Production of Diabetic Offspring Using Cryopreserved Epididymal Sperm by In Vitro Fertilization and Intrafallopian Insemination Techniques in Transgenic Pigs     

Kazuhiro UMEYAMA  Kasumi HONDA  Hitomi MATSUNARI  Kazuaki NAKANO  Tatsuro HIDAKA  Keito SEKIGUCHI  Hironori MOCHIZUKI  Yasuhiro TAKEUCHI  Tsukasa FUJIWARA  Masahito WATANABE  Masaki NAGAYA  Hiroshi NAGASHIMA 《The Journal of reproduction and development》2013,59(6):599-603

Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains that model human diseases. However, production of numerous cloned disease model pigs by SCNT for large-scale experiments is impractical due to its complexity and inefficiency. In the present study, we aimed to establish an efficient procedure for proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and treated with insulin to allow for normal growth to maturity, at which point epididymal sperm could be collected for cryopreservation. In vitro fertilization and intrafallopian insemination using the cryopreserved epididymal sperm resulted in diabetes model transgenic offspring. These results suggest that artificial reproductive technology using cryopreserved epididymal sperm could be a practical option for proliferation of genetically modified disease model pigs.  相似文献   

Application of highly differentiated SNPs between Japanese Black and Holstein to a breed assignment test between Japanese Black and F1 (Japanese Black x Holstein) and Holstein     

Shota NISHIMURA  Toshio WATANABE  Atsushi OGINO  Kazuhiro SHIMIZU  Mitsuo MORITA  Yoshikazu SUGIMOTO  Akiko TAKASUGA 《Animal Science Journal》2013,84(1):1-7

Two taurine breeds, Japanese Black and Holstein, established from geographically distant origins and selected for different uses, beef and dairy, were extensively genotyped using a genome‐wide single nucleotide polymorphism (SNP) chip with more than 1000 animals of each breed. The genetic structure was examined by principal component analysis, in which the first principal component clearly separated the two breeds and explained more than 15% of the variance. Highly differentiated SNPs were detected throughout the genome, some of which were clustered within small regions on BTA4 (79.2–79.7 Mb, Btau4.0) and BTA26 (22.2–23.6 Mb). A breed assignment test was developed using 18 highly differentiated SNPs to distinguish Japanese Black from F₁ (Japanese Black × Holstein) and Holstein. The error rate that an F₁ or Holstein animal is misjudged as Japanese Black was expected to be < 0.8%, while the error rate that a Japanese Black animal is misjudged as F₁ or Holstein was expected to be < 0.001%. This test provides a reliable and powerful method to detect breed label falsification in retail beef.  相似文献   

Biological nitrification inhibition by Brachiaria humidicola roots varies with soil type and inhibits nitrifying bacteria, but not other major soil microorganisms     

Subramaniam GOPALAKRISHNAN  Takashi WATANABE  Stuart J. PEARSE  Osamu ITO  Zakir A.K.M. HOSSAIN  Guntur V. SUBBARAO 《Soil Science and Plant Nutrition》2009,55(5):725-733

The tropical pasture grass Brachiaria humidiola (Rendle) Schweick releases nitrification inhibitory compounds from its roots, a phenomenon termed 'biological nitrification inhibition' (BNI). We investigated the influence of root exudates of B. humidicola on nitrification, major soil microorganisms and plant growth promoting microorganisms using two contrasting soil types, Andosol and Cambisol. The addition of root exudates (containing BNI activity that is expressed in Allylthiourea unit (ATU) was standardized in a bioassay against a synthetic inhibitor of nitrification, allylthiourea, and their function in soil was compared to inhibition caused by the synthetic nitrification inhibitor dicyandiamide. At 30 and 40 ATU g⁻¹soil, root exudates inhibited nitrification by 95% in fresh Cambisol after 60 days. Nitrification was also similarly inhibited in rhizosphere soils of Cambisol where B. humidicola was grown for 6 months. Root exudates did not inhibit other soil microorganisms, including gram-negative bacteria, total cultivable bacteria and fluorescent pseudomonads. Root exudates, when added to pure cultures of Nitrosomonas europaea , inhibited their growth, but did not inhibit the growth of several plant growth promoting microorganisms, Azospirillum lipoferum , Rhizobium leguminosarum and Azotobacter chroococcum. Our results indicate that the nitrification inhibitors released by B. humidicola roots inhibited nitrifying bacteria, but did not negatively affect other major soil microorganisms and the effectiveness of the inhibitory effect varied with soil type.  相似文献   

Successful blastocyst production by intracytoplasmic injection of sperm after in vitro maturation of follicular oocytes obtained from immature female squirrel monkeys (Saimiri boliviensis)     

Shinichi WATANABE  Megumi MIURA  Hiromi MORITA  Moeka NISHI  Shin-ichi YOKOTA  Shosaku HATTORI  Hiromichi MATSUMOTO  Emiko FUKUI  Ken Takeshi KUSAKABE  Masanori OCHI  Naomi NAKAGATA  Yasuo KISO  Chieko KAI  Midori YOSHIZAWA 《The Journal of reproduction and development》2021,67(4):265

Advanced reproductive technologies are being applied for the propagation of squirrel monkeys, to ensure their preservation as a genetic resource and the effective use of their gametes in the future. In the present study, oocytes and spermatozoa were collected from live squirrel monkeys, following which piezo intracytoplasmic sperm injection (ICSI) was performed using these gametes. Follicular development was induced by administering equine chorionic gonadotropin (eCG) containing inhibin antiserum to an immature squirrel monkey female. The unilateral ovary was excised after the administration of human chorionic gonadotropin (hCG), to induce ovulation, following which the larger developed follicular oocytes were collected. Follicular oocytes were prepared for ICSI using sperm from the epididymal tail of a unilateral testis extracted from a mature male. The embryos were continuously incubated in CMRL 1066 medium supplemented with 10% (v/v) fetal bovine serum. Embryo culture was performed with cumulus cells. Two experiments of ICSI carried out with three females resulted in 14 mature oocytes from the 49 cumulus-oocyte complexes collected and five embryos, three of which developed into blastocysts. These blastocysts were vitrified, thawed, and transferred to recipient monkeys, but no pregnancies resulted. In conclusion, the present study is the first to successfully produce ICSI-derived blastocysts from MII oocytes obtained by means of hormone administration (a combination of eCG+inhibin antiserum and hCG) and in vitro maturation in immature squirrel monkeys.  相似文献   

稻田环境植物多样性研究   总被引：10，自引：1，他引：9  

余柳青  渡边泰 《中国水稻科学》1998,12(3):149-154

 选择稻田灌溉水无工业废水污染的禹山坞和受工业废水污染的城北村两地进行稻田环境植物多样性调查。禹山坞稻田普遍使用化学除草剂,杂草种数明显少于城北村,水竹叶成为禹山坞稻田的优势种;城北村稻田未用除草剂及低洼积水环境保护了槐叶萍、满江红、轮藻和尖头丽藻等。禹山坞的畦畔植物种数显著高于城北村。城北村灌溉水质差有利于部分耐性植物蔓延形成优势种,如空心莲子草的强烈竞争,使部分竞争力弱的植物逐步消失,植物种数减少。两村的沟渠植物种类差异明显。  相似文献   

Development of a sandwich enzyme-linked immunosorbent assay (ELISA) to quantify γ-glutamyl-carboxylated clotting factor IX and assess redox susceptibility of anticoagulant chemicals     

Ryo SATO  Kanami WATANABE  Ryo KAMATA  Kazuki TAKEDA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(6):804

Anticoagulant chemicals (ACCs) such as warfarin are widely used in medical applications as well as for their rodenticide properties. Their efficacy is greatly influenced by polymorphisms in the gene encoding vitamin K epoxide reductase (VKOR). Evaluation of the activity of ACCs toward VKOR variants is essential to determine their proper use. Presently, this is achieved by co-expressing VKOR of Rattus Norvegicus and human clotting factor IX in cultured cells and measuring inhibition of vitamin K-dependent gamma-glutamyl carboxylation of factor IX (glaFIX) activity. However, glaFIX has only been quantified using indirect methods like blood coagulation assays. We have developed a sandwich enzyme-linked immunosorbent assay using a glaFIX-specific antibody to quantify glaFIX and used this to analyze inhibition of VKOR activity by warfarin.  相似文献   

Prolactin gene expression in mouse pancreatic islets   总被引：1，自引：0，他引：1  

Toshio HARIGAYA  Michiko KOMORI  Harumi WATANABE  Satoshi WATANABE  Takuji MATSUI 《Animal Science Journal》2002,73(1):29-34

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Age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle     

Sachi TANAKA  Kohtaro MIYAZAWA  Atsuko KUWANO  Kouichi WATANABE  Shyuichi OHWADA  Hisashi ASO  Shigeru NISHIDA  Takahiro YAMAGUCHI 《Animal Science Journal》2008,79(3):368-374

It is well known that the immune system changes with age during development and maturation in Holstein cattle. But age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle still remain unclear. The aim of the present study was to investigate comparative changes of leukocytes (granulocytes, monocytes, B cells and T cells) and T cell subsets (CD4⁺, CD8⁺, γδ, CD8⁺γδ and WC1⁺γδ T cells) in Japanese Black cattle aged 0.5, 1, 2, 6, 18 and 36–41 (adult) months on flow cytometry using specific monoclonal antibodies for the cell surface markers. T cell proportion was approximately 40% in 2-month-old cattle and decreased to 20.6% in adults. In contrast, B cell proportion significantly increased from 7.4% to 28.2% with age. In T cell subsets the percentage of CD4⁺ T cells significantly increased from 40.5% to 60%, but that of WC1⁺γδ T cell subset significantly decreased with age. The percentages of CD8⁺ and CD8⁺γδ T cells did not change. The present study details the proportional changes in leukocyte and T cell subsets with age in the peripheral blood of Japanese Black cattle and these findings are similar to those described for Holstein cattle.  相似文献   

Ultrastructural Study on Colocalization of Glucagon-Like Peptide (GLP)-1 with GLP-2 in Chicken Intestinal L-Cells     

Kei NISHIMURA  Kohzy HIRAMATSU  Mohammad M. MONIR  Chihiro TAKEMOTO  Takafumi WATANABE 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2013,75(10):1335-1339

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Cystic Rete Testis with Testicular Dysplasia in a Rabbit     

James K. CHAMBERS  Kazuyuki UCHIDA  Yousuke MURATA  Ken-ichi WATANABE  Kenichiro ISE  Yasutsugu MIWA  Hiroyuki NAKAYAMA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(5):751-755

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