mx2 protein in the Vero cells.

    DOI: 10.1007/s007050170189

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  • K Kimura, N Sasaki, A Asano, J Mizukami, S Kayahashi, T Kawada, T Fushiki, M Morimatsu, T Yoshida, M Saito .  Mutated human beta 3-adrenergic receptor (Trp64Arg) lowers the response to beta 3-adrenergic agonists in transfected 3T3-L1 preadipocytes .  HORMONE AND METABOLIC RESEARCH32 ( 3 ) 91 - 96   2000.3Mutated human beta 3-adrenergic receptor (Trp64Arg) lowers the response to beta 3-adrenergic agonists in transfected 3T3-L1 preadipocytesReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:GEORG THIEME VERLAG  

    Wild-type or mutated human beta 3-adrenergic receptor (Trp64Arg) cDNAs were stably expressed in mouse 3T3-L1 cells. Saturation binding study using a beta-adrenergic ligand revealed that there was no significant difference in the receptor density and the equilibrium dissociation constant between the two cell lines. However, the ability of the mutant beta 3-adrenergic receptor to accumulate cyclic AMP (cAMP) in response to isoproterenol was much reduced and Kact for cAMP accumulation was lowered as compared to the wild type receptor. The amount of alpha subunit of stimulatory GTP-binding protein (GS alpha) and adenylyl cyclase activity in response to forskolin were not different in the two cell lines. The responses of the mutant receptor to epinephrine, norepinephrine and L-755,507, a highly specific agonist for human beta 3-adrenergic receptor, were also reduced, but the reduction of Kact for L-755,507 was more evident than other agonists tested. The cAMP accumulation in response to some conventional beta 3 agonists was less than 10% of that to isoproterenol even in the cells expressing the wild type receptor. These results suggest that the Trp64Arg mutant beta 3-adrenergic receptor has less ability to stimulate adenylyl cyclase, and that lipolytic activity through the beta 3-adrenergic receptor by catecholamines in subjects carrying this mutation might be suppressed.

    DOI: 10.1055/s-2007-978597

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  • A Asano, K Kimura, M Saito .  Cold-induced mRNA expression of angiogenic factors in rat brown adipose tissue .  JOURNAL OF VETERINARY MEDICAL SCIENCE61 ( 4 ) 403 - 409   1999.4Cold-induced mRNA expression of angiogenic factors in rat brown adipose tissueReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC VET SCI  

    Brown adipose tissue (BAT) is the major site of non-shivering thermogenesis in rodents. Rapid angiogenesis is induced in association with adaptive hyperplasia of this tissue when the animal is exposed to cold. We demonstrated previously adrenergic activation of mRNA expression of vascular endothelial growth factor (VEGF) in rat BAT and its possible contribution to the cold-induced angiogenesis in this tissue. In the present study, we examined the effect of cold exposure on mRNA expression of other two angiogenic factors. VEGF-B and basic fibroblast growth factor (bFGF), in rats. Conventional Northern blot analysis revealed abundant mRNA expression of VEGF-B as well as VEGF, but not bFGF, in BAT. When rats were exposed to cold at 4 degrees C, the VEGF mRNA level was increased by 2.7-fold in 1-4 hr and returned to the basal level within 24 hr. In contrast, the VEGF-B mRNA level did not change throughout the course of cold exposure. A significant expression of bFGF mRNA was detected in BAT by reverse transcription polymerase chain reaction (RT-PCR). To evaluate the tissue bFGF mRNA level quantitatively, a competitive RT-PCR method was developed using a shorter RNA fragment as a competitor. The bFGF mRNA level in BAT was found to increase by 2.3-fold in 4 hr and decreased to the basal level within 24 hr after cold exposure. These results suggest that cold exposure leads to induce VEGF and bFGF rapidly and transiently in BAT, which in turn stimulate the proliferation of vascular endothelial cells in this tissue.

    DOI: 10.1292/jvms.61.403

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  • 浅野 淳 .  げっ歯類の褐色脂肪組織での血管内皮細胞増殖因子(VEGF)ファミリー発現の交感神経性調節 .      1999.3げっ歯類の褐色脂肪組織での血管内皮細胞増殖因子(VEGF)ファミリー発現の交感神経性調節

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    Language:Japanese   Publishing type:Doctoral thesis  

  • N Sakane, T Yoshida, T Umekawa, A Kogure, M Kondo, Y Nakamura, Y Sasaki, A Asano, M Saito .  Acute and chronic regulation of ob mRNA levels by beta(3)-adrenoceptor agonists in obese yellow KK mice .  ENDOCRINE JOURNAL45 ( 5 ) 647 - 651   1998.10Acute and chronic regulation of ob mRNA levels by beta(3)-adrenoceptor agonists in obese yellow KK miceReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN ENDOCRINE SOCIETY  

    The inhibitory effect of beta(3)-adrenoceptor agonists on the ob gene in brown adipose tissue (BAT) and white adipose tissue (WAT) is now well documented both in viva in lean animals and in vitro, but the reported effects of beta(3)-adrenoceptor agonists on ob gene expression in obese animals remain controversial. We investigated whether ob gene expression in BAT and WAT is reduced by acute and chronic administrations of a beta(3)-adrenoceptor agonist, CL316,243 (CL). The ob gene mRNA levels in BAT, perimetric and inguinal WAT of obese Yellow KK mice were about 4-fold higher than those of lean controls. Acute exposure (6 h) to CL decreased ob gene mRNA levels in three fat depots in both animals. Chronic exposure (10 days) to CL also decreased ob gene mRNA levels in BAT, perimetric, and inguinal WAT in both animals. We concluded that acute and chronic regulation by a beta(3)-adrenoceptor agonist suppressed ob gene expression in obese Yellow KK mice and lean cotrols.

    DOI: 10.1507/endocrj.45.647

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  • A Asano, M Morimatsu, H Nikami, T Yoshida, M Saito .  Adrenergic activation of vascular endothelial growth factor mRNA expression in rat brown adipose tissue: implication in cold-induced angiogenesis .  BIOCHEMICAL JOURNAL328   179 - 183   1997.11Adrenergic activation of vascular endothelial growth factor mRNA expression in rat brown adipose tissue: implication in cold-induced angiogenesisReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PORTLAND PRESS  

    Cold exposure produces adaptive hyperplasia and growth of brown adipose tissue (BAT), the major site of non-shivering thermogenesis in rodents, associated with increased angiogenesis in this tissue. Vascular endothelial growth factor (VEGF), one of the most potent angiogenic factors, was found to be expressed abundantly in BAT of the rat. When rats were exposed to cold at 4 degrees C, the VEGF mRNA level in BAT was increased by 2-3-fold in 1-4 h, but returned to the basal level within 24 h. VEGF expression in other tissues such as heart, kidney and lung did not change after cold exposure. The cold-induced increase in VEGF mRNA was abolished by surgical sympathetic denervation, but mimicked by administration of noradrenaline or a beta(3)-adrenoceptor agonist CL316,243, indicating the critical role of the beta-adrenergic pathway in VEGF expression in BAT. Among three isoforms of VEGF, the mRNA of a short form (VEGF120) lacking heparin-binding activity was preferentially increased after cold exposure and treatment with the adrenergic agonists. These results suggest that cold exposure activates the sympathetic nerves and leads to a rapid increase in synthesis of VEGF in BAT, which in turn stimulates the proliferation of surrounding vascular endothelial cells.

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  • A TERAO, H KITAMURA, A ASANO, M KOBAYASHI, M SAITO .  ROLES OF PROSTAGLANDINS D-2 AND E(2) IN INTERLEUKIN-1-INDUCED ACTIVATION OF NOREPINEPHRINE TURNOVER IN THE BRAIN AND PERIPHERAL ORGANS OF RATS .  JOURNAL OF NEUROCHEMISTRY65 ( 6 ) 2742 - 2747   1995.12ROLES OF PROSTAGLANDINS D-2 AND E(2) IN INTERLEUKIN-1-INDUCED ACTIVATION OF NOREPINEPHRINE TURNOVER IN THE BRAIN AND PERIPHERAL ORGANS OF RATSReviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:LIPPINCOTT-RAVEN PUBL  

    Possible roles of prostaglandins (PGs) in interleukin-l (Il-l)-induced activation of noradrenergic neurons were examined by assessing norepinephrine (NE) turnover in the brain and peripheral organs of rats, An intraperitoneal injection of human recombinant IL-1 beta accelerated NE turnover in the hypothalamus, spleen, lung, diaphragm, and pancreas. A similar increase in NE turnover was also observed after intracerebroventricular injection of corticotropin-releasing hormone (CRH), Pretreatment with indomethacin (cyclooxygenase inhibitor) abolished the IL-1-induced, but not the CRH-induced, increase in hypothalamic and splenic NE turnover. To elucidate which eicosanoid-cyclooxygenase product(s) is responsible for accelerating NE turnover, PGD(2), PGE(2), PGF(2 alpha), U-46619 (stable thromboxane A(2) analogue), or carbacyclin (stable prostacyclin analogue) was administered intracerebroventricularly. Among them, PGE, was the only eicosanoid effective in increasing NE turnover in spleen, whereas PGD(2) was effective in the hypothalamus. The stimulative effect of PGD(2) was abolished by pretreatment with intracerebroventricular injection of a CRH antiserum. These results suggest that the action of IL-1 is mediated through PGD, production to activate the noradrenergic neurons in the hypothalamus, and through PGE(2) production to increase sympathetic nerve activity in spleen.

    DOI: 10.1046/j.1471-4159.1995.65062742.x

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Books

MISC

  • 【AVMA動物の安楽死指針:2020年版の解説】

    浅野 淳, 高井 了

    実験動物技術   57 ( 2 )   71 - 77   2022.12

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    Language:Japanese   Publisher:(一社)日本実験動物技術者協会  

  • AVMA動物の安楽死指針 2020年版の解説

    岡村 匡史, 森松 正美, 浅野 淳

    LABIO 21   ( 87 )   27 - 30   2022.9

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    Language:Japanese   Publisher:(公社)日本実験動物協会  

Presentations

  • 浅野淳、瀬戸山健太郎   教育講演「安楽死の考え方」   Invited

    第105回日本獣医麻酔外科学会学術集会  2022.12  日本獣医麻酔外科学会

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    Event date: 2022.12

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:福岡国際会議場   Country:Japan  

  • 浅野淳、髙井了   米国獣医学会 動物の安楽死指針:2020 年版の概説  

    第56回 日本実験動物技術者協会総会  2022.10  日本実験動物技術者協会

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    Event date: 2022.10

    Language:Japanese   Presentation type:Symposium, workshop panel (public)  

    Venue:松本市  

  • 林原安里、割田克彦、浅野淳   マウス精巣におけるインクレチン受容体の発現とその作用に関する研究  

    第35回九州実験動物研究会総会  九州実験動物研究会

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    Event date: 2017.11

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:陽子線治療センター 指宿ベイヒルズ HOTEL & SPA 大会議室(3階)(指宿市)  

  • 林原安1、割田克彦、浅野淳   マウス精巣におけるインクレチン受容体の発現とその発現調節因子に関する研究  

    第160回日本獣医学会学術大会  日本獣医学会

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    Event date: 2017.9

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:鹿児島  

Intellectual Property

  • マウス肝炎ウイルス由来ポリペプチドおよび/またはセンダイウイルス由来ポリペプチド、これらを用いたマウス肝炎ウイルス感染および/またはセンダイウイルス感染検査キット、ならびにマウス肝炎ウイルス感染および/またはセンダイウイルス感染の検出方法

    安居院高志、佐々木宣哉、鳥越大輔、浅野淳

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    Application no:PCT/JP2010/057188  Date applied:2009.4

    Country of applicant:Domestic  

  • センダイウイルス由来ポリペプチドおよびこれを用いたセンダイウイルス感染検査キットならびにセンダイウイルス感染の検出方法

    安居院高志、佐々木宣哉、鳥越大輔、浅野淳

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    Application no:特願2009-104905  Date applied:2009.4

    Country of applicant:Domestic  

  • マウス肝炎ウイルス由来ポリペプチドおよびこれを用いたマウス肝炎ウイルス感染検査キットならびにマウス肝炎ウイルス感染の検出方法

    安居院高志、佐々木宣哉、鳥越大輔、浅野淳

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    Application no:特願2009-104906  Date applied:2009.4

    Country of applicant:Domestic  

Research Projects

  • 肥満・糖尿病モデル動物を用いたインクレチンによる雄性生殖機能の調節に関する研究

    2018.4 - 2021.3

    科学研究費補助金  基盤研究(C)

  • 精巣特異的ホメオドメインタンパク質による遺伝子発現制御機構の解析

    Grant number:25450465  2013 - 2015

    文部科学省  科学研究費補助金(基盤研究(C))  基盤研究(C)

    浅野 淳

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5330000 ( Direct Cost: \4100000 、 Indirect Cost:\1230000 )

  • Analysis of defense system regulated by interferon-induced GTP binding proteins

    Grant number:20780208  2008 - 2009

    Ministry of Education, Culture, Sports, Science and Technology  Grants-in-Aid for Scientific Research(若手研究(B))  若手研究(B)

    Atsushi ASANO

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3510000 ( Direct Cost: \2700000 、 Indirect Cost:\810000 )

    Interferon (IFN)-induced GTP binding proteins (GBPs) were thought to play a part in inhibition of amplification of infected viruses in animal. I identified two novel GBP genes, GBP2 and GBP3, of the chicken. The mRNA of these genes was induced by various IFNs such as IFNβ, IFNγ and IFNλ. Chicken GBP2 protein was located in the cytoplasm, which is similar to Mx, one of the antiviral GTP binding protein. Chicken GBP2 has the highest homology to human GBP1, which is an antiviral GBP, suggesting its antiviral property.

  • マカク属(ニホンザル、アカゲザル等)に存在するウイルス抵抗性遺伝子の検索

    Grant number:19650104  2007 - 2009

    日本学術振興会  科学研究費助成事業 挑戦的萌芽研究  挑戦的萌芽研究

    山本 博, 浅野 淳, 足立 伊佐夫, 大塚 哲, 足立 伊佐雄, 安居院 高志

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    Grant amount:\3200000 ( Direct Cost: \3200000 )

    1)H20年度につづき、マカク属サルのウイルス抵抗性遺伝子の探索の目的のうちアカゲザルLLC-MK_2細胞および不死化したBLCL(B Lymphoblostoid cell line)よりpoly(I)/(C)を用い細胞を刺激したものとpoly(I)/(C)を用いないノーマル細胞をトリゾールにてTotal RNAを抽出し、Mx遺伝子の探索を目標にアカゲザルのシークエンスを参考に作製したPrimer setを用いてPCR反応により出現するバンド(327bp)について検討を行い、シークエンス解析を行った。
    2)今年はPrimerに工夫を加え、ニホンサル、アフリカミドリザルについて検査を行った。
    3)ニホンサルについてはMx2遺伝子の確認とシークエンスを解析した。Mx1遺伝子については、ニホンサル、カニクイザルについて解析を行った。
    4)アフリカミドリザルについては、Mx遺伝子の存在を確認したが、シークエンス解析は今後の検討課題である。

  • Analysis of the system for osmosensation, drinking, and body fluid regulation at the cellular, molecular and whole-body levels

    Grant number:18380175  2006 - 2008

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    SHIBUYA Izumi, KITAMURA Naoki, YAMANO Yoshiaki, ASANO Atsushi

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    Grant amount:\16930000 ( Direct Cost: \15400000 、 Indirect Cost:\1530000 )

  • Analysis of the antigen epitopes of infectious pathogens in laboratory animals using peptide tips

    Grant number:15300138  2003 - 2005

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    AGUI Takashi, ARIKAWA Jiro, ASANO Atsushi, TAKAKURA Akira, KUNIMATU Mitoshi

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    Grant amount:\16700000 ( Direct Cost: \16700000 )

    We selected mouse hepatitis virus, Sendai virus, Mycoplasma plumonis, and hantavirus from the aspect of the frequency and danger upon infectious accidents. We determined antigenic epitopes by incubating polyclonal anti-sera with peptide tips produced according to the amino acid sequences reported previously elsewhere. Further, we tried to develop a preliminary ELISA kit using antigenic epitopes that we have determined. With respect to mouse hepatitis virus and Sendai virus, we succeeded to develop an ELISA system, which was more sensitive or equal to the commercially available ELISA system (paper in preparation). With respect to hantavirus, we succeeded to determine an antigenic epitope, EDVNGIRK (amino acid 166-173), to monoclonal antibody E5/G6, which recognizes nuclear protein of hantavirus. With respect to Mycoplasma plumonis, we first searched antigenic protein from the components of the bacteria, since there are a lot of proteins composing of bacteria. We found a protein showing a homology to P46 protein, which had been reported as an antigenic protein in other Mycoplasmas and named it P46L. We produced recombinant P46L protein conjugated with glutathione S-transferase and made an ELISA system with purified P46L. This ELISA system was more sensitive than the commercially available ELISA system.
    We further applied this method to the determination of antigenic epitopes of West Nile virus and SARS virus and succeeded it. We furthermore succeeded to apply this method to not only determining antigenic epitopes of infectious pathogens but also determining protein-protein interaction site such as binding sits of FAS-antigen and angiotensin II.

  • Production of a chicken resistant strain using antiviral Mx gene

    Grant number:14360161  2002 - 2004

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    WATANABE Tomomasa, MORI Tadashi, ASANO Atsushii, SUZUKI Keita, OHARA Mutsuo

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    Grant amount:\13400000 ( Direct Cost: \13400000 )

    We have reported the polymorphism of antiviral Mx gene in the chicken, and that the specific amino acid substitution at position 631 corresponds to the differential antiviral activity. Namely, the chicken Mx gene is a resistant character in the case of Ash at the position 631 against the infectious experiment using influenza virus and vesicular stomatitis virus, and is sensitive in the case of Ser. In this study, we have established the simple method of genetic diagnosis whether each chicken carry the resistant or sensitive Mx gene by PCR-RLFP (restriction enzyme length fragment polymorphism) using genomic DNA extracted from one-drop of blood. As the result of determination in 500 indivuduals of the chicken commercial breeds, about 50% of them carry the resistant Mx gene and also about 50% have the sensitive Mx gene. In addition, it was demonstrated by the nucleotide sequence analysis of last exon that the Mx gene from only a jungle fowl (red jungle fowl) coded the resistant type of Ash, but those from the all other jungle fowl (red, gray and green jungle fowl) did the sensitive type of Ser. Furthermore, another birds (Galliformes) such as Japanese quail and and turkey carry the sensitive type of Ser. Therefore, from these results, the sensitive Mx gene was considered as the ancestor type in the bird, and the resistant Mx gene might appear with domestication of chicken.

  • ニワトリにおける抗病性遺伝形質のDNA変異と品種改良への応用

    Grant number:14656108  2002 - 2004

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    渡辺 智正, 上田 純治, 森 匡, 浅野 淳, 大原 睦生

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    Grant amount:\3200000 ( Direct Cost: \3200000 )

    私たちは、すでに30品種以上のニワトリを用いてMx遺伝子の多様性と抗病性研究を行い、in vitro感染実験の結果、631番地のアミノ酸がAsnの場合抵抗性、Serの場合感受性であることを明らかにした。今回、このニワトリMx遺伝子における変異を、少量の血液からゲノムDNAを抽出し、ミスマッチPCR-RFLP法により簡易的に抵抗性あるいは感受性形質かを診断する方法を確立した。この簡易診断法を、(独)家畜改良センター岡崎牧場のニワトリ集団に応用することを試みた。とくに、卵用鶏の最も主要品種である白色レグホーン種のMK系統について集団調査を試みた。合計2210羽を調べ、1090羽が抵抗性ホモ、998羽がヘテロ、122羽が感受性ホモであった。その中から、抵抗性ホモ330羽を選抜し、次世代生産のための交配を開始した。また、それらの集団の産卵率と卵質検査などの各項目の経済形質では有為な差は認められず、実用鶏として有効であることが確認された。感染実験を行う際の対象群として、感受性ホモ35羽からも次世代を選抜した。今後は、抵抗性ホモあるいは感受性ホモの遺伝子型を確認しながら、実用鶏としての産卵性能の改良も行っていく予定である。次に、卵用鶏の主要品種であるロードアイランドレッド(RIR)については、調べた約100羽が全て感受性ホモであったため、北大の実験家系群の抵抗性ホモ型のニワトリを導入して、抵抗性遺伝子頻度を増していく必要があると考えられた。

  • 家畜のRNAウイルス抵抗Mx遺伝子に関する研究

    Grant number:12876065  2000 - 2001

    日本学術振興会  科学研究費助成事業 萌芽的研究  萌芽的研究

    渡辺 智正, 浅野 淳

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    Grant amount:\1700000 ( Direct Cost: \1700000 )

    動物はウイルスなどの病原体に攻撃された時、防御機構が働いて感染に打ち勝とうとする。マウスにおいてこの遺伝的要因が明らかにされつつある。例えば、Mx遺伝子は、インターフェロンにより発現が誘導され、特定のRNAウイルスに対して抑制作用を示す。ところが、家畜のMx遺伝子はcDNAクローニングされたものの、変異の検出については報告がない。家畜において抵抗性・感受性型遺伝子が検出された場合、病気に強い品種を選抜することが可能となり、その意義は大きい。家畜でMx遺伝子について変異の報告がないのは、一品種しか調べられていないことが理由と考えられた。そこで、今回多数のブタおよびニワトリ品種からMx遺伝子をクローニングした。ブタでは15品種の合計341頭のゲノムDNAから最終エクソン14の塩基配列を調べたところ、ランドレースを含めた3品種に11塩基の欠失が検出され、結果として8アミノ酸の置換と23アミノ酸の延長が推測された。感染実験の結果、明らかに11塩基欠失タイプの方がインフルエンザウイルスに対して感受性でその増殖を抑える力が弱かった。一方、ニワトリの場合、受精胚からMx遺伝子のcDNAをクローニングすることが容易であり、実用鶏、日本在来種を含めて15品種のMxcDNAの全塩基配列を決定した。ニワトリMx遺伝子は多型性に富み、25箇所の塩基置換と、そのうち14箇所はアミノ酸置換を伴った。アミノ酸置換を伴うニワトリMx遺伝子のウイルス感染に対する機能的差異を検討するために、文献の白レグの感受性遺伝子と遺伝的距離の遠いと推定されたシャモとナゴヤ種のMxcDNAをマウス培養細胞に遺伝子導入した後、インフルエンザと水疱性口内炎ウイルスを用いて感染実験を行ったところ、ナゴヤ種Mxは文献と同じウイルス増殖抑制のない感受性タイプであったが、シャモは明らかに増殖抑制のある抵抗性であった。また、このウイルス感染抵抗性・感受性を決定しているのは631番地のアミノ酸置換であり、Asnの場合抵抗性であるのに対して、Serの場合感受性タイプであった。

  • 体脂肪の調節における血管新生因子の役割に関する研究

    Grant number:98J02668  1998 - 1999

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    浅野 淳

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    Grant amount:\1800000 ( Direct Cost: \1800000 )

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