MF Researcher Database

研究者総覧

詳細

山本　大介 (ﾔﾏﾓﾄ ﾀﾞｲｽｹ)

  • 感染・免疫学講座（医動物学部門）　准教授
Last Updated :2023/05/31

研究者情報

通称等の別名

    YAMAMOTO,Daisuke S.

学位

  • 博士（理学）（神戸大学）

ホームページURL

科研費研究者番号

  • 90597189

J-Global ID

プロフィール

  • 昆虫を実験材料に、発生、生殖、病原体感染などの生命現象の研究を行っています。
    現在は主に、マラリア媒介昆虫であるハマダラカを実験材料に、ゲノム編集や遺伝子組換えを利用した有用系統の作製、個体レベルでの遺伝子機能解析を行っています。これらの成果を蚊の防除やマラリア制御の技術開発につなげることを目指しています。

研究キーワード

  • 発生生物学   応用昆虫学   昆虫   ゲノム編集   遺伝子組換え昆虫   害虫防除   吸血昆虫   ハマダラカ   マラリア   生殖細胞   性決定   減数分裂   

研究分野

  • ライフサイエンス / 遺伝学 / 発生遺伝
  • 環境・農学 / 昆虫科学 / 昆虫分子生物学
  • ライフサイエンス / 寄生虫学
  • ライフサイエンス / 発生生物学 / 生殖細胞

経歴

  • 2021年08月 - 現在  自治医科大学 感染・免疫学講座 医動物学部門 准教授
  • 2017年04月 - 2021年07月  自治医科大学 感染・免疫学講座 医動物学部門 講師
  • 2010年09月 - 2017年03月  自治医科大学感染・免疫学講座 医動物学部門助教
  • 2008年04月 - 2010年08月  自治医科大学感染・免疫学講座 医動物学部門ポストドクター
  • 2005年07月 - 2008年03月  独立行政法人 農業生物資源研究所特別研究員
  • 2005年04月 - 2005年06月  独立行政法人 農業生物資源研究所非常勤職員

学歴

  • 2002年04月 - 2005年03月   神戸大学大学院   自然科学研究科   生命科学専攻（博士後期課程）
  • 2000年04月 - 2002年03月   神戸大学大学院   自然科学研究科   生物学専攻（博士前期課程）
  • 1996年04月 - 2000年03月   神戸大学   理学部   生物学科

所属学協会

  • 日本応用動物昆虫学会   日本節足動物発生学会   日本動物学会   日本寄生虫学会   日本衛生動物学会   

研究活動情報

論文

  • Naoaki Shinzawa, Chisako Kashima, Hiroka Aonuma, Kei Takahashi, Masayuki Shimojima, Shinya Fukumoto, Erisha Saiki, Daisuke S. Yamamoto, Shigeto Yoshida, Hiroyuki Matsuoka, Yoshihiro Kawaoka, Hirotaka Kanuka
    Frontiers in Tropical Diseases 3 2022年05月 [査読有り][通常論文]
     
    Live microbe vaccines are designed to elicit strong cellular and antibody responses without developing the symptoms of the disease, and these are effective in preventing infectious diseases. A flying vaccinator (also known as a flying syringe) is a conceptual, genetically engineered hematophagous insect that is used to deliver vaccines such as an antigen from a parasite produced in mosquito saliva; bites from such insects may elicit antibody production by immunizing the host with an antigen through blood-feeding. In addition to a simple vaccine antigen, a flying vaccinator may potentially load a live attenuated microbe with an appropriate mechanism for sustaining its constitutive proliferation in the insect. In this study, a recombinant vesicular stomatitis virus (VSV) lacking the glycoprotein gene (VSV-G) was used to produce replication-restricted VSV (rrVSV) containing GFP. Transgenic Anopheles stephensi mosquitoes, in which the salivary glands expressed a VSV-G gene driven by an aapp salivary gland-specific promoter, were generated and injected intraperitoneally with rrVSV. The injected rrVSV entered the cells of the salivary gland and stimulated endogenous production of progeny rrVSV particles, as seen in rrVSV-infected Drosophila melanogaster expressing VSV-G. These data suggested the possibility of developing a valuable tool for delivering genetically attenuated virus vaccines via mosquito saliva, although efficient replication-restricted virus production is required.
  • Ahmed Tabbabi, Daiki Mizushima, Daisuke S. Yamamoto, Hirotomo Kato
    Parasitologia 2 2 71 - 87 2022年04月 [査読有り][通常論文]
     
    Sand flies are a significant public health concern in many parts of the world where they are known to transmit agents of several zoonotic diseases to humans, such as leishmaniasis. Vector control remains a key component of many anti-leishmaniasis programs and probably will remain so until an effective vaccine becomes available. The sand fly gut microbiota has recently emerged as an encouraging field for the exploration of vector-based disease control. In particular, the gut microbiome was previously reported to either enhance or inhibit parasite activity depending on the species of bacteria and, thus, has the potential to alter vector competence. Here, we describe the technological advances that are currently expanding our understanding of microbiota composition in sand flies. The acquisition and composition of microbiomes are influenced by several abiotic and biotic factors, including host immunity, genetics, and the environment. Therefore, the microbiomes of sand flies can vary substantially between individuals, life stages, species, and over geographical space, and this variation likely contributes to differences in host phenotypes, highlighting opportunities for novel vector control strategies.
  • Mohammad Shahnaij, Mitsuhiro Iyori, Hiroaki Mizukami, Mayu Kajino, Iroha Yamagoshi, Intan Syafira, Yenni Yusuf, Ken Fujiwara, Daisuke S Yamamoto, Hirotomo Kato, Nobuhiko Ohno, Shigeto Yoshida
    Frontiers in immunology 12 612910 - 612910 2021年 [査読有り]
     
    Hepatocyte infection by malaria sporozoites is a bottleneck in the life-cycle of Plasmodium spp. including P. falciparum, which causes the most lethal form of malaria. Therefore, developing an effective vaccine capable of inducing the strong humoral and cellular immune responses necessary to block the pre-erythrocytic stage has potential to overcome the spatiotemporal hindrances pertaining to parasite biology and hepatic microanatomy. We recently showed that when combined with a human adenovirus type 5 (AdHu5)-priming vaccine, adeno-associated virus serotype 1 (AAV1) is a potent booster malaria vaccine vector capable of inducing strong and long-lasting protective immune responses in a rodent malaria model. Here, we evaluated the protective efficacy of a hepatotropic virus, adeno-associated virus serotype 8 (AAV8), as a booster vector because it can deliver a transgene potently and rapidly to the liver, the organ malaria sporozoites initially infect and multiply in following sporozoite injection by the bite of an infected mosquito. We first generated an AAV8-vectored vaccine expressing P. falciparum circumsporozoite protein (PfCSP). Intravenous (i.v.) administration of AAV8-PfCSP to mice initially primed with AdHu5-PfCSP resulted in a hepatocyte transduction rate ~2.5 times above that seen with intramuscular (i.m.) administration. This immunization regimen provided a better protection rate (100% sterile protection) than that of the i.m. AdHu5-prime/i.m. AAV8-boost regimen (60%, p < 0.05), i.m. AdHu5-prime/i.v. AAV1-boost (78%), or i.m. AdHu5-prime/i.m. AAV1-boost (80%) against challenge with transgenic PfCSP-expressing P. berghei sporozoites. Compared with the i.m. AdHu5-prime/i.v. AAV1-boost regimen, three other regimens induced higher levels of PfCSP-specific humoral immune responses. Importantly, a single i.v. dose of AAV8-PfCSP recruited CD8+ T cells, especially resident memory CD8+ T cells, in the liver. These data suggest that boost with i.v. AAV8-PfCSP can improve humoral and cellular immune responses in BALB/c mice. Therefore, this regimen holds great promise as a next-generation platform for the development of an effective malaria vaccine.
  • Hirotomo Kato, Abraham G Cáceres, Eduardo A Gomez, Ahmed Tabbabi, Daiki Mizushima, Daisuke S Yamamoto, Yoshihisa Hashiguchi
    Frontiers in cellular and infection microbiology 11 625001 - 625001 2021年 [査読有り]
     
    Approximately 20 Leishmania species are known to cause cutaneous, mucocutaneous, and visceral disorders in humans. Identification of the causative species in infected individuals is important for appropriate treatment and a favorable prognosis because infecting species are known to be the major determinant of clinical manifestations and may affect treatments for leishmaniasis. Although Leishmania species have been conventionally identified by multilocus enzyme electrophoresis, genetic analysis targeting kinetoplast and nuclear DNA (kDNA and nDNA, respectively) is now widely used for this purpose. Recently, we conducted countrywide epidemiological studies of leishmaniasis in Ecuador and Peru to reveal prevalent species using PCR-RFLP targeting nDNA, and identified unknown hybrid parasites in these countries together with species reported previously. Furthermore, comparative analyses of kDNA and nDNA revealed the distribution of parasites with mismatches between these genes, representing the first report of mito-nuclear discordance in protozoa. The prevalence of an unexpectedly high rate (~10%) of genetically complex strains including hybrid strains, in conjunction with the observation of mito-nuclear discordance, suggests that genetic exchange may occur more frequently than previously thought in natural Leishmania populations. Hybrid Leishmania strains resulting from genetic exchanges are suggested to cause more severe clinical symptoms when compared with parental strains, and to have increased transmissibility by vectors of the parental parasite species. Therefore, it is important to clarify how such genetic exchange influences disease progression and transmissibility by sand flies in nature. In addition, our aim was to identify where and how the genetic exchange resulting in the formation of hybrid and mito-nuclear discordance occurs.
  • Ahmed Tabbabi, Shinya Watanabe, Daiki Mizushima, Abraham G Caceres, Eduardo A Gomez, Daisuke S Yamamoto, Longzhu Cui, Yoshihisa Hashiguchi, Hirotomo Kato
    Microorganisms 9 1 2020年12月 [査読有り]
     
    Differences in the gut microbial content of Lutzomyia (Lu.) ayacuchensis, a primary vector of Andean-type cutaneous leishmaniasis in Ecuador and Peru, may influence the susceptibility of these sand flies to infection by Leishmania. As a first step toward addressing this hypothesis, a comparative analysis of bacterial and fungal compositions from Lu. ayacuchensis populations with differential susceptibilities to Leishmania was performed. Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing approaches were used to characterize the bacterial composition in wild-caught populations from the Andean areas of Ecuador and southern Peru at which the sand fly species transmit Leishmania (Leishmania) mexicana and Leishmania (Viannia) peruviana, respectively, and a population from the northern Peruvian Andes at which the transmission of Leishmania by Lu. ayacuchensis has not been reported. In the present study, 59 genera were identified, 21 of which were widely identified and comprised more than 95% of all bacteria. Of the 21 dominant bacterial genera identified in the sand flies collected, 10 genera had never been detected in field sand flies. The Ecuador and southern Peru populations each comprised individuals of particular genera, while overlap was clearly observed between microbes isolated from different sites, such as the number of soil organisms. Similarly, Corynebacterium and Micrococcus were slightly more dominant bacterial genera in the southern Peru population, while Ochrobactrum was the most frequently isolated from other populations. On the other hand, fungi were only found in the southern Peru population and dominated by the Papiliotrema genus. These results suggest that variation in the insect gut microbiota may be elucidated by the ecological diversity of sand flies in Peru and Ecuador, which may influence susceptibility to Leishmania infection. The present study provides key insights for understanding the role of the microbiota during the course of L. (L.) mexicana and L. (V.) peruviana infections in this important vector.
  • Ahmed Tabbabi, Abraham G Cáceres, T Pershing Bustamante Chauca, Chisato Seki, Yanisa Choochartpong, Daiki Mizushima, Daisuke S Yamamoto, Yoshihisa Hashiguchi, Hirotomo Kato
    PLoS neglected tropical diseases 14 10 e0008797  2020年10月 [査読有り]
     
    Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mannose phosphate isomerase (mpi) gene was applied to 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru for identification of the infecting parasite at the species level, and the results were compared with those of cytochrome b (cyt b) gene sequencing obtained in previous studies. Although most results (121/134) including 4 hybrids of Leishmania (Viannia) braziliensis and L. (V.) peruviana corresponded to those obtained in the previous study, PCR-RFLP analyses revealed the distribution of putative hybrid strains between L. (V.) peruviana and L. (V.) lainsoni in two samples, which has never been reported. Moreover, parasite strains showing discordance between kinetoplast and nuclear genes (kDNA and nDNA), so-called mito-nuclear discordance, were identified in 11 samples. Of these, six strains had the kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) guyanensis, and three strains had the kDNAs of L. (V.) shawi and nDNAs of L. (V.) braziliensis. The rest were identified as mito-nuclear discordance strains having kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) lainsoni, and kDNAs of L. (V.) lainsoni and nDNAs of L. (V.) braziliensis. The results demonstrate that Leishmania strains in Peru are genetically more complex than previously considered.
  • Daiki Mizushima, Ahmed Tabbabi, Daisuke S Yamamoto, Le Trung Kien, Hirotomo Kato
    Acta tropica 210 105473 - 105473 2020年10月 [査読有り][通常論文]
     
    Salivary gland transcriptome analysis of the Asiatic Triatoma rubrofasciata was performed by high-throughput RNA sequencing. This analysis showed that the majority of reads accounting for 85.38% FPKM (fragments per kilobase of exon per million mapped fragments) were mapped with a secreted class. Of these, the most abundant subclass accounting for 89.27% FPKM was the lipocalin family. In the lipocalin family, the most dominant molecules making up 70.49% FPKM were homologues of procalin, a major allergen identified from T. protracta saliva, suggesting an important role in blood-sucking of T. rubrofasciata. Other lipocalins showed similarities to pallidipin and triplatin, inhibitors of collagen-induced platelet aggregation identified from T. pallidipennis and T. infestans, respectively, Td38 from T. dimidiata with unknown function, triatin-like lipocalin with unknown function, and triafestin, an inhibitor of the activation of the kallikrein-kinin system, identified from T. infestans saliva. Other than lipocalin family proteins, homologues of antigen-5 (3.38% FPKM), Kazal-type serine protease inhibitor (1.36% FPKM), inositol polyphosphate 5-phosphatase (1.32% FPKM), and apyrase/5'-nucleotidase (0.64% FPKM) were identified as abundant molecules in T. rubrofasciata saliva. Through this study, de novo assembly of 42,580,822 trimmed reads generated 35,781 trinity transcripts, and a total of 1,272 coding sequences for the secreted class were deposited in GenBank. The results provide further insights into the evolution of salivary components in blood-sucking arthropods.
  • Jan Philip Oeyen, Patrice Baa-Puyoulet, Joshua B Benoit, Leo W Beukeboom, Erich Bornberg-Bauer, Anja Buttstedt, Federica Calevro, Elizabeth I Cash, Hsu Chao, Hubert Charles, Mei-Ju May Chen, Christopher Childers, Andrew G Cridge, Peter Dearden, Huyen Dinh, Harsha Vardhan Doddapaneni, Amanda Dolan, Alexander Donath, Daniel Dowling, Shannon Dugan, Elizabeth Duncan, Elena N Elpidina, Markus Friedrich, Elzemiek Geuverink, Joshua D Gibson, Sonja Grath, Cornelis J P Grimmelikhuijzen, Ewald Große-Wilde, Cameron Gudobba, Yi Han, Bill S Hansson, Frank Hauser, Daniel S T Hughes, Panagiotis Ioannidis, Emmanuelle Jacquin-Joly, Emily C Jennings, Jeffery W Jones, Steffen Klasberg, Sandra L Lee, Peter Lesný, Mackenzie Lovegrove, Sebastian Martin, Alexander G Martynov, Christoph Mayer, Nicolas Montagné, Victoria C Moris, Monica Munoz-Torres, Shwetha Canchi Murali, Donna M Muzny, Brenda Oppert, Nicolas Parisot, Thomas Pauli, Ralph S Peters, Malte Petersen, Christian Pick, Emma Persyn, Lars Podsiadlowski, Monica F Poelchau, Panagiotis Provataris, Jiaxin Qu, Maarten J M F Reijnders, Björn Marcus von Reumont, Andrew J Rosendale, Felipe A Simao, John Skelly, Alexandros G Sotiropoulos, Aaron L Stahl, Megumi Sumitani, Elise M Szuter, Olivia Tidswell, Evangelos Tsitlakidis, Lucia Vedder, Robert M Waterhouse, John H Werren, Jeanne Wilbrandt, Kim C Worley, Daisuke S Yamamoto, Louis van de Zande, Evgeny M Zdobnov, Tanja Ziesmann, Richard A Gibbs, Stephen Richards, Masatsugu Hatakeyama, Bernhard Misof, Oliver Niehuis
    Genome biology and evolution 12 7 1099 - 1188 2020年07月 [査読有り][通常論文]
     
    The tremendous diversity of Hymenoptera is commonly attributed to the evolution of parasitoidism in the last common ancestor of parasitoid sawflies (Orussidae) and wasp-waisted Hymenoptera (Apocrita). However, Apocrita and Orussidae differ dramatically in their species richness, indicating that the diversification of Apocrita was promoted by additional traits. These traits have remained elusive due to a paucity of sawfly genome sequences, in particular those of parasitoid sawflies. Here, we present comparative analyses of draft genomes of the primarily phytophagous sawfly Athalia rosae and the parasitoid sawfly Orussus abietinus. Our analyses revealed that the ancestral hymenopteran genome exhibited traits that were previously considered unique to eusocial Apocrita (e.g., low transposable element content and activity) and a wider gene repertoire than previously thought (e.g., genes for CO2 detection). Moreover, we discovered that Apocrita evolved a significantly larger array of odorant receptors than sawflies, which could be relevant to the remarkable diversification of Apocrita by enabling efficient detection and reliable identification of hosts.
  • Daiki Mizushima, Ahmed Tabbabi, Daisuke S Yamamoto, Le Trung Kien, Hirotomo Kato
    Data in brief 30 105647 - 105647 2020年06月 [査読有り][通常論文]
     
    The dataset in this report is related to the research article entitled: "Salivary gland transcriptome of the Asiatic Triatoma rubrofasciata" [1]. Lipocalin family proteins were identified as the dominant component in T. rubrofasciata saliva, and phylogenetic analysis of the salivary lipocalins resulted in the formation of five major clades (clade I-V). For further characterization, each clade of T. rubrofasciata lipocalin was subjected to alignment and phylogenetic analyses together with homologous triatomine lipocalins: procalin, a major allergen in T. protracta saliva and its homologue Td04 from T. dimidiata (clade I), pallidipin and triplatin, inhibitors of collagen-induced platelet aggregation identified from T. pallidipennis and T. infestans, respectively, and their homologue Pc20 identified from Panstrongylus chinai (clade II), Td30 and Td38 from T. dimidiata with unknown functions (clade III), triatin-like salivary lipocalins, Pc58 and Pc226 identified from P. chinai and Td18 from T. dimidiata (clade IV), and triafestin, an inhibitor of the activation of the kallikrein-kinin system, identified from T. infestans saliva and its homologues, Td25 and Td40 from T. dimidiata and Pc64 from P. chinai (clade V).
  • Tanaka T, Tanaka N, Nagano N, Kanuka H, Yamamoto DS, Yamamoto N, Nanba E, Nishiuchi T
    Journal of Environment and Safety 11 2 31 - 35 2020年 [査読有り][通常論文]
  • Kato H, Cáceres AG, Seki C, Silupu García CR, Holguín Mauricci C, Castro Martínez SC, Moreno Paico D, Castro Muniz JL, Troyes Rivera LD, Villegas Briones ZI, Guerrero Quincho S, Sulca Jayo GL, Tineo Villafuerte E, Manrique de Lara, Estrada C, Arias FR, Passara FS, Ruelas Llerena N, Kubo M, Tabbabi A, Yamamoto DS, Hashiguchi Y
    PLoS neglected tropical diseases 13 6 e0007496  2019年06月 [査読有り][通常論文]
     
    To obtain further insight into geographic distribution of Leishmania species in Peru, a countrywide survey, including central to southern rainforest areas where information on causative parasite species is limited, was performed based on cytochrome b (cyt b) and mannose phosphate isomerase (mpi) gene analyses. A total of 262 clinical samples were collected from patients suspected of cutaneous leishmaniasis (CL) in 28 provinces of 13 departments, of which 99 samples were impregnated on FTA (Flinders Technology Associates) cards and 163 samples were Giemsa-stained smears. Leishmania species were successfully identified in 83 (83.8%) of FTA-spotted samples and 59 (36.2%) of Giemsa-stained smear samples. Among the 142 samples identified, the most dominant species was Leishmania (Viannia) braziliensis (47.2%), followed by L. (V.) peruviana (26.1%), and others were L. (V.) guyanensis, L. (V.) lainsoni, L. (V.) shawi, a hybrid of L. (V.) braziliensis and L. (V.) peruviana, and Leishmania (Leishmania) amazonensis. Besides the present epidemiological observations, the current study provided the following findings: 1) A hybrid of L. (V.) braziliensis and L. (V.) peruviana is present outside the Department of Huanuco, the only place reported, 2) Many cases of CL due to L. (V.) lainsoni, an uncommon causative species in Peru, were observed, and 3) L. (V.) shawi is widely circulating in southern Amazonian areas in Peru.
  • Yamamoto DS, Sumitani M, Kasashima K, Sezutsu H, Matsuoka H, Kato H
    Scientific reports 9 1 8160 - 8160 2019年06月 [査読有り][通常論文]
     
    Conditional cell death systems are useful for various aspects of basic science with a wide range of applications, including genetic pest control. We recently demonstrated that expression of the mammalian pro-apoptotic factor, B-cell leukaemia/lymphoma 2-associated X protein (Bax), can induce apoptosis in specific tissues by using tissue specific promoters in silkworm and mosquito. Here, we newly identified a functional promoter in the Asian malaria vector, Anopheles stephensi, which enables gene expression specifically in the testis. We produced a transgenic mosquito line that expresses mouse Bax under the control of this testis-specific promoter. Transgenic mosquito males exhibited aberrant testes without functional sperm and complete sterility, whereas transgenic females maintained normal fecundity. Despite their abnormal testes, the transgenic males maintained normal function of male accessory glands and typical mating behaviour. As a result of mating with these males, females showed refractoriness to further mating. These results suggest that transgenic males induce female sterility via mating. The mosquito is one of the most important disease vectors, and the control of their population benefits global public health. Thus, this Bax-mediated synthetic male-specific sterilization system could be applied to population control of mosquitoes.
  • Amelia F, Iyori M, Emran TB, Yamamoto DS, Genshi K, Otsuka H, Onoue Y, Yusuf Y, Islam A, Yoshida S
    Parasite immunology 41 5 e12624  2019年05月 [査読有り][通常論文]
     
    Plasmodium falciparum circumsporozoite protein (PfCSP) is the main target antigen in development of pre-erythrocytic malaria vaccines. To evaluate PfCSP vaccines in animal models, challenge by intravenous sporozoite injection is preferentially used. However, in clinical trials, vaccinated human volunteers are exposed to the bites of malaria-infected mosquitoes. In this study, we down-selected Escherichia coli-produced full-length PfCSP (PfCSP-F) and its three truncated PfCSPs based on their abilities to elicit immune response and protection in mice against two challenge models. We showed that immunization with three doses of PfCSP-F elicited high anti-PfCSP antibody titres and 100% protection against the bites of infected mosquitoes. Meanwhile, three-dose truncated PfCSP induced 60%-70% protection after immunization with each truncated PfCSP. Heterologous prime-boost immunization regimen with adenovirus-PfCSP-F and R32LR greatly induced complete protection against intravenous sporozoite injection. Our results suggest that Abs to both anti-repeat and anti-nonrepeat regions induced by PfCSP-F are required to confer complete protection against challenge by the bites of infected mosquitoes, whereas anti-repeat Abs play an important role in protection against intravenous sporozoite injection. Our findings provide a potential clinical application that PfCSP-F vaccine induces potent Abs capable of neutralizing sporozoites in the dermis inoculated by infected mosquitoes and subsequently sporozoites in the blood circulation.
  • Kato H, Gomez EA, Seki C, Furumoto H, Martini-Robles L, Muzzio J, Calvopiña M, Velez L, Kubo M, Tabbabi A, Yamamoto DS, Hashiguchi Y
    PLoS neglected tropical diseases 13 5 e0007403  2019年05月 [査読有り][通常論文]
     
    PCR-Restriction Fragment Length Polymorphism (RFLP) analyses targeting multiple nuclear genes were established for the simple and practical identification of Leishmania species without using expensive equipment. This method was applied to 92 clinical samples collected at 33 sites in 14 provinces of Ecuador, which have been identified at the species level by the kinetoplast cytochrome b (cyt b) gene sequence analysis, and the results obtained by the two analyses were compared. Although most results corresponded between the two analyses, PCR-RFLP analyses revealed distribution of hybrid strains between Leishmania (Viannia) guyanensis and L. (V.) braziliensis and between L. (V.) guyanensis and L. (V.) panamensis, of which the latter was firstly identified in Ecuador. Moreover, unexpected parasite strains having the kinetoplast cyt b gene of L. (V.) braziliensis and nuclear genes of L. (V.) guyanensis, L. (V.) panamensis, or a hybrid between L. (V.) guyanensis and L. (V.) panamensis were identified. This is the first report of the distribution of a protozoan parasite having mismatches between kinetoplast and nuclear genes, known as mito-nuclear discordance. The result demonstrated that genetically complex Leishmania strains are present in Ecuador. Since genetic exchanges such as hybrid formation were suggested to cause higher pathogenicity in Leishmania and may be transmitted by more species of sand flies, further country-wide epidemiological studies on clinical symptoms, as well as transmissible vectors, will be necessary.
  • Islam A, Emran TB, Yamamoto DS, Iyori M, Amelia F, Yusuf Y, Yamaguchi R, Alam MS, Silveira H, Yoshida S
    Scientific reports 9 1 3129 - 3129 2019年02月 [査読有り][通常論文]
     
    The saliva of hematophagous arthropods is enriched with a complex mixture of antihemostatic molecules, the biological functions of which are largely unknown. Anopheline antiplatelet protein (AAPP) from malaria vector mosquito exhibits strong antiplatelet activity when bound directly to host collagen by its C-terminus and through its N-terminus with Ca2+-binding activity. To investigate the biological functions of AAPP in blood feeding behavior and malaria transmission, we generated transgenic Anopheles stephensi mosquito lines expressing anti-AAPP antibody single-chain fragment (scFv) in their salivary glands. The AAPP-specific collagen-binding activity was completely abolished by AAPP-scFv complex formation in the saliva. Probing and prediuresis time, feeding success, blood meal size, and fecundity, which are all fitness characteristics, were significantly reduced in the transgenic mosquitoes. However, oocysts number in these mosquitoes were not significantly reduced following blood meal intake from Plasmodium berghei-infected mice. These results show that although AAPP plays an important role in mosquito blood feeding, its neutralizing activity did not affect sporogonic development in our laboratory model, but its high fitness cost would pose a survival risk for parasite-infected mosquitoes in nature.
  • Yusuf Y, Yoshii T, Iyori M, Mizukami H, Fukumoto S, Yamamoto DS, Emran TB, Amelia F, Islam A, Syafira I, Yoshida S
    Frontiers in immunology 10 2412 - 2412 2019年 [査読有り][通常論文]
     
    Malaria parasites undergo several stages in their complex lifecycle. To achieve reductions in both the individual disease burden and malaria transmission within communities, a multi-stage malaria vaccine with high effectiveness and durability is a more efficacious strategy compared with a single-stage vaccine. Here, we generated viral-vectored vaccines based on human adenovirus type 5 (AdHu5) and adeno-associated virus serotype 1 (AAV1) expressing a fusion protein of the pre-erythrocytic stage Plasmodium falciparum circumsporozoite protein (PfCSP) and the transmission-blocking sexual stage P25 protein (Pfs25). A two-dose heterologous AdHu5-prime/AAV1-boost immunization regimen proved to be highly effective for both full protection and transmission-blocking activity against transgenic P. berghei parasites expressing the corresponding P. falciparum antigens in mice. Remarkably, the immunization regimen induced antibody responses to both PfCSP and Pfs25 for over 9 months after the boosting and also maintained high levels of transmission-reducing activity (TRA: >99%) during that period, as evaluated by a direct feeding assay. If similar efficacies on P. falciparum can be shown following vaccination of humans, we propose that this multi-stage malaria vaccine regimen will be a powerful tool for malaria control, providing greater overall protection and cost-effectiveness than single-stage vaccines.
  • Yusuf Y, Yoshii T, Iyori M, Yoshida K, Mizukami H, Fukumoto S, Yamamoto DS, Alam A, Emran TB, Amelia F, Islam A, Otsuka H, Takashima E, Tsuboi T, Yoshida S
    Frontiers in immunology 10 730 - 730 2019年 [査読有り][通常論文]
     
    An ideal malaria vaccine platform should potently induce protective immune responses and block parasite transmission from mosquito to human, and it should maintain these effects for an extended period. Here, we have focused on vaccine development based on adeno-associated virus serotype 1 (AAV1), a viral vector widely studied in the field of clinical gene therapy that is able to induce long-term transgene expression without causing toxicity in vivo. Our results show the potential utility of AAV1 vectors as an extremely potent booster vaccine to induce durable immunity when combined with an adenovirus-priming vaccine in a rodent malaria model. We generated a series of recombinant AAV1s and human adenovirus type 5 (AdHu5) expressing either Plasmodium falciparum circumsporozoite protein (PfCSP) or P25 (Pfs25) protein. Heterologous two-dose immunization with an AdHu5-prime and AAV1-boost (AdHu5-AAV1) elicited robust and durable PfCSP- or Pfs25-specific functional antibodies over 280 days. Regarding protective efficacy, AdHu5-AAV1 PfCSP achieved high sterile protection (up to 80% protection rate) against challenge with transgenic Plasmodium berghei sporozoites expressing PfCSP. When examining transmission-blocking (TB) efficacy, we found that immunization with AdHu5-AAV1 Pfs25 maintained TB activity in vivo against transgenic P. berghei expressing Pfs25 for 287 days (99% reduction in oocyst intensity, 85% reduction in oocyst prevalence). Our data indicate that AAV1-based malaria vaccines can confer potent and durable protection as well as TB efficacy when administered following an AdHu5 priming vaccine, supporting the further evaluation of this regimen in clinical trials as a next-generation malaria vaccine platform.
  • Shimada M, Hirose Y, Shimizu K, Yamamoto DS, Hayakawa EH, Matsuoka H
    Tropical medicine and health 47 18  2019年 [査読有り][通常論文]
  • Emran TB, Iyori M, Ono Y, Amelia F, Yusuf Y, Islam A, Alam A, Tamura M, Ogawa R, Matsuoka H, Yamamoto DS, Yoshida S
    Journal of immunology (Baltimore, Md. : 1950) 201 8 2441 - 2451 2018年10月 [査読有り][通常論文]
     
    Baculovirus (BV), an enveloped insect virus with a circular dsDNA genome, possesses unique characteristics that induce strong innate immune responses in mammalian cells. In this study, we show that BV administration in BALB/c mice not only provides complete protection against a subsequent Plasmodium berghei sporozoite infection for up to 7 d after the injection but also eliminates existing liver-stage parasites completely. The elimination of sporozoites by BV was superior to that by primaquine, and this effect occurred in a TLR9-independent manner. At 6 h after BV administration, IFN-α and IFN-γ were robustly produced in the serum, and RNA transcripts of IFN-stimulated genes were markedly upregulated in the liver compared with control mice. The in vivo passive transfer of serum after BV administration effectively eliminated liver-stage parasites, and IFN-α neutralization abolished this effect, indicating that the BV liver-stage parasite-killing mechanism is downstream of the type I IFN signaling pathway. These findings provide evidence that BV-induced, fast-acting innate immunity completely kills liver-stage parasites and, thus, may lead to new malaria drug and vaccine strategies.
  • Yoshida K, Iyori M, Blagborough AM, Salman AM, Dulal P, Sala KA, Yamamoto DS, Khan SM, Janse CJ, Biswas S, Yoshii T, Yusuf Y, Tokoro M, Hill AVS, Yoshida S
    Scientific reports 8 1 3896  2018年03月 [査読有り][通常論文]
     
    With the increasing prevalence of artemisinin-resistant malaria parasites, a highly efficacious and durable vaccine for malaria is urgently required. We have developed an experimental virus-vectored vaccine platform based on an envelope-modified baculovirus dual-expression system (emBDES). Here, we show a conceptually new vaccine platform based on an adenovirus-prime/emBDES-boost heterologous immunization regimen expressing the Plasmodium falciparum circumsporozoite protein (PfCSP). A human adenovirus 5-prime/emBDES-boost heterologous immunization regimen consistently achieved higher sterile protection against transgenic P. berghei sporozoites expressing PfCSP after a mosquito-bite challenge than reverse-ordered or homologous immunization. This high protective efficacy was also achieved with a chimpanzee adenovirus 63-prime/emBDES-boost heterologous immunization regimen against an intravenous sporozoite challenge. Thus, we show that the adenovirus-prime/emBDES-boost heterologous immunization regimen confers sterile protection against sporozoite challenge by two individual routes, providing a promising new malaria vaccine platform for future clinical use.
  • Yamamoto DS, Sumitani M, Hatakeyama M, Matsuoka H
    Transgenic Research 27 1 51 - 60 2018年02月 [査読有り][通常論文]
     
    Anopheline mosquitoes are major vectors of malaria parasites. When the gametocytes of the malaria parasite are transferred from a vertebrate to mosquitoes, they differentiate into gametes, and are fertilized in the midguts of mosquitoes. Xanthurenic acid (XA), a waste product of the ommochrome synthesis pathway, has been shown to induce exflagellation during microgametogenesis in vitro however, it currently remains unclear whether endogenous XA affects the infectivity of anopheline mosquitoes to malaria parasites in vivo due to the lack of appropriate experimental systems such as a XA-deficient line. In the present study, we produced a XA-deficient line in Anopheles stephensi using transcription activator-like effector nuclease (TALEN)-mediated gene targeting (knockout) of the kynurenine 3-monooxygenase (kmo) gene, which encodes an enzyme that participates in the ommochrome synthesis pathway. The knockout of kmo resulted in the absence of XA, and oocyst formation was inhibited in the midguts of these XA-deficient mosquitoes, which, in turn, reduced sporozoite numbers in their salivary glands. These results suggest that endogenous XA stimulates exflagellation, and enhances the infectivity of anopheline mosquitoes to malaria parasites in vivo. The XA-deficient line of the anopheline mosquito provides a useful system for analyzing and understanding the associated factors of malaria gametogenesis in the mosquito midgut.
  • Iyori M, Yamamoto DS, Sakaguchi M, Mizutani M, Ogata S, Nishiura H, Tamura T, Matsuoka H, Yoshida S
    Malaria journal 16 1 390  2017年09月 [査読有り][通常論文]
     
    Background: Previous studies have shown that the baculovirus-vectored vaccine based on the "baculovirus dual expression system (BDES)" is an effective vaccine delivery platform for malaria. However, a point of weakness remaining for use of this vaccine platform in vivo concerns viral inactivation by serum complement. In an effort to achieve complement resistance, the gene encoding the human decay-accelerating factor (hDAF) was incorporated into the BDES malaria vaccine expressing the Plasmodium falciparum circumsporozoite protein (PfCSP). Results: The newly-developed BDES vaccine, designated BDES-sPfCSP2-Spider, effectively displayed hDAF and PfCSP on the surface of the viral envelope, resulting in complement resistance both in vitro and in vivo. Importantly, upon intramuscular inoculation into mice, the BDES-sPfCSP2-Spider vaccine had a higher protective efficacy (60%) than that of the control vaccine BDES-sPfCSP2-Spier (30%) against challenge with transgenic Plasmodium berghei sporozoites expressing PfCSP. Conclusion: DAF-shielded BDES-vaccines offer great potential for development as a new malaria vaccine platform against the sporozoite challenge.
  • Yamamoto DS, Sumitani M, Kasashima K, Sezutsu H, Matsuoka H
    PLoS Pathogens 12 9 e1005872  2016年09月 [査読有り][通常論文]
     
    Malaria is an important global public health challenge, and is transmitted by anopheline mosquitoes during blood feeding. Mosquito vector control is one of the most effective methods to control malaria, and population replacement with genetically engineered mosquitoes to block its transmission is expected to become a new vector control strategy. The salivary glands are an effective target tissue for the expression of molecules that kill or inactivate malaria parasites. Moreover, salivary gland cells express a large number of molecules that facilitate blood feeding and parasite transmission to hosts. In the present study, we adapted a functional deficiency system in specific tissues by inducing cell death using the mouse Bcl-2-associated X protein ( Bax) to the Asian malaria vector mosquito, Anopheles stephensi. We applied this technique to salivary gland cells, and produced a transgenic strain containing extremely low amounts of saliva. Although probing times for feeding on mice were longer in transgenic mosquitoes than in wild-type mosquitoes, transgenic mosquitoes still successfully ingested blood. Transgenic mosquitoes also exhibited a significant reduction in oocyst formation in the midgut in a rodent malaria model. These results indicate that mosquito saliva plays an important role in malaria infection in the midgut of anopheline mosquitoes. The dysfunction in the salivary glands enabled the inhibition of malaria transmission from hosts to mosquito midguts. Therefore, salivary components have potential in the development of new drugs or genetically engineered mosquitoes for malaria control.
  • Reza M, Yamamoto DS, Matsuoka H
    Medical Entomology and Zoology 65 3 147 - 150 2014年09月 [査読有り][通常論文]
  • Yamamoto DS, Yokomine T, Sumitani M, Yagi K, Matsuoka H, Yoshida S
    Insect molecular biology 22 6 685 - 693 6 2013年12月 [査読有り][通常論文]
     
    Mosquitoes inject saliva into a vertebrate host during blood feeding. The analysis of mosquito saliva in host skin is important for the elucidation of the inflammatory responses to mosquito bites, the development of antithrombotic drugs, and the transmission-blocking of vector-borne diseases. We produced transgenic Anopheles stephensi mosquitoes expressing the secretory luciferase protein (MetLuc) fused to a saliva protein (AAPP) in the salivary glands. The transgene product (AAPP-MetLuc) of transgenic mosquitoes exhibited both luciferase activity as a MetLuc and binding activity to collagen as an AAPP. The detection of luminescence in the skin of mice bitten by transgenic mosquitoes showed that AAPP-MetLuc was injected into the skin as a component of saliva via blood feeding. AAPP-MetLuc remained at the mosquito bite site in host skin with luciferase activity for at least 4h after blood feeding. AAPP was also suspected of remaining at the site of injury caused by the mosquito bite and blocking platelet aggregation by binding to collagen. These results demonstrated the establishment of visualization and time-lapse analysis of mosquito saliva in living vertebrate host skin. This technique may facilitate the analysis of mosquito saliva after its injection into host skin, and the development of new drugs and disease control strategies.
  • Yamamoto DS, Hatakeyama M, Matsuoka H
    The Journal of experimental biology 216 15 2960 - 2966 Pt 15 2013年08月 [査読有り][通常論文]
     
    In the past decade, many transgenic lines of mosquitoes have been generated and analyzed, whereas the maintenance of a large number of transgenic lines requires a great deal of effort and cost. In vitro fertilization by an injection of cryopreserved sperm into eggs has been proven to be effective for the maintenance of strains in mammals. The technique of artificial egg activation is a prerequisite for the establishment of in vitro fertilization by sperm injection. We demonstrated that artificial egg activation is feasible in the malaria vector mosquito, Anopheles stephensi (Diptera, Culicidae). Nearly 100% of eggs dissected from virgin females immersed in distilled water darkened, similar to normally oviposited fertilized eggs. It was revealed by the cytological examination of chromosomes that meiotic arrest was relieved in these eggs approximately 20 min after incubation in water. Biochemical examinations revealed that MAPK (mitogen-activated protein kinase)/ERK (extracellular signal-regulated protein kinase) and MEK (MAPK/ERK kinase) were dephosphorylated similar to that in fertilized eggs. These results indicate that dissected unfertilized eggs were activated in distilled water and started development. Injection of distilled water into body cavity of the virgin blood-fed females also induced activation of a portion of eggs in the ovaries. The technique of artificial egg activation is expected to contribute to the success of in vitro fertilization in A. stephensi.
  • Reza M, Yamamoto DS, Matsuoka H
    Medical Entomology and Zoology 64 2 67 - 71 2013年06月 [査読有り][通常論文]
  • Sumitani M, Kasashima K, Yamamoto DS, Yagi K, Yuda M, Matsuoka H, Yoshida S
    Insect molecular biology 22 1 41 - 51 1 2013年02月 [査読有り][通常論文]
     
    We have previously developed a robust salivary gland-specific expression system in transgenic Anopheles stephensi mosquitoes. To establish transgenic mosquito lines refractory to Plasmodium falciparum using this system, we generated a transgenic mosquito harbouring the gene encoding an anti-P.?falciparum circumsporozoite protein (PfCSP) single-chain antibody (scFv) fused to DsRed in a secretory form (mDsRed-2A10 scFv). Fluorescence microscopy showed that the mDsRed-2A10 scFv was localized in the secretory cavities and ducts of the salivary glands in a secreted form. To evaluate P.?falciparum transmission-blocking in a rodent malaria model, a transgenic Plasmodium berghei line expressing PfCSP in place of PbCSP (PfCSP/Pb) was constructed. The PfCSP/Pb parasites were able to bind to the mDsRed-2A10 scFv in the salivary glands of the transgenic mosquitoes. Importantly, the infectivity of the transgenic mosquitoes to mice was strongly impaired, indicating that the parasites had been inactivated. These results suggest that salivary gland-specific expression of antisporozoite molecules could be a promising strategy for blocking malaria transmission to humans.
  • Iyori M, Nakaya H, Inagaki K, Pichyangkul S, Yamamoto DS, Kawasaki M, Kwak K, Mizukoshi M, Goto Y, Matsuoka H, Matsumoto M, Yoshida S
    PloS one 8 8 e70819  8 2013年 [査読有り][通常論文]
     
    We have previously developed a new malaria vaccine delivery system based on the baculovirus dual expression system (BDES). In this system, expression of malaria antigens is driven by a dual promoter consisting of the baculovirus-derived polyhedrin and mammal-derived cytomegalovirus promoters. To test this system for its potential as a vaccine against human malaria parasites, we investigated immune responses against the newly developed BDES-based Plasmodium falciparum circumsporozoite protein vaccines (BDES-PfCSP) in mice and Rhesus monkeys. Immunization of mice with BDESPfCSP induced Th1/Th2-mixed type immune responses with high PfCSP-specific antibody (Ab) titers, and provided significant protection against challenge from the bites of mosquitoes infected with a transgenic P. berghei line expressing PfCSP. Next, we evaluated the immunogenicity of the BDES-PfCSP vaccine in a rhesus monkey model. Immunization of BDES-PfCSP elicited high levels of anti-PfCSP Ab responses in individual monkeys. Moreover, the sera from the immunized monkeys remarkably blocked sporozoite invasion of HepG2 cells. Taken together with two animal models, our results indicate that this novel vaccine platform (BDES) has potential clinical application as a vaccine against malaria.
  • Reza M, Yamamoto DS, Matsuoka H
    Medical Entomology and Zoology 63 3 217 - 222 2012年09月 [査読有り][通常論文]
  • Matsuoka H, Sano G, Hattori R, Tomita H, Yamamoto DS, Hirai M
    Tropical medicine and health 40 2 47 - 52 2 2012年06月 [査読有り][通常論文]
     
    It has been proposed that transgenic mosquitoes can be used as a "flying syringe" for infectious disease control. We succeeded in generating a transgenic (TG) mosquito, Anopheles stephensi, excreting and discharging DsRed in saliva. DsRed was deposited on the membrane where the TG mosquito probed with its proboscis. Repeated feeding by the TG mosquitoes induced anti-DeRed as well as anti-SG antibodies in mice. This indicates that the TG mosquitoes can immunize the animal. Moreover, in this report, we employed a pre-immunization method before exposing mice to the TG mosquitoes. We injected DsRed to mice to prepare memory B cells and exposed the mice to bites by the TG mosquitoes excreting DsRed. The mice produced a higher titer of antibody to DsRed, suggesting that the bites from TG mosquitoes act as a booster and that primary immunization with a vaccine protein and exposure to TG mosquitoes excreting the vaccine protein in the saliva produces a synergistic effect. © 2012 by The Japanese Society of Tropical Medicine.
  • Yamamoto DS, Sumitani M, Nagumo H, Yoshida S, Matsuoka H
    Insect molecular biology 21 2 223 - 233 2 2012年 [査読有り][通常論文]
     
    We produced a transgenic mosquito expressing a rodent malaria vaccine candidate antigen in the salivary gland. Three tandemly repeated amino acid units from the repeat region of circumsporozoite protein of Plasmodium berghei (PbCS3R) fused to red fluorescent protein (monomeric DsRed) was chosen as a vaccine candidate antigen. Immunoblot and fluorescence microscopic analyses showed the transgene expression in the female salivary gland. The transgene product was released from the proboscis as a component of saliva. The monomeric DsRed-fusion expression system could be suitable for transgene secretion in the saliva of female mosquitoes. Mice repeatedly bitten by transgenic mosquitoes raised antibodies against P. berghei sporozoites, and the sera had protective ability against sporozoite invasion of human hepatoma HepG2 cells. These results suggest that transgene products are immunogenically active in saliva, and induce the antibodies to malaria parasite. These findings indicate that this technology has the potential for production of a flying vaccinator for rodent malaria parasites.
  • Yamamoto DS, Nagumo H, Yoshida S
    Insect molecular biology 19 3 391 - 398 3 2010年06月 [査読有り][通常論文]
     
    'Flying vaccinator' is the concept of using genetically engineered hematophagous insects to deliver vaccines. Here we show the generation of a transgenic anopheline mosquito that expresses the Leishmania vaccine candidate, SP15, fused to monomeric red fluorescent protein (mDsRed) in its salivary glands. Importantly, mice bitten repeatedly by the transgenic mosquitoes raised anti-SP15 antibodies, indicating delivery of SP15 via blood feeding with its immunogenicity intact. Thus, this technology makes possible the generation of transgenic mosquitoes that match the original concept of a 'flying vaccinator'. However, medical safety issues and concerns about informed consent mitigate the use of the 'flying vaccinator' as a method to deliver vaccines. We propose that this expression system could be applied to elucidate saliva-malaria sporozoite interactions.
  • Hatakeyama, M, Sumitani, M, Yamamoto, DS, Lee, JM, Oka, K
    Proceedings of the Arthropodan Embryological Society of Japan 44 1 - 12 2009年09月 [査読有り][通常論文]
  • Yamamoto DS, Tachibana K, Sumitani M, Lee JM, Hatakeyama M
    Mechanisms of development 125 11-12 996 - 1008 11-12 2008年11月 [査読有り][通常論文]
     
    Extensive survey of meiotic metaphase II arrest during oocyte maturation in vertebrates revealed that the mitogen-activated protein kinase ( MAPK) pathway regulated by the c-mos proto-oncogene product, Mos, has an essential role in cytostatic activity, termed cytostatic factor (CSF). In contrast, little is known in invertebrates in which meiotic arrest occurs in most cases at metaphase I (MI arrest). A parthenogenetic insect, the sawfly Athalia rosae, in which artificial egg activation is practicable, has advantages to investigate the mechanisms of MI arrest. Both the MAPK/extracellular signal-regulated protein kinase kinase (MEK) and MAPK were phosphorylated and maintained active in MI-arrested sawfly eggs, whereas they were dephosphorylated soon after egg activation. Treatment of MI-arrested eggs with U0126, an inhibitor of MEK, resulted in dephosphorylation of MAPK and MI arrest was resumed. The sawfly c-mos gene orthologue encoding a serine/threonine kinase was cloned and analyzed. It was expressed in nurse cells in the ovaries. To examine CSF activity of the sawfly Mos, synthesized glutathione S-transferase (GST)fusion sawfly Mos protein was injected into MI-resumed eggs in which MEK and MAPK were dephosphorylated. Both MEK and MAPK were phosphorylated again upon injection. In these GST-fusion sawfly Mos-injected eggs subsequent mitotic ( syncytial) divisions were blocked and embryonic development was ceased. These results demonstrated that the MEK-MAPK pathway was involved in maintaining CSF arrest in sawfly eggs and Mos functioned as its upstream regulatory molecule. (c) 2008 Elsevier Ireland Ltd. All rights reserved.
  • Sumitani M, Yamamoto DS, Lee JM, Hatakeyama M
    Insect biochemistry and molecular biology 35 3 231 - 240 3 2005年03月 [査読有り][通常論文]
     
    We isolated and characterized the white gene orthologue of the sawfly, Athalia rosae (Hymenoptera). The A. rosae white (Ar white) cDNA cloned was 2058-bp long encoding 685 amino acids in a single open reading frame (ORF). Comparison of the cDNA sequence with the genomic DNA sequence revealed that the ORF was derived from 11 exons. Ar white was a single copy gene as evidenced by genomic Southern blotting and its cytological localization on the metaphase chromosomes. The deduced amino acid sequence aligned well with known insect white orthologous gene products sharing conserved regions such as the ATP-binding motif and the six transmembrane-spanning segments. Expression of Ar white was detected at embryonic and pupal stages by Northern blotting. In situ hybridization detected the embryonic expression in a pair of the lateral tips of protocephalic placodes from where optic organs are formed. Ar white function was examined using double-stranded RNA (dsRNA)-mediated interference. The synthesized dsRNA targeting Ar white transcripts caused a decrease in the level of the original mRNAs, and resulted in the white phenocopy in the embryonic eye pigmentation when microinjected into eggs from wild-type females. The effects occurred in a dose-dependent manner. (C) 2005 Elsevier Ltd. All rights reserved.
  • Yamamoto DS, Sumitani M, Tojo K, Lee JM, Hatakeyama M
    Development genes and evolution 214 3 128 - 133 3 2004年03月 [査読有り][通常論文]
     
    The cDNA of a decapentaplegic (dpp) orthologue from the sawfly, Athalia rosae (Hymenoptera), was cloned and characterized. The clone (Ar dpp) was 2,566 bp long and encoded 395 amino acids in a single open reading frame. Genomic Southern blotting showed that Ar dpp is a single copy gene. The deduced amino acid sequence can be aligned along its entire length with known insect DPPs. It shared common characteristics such as a signal sequence, a pro-domain region, and a ligand domain with seven cysteines at conserved locations. Ar dpp was expressed as a single 5.0-kb mRNA in embryos, larvae, pupae and adults. In situ hybridization showed that Ar dpp was expressed in the dorsal region proper in early embryonic stages and in the embryonic appendages of cephalic segments (labrum, antenna, mandible, maxilla, and labium), thoracic segments (thoracic legs), and all abdominal segments except the tenth segment (pleuropodia and proleg primordia). The present results indicate that Ar dpp expression reflects the primary determination of embryonic appendages.
  • Sumitani M, Yamamoto DS, Oishi K, Lee JM, Hatakeyama M
    Insect biochemistry and molecular biology 33 4 449 - 458 4 2003年04月 [査読有り][通常論文]
     
    A piggyBac construct carrying two green fluorescent protein (GFP)-coding sequences one driven by Bombyx mori actin gene promoter and the other by Drosophila melanogaster heat-shock protein 70 (hsp70) promoter were injected together with a nonautonomous helper plasmid containing an active piggyBac transposase gene into the posterior end of mature unfertilized eggs dissected from the ovaries of Athalia rosae (Hymenoptera: Symphyta). These injected eggs, which developed as haploid male embryos upon artificial activation, were cultured to adulthood. Of 278 injected eggs, 61 grew to G(0) haploid adult males. These G(0) haploid males were individually mated to diploid females. The progeny embryos (G(1) generation) were examined for GFP expression. Four GFP-positive embryos (from three independent G(0) matings) were obtained. Two eclosed as diploid adult G(1) females. Mature unfertilized eggs dissected from the GFP-positive G(1) diploid females were activated artificially, and the resultant embryos were examined for GFP expression, separated and cultured to adulthood (G(2) generation). The G(2) haploid embryos segregated to GFP-positive and negative individuals. By mating the G(2) adult haploid males individually to diploid females, stocks were established in which the piggyBac construct was stably integrated into the genome, as evidenced by GFP expression and Southern blot hybridization. The piggyBac transposition occurred at its canonical target TTAA sequence. These results, which demonstrate the first successful stable transposon-mediated germline transformation in Hymenoptera, will expand the usefulness of the piggyBac vector. (C) 2003 Elsevier Science Ltd. All rights reserved.
  • Matsumoto K, Yamamoto DS, Sumitani M, Lee JM, Hatakeyama M, Oishi K
    Archives of insect biochemistry and physiology 49 1 34 - 40 1 2002年01月 [査読有り][通常論文]
     
    Mitotic metaphase chromosomes of Athalia rosae (Hymenoptera) haploid males were subjected to fluorescence in situ hybridization (FISH) analysis using an rDNA probe and two vitellogenin (Vg) cDNA probes (one representing the 5' half and the other the 3' half of the gene, each about 3 kb long, and together covering the entire coding region). The rDNA probe produced signals in four chromosomes, all in pericentromeric regions (haploid chromosome number = 8), and the Vg probes, either the combined probes or the 3' region alone, produced a twin signal in the middle of a chromosome arm of a single chromosome. Arch. Insect Biochem. Physiol. 49:34-40, 2002. (C) 2002 Wiley-Liss, Inc.

書籍

講演・口頭発表等

  • ハマダラカにおけるアミノペプチダーゼN様遺伝子のノックアウト解析  [通常講演]
    山本大介, 水島大貴, 加藤大智
    第91回日本寄生虫学会大会 2022年05月
  • マダニ唾液中に含まれる免疫抑制物質の探索  [通常講演]
    佐々木萌可, 川田 逸人, 小池 優貴, 関口 智也, 佐々木 紗己, 中島 政斗, Emmanuel Pacia Hernandez, Kofi Dadzie Kwofie, Danielle Ladzekpo, 井上 貴裕, 神保 恵, 三上 房子, 山本 大介, 加藤 大智, 辻 尚利, 八田 岳士
    第91回日本寄生虫学会大会 2022年05月
  • マダニ唾液物質HISG-g22の血管内皮細胞に及ぼす影響  [通常講演]
    佐々木紗己, 川田 逸人, 小池 優貴, 関口 智也, 佐々木 萌可, 中島 政斗, Emmanuel Pacia Hernandez, Kofi Dadzie Kwofie, Danielle Ladzekpo, 井上 貴裕, 神保 恵, 三上 房子, 山本 大介, 加藤 大智, 辻 尚利, 八田 岳士
    第91回日本寄生虫学会大会 2022年05月
  • マダニ刺咬部位の病理学的解析  [通常講演]
    小池 優貴, 川田 逸人, 朽津 有紀, 佐々木 紗己, 佐々木 萌可, 中島 政斗, 関口 智也, Emmanuel Pacia Hernandez, Kofi Dadzie, Danielle Ladzekpo, 井上 貴裕, 神保 恵, 三上 房子, 久保 誠, 長塩 亮, 山本 大介, 加藤 大智, 辻 尚利, 八田 岳士
    第91回日本寄生虫学会大会 2022年05月
  • エクアドルに分布するリーシュマニア原虫の核遺伝子とキネトプラスト遺伝子の比較解析  [通常講演]
    加藤大智, Eduardo A. Gomez, 関千里・Ahmed Tabbabi, 山本大介, 橋口義久
    第91回日本寄生虫学会大会 2022年05月
  • キサンチンデヒドロゲナーゼ（XDH）は希少糖アロースのハマダラカ中腸におけるマラリア原虫の発達抑制機構に関与する  [通常講演]
    水島 大貴, Ahmed Tabbabi, 山本 大介, 新井 明治, 加藤 大智
    第91回日本寄生虫学会大会 2022年05月
  • マダニ吸血必須因子 HlSG-g22は VE-cadhelinを介して血管透過性を亢進する  [通常講演]
    川田 逸人, 小池優貴, 佐々木沙己, 朽津有紀, 関口智也, 佐々木 萌可, 中島 政斗, 山本 大介, 加藤 大智, 久保 誠, 長塩亮, 辻 尚利, 八田 岳士
    第74回日本衛生動物学会大会 2022年04月
  • ハマダラカAnopheles stephensi におけるdoublesex遺伝子雄型産物のノックアウト解析
    山本大介, 水島大貴, 加藤大智
    第66回日本応用動物昆虫学会大会 2022年03月
  • A mosquito control technique using a synthetic cell ablation system  [招待講演]
    山本大介
    北里大学 寄生虫学・熱帯医学/環境衛生学 合同セミナー 2021年12月
  • フタトゲチマダニ刺咬部位における唾液分子HISG-g22の役割
    川田 逸人, 小池優貴, 佐々木沙己, 朽津有紀, 横井愛香, 関口智也, 三上房子, Hernandez Emmanuel Pacia, Kwofie Kofi, Dadzie・Danielle Ladzekpo, 山本 大介, 加藤 大智, 長塩亮, 辻 尚利, 八田 岳士
    第72回日本衛生動物学会東日本支部大会 2021年11月
  • 南米アンデス地域で異なるリーシュマニア原虫媒介能をもつサシチョウバエLutzomyia ayacuchensisの遺伝子多型解析および腸内細菌叢の比較
    Ahmed Tabbabi, 水島 大貴, Abraham, G. Caceres, Eduardo A. Gomez, 山本大介, 橋口義久, 加藤大智
    第72回日本衛生動物学会東日本支部大会 2021年11月
  • ハマダラカAnopheles stephensiの中腸で発現するアミノペプチダーゼN様遺伝子apnlの遺伝子ノックアウト
    山本大介, 水島大貴, 加藤大智
    第72回日本衛生動物学会東日本支部大会 2021年11月
  • ステフェンスハマダラカ雌の生殖におけるアミノペプチダーゼN遺伝子の機能解析  [通常講演]
    山本大介, 水島大貴, 加藤大智
    第57回日本節足動物発生学会大会 2021年07月
  • 精子ができないマラリア媒介蚊の作出 ―遺伝子操作による蚊の生殖制御技術―  [招待講演]
    山本大介
    昆虫DNA研究会第17回研究集会 2021年05月
  • フタトゲチマダニにおける吸血部位微小環境の病理学的解析  [通常講演]
    小池 優貴, 川田 逸人, 佐々木 紗己, 横井 愛香, 朽津 有紀, 長塩 亮, 久保 誠, 山本 大介, 加藤 大智, 辻 尚利, 八田 岳士
    第90回日本寄生虫学会・第32回日本臨床寄生虫学会合同大会 2021年04月
  • Insight into the embryogenesis of the parthenogenetic ixodid tick, Haemaphysalis longicornis based on a classic transcriptomic study  [通常講演]
    Kwofie Kofi, Dadzie、Hernandez Emmanuel Pacia, Kawada Hayato, Dadzie Samuel, Ladzekpo Danielle, Matsubayashi Makoto, Yamamoto Daisuke S, Kato Hirotomo, Iwanaga Shiroh, Tsuji Naotoshi, Hatta Takeshi
    第90回日本寄生虫学会・第32回日本臨床寄生虫学会合同大会 2021年04月
  • Promoter analysis of elongation factor 1-alpha (EF1α) derived from Haemaphysalis longicornis  [通常講演]
    Hernandez Emmanuel Pacia, 川田 逸人, 山本 大介, 加藤 大智, 松林 誠, 田仲 哲也, 辻 尚利, 八田 岳士
    第90回日本寄生虫学会・第32回日本臨床寄生虫学会合同大会 2021年04月
  • ハマダラカの中腸環境を変化させる糖類のスクリーニング  [通常講演]
    水島 大貴, Tabbabi Ahme, 山本 大介, 新井 明治, 加藤 大智
    第90回日本寄生虫学会・第32回日本臨床寄生虫学会合同大会 2021年04月
  • ハマダラカにおける性分化遺伝子の制御を受ける中腸遺伝子の解析  [通常講演]
    山本大介, 炭谷めぐみ, 加藤大智
    第72回日本衛生動物学会大会 2020年04月
  • ハマダラカにおける性分化制御の研究  [招待講演]
    山本大介
    第75回日本寄生虫学会西日本支部大会 2019年09月
  • マウスBax遺伝子を利用した新しい細胞死誘導システム  [通常講演]
    炭谷めぐみ, 山本大介, 笠嶋克巳, 山田信人, 坪田拓也, 飯塚哲也, 瀬筒秀樹
    第90回日本動物学会大会 2019年09月
  • ハマダラカ雌におけるdoublesex遺伝子欠損の吸血・血液消化への影響  [通常講演]
    山本大介, 炭谷めぐみ, 加藤大智
    第90回日本動物学会大会 2019年09月
  • 性分化遺伝子の制御を受けるハマダラカ血液消化関連遺伝子の探索  [通常講演]
    山本大介, 炭谷めぐみ, 加藤大智
    第55回日本節足動物発生学会大会 2019年05月
  • 山本大介
    第71回日本衛生動物学会大会（シンポジウム） 2019年04月
  • 山本大介, 炭谷めぐみ, 加藤大智
    第63回日本応用動物昆虫学会大会 2019年03月
  • Efforts for safe use of gene drive technology in Japan  [通常講演]
    Tanaka T, Tanaka N, Nagano Y, Kanuka H, Yamamoto DS, Yamamoto N, Nanba E, Nishiuchi T
    The 5th Asian Conference on Safety and Education in Laboratory 2018年11月
  • 感染症媒介蚊におけるゲノム編集研究の現状  [招待講演]
    山本大介
    農業・食品産業技術総合研究機構 生物機能利用研究部門シンポジウム「機能改変昆虫の産業利用の可能性」 2018年11月
  • ハマダラカにおける性分化制御による吸血・血液消化に関わる遺伝子の探索  [招待講演]
    山本大介
    ROIS-DS-JOINT共同研究集会 [昆虫のゲノムデータベースとそれを活用したデータ解析II ] 2018年08月
  • 山本大介, 炭谷めぐみ, 加藤大智
    第70回日本衛生動物学会大会 2018年05月
  • ステフェンスハマダラカ doublesexホモログの雌特異的アイソフォーム欠損系統の解析  [通常講演]
    山本大介, 炭谷めぐみ, 加藤大智
    第54回日本節足動物発生学会大会 2018年05月
  • 山本大介
    平成30年度蚕糸・昆虫機能利用学術講演会（第88回日本蚕糸学会大会）シンポジウム 2018年03月
  • 山本大介, 炭谷めぐみ, 加藤大智
    第62回日本応用動物昆虫学会大会 2018年03月
  • 山本大介, 炭谷めぐみ
    グローバルヘルス合同大会2017（第58回日本熱帯医学会大会）シンポジウム 2017年11月
  • 病原体媒介蚊におけるゲノム編集研究の状況 ー遺伝子解析から Gene Drive 技術までー  [招待講演]
    山本大介
    第9回 遺伝子組換え実験安全研修会（主催：全国大学等遺伝子研究支援施設連絡協議会） 2017年07月
  • 細胞死誘導法によるハマダラカの雄不妊化の効果  [通常講演]
    山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之, 加藤大智
    第53回日本節足動物発生学会 2017年05月
  • 山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之, 加藤大智
    第69回日本衛生動物学会大会 2017年04月
  • 山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之, 加藤大智
    第61回日本応用動物昆虫学会大会 2017年03月
  • ゲノム編集技術を利用したハマダラカにおけるマラリア寄生関連遺伝子の解析  [招待講演]
    山本大介
    第87回日本動物学会大会 シンポジウム 2016年11月 口頭発表（招待・特別）
  • Conservation of Mos/MAPK signaling pathway during insect oocyte maturation: a case study in the silkworm, Bombyx mori.  [通常講演]
    Megumi Sumitani, Daisuke S. Yamamoto, Hideki Sezutsu, Masatsugu Hatakeyama
    XXV International Congress of Entomology (ICE) 2016年09月 口頭発表（招待・特別）
  • Adaptation to an inducible tissue-specific cell death system in the malaria vector, Anopheles stephensi: A case of the salivary gland  [通常講演]
    Daisuke S. Yamamoto, Megumi Sumitani, Katsumi Kasashima, Hideki Sezutsu, Hiroyuki Matsuoka
    XXV International Congress of Entomology (ICE) 2016年09月 口頭発表（一般）
  • マウス細胞死誘導因子を利用したハマダラカの組織機能解析  [招待講演]
    山本大介
    昆虫デザイン研究会 2016年07月
  • 細胞死誘導系を利用したハマダラカにおける唾液腺機能解析  [通常講演]
    山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之
    第52回日本節足動物発生学会 2016年06月 口頭発表（一般）
  • 山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之
    第68回日本衛生動物学会大会 2016年04月 口頭発表（一般）
  • Analysis of the salivary gland function of anopheline mosquito using transgenesis.  [招待講演]
    Daisuke S. Yamamoto
    2nd Tokyo Vector Encounter 2016年03月 口頭発表（招待・特別）
  • 山本大介, 炭谷めぐみ, 畠山正統, 松岡裕之
    第85回日本寄生虫学会大会 2016年03月
  • ハマダラカにおける遺伝子ノックアウト解析  [通常講演]
    山本大介, 炭谷めぐみ, 畠山正統, 松岡裕之
    第36回菅平動物学セミナー 2015年12月
  • 炭谷めぐみ, 櫻井健志, 山本大介, 笠嶋克巳, 神崎亮平, 瀬筒秀樹
    日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集 2015年09月
  • 和田良樹, 山本大介, 炭谷めぐみ, 松岡裕之, 太田伸生, 鈴木高史
    第67回日本衛生動物学会大会 2015年03月
  • 山本大介, 炭谷めぐみ, 畠山正統, 松岡裕之
    第67回日本衛生動物学会大会 2015年03月 口頭発表（一般）
  • 山本大介, 炭谷めぐみ, 笠嶋克巳, 瀬筒秀樹, 松岡裕之
    第84回日本寄生虫学会大会 2015年03月
  • Identification of the beta2-tubulin gene promoter regulating testis-specific expression from Anopheles stephensi  [通常講演]
    Yamamoto DS, Sumitani M, Matsuoka H
    第50回日本節足動物発生学会大会 2014年07月
  • ルシフェラーゼを利用した皮膚注入後のハマダラカ唾液の可視化  [通常講演]
    山本大介, 横峰隆志, 炭谷めぐみ, 八木馨太, 松岡裕之, 吉田栄人
    第66回日本衛生動物学会 2014年03月
  • ハマダラカ（Anopheles stephensi）の生殖巣特異的に機能するプロモーターの探索  [通常講演]
    山本大介, 松岡裕之
    第34回菅平動物学セミナー 2013年12月
  • 山本大介, 畠山正統, 松岡裕之
    第84回日本動物学会大会 2013年09月
  • 唾液腺での抗菌ペプチド遺伝子過剰発現によるハマダラカへのマラリア原虫寄生への影響について  [通常講演]
    山本大介, 渡辺尚樹, 炭谷めぐみ, 鈴木高史, 松岡裕之
    第49回日本節足動物発生学会大会 2013年06月
  • 渡邊尚樹, 山本大介, 炭谷めぐみ, 鈴木高史, 松岡裕之
    第57回日本応用動物昆虫学会大会 2013年03月
  • ハマダラカにおける人為的卵付活技術開発の研究〜人工授精法開発へ向けた取り組み〜  [通常講演]
    山本大介, 畠山正統, 松岡裕之
    第72回日本寄生虫学会東日本支部大会、第10回分子寄生虫・マラリア研究フォーラム合同大会 2012年10月
  • 分泌型ルシフェラーゼ融合AAPPタンパク質を発現する遺伝子組換えハマダラカ  [通常講演]
    山本大介, 横峰隆志, 八木馨太, 松岡裕之, 吉田栄人
    第81回日本寄生虫学会大会 2012年03月
  • ハマダラカ卵の人為的付活法確立への取り組み  [通常講演]
    山本大介, 畠山正統, 松岡裕之
    昆虫ワークショップ2011東北 2011年10月 口頭発表（一般）
  • ハマダラカにおける唾液腺での遺伝子組換え技術の利用〜マラリア抗原媒介蚊によるマウスでの抗体誘導〜  [通常講演]
    山本大介, 炭谷めぐみ, 吉田栄人, 松岡裕之
    2011年昆虫ポストゲノム研究会 2011年09月
  • ステフェンスハマダラカにおける成熟未受精卵の人為的賦活  [通常講演]
    山本大介, 畠山正統, 松岡裕之
    第47回日本節足動物発生学会大会 2011年06月 口頭発表（一般）
  • 唾液腺にDsRed-CSP融合タンパク質を発現するトランスジェニックハマダラカの解析  [通常講演]
    山本大介, 南雲浩志, 炭谷めぐみ, 吉田栄人, 松岡裕之
    第63回日本衛生動物学会大会 2011年04月
  • ハマダラカ唾液腺における遺伝子発現制御システムの有用性  [招待講演]
    山本大介
    第19回病害動物の生理分子生物談話会 2011年04月
  • マラリアワクチン候補抗原を媒介する遺伝子組換えハマダラカの作製  [通常講演]
    山本大介, 炭谷めぐみ, 吉田栄人, 松岡裕之
    第55回日本応用動物昆虫学会大会 2011年03月
  • ハマダラカ唾液腺での遺伝子発現制御システムの利用法〜マラリアワクチン抗原を媒介する蚊〜  [通常講演]
    山本大介, 炭谷めぐみ, 吉田栄人, 松岡裕之
    第31回菅平動物学セミナー 2010年12月
  • CSPを唾液として分泌する遺伝子組換えハマダラカ〜flying syringeの作製をめざして〜  [通常講演]
    山本大介, 炭谷めぐみ, 吉田栄人, 松岡裕之
    第9回分子寄生虫・マラリア研究フォーラム 2010年10月
  • マラリアCSタンパク質を吸血時に唾液として分泌するトランスジェニックハマダラカ  [通常講演]
    山本大介, 炭谷めぐみ, 吉田栄人, 松岡裕之
    第62回日本衛生動物学会東日本支部大会 2010年10月
  • ワクチン抗原伝播ハマダラカの作製—蛍光タンパク質による唾液の可視化を用いた解析—  [招待講演]
    山本大介, 南雲浩志, 吉田栄人, 松岡裕之
    第79回日本寄生虫学会大会シンポジウム 2010年05月
  • ワクチン候補抗原を吸血時に媒介する遺伝子組換え蚊  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第54回日本応用動物昆虫学会大会 2010年03月
  • ワクチン抗原を唾液として分泌する遺伝子組換え蚊  [通常講演]
    山本大介, 吉田栄人
    第30回菅平前口動物学セミナー 2009年12月
  • リーシュマニアワクチン候補タンパク質を唾液として分泌する遺伝子組換えハマダラカの解析  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第50回日本熱帯医学会大会 2009年10月
  • 唾液中に導入遺伝子産物を発現するハマダラカの作製  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    2009年昆虫ポストゲノム研究会 2009年09月
  • 遺伝子組換えハマダラカにおける導入遺伝子産物を唾液成分として発現させるシステム  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第45回日本節足動物発生学会大会 2009年06月
  • サシチョウバエ唾液タンパク質を発現するトランスジェニックハマダラカの作製  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第78回日本寄生虫学会大会 2009年03月
  • 遺伝子組換えハマダラカにおける導入遺伝子産物を唾液成分として発現させるシステム  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第53回日本応用動物昆虫学会大会 2009年03月
  • 唾液腺にワクチン候補タンパク質を発現するトランスジェニックハマダラカの作製  [通常講演]
    山本大介, 南雲浩志, 吉田栄人
    第29回菅平前口動物学セミナー 2008年12月
  • 岡和希, 山本大介, 町田龍一郎, 畠山正統
    日本動物学会大会第79回大会 2008年09月
  • Involvement of Mos-MEK-MAPK pathway in MI arrest of the sawfly, Athalia rosae  [通常講演]
    Hatakeyama M, Yamamoto DS, Tachibana K
    International Workshop “Oocyte Maturation and the Cell Cycle” 2008年03月
  • カブラハバチ卵において卵賦活により消失する60 kDaタンパク質  [通常講演]
    山本大介, 李載旼, 畠山正統
    第28回菅平前口動物学セミナー 2007年12月
  • 山本大介, LEE Jae Min, 立花和則, 畠山正統
    第78回日本動物学会大会 2007年09月
  • カブラハバチにおける CSF機能の細胞学的解析  [通常講演]
    山本大介, 李載旼, 畠山正統
    第43回日本節足動物発生学会大会 2007年07月 口頭発表（一般）
  • カブラハバチにおける卵減数分裂を人為的に制御できる形質転換系統の作製  [通常講演]
    山本大介, 岡和希, 李載旼, 畠山正統
    第27回菅平前口動物学セミナー 2006年12月
  • カブラハバチにおけるTet-Off系を利用した遺伝子発現制御  [通常講演]
    畠山正統, 新美輝幸, 瀬筒秀樹, 山本大介, 李載旼
    日本動物学会第77回大会 2006年09月
  • カブラハバチ胚における腹脚形成過程の観察とDistal-less遺伝子の発現解析  [通常講演]
    岡和希, 山本大介, 李載旼, 東城幸治, 畠山正統, 町田龍一郎
    日本動物学会第77回大会 2006年09月
  • 膜翅目昆虫カブラハバチの卵減数分裂周期におけるMos-MEK-MAPK経路の機能解析  [通常講演]
    山本大介, 李載旼, 立花和則, 畠山正統
    第77回日本動物学会大会 2006年09月 口頭発表（一般）
  • トランスジェニックカブラハバチにおける遺伝子発現制御システム  [通常講演]
    畠山正統, 新美輝幸, 瀬筒秀樹, 山本大介, 李載旼
    日本節足動物発生学会第42回大会 2006年06月
  • カブラハバチにおける形質転換系を利用したMos機能の遺伝学的解析への取り組み  [通常講演]
    山本大介, 李載旼, 畠山正統
    第26回菅平前口動物学セミナー 2005年11月
  • カブラハバチの卵減数分裂におけるMos-MEK-MAPK経路活性化様式の解析  [通常講演]
    山本大介, 李載旼, 立花和則, 畠山正統
    第76回日本動物学会大会 2005年10月 ポスター発表
  • カブラハバチの卵減数分裂周期制御に関する研究  [通常講演]
    山本大介, 李載旼, 畠山正統
    第25回菅平前口動物学セミナー 2004年11月
  • カブラハバチ卵成熟過程におけるMosの機能  [通常講演]
    山本大介, 李載旼, 立花和則, 畠山正統
    第75回日本動物学会大会 2004年09月 ポスター発表
  • カブラハバチc-mos遺伝子の同定と発現解析  [通常講演]
    山本大介, 李載旼, 畠山正統
    第40回日本節足動物発生学会大会 2004年06月 口頭発表（一般）
  • 昆虫の卵減数分裂が停止するメカニズムを探る —カブラハバチを用いた研究—  [通常講演]
    山本大介, 李載旼, 畠山正統
    第24回菅平前口動物学セミナー 2003年11月
  • RNAi法によるカブラハバチwhite遺伝子の機能解析  [通常講演]
    炭谷めぐみ, 山本大介, 李載旼, 畠山正統
    日本動物学会第74回大会 2003年09月
  • 昆虫の卵減数分裂MI停止機構の解析をめざして −膜翅目昆虫カブラハバチを用いたアプローチ−  [通常講演]
    畠山正統, 山本大介
    日本動物学会第74回大会 2003年09月
  • カブラハバチ卵減数分裂における第一減数分裂中期制御機構の解析  [通常講演]
    山本大介, 李載旼, 立花和則, 畠山正統
    第74回日本動物学会大会 2003年09月 口頭発表（一般）
  • Stable germline transformation of the Hymenopteran insect, Athalia rosae with the piggyBac transposon vector.  [通常講演]
    Hatakeyama M, Sumitani M, Lee JM, Yamamoto DS, Oishi K
    Fourth International Workshop on Transgenesis and Genomics of Invertebrate Organisms 2003年
  • カブラハバチにおける減数分裂機構の研究  [通常講演]
    山本大介, 李載旼, 立花和則, 畠山正統
    第23回菅平前口動物学セミナー 2002年11月
  • カブラハバチ（膜翅目）の胚発生におけるdpp 遺伝子の解析  [通常講演]
    山本大介, 李載旼, 畠山正統, 大石陸生
    第38回日本節足動物発生学会大会 2002年07月 口頭発表（一般）
  • カブラハバチの胚発生におけるdpp遺伝子  [通常講演]
    山本大介, 炭谷めぐみ, 畠山正統, 大石陸生
    第22回菅平前口動物学セミナー 2001年12月
  • 膜翅目昆虫カブラハバチのdoublesexホモログの単離と構造機能  [通常講演]
    畠山正統, 松本耕平, 山本大介, 炭谷めぐみ, 李載旼
    日本動物学会第72回大会 2001年10月
  • カブラハバチ初期胚におけるdecapentaplegic遺伝子ホモログの機能  [通常講演]
    山本大介, 炭谷めぐみ, 李載旼, 畠山正統, 大石陸生
    第72回日本動物学会大会 2001年10月 口頭発表（一般）
  • 炭谷めぐみ, 山本大介, LEE J‐M, 畠山正統, 大石陸生
    第73回日本遺伝学会大会 2001年09月
  • decapentaplegic遺伝子の局所的発現によって起こるカブラハバチ初期胚の異常  [通常講演]
    山本大介, 李載旼, 畠山正統, 大石陸生
    第37回日本節足動物発生学会大会 2001年06月 口頭発表（一般）
  • カブラハバチにおけるdecapentaplegicホモログの発現解析  [通常講演]
    山本大介, 畠山正統, 大石陸生
    第21回菅平前口動物学セミナー 2000年12月
  • カブラハバチdecapentaplegicホモログcDNAのクローニング  [通常講演]
    山本大介, 李載旼, 畠山正統, 大石陸生
    第72回日本遺伝学会大会 2000年11月 口頭発表（一般）
  • カブラハバチにおけるdecapentaplegic cDNAの構造解析  [通常講演]
    山本大介, 李載旼, 畠山正統, 大石陸生
    日本節足動物発生学会第36回大会 2000年06月 口頭発表（一般）
  • カブラハバチにおけるdecapentaplegicホモログの検索と同定  [通常講演]
    山本大介, 畠山正統, 大石陸生
    第20回菅平前口動物学セミナー 1999年11月

MISC

受賞

  • 2017年 平成29年度自治医科大学優秀論文賞
     
    受賞者: 山本 大介
  • 2013年 平成25年度自治医科大学優秀論文賞
     
    受賞者: 山本大介

共同研究・競争的資金等の研究課題

  • 吸血昆虫の囲食膜の機能解明とバリア機能強化による病原体伝播阻止法の開発
    日本学術振興会：科学研究費助成事業 挑戦的研究(萌芽)
    研究期間 : 2021年07月 -2024年03月 
    代表者 : 加藤 大智, 山本 大介, 水島 大貴
  • 吸血昆虫における雌特異的な吸血を誘導する分子機構の解明
    日本学術振興会：科学研究費助成事業 基盤研究(C)
    研究期間 : 2021年04月 -2024年03月 
    代表者 : 山本 大介, 加藤 大智
  • 吸血昆虫による病原体媒介の分子機構の解明
    日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 2020年04月 -2024年03月 
    代表者 : 加藤 大智, 新井 明治, 水島 大貴, 山本 大介
     
    病原体媒介性吸血昆虫（ベクター）の体内では、どのような状況で病原体の発育や増殖が促進され、どのような状況では阻害されるのか、病原体媒介の成否に直結するこのイベントの分子機構は明らかではない。本研究では、抗生物質や希少糖の投与といった簡便な方法で病原体の発育・増殖能力を攪乱したベクターを用いて、ベクター内での病原体の発育や増殖の場となる中腸に発現する分子を比較解析し、病原体媒介能を規定するベクター中腸分子の解明を目指す。本年度の研究実績の概要が次の通りである。1）ルシフェラーゼ導入原虫を用いてハマダラカに感染するマラリア原虫およびサシチョウバエに感染するリーシュマニア原虫の簡易定量法を確立した。2）吸血によって感染したハマダラカの体内でのマラリア原虫の発育・増殖に影響を及ぼす糖類をスクリーニングした。3）希少糖によるハマダラカ体内でのマラリア原虫の発育阻害がどのステージで起きているかについて培養条件下で検討した。4）希少糖を投与してマラリア原虫媒介能が低下したハマダラカと通常のフルクトースを投与したハマダラカの腸内細菌叢を比較解析した。5）エクアドルとペルーに分布するリーシュマニア原虫媒介能の異なる同種のサシチョウバエの腸内細菌叢の比較解析を行い、その成果を国際誌に発表した。6）同じエサを与えた異なるサシチョウバエ種の腸内細菌叢の解析を行った。7）異なるエサを与えた同種のサシチョウバエで、病原体媒介能が変化するのか検討した。
  • 細胞死誘導システムを用いた病害虫・有用昆虫の不妊化系統の作出
    日本学術振興会：科学研究費助成事業 基盤研究(C)
    研究期間 : 2017年04月 -2022年03月 
    代表者 : 笠嶋 めぐみ, 山本 大介
     
    バイナリシステムあるいは組織特異的発現プロモーターを利用した細胞死誘導は、基礎研究および昆虫の遺伝的防除方法への応用に有用である。我々のグループは、哺乳動物のプロアポトーシス因子であるBax（B-cell leukemia/lymphoma 2-associated X protein）を組織特異的に発現誘導することによって限定的にアポトーシスを誘導できることを示した。 令和2年度はカイコメス特異的部位へのGal4のノックイン実験を実施した。カイコの性染色体はZW型と呼ばれるメスヘテロ型で、W染色体にメス決定因子がコード されている。しかしながら、その他の領域には、タンパク質をコードする遺伝子領域はなく、トランスポゾンが大量に挿入された繰り返し配列が散在しており、部位特異的ノックインのための標的領域を設定することが難しかった。そこで、すでに樹立されているW染色体にpiggyBacベクターが挿入されている系統に対してノックインを実施することにした。このpiggyBacには、マーカーとして3xP3プロモーターDsRed遺伝子があり、この領域を標的することで、眼のDsRedマーカーがノックインのマーカーとなる。すなわち、眼のDsRedの蛍光が消えた個体はノックイン個体として単離できる。この系統に、Gal4あるいは細胞死誘導エフェクター因子をノックインし、雌を致死する、あるいは性分化の撹乱を行うことを計画した。DsRedを標的とするノックインのためのTALENベクターを構築し、導入遺伝子を含むドナープラスミドとともにTALEN mRNAをカイコ胚にに注入し、ノックイン個体作成実験を実施した。現在、注射当代の個体をすべて飼育して、導入遺伝子を持つ個体（ノックイン個体）のスクリーニングを行っているところである。
  • 日本学術振興会：科学研究費助成事業 挑戦的研究(萌芽)
    研究期間 : 2018年06月 -2021年03月 
    代表者 : 加藤 大智, 崔 龍洙, 渡邊 真弥, 山本 大介, 水島 大貴
     
    本研究では、吸血昆虫の中腸の環境形成に重要な役割を担う腸内フローラが、病原体の生存・発育に影響を及ぼし、吸血昆虫の病原体媒介能を決定する因子となるのかについて検討した。1）エクアドルとペルーに分布するリーシュマニア原虫媒介能の異なる同種のサシチョウバエの腸内フローラを比較解析し、サシチョウバエの生息地域と腸内フローラの相関について明らかにすることができた。2）希少糖を投与してマラリア原虫媒介能が低下したハマダラカと通常のフルクトースを投与したハマダラカの腸内フローラを経時的に比較解析したところ、希少糖投与群でいくつかのユニークな腸内細菌の増殖が確認された。
  • 日本学術振興会：科学研究費助成事業 若手研究(B)
    研究期間 : 2016年04月 -2020年03月 
    代表者 : 山本 大介
     
    ハマダラカにおける吸血や血液消化に重要な遺伝子を明らかにするため、性分化遺伝子のノックアウトに伴いメスで血液消化が出来ないハマダラカ系統において中腸や唾液腺の遺伝子発現を解析した。中腸においては野生型との網羅的な比較解析を行い、野生型と比べて吸血後の発現量に差がある遺伝子群を明らかにした。それらの遺伝子のうち、アミノ構造から消化酵素と予想されるアミノペプチダーゼ遺伝子についてCRISPR/Cas9によるノックアウト解析を行い、雌の妊性に関与することを明らかにした。
  • 蚊における簡便で安定した系統保存法開発に向けた人工授精技術の確立
    大下財団：研究助成
    研究期間 : 2018年04月 -2019年03月 
    代表者 : 山本大介
  • マラリア媒介昆虫ハマダラカにおける組織特異的細胞死誘導系の利用：唾液腺への利用
    日本科学協会：平成２８年度海外発表促進助成
    研究期間 : 2016年 
    代表者 : 山本 大介
  • 唾液腺での細胞死誘導系を用いた吸血・マラリア伝播に関わるハマダラカ由来唾液物質の解析
    武田科学振興財団：医学系研究奨励
    研究期間 : 2015年 
    代表者 : 山本大介
  • 唾液腺遺伝子のゲノム編集によるマラリアベクター制御
    日本私立学校振興・共済事業団：学術研究振興基金（若手研究者奨励金）
    研究期間 : 2014年 
    代表者 : 山本大介
  • 膜翅目昆虫をモデル系とした昆虫の卵減数分裂周期制御機構の研究
    日本科学協会：笹川科学研究助成
    研究期間 : 2004年 
    代表者 : 山本大介

委員歴

  • 2018年05月 - 現在   日本節足動物発生学会   学会広報ワーキンググループ グループ員
  • 2016年12月 - 現在   全国大学等遺伝子研究支援施設連絡協議会   Gene Drive ワーキンググループ グループ員
  • 2012年06月 - 現在   日本節足動物発生学会   編集委員
  • 2012年06月 - 2018年05月   日本節足動物発生学会   庶務幹事

担当経験のある科目

  • 人体機能学自治医科大学大学院
  • 生命科学概論 I・II東京大学大学院
  • 医動物学実習自治医科大学
  • 医動物学講義自治医科大学

メディア報道

  • 雷鳴抄
    報道 : 2018年07月03日
    番組・新聞雑誌 : 下野新聞
    　新聞・雑誌
  • あさラジ
    報道 : 2016年08月17日
    番組・新聞雑誌 : ニッポン放送
    　テレビ・ラジオ番組
  • 蚊の退治 遺伝子狙え
    報道 : 2016年07月17日
    番組・新聞雑誌 : 日本経済新聞
    　新聞・雑誌
  • 病を運ぶ蚊 退治せよ
    報道 : 2016年05月08日
    番組・新聞雑誌 : 読売新聞
    　新聞・雑誌
  • OXITEC社の遺伝子組換え蚊（GMM）放出計画について：専門家コメント
    報道 : 2015年02月12日
    発行元・放送局 : サイエンス・メディア・センター

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