2024/04/18 更新

写真a

スギヤマ トシエ
杉山 稔恵
SUGIYAMA Toshie
所属
教育研究院 自然科学系 農学系列 教授
農学部 教授
職名
教授
通称等の別名
杉山 稔恵
外部リンク

学位

  • 博士(学術) ( 1994年3月   新潟大学 )

研究キーワード

  • 骨髄骨

  • カルシウム代謝

  • 脚弱

  • 卵殻質

  • 骨代謝

  • 鳥類

  • 産業動物科学

研究分野

  • ライフサイエンス / 応用分子細胞生物学

  • ライフサイエンス / 獣医学

  • ライフサイエンス / 動物生産科学

経歴(researchmap)

  • 日本学術振興会 特別研究員(PD)

    1994年 - 1995年

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経歴

  • 新潟大学   農学部   教授

    2018年4月 - 現在

  • 新潟大学   農学部 農学科   准教授

    2017年4月 - 2018年3月

  • 新潟大学   自然科学研究科 生命・食料科学専攻   准教授

    2010年7月 - 2018年3月

  • 新潟大学   自然科学研究科 生命・食料科学専攻   准教授

    2010年7月 - 2018年3月

  • 新潟大学   農業生産科学科   准教授

    2010年7月 - 2017年3月

  • 新潟大学   農業生産科学科   助教

    2004年4月 - 2010年6月

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学歴

  • 新潟大学   大学院自然科学研究科   生命システム科学

    - 1994年

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    国名: 日本国

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  • 新潟大学   農学研究科

    - 1991年

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  • 新潟大学   農学部   畜産学科

    - 1989年

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    国名: 日本国

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所属学協会

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論文

  • Mating in the cold. Prolonged sperm storage provides opportunities for forced copulation by male bats during winter

    Takahiro Sato, Toshie Sugiyama, Tsuneo Sekijima

    Frontiers in Physiology   14   1241470   2023年9月

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.3389/fphys.2023.1241470

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  • Reduced bone formation and increased bone resorption drive bone loss in Eimeria infected broilers

    Yuguo Hou Tompkins, Janghan Choi, Po-Yun Teng, Masayoshi Yamada, Toshie Sugiyama, Woo Kyun Kim

    SCIENTIFIC REPORTS   13 ( 1 )   2023年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PORTFOLIO  

    Coccidiosis is an economically significant disease in the global poultry industry, but little is known about the mechanisms of bone defects caused by coccidiosis; thus, the study focused on effects of coccidiosis on the bone homeostasis of young broiler chickens. A total of 480 male Cobb500 broilers were randomly allocated into four treatment groups, including an uninfected control consuming diet ad libitum, two infected groups were orally gavaged with two different concentrations of sporulated Eimeria oocysts, and an uninfected pair-fed group fed the same amount of feed as the high Eimeria-infected group consumed. Growth performance and feed intake were recorded, and samples were collected on 6 days post infection. Results indicated that coccidiosis increased systemic oxidative status and elevated immune response in bone marrow, suppressing bone growth rate (P < 0.05) and increasing bone resorption (P < 0.05) which led to lower bone mineral density (P < 0.05) and mineral content (P < 0.05) under Eimeria infection. With the same amount of feed intake, the uninfected pairfed group showed a distinguished bone formation rate and bone resorption level compared with the Eimeria infected groups. In conclusion, inflammatory immune response and oxidative stress in broilers after Eimeria infection were closely associated with altered bone homeostasis, highlighting the role of inflammation and oxidative stress in broiler bone homeostasis during coccidiosis.

    DOI: 10.1038/s41598-023-27585-5

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  • Overexpression of miR-125b in Osteoblasts Improves Age-Related Changes in Bone Mass and Quality through Suppression of Osteoclast Formation. 国際誌

    Shota Ito, Tomoko Minamizaki, Shohei Kohno, Yusuke Sotomaru, Yoshiaki Kitaura, Shinsuke Ohba, Toshie Sugiyama, Jane E Aubin, Kotaro Tanimoto, Yuji Yoshiko

    International journal of molecular sciences   22 ( 13 )   2021年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We recently reported an unexpected role of osteoblast-derived matrix vesicles in the delivery of microRNAs to bone matrix. Of such microRNAs, we found that miR-125b inhibited osteoclast formation by targeting Prdm1 encoding a transcriptional repressor of anti-osteoclastogenesis factors. Transgenic (Tg) mice overexpressing miR-125b in osteoblasts by using human osteocalcin promoter grow normally but exhibit high trabecular bone mass. We have now further investigated the effects of osteoblast-mediated miR-125b overexpression on skeletal morphogenesis and remodeling during development, aging and in a situation of skeletal repair, i.e., fracture healing. There were no significant differences in the growth plate, primary spongiosa or lateral (periosteal) bone formation and mineral apposition rate between Tg and wild-type (WT) mice during early bone development. However, osteoclast number and medial (endosteal) bone resorption were less in Tg compared to WT mice, concomitant with increased trabecular bone mass. Tg mice were less susceptible to age-dependent changes in bone mass, phosphate/amide I ratio and mechanical strength. In a femoral fracture model, callus formation progressed similarly in Tg and WT mice, but callus resorption was delayed, reflecting the decreased osteoclast numbers associated with the Tg callus. These results indicate that the decreased osteoclastogenesis mediated by miR-125b overexpression in osteoblasts leads to increased bone mass and strength, while preserving bone formation and quality. They also suggest that, in spite of the fact that single miRNAs may target multiple genes, the miR-125b axis may be an attractive therapeutic target for bone loss in various age groups.

    DOI: 10.3390/ijms22136745

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  • Effects of early high nutrition related to metabolic imprinting events on growth, carcass characteristics, and meat quality of grass-fed Wagyu (Japanese Black cattle). 国際誌

    Sithyphone Khounsaknalath, Kotaro Etoh, Kaori Sakuma, Kunihiko Saito, Akira Saito, Tsuyoshi Abe, Fumio Ebara, Toshie Sugiyama, Eiji Kobayashi, Takafumi Gotoh

    Journal of animal science   99 ( 6 )   2021年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The study was conducted to clarify how early high plane of nutrition related to metabolic imprinting affected growth, carcass characteristics, and meat quality of grass-fed Wagyu (Japanese Black cattle). Wagyu steers were allocated randomly into 2 dietary groups: (1) steers fed milk replacer (crude protein 26.0%, crude fat 25.5%; maximum intake 0.6 kg/d) until 3 mo of age and then fed roughage (orchard grass hay) ad libitum from 4 to 10 mo of age (roughage group, RG; n = 11); (2) steers fed milk replacer (maximum intake of 1.8 kg/d) until 3 mo of age and then fed a high-concentrate diet from 4 to 10 mo of age (early high nutrition, EHN; n = 12). After 11 mo of age, all steers were fed roughage ad libitum until 31 mo of age and then slaughtered. Growth performance, carcass traits, longissimus muscle (LM) meat quality and intramuscular fat (IMF) content, plasma insulin-like growth factor I (IGF-I) concentration, and bone mineral density were measured. Body weight was greater in EHN steers (571 kg) than RG steers (520 kg; P < 0.01). Plasma IGF-I levels were higher in EHN steers than in RG steers at 3, 10, and 14 mo of age (P < 0.01, P < 0.005, P < 0.001, respectively); however, plasma IGF-I levels were lower in EHN steers compared with RG steers at 30 mo of age (P < 0.01). The total weight of the muscles and bones of the left half of the carcass was not different between the 2 groups (P = 0.065). Five of the 19 muscles investigated (semimembranosus, P = 0.036; infraspinatus, P = 0.024; supraspinatus, P = 0.0019; serratus ventralis cervicis, P = 0.032; serratus ventralis thoracis, P = 0.027) were heavier in EHN steers. Total fat weight in the left half of the carcass was 30% greater (P = 0.025) in HNE carcasses. Subcutaneous and perirenal fat weights were 53% and 84% greater (P = 0.008, P = 0.002, respectively) in EHN carcasses. The LM IMF content was greater in EHN loins (13.2%) compared with RG loins (9.4%) at 31 mo of age (P = 0.038); however, no differences were found for shear force, tenderness, and cook loss. These results suggested early high-nutrition affected the growth and meat quality of livestock.

    DOI: 10.1093/jas/skab123

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  • Effect of Age on Bone Structure Parameters in Laying Hens

    Masayoshi Yamada, Chongxiao Chen, Toshie Sugiyama, Woo Kyun Kim

    ANIMALS   11 ( 2 )   2021年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI  

    Simple SummaryLaying hens supply calcium to their eggshells via a process of dynamic bone formation and resorption called remodeling. As hens age, their bone becomes weak and brittle due to an imbalance in remodeling, resulting in bone disorders. Osteoporosis, characterized by bone mass loss and increased risk of fractures, is a serious problem in the egg industry, affecting both productivity and animal welfare. Nonetheless, the mechanisms of age-related osteoporosis remain unknown. We evaluated differences in bone structure between age groups, using microscopy and 3D imaging. Bone quality was reduced by an increase in porosity, with loss of volume, contributing to age-related osteoporosis.Changes in medullary and cortical bone structure with age remain unclear. Twenty Hy-Line W36 hens, 25 or 52 weeks of age, were euthanized, and both tibiae were collected when an egg was present in the magnum. Serial cross sections of the tibiae were stained with Alcian blue. The bones were scanned using micro-computed tomography. Trabecular width (Tb.Wi) was significantly higher (p < 0.05) in 25-week-old hens, whereas medullary bone tissue volume (TV) was significantly higher (p < 0.01) in 52-week-old hens. 25-week-old hens had significantly higher (p < 0.01) bone volume fraction (BVF = calcified tissue / TV). Moreover, the cortical bone parameters were significantly higher (TV and bone mineral content (BMC) at p < 0.05, and bone volume (BV) and BVF at p < 0.01) in younger hens. Open porosity and total porosity, which indicate less density, were significantly higher (p < 0.01) in older hens. Older hens showed significantly higher (p < 0.01) tibial diaphysis TV than younger hens. Younger hens had significantly higher (p < 0.01) BV, BVF and bone mineral density (BMD) of the tibial diaphysis. These findings reveal that reductions in medullary bone quality might be associated with age-related low estrogen levels and stimulation of osteoclastic bone resorption by parathyroid hormone. Cortical bone quality decreased with enlargement of the Haversian canals and loss of volume, with a longer egg-laying period leading to osteoporosis.

    DOI: 10.3390/ani11020570

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  • The matrix vesicle cargo miR-125b accumulates in the bone matrix, inhibiting bone resorption in mice. 国際誌

    Tomoko Minamizaki, Yuko Nakao, Yasumasa Irie, Faisal Ahmed, Shota Itoh, Nushrat Sarmin, Hirotaka Yoshioka, Asako Nobukiyo, Chise Fujimoto, Shumpei Niida, Yusuke Sotomaru, Kotaro Tanimoto, Katsuyuki Kozai, Toshie Sugiyama, Edith Bonnelye, Yuichiro Takei, Yuji Yoshiko

    Communications Biology   3 ( 1 )   30 - 30   2020年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Communication between osteoblasts and osteoclasts plays a key role in bone metabolism. We describe here an unexpected role for matrix vesicles (MVs), which bud from bone-forming osteoblasts and have a well-established role in initiation of bone mineralization, in osteoclastogenesis. We show that the MV cargo miR-125b accumulates in the bone matrix, with increased accumulation in transgenic (Tg) mice overexpressing miR-125b in osteoblasts. Bone formation and osteoblasts in Tg mice are normal, but the number of bone-resorbing osteoclasts is reduced, leading to higher trabecular bone mass. miR-125b in the bone matrix targets and degrades Prdm1, a transcriptional repressor of anti-osteoclastogenic factors, in osteoclast precursors. Overexpressing miR-125b in osteoblasts abrogates bone loss in different mouse models. Our results show that the MV cargo miR-125b is a regulatory element of osteoblast-osteoclast communication, and that bone matrix provides extracellular storage of miR-125b that is functionally active in bone resorption.

    DOI: 10.1038/s42003-020-0754-2

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  • Salt Tolerance Improvement in Rice through Efficient SNP Marker-Assisted Selection Coupled with Speed-Breeding

    Md Masud Rana, Takeshi Takamatsu, Marouane Baslam, Kentaro Kaneko, Kimiko Itoh, Naoki Harada, Toshie Sugiyama, Takayuki Ohnishi, Tetsu Kinoshita, Hiroki Takagi, Toshiaki Mitsui

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   20 ( 10 )   2019年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI  

    Salinity critically limits rice metabolism, growth, and productivity worldwide. Improvement of the salt resistance of locally grown high-yielding cultivars is a slow process. The objective of this study was to develop a new salt-tolerant rice germplasm using speed-breeding. Here, we precisely introgressed the hst1 gene, transferring salinity tolerance from Kaijin into high-yielding Yukinko-mai (WT) rice through single nucleotide polymorphism (SNP) marker-assisted selection. Using a biotron speed-breeding technique, we developed a BC3F3 population, named YNU31-2-4, in six generations and 17 months. High-resolution genotyping by whole-genome sequencing revealed that the BC3F2 genome had 93.5% similarity to the WT and fixed only 2.7% of donor parent alleles. Functional annotation of BC3F2 variants along with field assessment data indicated that YNU31-2-4 plants carrying the hst1 gene had similar agronomic traits to the WT under normal growth condition. YNU31-2-4 seedlings subjected to salt stress (125 mM NaCl) had a significantly higher survival rate and increased shoot and root biomasses than the WT. At the tissue level, quantitative and electron probe microanalyzer studies indicated that YNU31-2-4 seedlings avoided Na+ accumulation in shoots under salt stress. The YNU31-2-4 plants showed an improved phenotype with significantly higher net CO2 assimilation and lower yield decline than WT under salt stress at the reproductive stage. YNU31-2-4 is a potential candidate for a new rice cultivar that is highly tolerant to salt stress at the seedling and reproductive stages, and which might maintain yields under a changing global climate.

    DOI: 10.3390/ijms20102585

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  • An estimation of number of birds to be consecutively released in the reintroduction of Japanese Crested Ibises (Nipponia nippon)

    Yuichi Wajiki, Yoshinori Kaneko, Toshie Sugiyama, Takahisa Yamada, Hiroaki Iwaisaki

    WILSON JOURNAL OF ORNITHOLOGY   130 ( 4 )   874 - 880   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILSON ORNITHOLOGICAL SOC  

    The Japanese captive population of Crested Ibis (Nipponia nippon) has rapidly increased, and reintroduction programs were initiated on Sado Island in 2008. The Japanese captive population, however, was founded by only 5 individuals originating from the Chinese captive population. Because efforts were directed toward preserving genetic diversity in the captive population, it was appropriate to consider genetic diversity in the reintroduced population to evaluate its potential to survive in the wild, where environmental changes are more significant than in captivity. The purpose of this study was to determine the number of individuals to release in the future to preserve genetic diversity of the reintroduced population. We simulated the probability that the reintroduced population after 50 years would retain a rare allele existing in the captive population, based on demographic parameters obtained from either the Japanese captive or reintroduced populations, or Chinese wild populations. As a result, the 10-15 individuals currently released in the annual reintroduction would be the number needed to be released each year over 50 years to retain the rare allele with a frequency in the captive population of 0.05 with a >90% probability. Therefore, the current reintroduction program seems reasonable and should be continued.

    DOI: 10.1676/1559-4491.130.4.874

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  • Short communication: Temporal changes in the skin morphology of dairy cows during the periparturient period

    Banri Suzuki, Toshie Sugiyama, Chikako Yoshida, Toshihiko Nakao

    JOURNAL OF DAIRY SCIENCE   101 ( 7 )   6616 - 6621   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    Management of dairy cow productivity requires monitoring of their nutritional status by visual observation. It has been suggested that changes in hair coat appearance are among the indicators of nutritional state in dairy cows. Temporal changes in the skin morphology in cows, however, have riot been reported. In this study, we examined the changes in the skin of dairy cows that occur during the peripartum period. Seven pluriparous cows were used. Skin samples were collected at 28 d before the due date and 28 d and 56 d after calving for morphological examination. Hair follicle width was 108.8 +/- 5.9 mu m (+/- SD) in the dry period, 95.5 +/- 5.5 at 28 d after calving, and 104.2 +/- 5.3 mu m at 56 d postpartum. The percentages of anagen hair follicles during these 3 periods were 41.4 +/- 3.4, 18.5 +/- 3.4, and 32.3 +/- 3.3%, respectively. The corresponding sebaceous gland sizes were 8,362.0 +/- 707.6, 7,800.0 +/- 831.4, and 9,186.8 +/- 962.6 mu m(2), respectively. Hair follicle width was positively correlated with percentage of anagen hair follicles. The thickness of epidermal and proliferation rate of epidermal cell were also correlated. However, the hair follicle width, sebaceous gland size arid cell proliferation rate, and thickness and proliferation rate of epidermal cells did not show any marked changes.

    DOI: 10.3168/jds.2017-13831

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  • 鶏脛骨軟骨異形成症におけるコラーゲンおよびグリコサミノグリカンの局在に関する免疫組織化学的観察

    池田 祥, 杉山稔恵

    2018年4月

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  • Chemistry supports the identification of gender-specific reproductive tissue in Tyrannosaurus rex 国際誌

    Mary Higby Schweitzer, Wenxia Zheng, Lindsay Zanno, Sarah Werning, Toshie Sugiyama

    SCIENTIFIC REPORTS   6   23099 - 23099   2016年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Medullary bone (MB), an estrogen-dependent reproductive tissue present in extant gravid birds, is texturally, histologically and compositionally distinct from other bone types. Phylogenetic proximity led to the proposal that MB would be present in non-avian dinosaurs, and recent studies have used microscopic, morphological, and regional homologies to identify this reproductive tissue in both theropod and ornithischian dinosaurs. Here, we capitalize on the unique chemical and histological fingerprint of MB in birds to characterize, at the molecular level, MB in the non-avian theropod Tyrannosaurus rex (MOR 1125), and show that the retention of original molecular components in fossils allows deeper physiological and evolutionary questions to be addressed.

    DOI: 10.1038/srep23099

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  • Genetic Analyses in the Japanese Captive Population of Japanese Crested Ibis (Nipponia nippon) using Pedigree Information

    Yuichi Wajiki, Yoshinori Kaneko, Toshie Sugiyama, Takahisa Yamada, Hiroaki Iwaisaki

    JOURNAL OF POULTRY SCIENCE   53 ( 1 )   1 - 7   2016年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN POULTRY SCIENCE ASSOC  

    The Japanese captive population of Japanese crested ibis (Nipponia nippon) was established using 5 founders derived from the Chinese captive population. Its size has increased rapidly, and the maintenance phase is about to start. Thus, this study was designed to perform genetic analyses in this population with pedigree information, considering the adoption of mean kinship strategy as the breeding strategy suited to the maintenance phase. Because the relationships among the 5 founders were unknown, different assumptions were set up ranging from 0 to 0.25 of kinship coefficients between the 5 founders. Assuming that the 5 founders were non-inbred in all the assumptions, the results showed that the gene diversity and the mean inbreeding coefficient would fluctuate largely from similar to 65% to similar to 82% and from similar to 0.07 to similar to 0.29, respectively. Moreover, the genetic importance of individuals based on mean kinship shifted largely. This study suggested that the Japanese captive population had low gene diversity and high mean inbreeding coefficient even under the assumption that the 5 founders were unrelated and non-inbred. In addition, the study also suggested that it became more effective to analyze the genetic status and to introduce mean kinship strategy into this population with more credible molecular evaluation of the relationships among founders.

    DOI: 10.2141/jpsa.0150040

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  • Alligator osteoderms as a source of labile calcium for eggshell formation

    C. G. Dacke, R. M. Elsey, P. L. Trosclair, T. Sugiyama, J. G. Nevarez, M. H. Schweitzer

    JOURNAL OF ZOOLOGY   297 ( 4 )   255 - 264   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    The calcium (Ca) demand on alligators in active reproduction is significant, yet the source of this additional Ca is unclear. Three possible sources for Ca mobilization are endolymphatic deposits, as in anurans and some lizards; short-lived skeletal medullary bone or analogous deposits, as in birds; or some other source such as the osteoderm layer of the integument or simply mobilization of structural bone. Here, we investigate possible extra-skeletal sources for labile Ca in the reproducing alligator, including endolymphatic Ca deposits, by analogy with anuran amphibian and some reptiles and integumentary osteodermal (scale) Ca deposits. We conducted X-ray image analyses of skulls for the presence of significant endolymphatic Ca deposits. We also examined dermal bone of scutes (osteoderm, scales) from the dorsal integument using both X-ray and histological analyses. Tissues from reproducing females containing mature but unovulated follicles were compared with those from specimens that had nested (laid eggs) or contained eggs within the oviduct at advanced stages of calcification. A small number of immature specimens and an adult male were also compared. No clear differences were observed in endolymphatic deposits between pre- and post-ovulatory specimens. Scute (osteoderm) X-ray density was significantly greater in females with ripe ovarian follicles compared with those that had recently laid (nested) or contained heavily calcified eggs within their oviducts. The latter groups also showed histological evidence of scute resorption compared with the former, suggesting that the scutes play a role in Ca storage during egglay.

    DOI: 10.1111/jzo.12272

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  • Bone morphological analyses in Spontaneously Diabetic Toni (SDT) fatty rats

    Takeshi Ohta, Shuichi Kimura, Masaya Hirata, Takahisa Yamada, Toshie Sugiyama

    JOURNAL OF VETERINARY MEDICAL SCIENCE   77 ( 10 )   1327 - 1330   2015年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN SOC VET SCI  

    The Spontaneously Diabetic Toni (SDT) fatty rat, a model for obese type 2 diabetes, shows bone quantitative abnormalities, namely low bone mineral density (BMD). The objective of this study was to evaluate bone morphological changes, in particular identifying the bone qualitative abnormalities, in the SDT fatty rat. Male SDT fatty rats showed increases in total trabecular area and trabecular number and decreases in trabecular thickness in cancellous bones of the proximal tibia, indicating trabecular miniaturization. The SDT fatty rat is useful for investigation of pathophysiological changes in bone quality in diabetic osteoporosis.

    DOI: 10.1292/jvms.15-0154

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  • ASSOCIATION OF THE EXPRESSION LEVEL OF THE MYBPC1 GENE IN SKELETAL MUSCLE WITH MARBLING TRAIT IN JAPANESE BLACK BEEF CATTLE

    Bin Tong, Youji Muramatsu, Takeshi Ohta, Hiroyuki Kose, Hideaki Yamashiro, Toshie Sugiyama, Takashisa Yamada

    Annals of Animal Science   15 ( 2 )   349 - 358   2015年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:DE GRUYTER OPEN LTD  

    Marbling characterized by the amount and distribution of intramuscular fat (IMF) in longissimus muscle (LM) and measured as beef marbling score (BMS), is an economically important trait of beef cattle in Japan. The myosin binding protein C, slow type (MYBPC1) gene, involved in efficient energy metabolism and homeostasis during muscle contraction in slow skeletal muscle, has been previously shown to be expressed at different levels in the LM between high-marbled and low-marbled steer groups using differential-display PCR (ddPCR). In this study, we found that IMF area (%) in the sacrococcygeus muscle (SM) was positively correlated with BMS in the LM in Japanese Black steers (n=22, r=0.941, P<0.0001). This suggested that the IMF area (%) in the SM tends to equate marbling level in the LM. Furthermore, we showed that the MYBPC1 expression level in SM was significantly higher in the Japanese Black steers (n=5) with high BMS than in the Japanese Black steers (n=5) with low BMS (P<0.001). Moreover, correlation analyses showed that the expression level of the MYBPC1 gene was positively correlated with IMF area (%) (n=22, r=0.858, P<0.0001) and BMS (n=22, r=0.769, P<0.0001), indicating the association of MYBPC1 expression level with marbling trait. These results, together with the previous ddPCR result, suggested that high level of MYBPC1 expression may be associated with the development of marbling in Japanese Black beef cattle.

    DOI: 10.1515/aoas-2015-0014

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  • Structure and Polymorphism of the Major Histocompatibility Complex Class II Region in the Japanese Crested Ibis, Nipponia nippon

    Yukio Taniguchi, Keisuke Matsumoto, Hirokazu Matsuda, Takahisa Yamada, Toshie Sugiyama, Kosuke Homma, Yoshinori Kaneko, Satoshi Yamagishi, Hiroaki Iwaisaki

    PLOS ONE   9 ( 9 )   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    The major histocompatibility complex (MHC) is a highly polymorphic genomic region that plays a central role in the immune system. Despite its functional consistency, the genomic structure of the MHC differs substantially among organisms. In birds, the MHC-B structures of Galliformes, including chickens, have been well characterized, but information about other avian MHCs remains sparse. The Japanese Crested Ibis (Nipponia nippon, Pelecaniformes) is an internationally conserved, critically threatened species. The current Japanese population of N. nippon originates from only five founders; thus, understanding the genetic diversity among these founders is critical for effective population management. Because of its high polymorphism and importance for disease resistance and other functions, the MHC has been an important focus in the conservation of endangered species. Here, we report the structure and polymorphism of the Japanese Crested Ibis MHC class II region. Screening of genomic libraries allowed the construction of three contigs representing different haplotypes of MHC class II regions. Characterization of genomic clones revealed that the MHC class II genomic structure of N. nippon was largely different from that of chicken. A pair of MHC-IIA and -IIB genes was arranged head-to-head between the COL11A2 and BRD2 genes. Gene order in N. nippon was more similar to that in humans than to that in chicken. The three haplotypes contained one to three copies of MHC-IIA/IIB gene pairs. Genotyping of the MHC class II region detected only three haplotypes among the five founders, suggesting that the genetic diversity of the current Japanese Crested Ibis population is extremely low. The structure of the MHC class II region presented here provides valuable insight for future studies on the evolution of the avian MHC and for conservation of the Japanese Crested Ibis.

    DOI: 10.1371/journal.pone.0108506

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  • Use of In Ovo Chorioallantoic Membrane Engraftment to Culture Testes from Neonatal Mice

    Emi Uematsu, Sachio Takino, Hidemi Okajima, Bin Tong, Toshie Sugiyama, Takahisa Yamada, Sueo Niimura, Hideaki Yamashiro

    COMPARATIVE MEDICINE   64 ( 4 )   264 - 269   2014年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ASSOC LABORATORY ANIMAL SCIENCE  

    Many attempts have been made to culture germ cells in vitro by mimicking their development in vivo. The objective of this study was to establish an alternative method of xenotransplantation by developing a new approach for the rapid induction of spermatogenesis by using the chorioallantoic membrane of developing chicken embryos. Fertilized chicken eggs were incubated for 7 d, after which a small window was cut into the shell of the egg. We then transplanted testes from 7- to 8-d-old B6D2F1 mice onto the vessels of the chorioallantoic membrane and incubated them at 35.0 degrees C for 14 d or 37.5 degrees C for 12 d. After this in ovo CAM (iCAM) culture, the survival rates of the eggs and testes were assessed histologically and immunohistologically. The transplanted testes in the chicken embryos that survived were supported by the CAM, with an associated chronic vascularization response. The testes cultured at 35.0 degrees C had lower rates of generation and higher rates of death than did those cultured at 37.5 degrees C. Histologic examination of the testes cultured at 37.5 degrees C revealed the presence of spermatogonia and primary spermatocyte-like germ cells in the seminiferous tubules. The number of cells positive for synaptonemal complex protein 3 in the seminiferous tubules was significantly higher than that in the noniCAM-cultured testes from control mice. These results suggest that iCAM culturing of neonatal donor testis induces androcyte development. This method could be the foundation for a method that would enable in vitro spermatogenesis.

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  • Identification of novel genetic markers and evaluation of genetic structure in a population of Japanese crested ibis.

    Kanako Tsubono, Yukio Taniguchi, Hirokazu Matsuda, Takahisa Yamada, Toshie Sugiyama, Kosuke Homma, Yoshinori Kaneko, Satoshi Yamagishi, Hiroaki Iwaisaki

    ANIMAL SCIENCE JOURNAL   85 ( 4 )   356 - 364   2014年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    Japanese population of the Japanese crested ibis Nipponia nippon was founded by five individuals gifted from the People's Republic of China. In order to exactly evaluate genetic structure, we first performed development of novel genetic makers using 89 microsatellite primer pairs of related species for cross-amplification. Of these, only three primer pairs were useful for the genetic markers. Additionally, we sequenced allelic PCR products of these three markers together with 10 markers previously identified. Most markers showed typical microsatellite repeat units, but two markers were not simple microsatellites. Moreover, over half of the markers did not have the same repeat units as those of the original species. These results suggested that development of novel genetic markers in this population by cross-amplification is not efficient, partly because of low genetic diversity. Furthermore, the cluster analysis by STRUCTURE program using 17 markers showed that the five founders were divided into two clusters. However, the genetic relationships among the founders indicated by the clustering seemed to be questionable, because the analysis relied largely on a small number of triallelic markers, in spite of the addition of the three useful markers. Therefore, more efficient methods for identifying large numbers of single nucleotide polymorphisms are desirable.

    DOI: 10.1111/asj.12155

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  • Polymorphisms of the dopamine receptor D4 gene in the Sado population of the Japanese crested ibis 査読

    Yamashita M, Komatsu Y, Yamada T, Tong B, Ohta T, Sugiyama T, Yamashiro H, Kaneko Y, Nagata H, Iwaisaki H

    J. Anim. Genet.   42 ( 2 )   65 - 69   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.5924/abgri.42.65

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  • The nucleotide sequences for the dopamine receptor D4 gene in the Japanese crested ibis 査読

    Yuuta KOMATSU, Mai YAMASHITA, Takahisa YAMADA, Bin TONG, Takeshi OHTA, Toshie SUGIYAMA, Hideaki YAMASHIRO, Yoshinori KANEKO, Hisashi NAGATA, Hiroaki IWAISAKI

    The Journal of Animal Genetics   42 ( 1 )   3 - 9   2014年

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.5924/abgri.42.3

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  • Genome-Wide SNP and STR Discovery in the Japanese Crested Ibis and Genetic Diversity among Founders of the Japanese Population

    Yukio Taniguchi, Hirokazu Matsuda, Takahisa Yamada, Toshie Sugiyama, Kosuke Homma, Yoshinori Kaneko, Satoshi Yamagishi, Hiroaki Iwaisaki

    PLOS ONE   8 ( 8 )   2013年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    The Japanese crested ibis is an internationally conserved, critically threatened bird. Captive-breeding programs have been established to conserve this species in Japan. Since the current Japanese population of crested ibis originates only from 5 founders donated by the Chinese government, understanding the genetic diversity between them is critical for an effective population management. To discover genome-wide single nucleotide polymorphisms (SNPs) and short tandem repeats (STRs) while obtaining genotype data of these polymorphic markers in each founder, reduced representation libraries were independently prepared from each of the founder genomes and sequenced on an Illumina HiSeq2000. This yielded 316 million 101-bp reads. Consensus sequences were created by clustering sequence reads, and then sequence reads from each founder were mapped to the consensus sequences, resulting in the detection of 52,512 putative SNPs and 162 putative STRs. The numbers of haplotypes and STR alleles and the investigation of genetic similarities suggested that the total genetic diversity between the founders was lower, although we could not identify a pair with closely related genome sequences. This study provided important insight into protocols for genetic management of the captive breeding population of Japanese crested ibis in Japan and towards the national project for reintroduction of captive-bred individuals into the wild. We proposed a simple, efficient, and cost-effective approach for simultaneous detection of genome-wide polymorphic markers and their genotypes for species currently lacking a reference genome sequence.

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  • Genetic Diversity and Structure in the Sado Captive Populationof the Japanese Crested Ibis

    Kensuke Urano, Kanako Tsubono, Yukio Taniguchi, Hirokazu Matsuda, Takahisa Yamada, Toshie Sugiyama, Kosuke Homma, Yoshinori Kaneko, Satoshi Yamagishi, Hiroaki Iwaisaki

    ZOOLOGICAL SCIENCE   30 ( 6 )   432 - 438   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ZOOLOGICAL SOC JAPAN  

    The Japanese crested ibis Nipponia nippon is a critically threatened bird. We assessed genetic diversity and structure in the Sado captive population of the Japanese crested ibis based on 24 and 50 microsatellite markers developed respectively for the same and related species. Of a total of 74 loci, 19 showed polymorphisms in the five founder birds of the population, and therefore were useful for the analysis of genetic diversity and structure. Genetic diversity measures, A, ne, He, Ho and PIC, obtained by genotyping of the 138 descendants were similar to those of other species with population bottlenecks, and thus considerably low. The low level of genetic diversity resulting from such bottlenecks was consistent with the results of lower genetic diversity measures for the Sado captive relative to the Chinese population that is the source population for the Sado group as determined using previously reported data and heterozygosity excess by Hardy-Weinberg equilibrium tests. Further, individual clustering based on the allele-sharing distance and Bayesian model-based clustering revealed that the founder genomes were equally at population in total, and with various admixture patterns at individual levels inherited by the descendants. The clustering results, together with the result of inheritance of all alleles of the microsatellites from the founders to descendants, suggest that planned mating in captive-breeding programs for the population has succeeded in maintaining genetic diversity and minimizing kinship. In addition, the Bayesian model-based clustering assumed two different components of genomes in the Sado captive Japanese crested ibis, supporting a considerably low level of genetic diversity.

    DOI: 10.2108/zsj.30.432

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  • Effects of 25-hydroxy-cholecalciferol on the development of osteochondrosis in swine

    Toshie Sugiyama, Seiji Kusuhara, Thau Kiong Chung, Hiroshi Yonekura, Elisabeth Azem, Takehiko Hayakawa

    ANIMAL SCIENCE JOURNAL   84 ( 4 )   341 - 349   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    The principal objective of this experiment was to evaluate the effect of 25-hydroxy-cholecalciferol (25-OH-D3) on the development of osteochondrosis in 6- to 110-kg castrated male pigs. The growth rate and serum calcium and inorganic phosphate levels neither increased nor decreased in response to supplementation of 25-OH-D3. However, supplemental 25-OH-D3 significantly increased serum levels of 25-OH-D3 and 1,25-hydroxy-cholecalciferol without any influence on bone mineral density. The 25-OH-D3-treated group had significant (P<0.05) reduced incidence of osteochondrotic lesions compared to the control group as evidenced by macroscopically examining the articular cartilage of the distal humerus (32.4% vs. 59.3%) and distal femur (47.1% vs. 87.5%). Likewise, supplemental 25-OH-D3 significantly reduced osteochondrotic lesions over the control when histologically examining humerus (20.6% vs. 43.8%) and femur (52.9% vs. 87.5%). The results of this experiment suggested that 25-OH-D3 supplementation in pig diets had a tendency to promote normal endochondral ossification, inhibit osteochondrosis progression and possibly regenerate destroyed cartilage tissue.

    DOI: 10.1111/asj.12000

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  • High ambient temperature influences eggshell quality and calbindin-D28k localization of eggshell gland and all intestinal segments of laying hens. 国際誌

    T A Ebeid, T Suzuki, T Sugiyama

    Poultry science   91 ( 9 )   2282 - 7   2012年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Eggshell quality deterioration associated with heat stress is a well-known phenomenon. The involvement of the 28-kDa calcium-binding protein (calbindin, CaBP-D28k) localization in this failure is not clearly understood. To test a possible direct effect of ambient temperature on calbindin-D28k localization, 40 White Leghorn laying hens were housed in individual cages and exposed to high ambient temperature (30-33°C) and thermoneutral temperature (20-22°C) which served as a control. Eggshell quality characteristics and immunohistochemical localization of all intestinal segments and eggshell gland calbindin-D28k were performed under both environmental conditions. As expected, egg weight, eggshell thickness, eggshell percentage, and eggshell density were negatively affected by high ambient temperature (P ≤ 0.01). Immunohistochemistry showed that calbindin was localized in the intestinal enterocyte cytoplasm and glandular cell cytoplasm under thermoneutral conditions. However, the calbindin intensity was prominently decreased in ileum, cecum, colon, and eggshell gland under heat stress conditions. Therefore, it could be concluded that calbindin-D28k localization in intestinal segments and eggshell gland is negatively affected by high ambient temperature which might be related to the deterioration of eggshell quality characteristics under heat stress conditions.

    DOI: 10.3382/ps.2011-01898

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  • Evidence for estrogen receptor expression during medullary bone formation and resorption in estrogen-treated male Japanese quails (Coturnix coturnix japonica) 査読

    Shinji Hiyama, Toshie Sugiyama, Seiji Kusuhara, Takashi Uchida

    JOURNAL OF VETERINARY SCIENCE   13 ( 3 )   223 - 227   2012年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:KOREAN SOC VETERINARY SCIENCE  

    The temporal expression of estrogen receptor (ER)-alpha and ER-beta mRNA was examined in male Japanese quails. Femurs of quails receiving 17 beta-estradiol underwent RT-PCR and histochemical analysis 1 to 15 days after treatment. Untreated quails were used as controls (day 0). Between days 0 and 5, cells lining the bone endosteal surface differentiated into osteoblasts, which in turn formed medullary bone. Expression of ER-alpha was already observed on day 0 and increased slightly during bone formation whereas ER-beta was hardly detected throughout this process. After osteoclasts appeared on the medullary bone surface, this type of bone disappeared from the bone marrow cavity (days 7 similar to 15). ER-alpha expression simultaneously decreased slightly and ER-beta levels remained very low. These results suggest that estrogen activity mediated by ER-alpha not only affects medullary bone formation but also bone resorption.

    DOI: 10.4142/jvs.2012.13.3.223

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  • Sequential Expression of Osteoblast PhenotypicGenes During Medullary Bone Formation and Resorption in Estrogen-Treated Male Japanese Quails 査読

    Shinji Hiyama, Toshie Sugiyama, Seiji Kusuhara, Takashi Uchida

    JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION   318B ( 5 )   344 - 352   2012年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Medullary bone is formed reticularly in the bone marrow cavity of the long bones of female birds. Although this bone matrix contains fewer collagen fibers and more acid mucopolysaccharides than cortical bone, it is not clear that the expression pattern of osteoblast phenotypic genes during bone remodeling. Therefore, 17 beta-estradiol (E2)-treated male Japanese quails were used to examine the temporal expression patterns of osteoblast phenotypic genes, and to simultaneously confirm the morphological changes occurring in the bone marrow cavity during medullary bone formation and resorption. After E2 treatment, bone lining cells proliferated and developed into mature osteoblasts that had intense alkaline phosphatase (ALP) activity. These cells began to form medullary bone that contained acid mucopolysaccharides and tartrate-resistantacid phosphatase. Runt-related gene 2 (Runx2) mRNA was stably expressed throughout the process. The expression of both ALP and type I collagen mRNAs increased initially, and then rapidly decreased after day 7, while osteoclasts began to resorb medullary bone at day 5. The expression of bone matrix-related genes peaked at day 5, and suddenly decreased at day 7, except for osteopontin. Taken together with these results, the expression patterns of bone matrix-related genes during the later stages might be related to osteoclast activity. Additionally, the constant expression of Runx2 during bone formation and resorption suggested that osteoprogenitor cells always exist in the bone marrow cavity. Therefore, the expression patterns of these genes and the characteristics of bone matrix might extremely be related to the quick remodeling of medullary bone. J. Exp. Zool. (Mol. Dev. Evol.) 318B:344-352, 2012. (c) 2012 Wiley Periodicals, Inc.

    DOI: 10.1002/jez.b.22451

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  • The W- and Z-linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island. 国際誌

    Kaori Kasuga, Maho Higashi, Takahisa Yamada, Toshie Sugiyama, Yukio Taniguchi, Hiroaki Iwaisaki, Kosuke Homma, Yuuichi Wajiki, Yoshinori Kaneko, Satoshi Yamagishi

    Animal science journal = Nihon chikusan Gakkaiho   83 ( 1 )   83 - 7   2012年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR-based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W- and Z-linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR-based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR-based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.

    DOI: 10.1111/j.1740-0929.2011.00971.x

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  • Polymorphism in Promoter Region of CDC10 Gene Showing Marbling-Associated Expression Changes

    Bin Tong, Youji Muramatsu, Takuji Yamamoto, Hideki Tanomura, Takeshi Ohta, Hiroyuki Kose, Toshie Sugiyama, Takahisa Yamada

    JOURNAL OF ANIMAL AND VETERINARY ADVANCES   11 ( 11 )   1930 - 1933   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MEDWELL ONLINE  

    Researchers have previously showed that the septin 7 (CDC10) gene involved in cellular proliferation possesses expression differences in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (BM6437) close to the CDC10 was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the promoter region of the CDC10 gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distribution of the SNP, referred to as g.63629097G>C in the CDC10 was indistinguishable between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the BM6437 marker but not the g.636290970>C SNP may be related to changes in CDC10 gene expression and/or marbling.

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  • Effects of beta-Cryptoxanthin on Bone Metabolism in a Rat Model of Osteoporosis 査読

    Noriko Ikeda, Toshie Sugiyama, Toshiko Suzuki, Katsuyuki Mukai, Seiji Kusuhara

    JOURNAL OF ANIMAL AND VETERINARY ADVANCES   11 ( 1 )   30 - 35   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MEDWELL ONLINE  

    Osteoporosis is a bone disease in which Bone Mineral Density (BMD) is reduced with a consequent increase in the risk of bone fractures. P-cryptoxanthin (beta-CRP) is present in large amounts in Satsuma mandarins and was recently reported to stimulate bone formation. In this study, researchers investigated the effects of beta-CRP in Satsuma Mandarin. Pulp (SW; 2 mg g(-1) beta-CRP) on bone metabolism in an Ovariectomized (OVX) rat model of osteoporosis. Female rats (12 weeks of age) were ovariectomized and orally administered vehicle, 0.03 g day(-1) SMP or 0.3 g day(-1) SMP for 5 weeks. After that serum concentrations of osteocalcin (an osteoblastic bone formation marker) tended to be higher in the SMT groups than in the OVX vehicle group while those of collagen type I degradation products an osteoclastic bone resorption marker) tended to be lower in the SMP groups. By bone histomorphometry, bone trabecular volume/tissue volume ratios and trabecular numbers were significantly higher in the SNIP groups than in the OVX vehicle group while trabecular separation and osteoclast number/bone surface ratios were significantly lower in the SNIP groups. By immunohistochemistry, percentage areas of osteocalcun immunoreactivity on trabecular surface were significantly greater in the SNIP groups than in the OVX vehicle group. Dual-energy X-ray absorptiometry analyses revealed that BMDs of the lumbar vertebrae and femora, tibiae tended to increase as the dose of SMP increased in the OVX rats. In conclusion, oral Styli) administration stimulated osteoblastic bone formation and inhibited osteoclastic bone resorption in OVX rats, thereby preventing the bone loss associated with osteoporosis.

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  • Preliminary Association Study of SNPs in MFN2 Gene Showing Marbling-Associated Expression Changes

    Bin Tong, Youji Muramatsu, Takuji Yamamoto, Hideki Tanomura, Takeshi Ohta, Hiroyuki Kose, Toshie Sugiyama, Takahisa Yamada

    JOURNAL OF ANIMAL AND VETERINARY ADVANCES   11 ( 15 )   2796 - 2800   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MEDWELL ONLINE  

    Researchers have previously showed that the mitofusin 2 (MFN2) gene involved in energy balance through mitochondrial fusion and expressed in slow-twitch oxidative fiber that is observed in high-marbled muscle, possesses expression differences in muscles in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (IDVGA-49) close to the MFN2 was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the MFN2 gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distributions of the 6 SNPs in the MFN2 were indistinguishable between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the IDVA-49 marker but not the 6 SNPs may be related to changes in MFN2 gene expression and or marbling. The IDVA-49 marker may be useful for effective marker-assisted selection to increase the levels of marbling.

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  • Useful Marker Around TRDN Gene Showing Marbling-Associated Expression Changes

    Bin Tong, Youji Muramatsu, Takuji Yamamoto, Hideki Tanomura, Takeshi Ohta, Hiroyuki Kose, Toshie Sugiyama, Takahisa Yamada

    JOURNAL OF ANIMAL AND VETERINARY ADVANCES   11 ( 15 )   2792 - 2795   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MEDWELL ONLINE  

    Researchers have previously showed that the triadin (TRDN) gene involved in muscle contraction possesses expression differences in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (BMS817) close to the TRDN was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the TRDN gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distribution of the SNP referred to as g.3834941C>T in the TRDN was indistinguishable between Japanese Black sires with extremely high predicted breeding value for marble and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the BMS17 marker but not the g.3834941C>T SNP may be related to changes in TRDN gene expression and/or marbling. The BMS817 marker may be useful for effective marker-assisted selection to increase the levels of marbling.

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  • Mechanism of Visceral Fat Reduction in Tsumura Suzuki Obese, Diabetes (TSOD) Mice Orally Administered beta-Cryptoxanthin from Satsuma Mandarin Oranges (Citrus unshiu Marc) 国際誌

    Katsuhiko Takayanagi, Sho-ichi Morimoto, Yoshiyuki Shirakura, Katsuyuki Mukai, Toshie Sugiyama, Yoshihiko Tokuji, Masao Ohnishi

    JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY   59 ( 23 )   12342 - 12351   2011年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    The carotenoid beta-cryptoxanthin (beta-CRX) is abundant in Satsuma mandarins (Citrus unshiu Marc). Several studies have shown a relationship between Satsuma mandarin consumption and a low risk of several diseases, for example, diabetes, gout, and hypertension, suggesting beta-CRX involvement in disease prevention. We investigated the effect of beta-CRX on mildly obese males. beta-CRX administration reduced visceral adipose tissue, body weight, and abdominal circumference. However, the detailed mechanism by which beta-CRX mediates these changes remains unknown. To identify this mechanism, we used an obese model mouse (TSOD). Oral beta-CRX administration repressed body weight, abdominal adipose tissue weight, and serum lipid concentrations in TSOD; these results are identical to previous human trial results. beta-CRX administration significantly repressed adipocyte hypertrophy. Gene expression analysis strongly indicated that beta-CRX can alter cytokine secretion and cell proliferation. These results suggest that beta-CRX derived from Satsuma mandarins can help prevent obesity by repressing hypertrophy of abdominal adipocytes.

    DOI: 10.1021/jf202821u

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  • Non-invasive sampling technique for DNA extraction from captive Japanese Crested Ibis on Sado Island

    Kensuke Urano, Takahisa Yamada, Yukio Taniguchi, Hiroaki Iwaisaki, Toshie Sugiyama, Kosuke Homma, Yoshinori Kaneko, Satoshi Yamagishi

    ANIMAL SCIENCE JOURNAL   82 ( 4 )   616 - 619   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The Japanese Crested Ibis Nipponia nippon is a critically threatened bird. The post-hatch eggs of the current captive population of this species on Sado Island have been stored at room temperature for the long-term. In this study, we investigated the suitability of the vascularized chorioallantois membrane from the eggs as a non-invasive DNA source. Using microsatellite loci developed for the Japanese Crested Ibis, we performed three experiments for comparison of genotypes obtained among DNA. First, DNA from five different sites of the identical membrane showed the same genotypes at either of two loci examined. Second, DNA from the membrane of each full-sibling birds and blood of their parents showed the genotypes that were consistent with Mendelian parent-offspring relationships at any of eight loci examined. Third, DNA from the membrane and blood of the same bird showed the matched genotypes at any of eight loci examined. These results indicate that the vascularized chorioallantois membrane from post-hatch eggs stored at room temperature for the long-term can be used as a reliable DNA source of offspring that had hatched from the egg. This study will promote a molecular genetics study on genetic diversity of the current captive Japanese Crested Ibis population on Sado Island.

    DOI: 10.1111/j.1740-0929.2011.00902.x

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  • Intensified nursing dramatically accelerates growth performance and the size of the body frame in Japanese black and holstein crossbred steers 査読

    Sunao Inada, Fumio Ebara, Sohei Asaoka, Kenichi Asada, Yoshihiro Isozaki, Akira Saito, Toshie Sugiyama, Takafumi Gotoh

    Journal of Animal and Veterinary Advances   9 ( 6 )   1037 - 1047   2010年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Crossbred (Japanese Black male and Holstein female) steers were used to investigate the effects of feeding a large amount of high-protem milk replacer on growth performance, morphological change and myogenic regulation in skeletal muscle. Group HP (n = 7) was fed high-protein milk replacer (Crude Protein (CP): 29%, maximum intake 2.0 kg day-1) and group C (n = 14) was fed a standard milk replacer (CP: 26%, 0.5 kg day-1) from 56-147 days of age. Changes in Body Weight (BW), body frame measurements, plasma Insulin-like Growth Factor-1 (IGF-1) and plasma Alkaline Phosphatase (ALP) were investigated. M. Longissimus Thoracis (LT) samples were obtained by biopsy to investigate the myofiber type composition, diameter and the mRNA expression of Myogenic Regulatory related genes by quantitative real-time RT-PCR. A large amount of high protein milk replacer (Intensified Nursing) in group HP improved their synthesis of IGF-1 in the liver and accelerated skeletal development by maintaining high levels of plasma IGF-1 and ALP concentration, which was highly controlled by CP intake from diet. The myofiber type composition and size in the LT was not influenced by nutritional condition and aging during the nursing period. Intensified Nursing extends myofiber length rather than increasing the thickness of the myofiber, according to the increase in the size of the body frame during this phase. The expression of MyoD and IGF-1 receptors in the LT decreased at the early stage of the nursing period without the influence of nutritional condition. This suggested the change would control the level of differentiation from the myoblast to the myocyte. © Medwell Journals, 2010.

    DOI: 10.3923/javaa.2010.1037.1047

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  • Cartilage Pathology of Intra-articular Papain-induced Leg Weakness in a Pig Model

    Sachiko ARAI, Seigo ITOH, Toshie SUGIYAMA, Seiji KUSUHARA, Fujiko SUNAGA

    47 ( 3 )   136 - 138   2010年

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    掲載種別:研究論文(学術雑誌)  

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  • Clinical model of leg weakness induced by intraarticular administration of papain in pigs

    S Arai, S Itoh, M Kachi, T Sugiyama, S Kusuhara

    Japanese Journal of Swine Science   46 ( 4 )   183 - 189   2009年12月

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  • トキの頚部筋系の形態学的観察 査読

    金子良則, 遠藤秀紀, 成島悦雄, 橋崎文隆, 杉山稔恵, 楠原征治

    動物園水族館雑誌   50 ( 3-4 )   79 - 87   2009年8月

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  • トキの頚椎の形態学的観察

    金子良則, 遠藤秀紀, 成島悦雄, 橋崎文隆, 杉山稔恵, 楠原征治

    動物園水族館雑誌   50 ( 2 )   47 - 55   2009年5月

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  • Evidence for the expression of estrogen receptors in osteogenic cells isolated from hen medullary bone 査読

    Shinji Hiyama, Toshie Sugiyama, Seiji Kusuhara, Takashi Uchida

    ACTA HISTOCHEMICA   111 ( 6 )   501 - 507   2009年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER GMBH, URBAN & FISCHER VERLAG  

    Medullary bone is a unique tissue in female birds and forms in the cavity of tong bones. This bone displays rapid remodeling in response to circulating estrogen levels, suggesting that the osteoblasts in this bone are highly sensitive to estrogen. The present study examined expression of two estrogen receptor (ER) mRNAs in osteogenic cells of medullary bone of white Leghorn hens in vitro. At day 3, isolated cells from the hen medullary bone expressed alkaline phosphatase activity. Using immunocytochemistry, ER protein was demonstrated in the nuclei of these cells. RT-PCR analysis revealed that ER-alpha mRNA was constantly expressed from day 3 to day 15 of culture, white ER-beta mRNA was not detected throughout the culture period. These results indicate that estrogen may act via ER-alpha, but not ER-beta, on osteogenic cells of the avian medullary bone. (C) 2008 Elsevier GmbH. All rights reserved.

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  • ニワトリ脛骨軟骨異形成誘発病変における軟骨細胞および軟骨管の微細構造学的観察 査読

    可知 真奈美, 杉山 稔恵, 楠原 征治

    家畜衛生学雑誌   33 ( 4 )   143 - 149   2008年3月

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    記述言語:日本語   出版者・発行元:日本家畜衛生学会  

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  • Endochondral ossification of chick embryonic femora in vitro and on chorioallantoic membrane 査読

    Manami Kachi, Toshie Sugiyama, Seiji Kusuhara

    Journal of Poultry Science   45 ( 1 )   51 - 56   2008年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We observed angiogenesis and endochondral ossification of the femora of chick embryos in vitro, on chorioallantoic membrane (CAM), and in vivo to clarify the cellular processes of avian endochondral ossification. We found that the inside of the in vitro femora was still filled with chondrocytes, despite being cultured for 10 days, while calcification of the diaphysis was not observed. As well, only the cartilage tissue of the epiphysis was enlarged. By contrast, blood vessels invaded into the diaphysis and the marrow cavity was formed in CAM-cultured femora. Cartilage canals extended from the marrow cavity and reached to the resting chondrocyte zone, with normal endochondral ossification occurring as in vivo. This study demonstrates that endochondral ossification occurs in femora in CAM culture similar to that in in vivo femora, but not in in vitro femora. The ossification is dependent on vascular invasion into the embryonic femora. In conclusion, for the endochondral ossification of long bones it is essential to supply the embryo with blood vessels. CAM culture system was found to be a superior endochondral ossification model of the embryonic femur. Copy right © 2008, Japan Poultry Science Association.

    DOI: 10.2141/jpsa.45.51

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  • ブロイラーにおける脛骨軟骨異形成(Tibial Dyschondroplasia: TD)誘発病変の形態学的観察 査読

    可知 真奈美, 杉山 稔恵, 楠原 征治

    家畜衛生学雑誌   33 ( 3 )   71 - 78   2007年11月

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    記述言語:日本語   出版者・発行元:日本家畜衛生学会  

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  • Changes in localization of lysosomal enzymes in muscle induced by conditioning or high pressure treatment. 査読

    Fukumura, M, Sugiyama, T, Nishiumi, T, Suzuki, A

    High Pressure Bioscience and Biotechnology   1   239 - 244   2007年8月

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)  

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  • Expression and localisation of calbindin D28k in all intestinal segments of the laying hen. 国際誌

    T Sugiyama, H Kikuchi, S Hiyama, K Nishizawa, S Kusuhara

    British poultry science   48 ( 2 )   233 - 8   2007年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. The aim of the present study was to investigate expression and localisation of a 28-kDa calcium-binding protein (CaBP-D28k) related to active calcium (Ca) absorption, in the entire intestine of egg-laying hens. 2. Western blotting analysis showed that the entire intestine expressed CaBP-D28k to the following degree: duodenum > jejunum > caecum > ileum > colon. Immunohistochemistry showed strong CaBP-D28k localisation in enterocytes along the villus tip-crypt axis in the duodenum and in villus tips in the caecum and colon. The jejunum and ileum had moderate localisation with respect to the number of immunoreactive cells and staining intensity. 3. These results suggest that laying hens actively absorb Ca in both the large and small intestines.

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  • Changes in the Outer Shape, Inner Structure and Bone Mineral Density of the Tibia in Growing Japanese Quail, Coturnix japonica

    Kouichi Nishimura, Susumu Miyamoto, Takao Takeda, Eishi Ayukawa, Toshie Sugiyama, Seiji Kusuhara

    Journal of Poultry Science   44 ( 4 )   426 - 432   2007年

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  • Changes in the immunogold electron-microscopic localization of calpain in bovine skeletal muscle induced by conditioning and high-pressure treatment 査読

    Gerelt Borjigin, Shuhei Yamamoto, Toshie Sugiyama, Tadayuki Nishiumi, Atsushi Suzuki

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 5 )   1249 - 1254   2006年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Changes in the localization of calpain in conditioned and pressurized bovine skeletal muscles were investigated by immunogold electron-microscopy. In the muscle immediately after thawing (control), the relative distributions of colloidal particles statistically calculated by counting the colloidal particles were about 65% and 35% in the I-band/Z-disk and A-band regions, respectively. In the muscle conditioned for 7 days, distribution of colloidal particles was more than two times greater in both the I-band/Z-disk and A-band regions than in the control muscle. Almost no change in either the absolute concentration or relative distribution of the colloidal particles was detectable during further storage. In the muscle exposed to a pressure of 100 MPa or, 200 MPa, slightly more immunogold was detected in the sarcomere than in that of the control muscle. Increasing pressure up to 300 MPa enabled high-density particles to be seen throughout the sarcomere. Conversely, few particles were detected in the sarcomere of the muscle exposed to 400 MPa. These electron-microscopic observations were confirmed from the statistical analysis as with the conditioned muscles. It was clear from the results obtained that the pressure-induced changes in calpain localization were much;more pronounced than those from conditioning.

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  • Expression and localization of estrogen receptors α and β mRNA in medullary bone of laying hens 査読

    Tomohiko Imamura, Toshie Sugiyama, Seiji Kusuhara

    Animal Science Journal   77 ( 2 )   223 - 229   2006年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The aim of this study was to observe the expression and localization of estrogen receptor (ER) α and ER β mRNA in the medullary bone of laying hens. First, medullary bone, liver, kidney, and shell gland of the oviduct tissues were dissected from laying hens. Then, the total cellular RNA was isolated from each tissue specimen, and the ER α and ER β mRNA expression was observed using semiquantitative RT-PCR. Second, the localization of ER α mRNA in the medullary bone was detected with in situ hybridization using digoxigenin-11-UTP-labeled cRNA probes. As a result, the expression of ER α mRNA was higher than that of ER β mRNA in the medullary bone, liver, and shell gland of the oviduct from laying hens. In the kidney, ER α mRNA expression was lower than that of ER β mRNA. The expression pattern of ER α and ER β mRNA of the medullary bone was similar to that of the shell gland of the oviduct. Moreover, ER α mRNA was intensively expressed in osteoblasts on the medullary bone surface and bone marrow stromal cells but was not expressed in osteoclasts. These results suggest that in medullary bone, estrogen action may be regulated not by ER β but by ER α. © 2006 Japanese Society of Animal Science.

    DOI: 10.1111/j.1740-0929.2006.00341.x

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  • Cellular and molecular phenotypes of osteogenic cells isolated from the medullary bone of the hen in vitro 査読

    Shinji Hiyama, Toshie Sugiyama, Seiji Kusuhara, Takashi Uchida

    Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology   142 ( 4 )   419 - 425   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In this study, cells isolated from hen medullary bone were cultured to examine their matrix formation. Furthermore, we compared medullary bone cells with rat bone marrow cells regarding the temporal changes in osteoblast developmental markers. Medullary bone cells were positive for alkaline phosphatase (ALP) activity and formed bone nodules, apparent with Alcian blue and von Kossa staining. The intensity of these stains became stronger with the maturation of those bone nodules. In this developmental process, the expression patterns of osteoblast phenotypes of medullary bone cells differed from those of rat bone marrow cells. ALP mRNA was expressed at the maximum level in the proliferation stage and gradually decreased in medullary bone cells, but that expression showed the opposite pattern in rat bone marrow cells. Medullary bone cells strongly expressed two non-collagenous protein mRNAs from the early stages, but the expression of these mRNAs in rat bone marrow cells increased only in the later stages. These results suggest that the features of medullary bone osteoblasts differ from those of mammalian osteoblasts and are reflected in the characteristics of medullary bone in vivo. © 2005 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.cbpb.2005.09.006

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  • 鳥類カルシウム代謝における骨髄骨の形成と吸収

    杉山 稔恵

    日本家禽学会誌   42 ( 4 )   J197 - J208   2005年11月

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    記述言語:日本語   出版者・発行元:日本家禽学会  

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  • Effect of calcium regulating hormones on osteoclast-like cell formation in hen medullary bone marrow culture

    T. Sugiyama, M. Sakurai, S. Hiyama, S. Kusuhara

    Journal of Poultry Science   41 ( 4 )   307 - 314   2004年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.2141/jpsa.41.307

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  • ブタの品種間における骨軟骨症の発生実態

    大畑 博義, 杉山 稔恵, 楠原 征治

    家畜衛生学雑誌   28 ( 2 )   65 - 70   2002年12月

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    記述言語:日本語   出版者・発行元:日本家畜衛生学会  

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  • Osteoclast-like cell formation in medullary bone marrow cell culture of laying hens

    SUGIYAMA Toshie, SAKURAI Midori, HIYAMA Shinji, KUSUHARA Seiji

    Journal of Poultry Science   39 ( 4 )   256 - 265   2002年10月

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    記述言語:英語   出版者・発行元:Japan Poultry Science Association  

    DOI: 10.2141/jpsa.39.256

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  • Changes in immunoelectron microscopic localization of cathepsin D in muscle induced by conditioning or high-pressure treatment

    T Kubo, B Gerelt, GD Han, T Sugiyama, T Nishiumi, A Suzuki

    MEAT SCIENCE   61 ( 4 )   415 - 418   2002年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    Changes in the localization of cathepsin D in postmortem and pressurized rabbit muscles were investigated by immunoelectron microscopy. The anti-cathepsin D monoclonal antibody strongly labeled a large vesicle in a subsarcolemmal part of the cell, which strongly suggests that this is the primary lysosome. The liberation of the cathepsin D entrapped in the lysosomes in the muscle prepared immediately after death proceeded with the progress of the conditioning. The release of almost all cathepsin D from the lysosomes and its absorption on the myofibrils were observed in the muscle conditioned for 14 days. The accumulation of lysosomes having various volumes and shapes accompanied with the disruption of myofibrillar structure was also observed. The liberation of cathepsin D from the lysosomes can be attributed to the modification of membranes permeability of the lysosomes during conditioning. When the muscle was pressurized at 100 MPa, the modification of the round shape of the lysosome was observed. With the increase of the pressure applied to the muscle, the release of cathepsin D from the lysosome due to the disruption of membrane was accelerated, and absorption of the released cathepsin D on the myofibrils was observed. From the results obtained, it was clear that the changes in the localization of cathepsin D accompanied with the modification of lysosomes induced by the brief exposure to high pressure were drastic in comparison with that in the muscle during conditioning. (C) 2002 Elsevier Science Ltd. All rights reserved.

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  • 放牧牛から分離したTrypanosoma theileriの継代培養株の形態学的比較

    石田秀史, 杉山稔恵, 楠原征治

    家畜衛生研究会報   27 ( 2 )   65 - 70   2002年3月

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  • Immunohistochemical study of matrix metalloproteinase-3 (MMP-3) at the articular cartilage in oseteochondrotic pigs

    H. OHATA, T. SUGIYAMA, S. KUSUHARA

    Animal Science Journal   73, 517-522   2002年

  • Avian calcium metabolism and bone function 招待 査読

    T Sugiyama, S Kusuhara

    ASIAN-AUSTRALASIAN JOURNAL OF ANIMAL SCIENCES   14   82 - 90   2001年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ASIAN-AUSTRALASIAN ASSOC ANIMAL PRODUCTION SOCIETIES  

    Avian calcium metabolism in an egg-laying period is extraordinary when compared with all other classes of vertebrates. The hard eggshell consists of calcium carbonate (CaCo3). About 60-75% of the calcium in the eggshell is derived from dietary sources and the remaining 25-40% from skeletal stores, called "medullary bone". Medullary bone is specifically developed in marrow cavities of long bones and plays an important role as a calcium reservoir for eggshell formation. An onset of medullary bone formation coincides with initiation of ovarian follicle maturation and the secretion of gonadal hormones, estrogen and androgen. Therefore their formation requires the combined influence of both estrogen and androgen. However, not only gonadal hormones but 1,25-dihydroxy vitamin D-3 is necessary for the formation of fully mineralized medullary bone. There are osteoblasts, bone-forming cells, and osteoclasts, bone-resorbing cells, on the hen medullary bone surface. In the domestic hen, medullary bone formation and resorption alternately occur during the 24-hour egg-laying cycle. These cyclic changes in medullary bone metabolism depend on the plasma levels of endogenous hormones fluctuating during the egg-laying cycle. Namely, when an egg is in the infundibulum, magnum or isthmus of the oviduct, osteoblasts actively form medullary bone with the induction of estrogen secreted by matured follicles. On the other hand, when an egg enters into the shell gland and begins to be calcified, the mobilization of calcium for eggshell formation causes a decrease of plasma ionized calcium concentrations. Consequently, to compensate the plasma calcium concentrations to be normal, parathyroid hormone is secreted from parathyroid glands to accelerate the osteoclastic bone resorption. With the end of eggshell formation, calcitonin is secreted from ultimobranchial glands thus inhibiting the osteoclastic bone resorption. In recent years, a number of manmade chemicals have shown to be able to mimic estrogen and androgen. It has been hypothesized that alterations in the normal pattern of endocrine systems, seen in some populations of wildlife, are linked with exposure to these chemicals. Interestingly, the medullary bone formation is induced by both estrogen and androgen, and the receptors of endocrine disruptors such as estrogen, androgen and aryl hydrocarbon (dioxin) receptors are highly expressed in the medullary bone. Therefore, it is sure that medullary bone would be a useful model to elucidate the effects of endocrine disruptors on wildlife and humans.

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  • Pressure-induced changes in the connectin/titin localization in the myofibrils revealed by immunoelectron microscopy

    A. Suzuki, Y. Homma, K. Kim, Y. Ikeuchi, T. Sugiyama, M. Saito

    Meat Science   59 ( 2 )   193 - 197   2001年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Model selection and genetic parameter estimation for performance traits, body measurement traits and leg seore traits in a closed population of Duroc pigs

    K. Fukawa, T. Sugiyama, S. Kusuhara, O. Kudoh, K. Kameyama

    Animal Science Journal   72 ( 2 )   97 - 106   2001年3月

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  • 家禽における産卵とカルシウム代謝

    杉山稔恵

    北信越畜産学会誌   83   17 - 24   2001年

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  • Estimation of Genetic Parameters on Leg Score and Joint Cartilage Lesion Scores in a Closed Population of Duroc Pig

    FUKAWA Kazuo, SUGIYAMA Toshie, KUSUHARA Seiji, KUDOH Osamu, KAMEYAMA Kenji

    Animal Science Journal   71 ( 4 )   353 - 362   2000年7月

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    記述言語:英語   出版者・発行元:Japanese Society of Animal Science  

    Closed population of 6 generations Duroc pigs was selected for higher productivity and better leg soundness. The leg joint lesions of 174 slaughtered pigs at approximately 90kg live weight were scored from 0 (no lesion) to 4 (very severe lesion) in optical, radiographical and histological standpoints throughout the six generations. Moreover, one thousand and three hundred eighty three pigs were scored from 1 (very good) to 5 (very weak) for leg weakness. Significant differences in the leg weakness score and joint lesion scores were observed among selected generations. Significant differences were observed between gilt and boar/barrow group in 9 traits. The heritabilities of joint lesion scores were low through intermediate. The heritabilities of histological joint scores were also low through intermediate. The phenotypic and genetic correlations between the scores of front (humerus and ulna) and rear legs (femur) were low. The genetic correlations between optical, radiographical and histological scores of the same joint were generally high. The phenotypic correlations between the leg weakness score and joint lesion scores were positive although statistically not significant from zero. The genetic correlations between the leg weakness score and joint lesion scores were generally intermediately or highly positive. The result suggested that selection based on joint lesion scores could lead to a better leg joint condition optically and histologically. It also suggested that selection based on leg weakness score could lead to the reduction of leg joint lesion according to the correlated response.

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  • Perspectives on osteoblast and osteoclast function

    CV Gay, VR Gilman, T Sugiyama

    POULTRY SCIENCE   79 ( 7 )   1005 - 1008   2000年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:POULTRY SCIENCE ASSOC INC  

    We present evidence that the polar, matrix-forming osteoblasts are connected laterally to form an impervious layer of cells. Next, the possible mechanisms by which calcium ions are translocated across the layer of cells into sites of mineralization are analyzed. Finally, mechanisms of attachment of bone-resorbing osteoclasts are considered. Osteoclasts adhere to matrix, in part, though an arginine-glycine-aspartic acid (RGD)-dependent mechanism. Adherence is under control of parathyroid hormone and 17 beta-estradiol.

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  • エストロジェンおよびアンドロジェンの投与量がニワトリ骨髄骨の発達に及ぼす影響

    杉山稔恵, 布村夏子, 楠原征治, 對馬宣道

    日本畜産学会北陸支部会報   79   42 - 45   1999年

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  • Characteristics of the Bone Matrix Formed by Osteogenic Cells Isolated from Embryonic Chick Calvaria and Hen Medullary Bones.

    Shinji Hiyama, Toshie Sugiyama, Seiji Kusuhara

    Japanese Poultry Science   35 ( 4 )   228 - 233   1998年7月

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    記述言語:英語   出版者・発行元:Japan Poultry Science Association  

    本研究では,鶏胚頭蓋冠および産卵鶏骨髄骨から分離された骨形成系細胞が形成した骨基質の特徴を検討した。頭蓋冠から分離された骨形成系細胞の細胞小器官は,骨髄骨から分離された骨形成系細胞と比較して,著しく発達していた。これらの細胞はいずれもアルカリ性ホスファターゼ活性を有し,頭蓋冠から分離された骨形成系細胞が形成した基質では,酸性粘液多糖類が少なく,太いコラーゲン線維が密に存在していた。一方,骨髄骨から分離された骨形成系細胞が形成した基質では,酸性粘液多糖類が豊富で,細いコラーゲン線維がまばらに分布していた。さらに,これらの基質にはいずれも石灰化像を示す塊状の結晶構造物が観察された。以上の結果から,鶏胚頭蓋冠および産卵鶏骨髄骨から分離された細胞は,骨形成能を有し,皮質骨および骨髄骨に類似した骨基質をそれぞれ形成することが示唆された。

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  • Adhesive and Bone Resorptive Activities of Isolated Osteoclasts from Hen Medullary Bone.

    SUGIYAMA Toshie, KUSUHARA Seiji

    Journal of Veterinary Medical Science   60 ( 5 )   573 - 578   1998年

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    記述言語:英語   出版者・発行元:JAPANESE SOCIETY OF VETERINARY SCIENCE  

    In the present study, osteoclasts were isolated from hen medullary bones at the formative and resorptive phases. The cells were cultured on glass culture dishes and bone slices. After culturing, the adhesion activity of the isolated osteoclasts with the substrates was estimated with a light microscope, and the surfaces of the bone slices were observed with a scanning electron microscope. The results showed that the adhesion activity of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts is higher at the bone resorptive phase than at the bone formative phase, and this tendency in isolated osteoclasts was observed more frequently on the bone slices than on the glass culture dishes. Furthermore, scanning electron microscopy showed that the isolated osteoclasts in the bone resorptive phase adhered to the bone surface with developed-cytoplasmic projections and formed broad pits where collagen fibrils were exposed. On the other hand, isolated osteoclasts in the bone formative phase adhered to the bone slice with board-shaped cytoplasmic projections and did not form any pits. These results suggest that isolated osteoclasts in the bone resorptive phase have a high level of adhesion activity and actively resorb the bone, whereas isolated osteoclasts in the bone formative phase have a low level of adhesion activity and cease bone resorption. The procedure reported here is useful for studying the bone-resorptive mechanism of authentic osteoclasts.

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  • 産卵鶏の卵管殻腺部におけるCa<sup>2+</sup>-ATPaseおよびcarbonic anhyrase IIの免疫組織化学的研究

    新井 達弥, 杉山 稔恵, 楠原 征治

    日本家禽学会誌   33 ( 6 )   371 - 376   1996年11月

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    記述言語:日本語   出版者・発行元:日本万国家禽学会  

    本研究では,放卵後3時間および12時間の産卵鶏を用い,卵管の卵殻腺部におけるcalcium-adeno-sine triphosphatase(Ca<sup>2+</sup> -ATPase)およびcarbonic anhydrase II (CAII)の局在について免疫組織化学的に検出し,卵殻腺部の粘膜上皮および管状腺におけるCa<sup>2+</sup>とHCO<sub>3</sub>の分泌について検討した。<br>卵殻腺部におけるCa<sup>2+</sup> -ATPaseの局在を観察したところ,粘膜上皮においては,免疫反応は認められなかった。管状腺においては,管状腺細胞の腺腔面および導管部において強い免疫反応が認められた。また,放卵後3時間および12時間の産卵鶏における免疫反応の間に違いは認められなかった。<br>卵殻腺部におけるCAIIの局在を観察したところ,粘膜上皮において強いCAIIの免疫反応が認められ,管状腺においては,粘膜上皮と比較して幾分弱いCAIIの免疫反応が認められた。また,放卵後3時間および12時間の産卵鶏で差はみられなかった。<br>これらのことから,卵殻腺部におけるCa<sup>2+</sup>の分泌は,管状腺に存在するCa<sup>2+</sup>-ATPaseによる能動輸送によって行なわれていることが推察された。また,卵殻形成に必要とされるHCO<sub>3</sub>の産生と分泌には,管状腺よりもむしろ粘膜上皮の方がより強く関与していることが推察された。

    DOI: 10.2141/jpsa.33.371

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  • Effects of parathyroid hormone and calcitonin on kinetics of actin filaments in chicken osteoclasts of cultured medullary bone

    T. Sugiyama, S. Kusuhara

    Animal Science and Technology   67 ( 6 )   526 - 532   1996年6月

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    記述言語:英語   出版者・発行元:Japanese Society of Animal Science  

    放卵後3時間で骨吸収が停止している時期および放卵後12時間で骨吸収が促進している時期の産卵鶏骨髄骨を合成ヒト副甲状腺ホルモン(PTH)および合成ウナギカルシトニン(CT)添加培地でそれぞれ培養し,破骨細胞におけるアクチンフィラメント分布におよぼす影響について検討した.放卵後3時間における培養前の骨髄骨では,破骨細胞において骨基質側の明帯に相当する部分に,帯状を呈したアクチンフィラメントが強く認められた.これらをPTH無添加培地で24時間培養したところ,アクチンフィラメント分布は培養前と変化しなかったものの,PTH添加培地ではアクチンフィラメントの帯状の中心部は不明瞭となり,骨基質に対して垂直な線状を呈したアクチンフィラメントが観察された.一方,放卵後12時間における培養前の骨髄骨では,破骨細胞において骨基質側の波状縁部分に,骨基質に対して垂直な線状を呈したアクチンフィラメントが認められた.これらをCT無添加培地で24時間培養したところ,アクチンフィラメント分布は培養前と変化しなかったものの,CT添加培地では線状を呈したアクチンフィラメントは消失し,帯状を呈したアクチンフィラメントが観察された.以上の結果から,PTHおよびCTは,アクチンフィラメントの分布を改変することにより破骨細胞による骨吸収を調節していることが示唆された.

    DOI: 10.2508/chikusan.67.526

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  • ウズラ大腿骨器官培養の骨髄骨形成に及ぼす性ホルモンの影響

    加藤久美子, 杉山稔恵, 楠原征治

    日本畜産学会報   67 ( 1 )   64 - 68   1996年1月

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  • THE KINETICS OF ACTIN-FILAMENTS IN OSTEOCLASTS ON CHICKEN MEDULLARY BONE DURING THE EGG-LAYING CYCLE

    T SUGIYAMA, S KUSUHARA

    BONE   15 ( 3 )   351 - 353   1994年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    Distributions of actin filaments in osteoclasts on chicken medullary bone were examined during the egg-laying cycle. When an egg was in the magnum or isthmus of the oviduct at 6 h after oviposition, the osteoclasts lacked ruffled borders and attached to medullary bone surface via clear zones. Actin filaments were localized as amorphous bands in the bone site of the cytoplasm, corresponding to clear zones in the osteoclasts. When an egg was in the shell gland of the oviduct at 15 h after oviposition, the osteoclasts had long and slender, well-developed ruffled borders. Actin filaments were localized as lines perpendicular to the bone surface at the bone site of the cytoplasm. In medullary bone osteoclasts, actin filaments changed during the egg-laying cycle. The relationship between the actin filaments and the ultrastructure of the osteoclasts are discussed in this report.

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  • Effect of parathyroid hormone on osteoclasts in organ-cultured medullary bone

    T. SUGIYAMA, S. KUSUHARA

    Japanese Poultry Science   31 ( 6 )   392 - 399   1994年1月

  • ULTRASTRUCTURAL-CHANGES OF OSTEOCLASTS ON HEN MEDULLARY BONE DURING THE EGG-LAYING CYCLE

    T SUGIYAMA, S KUSUHARA

    BRITISH POULTRY SCIENCE   34 ( 3 )   471 - 477   1993年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CARFAX PUBL CO  

    1. The ultrastructure of osteodasts on hen medullary bone during the egg-laying cycle was observed by electron microscopy.
    2. At 0 to 6 h after oviposition, osteoclasts lacking ruffled borders were attached to the bone via the clear zone and appeared to have ceased bone resorption. Small vacuoles were scattered throughout this cytoplasm.
    3. At 9 to 21 h after oviposition, most of the osteoclasts had ruffled borders and appeared to be resorbing bone. The ruffled borders at 15 h were well developed, whereas at 9, 18 and 21 h they were poorly developed or showed similarities in structure to the clear zone. The small vacuoles were concentrated under the ruffled borders at 9 h and then decreased.
    4. These results demonstrate cyclic changes in osteoclasts during the egg-laying cycle, indicate that ruffled borders form at the beginning of bone resorption and suggest that they are derived from the clear zone and the small vacuoles. The results also indicate that the ruffled borders fuse into the clear zone and disappear at the completion of bone resorption.

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  • Inhibition of osteoclastic bone resorption by calcitonin in the cultured medullary bone of laying hens

    Toshie SUGIYAMA, Tomoo OHASHI, Seiji KUSUHARA

    Japanese Poultry Science   30 ( 1 )   16 - 23   1993年1月

  • Electron microscopic observations of osteoclasts and osteoblasts in cultured medullary bones of laying hens

    SUGIYAMA Toshie, OHASHI Tomoo, KUSUHARA Seiji

    Animal Science and Technology   65 ( 5 )   468 - 473   1992年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Japanese Society of Animal Science  

    産卵鶏の骨髄骨は,卵が卵管の卵白分泌部に存在する時期には骨形成期,卵殻腺部に存在する時期には骨吸収期に区別される,これらの骨形成期および骨吸収期の骨髄骨をそれぞれ器官培養し,破骨細胞および骨芽細胞を電子顕微鏡学的に観察して器官培養の可能性を検討した.骨形成期の骨髄骨では,48時間の培養後でも培養前と同様に破骨細胞は,豊富な細胞質を有していたが,ruffled borderは認められず,不活発な骨吸収像を呈していた.骨芽細胞も同様に変化せず,48時間の培養後まで粗面小胞体が発達した活発な骨形成像を示していた.一方,骨吸収期の骨髄骨では,72時間の培養後において破骨細胞は指状細胞質突起であるruffled borderを有し,培養前と同様に活発な骨吸収像を示していた.骨芽細胞は,24時間の培養後まで培養前と同様な形態を示し,一般に発達の悪い細胞質を有する不活発な骨形成像を呈していた.これらの結果から,骨形成期および骨吸収期における骨髄骨の破骨細胞および骨芽細胞の機能は,培養後短時間では維持されていることが示唆された.

    DOI: 10.2508/chikusan.63.468

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書籍等出版物

  • 獣医組織学

    日本獣医解剖学会

    学窓社  2017年3月  ( ISBN:4873627540

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    総ページ数:361  

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  • Sturkie's avian physiology

    Scanes, C. G., Sturkie, Paul D.

    Academic Press  2015年  ( ISBN:9780124071605

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    総ページ数:xxv, 1028 p.   記述言語:英語

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  • ニワトリの科学 (シリーズ・家畜の科学)

    古瀬 充宏

    朝倉書店  2014年7月  ( ISBN:4254455046

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    総ページ数:202  

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  • 獣医組織学

    日本獣医解剖学会

    学窓社  2014年3月  ( ISBN:4873627443

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    総ページ数:364  

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  • 生物科学・食品科学への高圧利用

    山本 克博, 林 力丸

    さんえい出版  2003年8月  ( ISBN:4915592766

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    総ページ数:142  

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  • Calcium Metabolism : Comparative Endocrinology

    Bio Scientifica  1999年 

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  • The Comparative Endocrinology of Calcium Regulation

    Journal of Endocrinology Limited  1996年 

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MISC

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受賞

  • 日本家禽学会奨励賞

    2004年  

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    受賞国:日本国

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共同研究・競争的資金等の研究

  • 家禽の強健な骨格構築を可能とする細胞外小胞による骨と筋のクロストーク機構の解明

    研究課題/領域番号:23K05518

    2023年4月 - 2026年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵

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    配分額:4810000円 ( 直接経費:3700000円 、 間接経費:1110000円 )

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  • イリシンとオステオカルシンのクロストーク機構を応用した家禽の強健な骨格構築

    研究課題/領域番号:20K06361

    2020年4月 - 2023年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵

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    配分額:4290000円 ( 直接経費:3300000円 、 間接経費:990000円 )

    現在、肉用鶏において脚弱が、卵用鶏においては破卵が頻発し、養鶏産業に多大な経済的損失をもたらしている。これらは、成長期において十分な骨量や骨密度を有した骨格が構築されず、急速な体重増加や高い産卵性により発症する。したがって、脚弱や破卵を防止するには、強健な骨格を構築することが不可欠である。本研究では、骨格筋より分泌されるマイオカイン(イリシン)と骨より分泌されるオステオカイン(オステオカルシン)の作用機序を明らかにするとともに、この骨と筋のクロストーク機構を利用して、強健な骨格の構築し、脚弱の防止と卵殻質の改善を図る。
    本年度は、オステオカルシンの作用機序を明らかにすることを目的とし、以下の知見が得られた。
    1.孵卵3-15日の肉用鶏胚におけるオステオカイン(オステオカルシン)ならびにオステオカルシン受容体GPRC6Aの相対的発現量について、経時的に検討した。その結果、孵卵の経過とともに、オステオカルシンの発現量は増加し、孵卵13、15日では孵卵11日と比較して発現量が有意に高かった。また、GPRC6Aの発現が、孵卵3-15日の全てにおいて観察された。アリザリンレッド染色を用いた透明全身骨格標本では、孵卵8日に石灰化領域が始めて観察され、その後、拡大した。以上のことから、骨石灰化に伴うオステオカルシンの発現開始とその後の発現量の増加により、筋線維の形成が促進されることが示唆された。
    2.孵卵10日に、大腿骨皮質骨内側表面に破骨細胞がみられ、骨吸収の開始がうかがえた。その後、破骨細胞数は増加し、孵卵12-15日の皮質骨においては破骨細胞のモデリングによる空胞がみられた。以上のことから、肉用鶏胚発生過程において、オステオカルシンは骨形成により骨基質に沈着した後、骨吸収による脱カルボキシル化が行われ、内分泌因子として機能していることが示唆された。

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  • 家禽の強固な骨格構築を目指した骨と筋のクロストーク機構の解明

    研究課題/領域番号:17K08041

    2017年4月 - 2020年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵, 二宮 禎

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    家禽では起立不能の脚弱あるいは卵殻の薄化による破卵が頻発し、多大な経済的損失を被っている。これらは、十分な骨量や骨密度を有した骨格が構築されないため発症する。
    本研究では、骨からのオステオカインであるオステオカルシンが、産卵鶏胚と比較して肉用鶏胚の胸骨において高い発現を示すことを明らかにした。また、筋からのマイオカインであるイリシンが、肉用鶏において胸筋で最も高い発現を示した。このことから、家禽における骨と筋のクロストーク機構を示唆し、これらを活用した破卵や脚弱の発生防止を目指した強固な骨格構築の可能性を示した。

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  • 家禽の破卵および脚弱を制御するリン代謝調節機構の解明

    研究課題/領域番号:26450374

    2014年4月 - 2017年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵

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    配分額:5070000円 ( 直接経費:3900000円 、 間接経費:1170000円 )

    鶏では起立不能の脚弱あるいは卵殻の薄化による破卵が頻発し、多大な経済的損失を被っている。これらは、いずれも、リンとカルシウムからなる骨の代謝異常に起因する。
    本研究では、リン輸送を司るナトリウム依存性リン酸トランスポーター(NaPi-II)が腎臓ならび腸管で発現しており、成熟とともにその発現量は増加しリン吸収が増加していることを明らかにした。また、これらリン代謝はエストロジェンによって生成された活性型ビタミンD3によって調節されていることを示唆した。

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  • 鳥類の骨髄骨代謝と卵殻腺における石灰化の同調機構

    研究課題/領域番号:24659813

    2012年4月 - 2015年3月

    制度名:科学研究費助成事業

    研究種目:挑戦的萌芽研究

    提供機関:日本学術振興会

    山本 敏男, 池亀 美華, 河井 まりこ, 杉山 稔惠

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    配分額:3640000円 ( 直接経費:2800000円 、 間接経費:840000円 )

    卵殻腺における卵殻石灰化と骨髄骨の同調機構について,プロゲステロン並びに殻腺に加わる張力に着目し,これらと骨髄骨破骨細胞の動態について調べた。プロゲステロン投与は卵殻石灰化の活発な時期の破骨細胞の活性を抑制した。また,卵殻石灰化時期の卵を卵殻腺から摘出し張力を除去すると,活発な時期の破骨細胞は不活発な様相呈するようになり,抑制を受けた。したがって,産卵周期に伴う骨髄骨破骨細胞の活性周期にはプロゲステロン並びに卵殻腺に加わる張力が重要であることが強く示唆された。

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  • 家禽の脚弱および破卵の防止に向けたビタミンD新規代謝の解明

    研究課題/領域番号:23580386

    2011年 - 2013年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵, 山田 宜永

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    配分額:5330000円 ( 直接経費:4100000円 、 間接経費:1230000円 )

    鶏では脚弱あるいは卵殻の薄化による破卵が頻発し、多大な経済的損失を被っている。これらの原因として、ビタミンD代謝が関与していると考えられる。
    本研究では、25-ヒドロキシビタミンDより活性型ビタミンDを産生する1α水酸化酵素が、腎臓以外のビタミンD標的組織である腸管ならびに卵管に存在することを示した。また、この酵素の発現は、エストロジェンによって増加し、加齢に伴い減少することを示した。以上のことから、標的組織で局所的に活性型ビタミンDが産生され、カルシウム代謝を調節していることが考えられた。また、25-ヒドロキシビタミンD給与により、卵殻質が改善されることが示唆された。

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  • 超高圧による筋肉タンパク質の特性制御

    研究課題/領域番号:15208025

    2003年 - 2005年

    制度名:科学研究費助成事業

    研究種目:基盤研究(A)

    提供機関:日本学術振興会

    鈴木 敦士, 西海 理之, 原 崇, 杉山 稔恵, 池内 義秀

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    配分額:27040000円 ( 直接経費:20800000円 、 間接経費:6240000円 )

    筋肉内在性酵素に焦点をあてながら、超高圧処理による筋肉タンパク質の制御を研究した。また、食肉アレルゲン性の低減化や、呈味物質生成に関与する酵素への影響についても検討した。
    1.トロポミオシンおよびα-アクチニンに及ぼす影響
    (1)α-アクチニンのヘリックス含量は処理圧力が増加するにつれて60%から40%に低下したがトロポミオシンは圧力に対して抵抗性を示した,(2)圧力によるα-アクチニンの構造変化は300MPaまでは可逆的であったが、それ以上の圧力になると不可逆的なものとなった,(3)α-アクチニンの超沈殿誘起能はATPアーゼ促進能よりもアクチンゲル化能と相関を示した。
    2.高圧と熱の併用効果
    (1)高圧と熱の併用は硬い肉の軟化に有効であった,(2)剪断力価は筋肉内結合組織と筋原線維の変形に何らかの関係を持っているかもしれない。
    3.筋肉内結合組織に及ぼす影響
    (1)筋線維だけでなく結合組織も超高圧処理によって柔らかくなった,(2)圧力依存性の筋肉内コラーゲンの熱安定性の低下と構造の脆弱化が認められた。
    4.筋肉内在性タンパク質分解酵素に及ぼす影響
    (1)超高圧処理はプロテアソームの活性化を引き起こし、3次構造の不可逆的変化をもたらしたが、熱処理に比べれば2次構造の変化は小さかった。それゆえ、圧力によるプロテアソームの活性化は、プロテアソームの活性部位におけるわずかなunfoldingによるのではないかと思われる,(2)熟成および圧力処理した牛肉内カルパインの局在性の変化を免疫電子顕微鏡により観察した結果、カルパインの局在性に及ぼす高圧処理の影響は熟成の影響に比べて激しいことが判明した。
    5.ウシ血清アルブミン(BSA)のアレルゲン性に及ぼす影響
    (1)BSAのアレルゲン性低下に及ぼす影響をアレルギー患者血清を感作させたヒト好塩基球様KU812F細胞からのヒスタミン遊離によって研究した,(2)アレルゲン性はBSAに加えられる圧力が高くなるにつれて徐々に減少した,(3)圧力によるBSAのアレルゲン性低下はBSAの3次構造の変化に関係する様に思える。
    6.食肉の呈味物質の生成に関与する酵素に及ぼす影響
    呈味物質の生成に関与する酵素活性は高圧処理を加えた肉中でも保持されているために、高圧処理を加えた肉中においてもイノシン酸は生成されていた。

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  • 家禽における内分泌攪乱化学物質に対する感受性とバイオアッセイ法の確立

    研究課題/領域番号:15580232

    2003年 - 2005年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉山 稔恵, 楠原 征治, 西村 宏一

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    配分額:3600000円 ( 直接経費:3600000円 )

    1.日齢に伴うダイオキシン受容体(AhR) mRNAの発現
    孵化後から390日齢にいたるニワトリ雌におけるダイオキシン受容体(AhR)の発現をRT-PCRにより検討した。その結果、筋胃、肝臓、肺、腎臓および心臓においては、全日齢を通してAhR mRNAの発現が確認された。しかしながら、卵管および骨髄では孵化直後ではAhR mRNAの発現は観察されず、30-90日齢でもその発現は低かった。これらの結果より、ニワトリ雌では卵管および骨髄でダイオキシンに対する感受性が日齢で変化することが示唆された。
    2.産卵鶏雌の卵管におけるAhRの局在
    卵管卵白分泌部、峡部および卵殻腺部にAhRがタンパク質レベルで発現していることが確認され、その局在は峡部ならびに卵殻腺部の粘膜上皮細胞細胞質に強くみられた。このことは、ダイオキシンは峡部および卵殻腺部の上皮細胞に作用して卵殻の脆弱化を引き起こすものと推測される。
    3.骨髄骨を用いたバイオアッセイ法の確立
    雄ウズラへエストロジェン様内分泌撹乱化学物質であるDDE (10mg/day)を7日間投与したところ、大腿骨骨髄腔内に骨髄骨の形成が観察された。また、これらの形成に伴い血中ビテロゲニン濃度も上昇した。このことは、鳥類における骨髄骨形成が感度の高い内分泌撹乱化学物質に対する有用なバイオアッセイ法となることを示している。
    4.内分泌撹乱化学物質受容体の発現と局在
    産卵鶏骨髄骨の骨芽細胞に、内分泌撹乱化学物質受容体であるアンドロジェン受容体、エストロジェン受容体αが発現していることが観察された。このことより、内分泌撹乱化学物質は骨形成に作用していることが示唆された。
    5.ウズラ肝臓でのビテロゲニン(Vt) mRNAアッセイ法の確立
    ウズラ肝臓におけるVt mRNAのRT-PCR法によるアッセイ法を確立した。また、Vt mRNAの発現は、エストロジェン受容体αを介していることが明らかとなった。

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  • 鶏カルシトニン受容体の遺伝子解析と発現に関する研究

    研究課題/領域番号:12760185

    2000年 - 2001年

    制度名:科学研究費助成事業

    研究種目:奨励研究(A)

    提供機関:日本学術振興会

    杉山 稔恵

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    配分額:2300000円 ( 直接経費:2300000円 )

    (1)鶏総RNAからのカルシトニン受容体cDNAの検出
    成熟した鶏(白色レグホーン種)雌の腎臓から総RNAを抽出し、逆転写反応(RT)によりcDNAを作製した。ブタ、ヒト、ラット、マウスおよびモルモットに共通して保存されているカルシトニン受容体アミノ酸配列を基として、degenerateプライマー5'-TAYTGYAAYMGIACITGGGAYGGITGG-3'(上流配列特異的プライマー)および5'-TAIATRACRAARACRTTRKTRCTYCAIGTY-37(下流配列特異的プライマー)を作製し、40サイクル(95℃1分間、45℃1分間、72℃1分30秒間)のポリメラーゼ連鎖反応(PCR)を行った。その結果、約1100bpの鶏カルシトニン受容体cDNAが得られた。このcDNAは、鶏カルシトニン受容体cDNAの一部分を示している。
    (2)カルシトニン受容体cDNAのプラスミドの作製
    得られた鶏カルシトニン受容体cDNAをTAクローニング法でベクターに挿入し、大量のカルシトニン受容体cDNAを得ることが可能になった。
    (3)カルシトニン受容体cDNAの塩基配列の決定
    得られたベクターについて、T7プライマーおよびSP6プライマーを用いてPCRを行い、PCR direct sequence法により鶏カルシトニン受容体cDNAの部分塩基配列を解析した。また、鶏心臓についてRT-PCR法によりカルシトニン受容体の発現を確認した所、哺乳類と同様に受容体の発現は認められなかった。

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  • 超高圧下での筋肉タンパク質変性機構の解析

    研究課題/領域番号:10460118

    1998年 - 2000年

    制度名:科学研究費助成事業

    研究種目:基盤研究(B)

    提供機関:日本学術振興会

    鈴木 敦士, 中原 勝, 池内 義秀, 杉山 稔恵

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    配分額:13700000円 ( 直接経費:13700000円 )

    超高圧下での筋肉構成タンパク質の変性機構を、組織学や生化学的手法に加えて、物理化学的手法を加えることにより研究した。
    (アクチン)
    1.ε-ATPのアクチンからの解離速度と,メチオニン由来プロトンNMRシグナルの消失速度がほぼ一致することから、一定以上の圧力が加わると、ATPがアクチン分子から解離し、それに伴ってDNaseI及びアクチン結合部位であるメチオニン領域が急速に崩壊して、結果的にATP交換能、DNaseI阻害活性能あるいは重合能を失うのではないかと推測された。
    2.処理圧力の増加に伴ってα-ヘリックス含量が減少し、300MPaを超えると不可逆的な変性を起こすことが明らかになった。
    (ミオシンおよびトロボミオシン)
    1.ミオシンの圧力下でのATPase活性は、加圧とともに増加し、100MPaの加圧下で最大活性を示した。さらに圧力を加えると活性は低下した。加圧後、圧力を開放し、ATPaseとCDスペクトルを測定した結果、200MPa^〜400MPaにかけて、活性の著しい低下とα-ヘリックス含量の減少が認められた。
    2.トロポミオシンに圧力を加え、解放後にα-ヘリックス含量を算出した結果、トロポミオシンが酸や熱と同様に、超高圧に対しても安定なタンパク質であることが判明した。
    (ミトコンドリアと結合組織)
    1.ミトコンドリアのCa^<2+>取り込み能の低下は、圧力によって誘起される内膜成分のATPase活性部位の遊離と相関した。
    2.筋肉内結合組織の筋周膜の構造は、処理圧力の増加につれて破壊されるが、プロテオグリカンの抽出性の変化は認められなかった。
    (プロテアソーム)
    筋肉内在性タンパク質分解酵素,プロテアソームの圧力下での活性を測定した結果、50MPaまでの圧力では加圧に伴い活性は増加したが、その後は徐々に低下し、200MPaの圧力ではほぼ失活した。構造の緩やかな変化が活性の増加につながると考えられる。
    (β-ラクトグロブリンおよび高圧解凍)
    1.NMRスペクトルから、乳タンパク質β-ラクトグロブリンは中性域よりも酸性域の方が圧力に対して強く,柔軟であるが,酸性pHにおいても加圧処理により天然状態の構造は一部失われるものと結論した。
    2.凍結肉の流水解凍と高圧解凍を比較した結果、200MPa以下の高圧解凍は豚肉からのドリップの減少をもたらすと同時に肉の軟化を生じ、肉製品の加工に有力な方法であることが明らかになった。

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