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Freeze-drying Spermatozoa in Mice OAK
鈴木, 宏志; 川瀬, 洋介.
Cryopreservation of mouse spermatozoa has been widely applied for maintenance of genetically modified mouse strains. Although cryopreservation of mouse spermatozoa is simpler, less time-consuming, and less costly than of embryos for preservation of the genetic materials, maintenance of cryopreserved spermatozoa still has a high running cost because of the need for a constant supply of liquid nitrogen. In 1998, it has been reported that freeze-dried mouse spermatozoa are capable of participating in normal embryonic development after injection into oocytes, and so they have attracted a great deal of attention as storable gene resources. However, attempts of freeze-dry spermatozoa are not new. Successful conception and full-term development using freeze-dried...
Palavras-chave: Mice; Freese-drying; Spermatozoa; マウス; 凍結乾燥; 精子.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4510
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TRANSIENT EXPRESSION OF A GREEN FLUORESCENT PROTEIN GENE IN BABESIA BOVIS OAK
Fujii, Kei; Yokoyama, Naoaki; Takabatake, Noriyuki; Suzuki, Hiroshi; Xuan, Xuenan; Igarashi, Ikuo; 横山, 直明; 鈴木, 宏志; 玄, 学南; 五十嵐, 郁男.
In hemoprotozoa,gene transfer technology provides an important tool to aid in the functional study of parasite genes. In this study,a transfer vector containing the enhanced green fluorescent protein (EGFP) gene laid between the Toxoplasma gondii GRAI promoter region and the GRA2 polyA signal region was constructed and transfected into the in vitro culture of Babesia boνis by the electroporation method.On the first and second days post-transfection,clear positive fluorescences were detected in some parasite bodies, indicating the successful expression of the EGFP gene controlled by the GRA-1 promoter in B. bovis. However, the positive parasites were not ubiquitous and finally disappeared until the forth day post-transfection. This is the first time that...
Ano: 2003 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/631
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Expression of Uterine Sensitization-Associated Gene-1 (USAG-1) in the Mouse Uterus During the Peri-Implantation Period OAK
MAEDA, Kanami; LEE, Dong-Soo; YANAGIMOTO UETA, Yoshiko; SUZUKI, Hiroshi; 鈴木, 宏志.
Rat uterine sensitization-associated gene-1 (USAG-1) mRNA is expressed in the uterus during the peri-implantation period, and its mRNA expression in uterine epithelial cells is highest on day 5 of pregnancy. On the other hand, since changes in USAG-1 mRNA expression in the mouse uterus are not seen during the estrous cycle, USAG-1 expression might be specifically regulated by embryonic factors rather than by the maternal environment. However, the expression pattern and function of USAG-1 in the mouse uterus have not been determined. Thus, we examined the tissue-specific USAG-1 mRNA expression in the uteri of ICR mice during peri-implantation using real-time quantitative PCR. Uterine tissues, such as the myometrium, luminal epithelium, and stroma, were...
Palavras-chave: Implantation; Mouse; MRNA; Uterine sensitization-associated gene-1 (USAG-1); Uterus.
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3024
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遺伝子の役割を明らかにする発生工学 OAK
鈴木, 宏志.
Ano: 2004 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/876
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Identification of a novel gene encoding a secreted antigen 1 of Babesia gibsoni and evaluation of its use in serodiagnosis OAK
Jia, Honglin; Zhou, Jinlin; Ikadai, Hiromi; Matsuu, Aya; Suzuki, Hiroshi; Fujisaki, Kozo; Xuan, Xuenan; 鈴木, 宏志; 五十嵐, 郁男; 玄, 学南.
Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti-BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These...
Palavras-chave: Babesia gibsoni; Secreted Antigen 1; Identification; Serodiagnosis.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/1655
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優良盲導犬の育成に関する発生・生殖工学的研究 : 遺伝資源の凍結保存 OAK
鈴木, 宏志.
Palavras-chave: 盲導犬; 凍結保存; 受精卵; 伴侶動物.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2889
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Freeze-dried Spermatozoa and Freeze-dried Sperm Injection into Oocytes OAK
SUZUKI, Hiroshi; 鈴木, 宏志.
Attempts to freeze-dry spermatozoa are not new. Successful conception and full-term development using freeze-dried spermatozoa were reported in the cow and rabbit prior to 1960. However, these preliminary results have not been confirmed. Spermatozoa become defective in motility after freeze-drying and are unable to fertilize eggs both in vivo and in vitro. However, in 1998, freeze-drying of mammalian spermatozoa was demonstrated without loss of genetic or reproductive potential. Freeze-dried spermatozoa support normal development when injected directly into oocytes by means of an intracytoplasmic sperm injection (ICSI) technique. Recent investigations of freeze-dried sperm have focused on the factors affecting freeze-drying in spermatozoa in an effort to...
Palavras-chave: Spermatozoa; Freeze-drying; ICSI; Preservation.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4189
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Beneficial Effect of Hyperbaric Oxygen Therapy on the Follicular Survival in the Mouse Ovary after Transplantation OAK
HAYASHI, Satomi; ASANO, Tomoyoshi; KAKIZAKI, Ryujiro; SUZUKI, Hiroshi; 鈴木, 宏志.
A large proportion of follicles are lost during the initial ischemia that occurs after transplantation of ovarian tissues. Thus, the effect of hyperbaric oxygen therapy (HBO) on the follicular loss of ovarian tissues after transplantation was examined in mice. Ovarian slices from ICR mice were transplanted under the kidney capsule in ovariectomized ICR. Hyperbaric oxygen with 100% oxygen was initiated for 30 min at 2.5 atmospheres absolute immediately after transplantation, and this treatment was repeated at 48-h intervals for 2 weeks. The number of follicles was dramatically reduced at 2 weeks post transplantation. However, HBO was significantly effective in enhancing the survival of transplanted ovarian follicles. The survival rates of primordial and...
Palavras-chave: Hyperbaric oxygen; Mice; Ovary; Transplantation.
Ano: 2012 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3857
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Sexing of Dog Sperm by Fluorescence In Situ Hybridization OAK
OI, Maya; YAMADA, Keisuke; HAYAKAWA, Hiroyuki; SUZUKI, Hiroshi; 鈴木, 宏志.
Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion...
Palavras-chave: Dog; Fluorescence in situ hybridization (FISH); Sexing; Sperm.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3858
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Effect of Zona Incision by Piezo-Micromanipulator (ZIP) on In Vitro Fertilization in 21 Transgenic Mice Lines OAK
KAWASE, Yosuke; TACHIBE, Takanori; HANI, Toshio; TATEISHI, Hiromi; JISHAGE, Kou-ichi; SUZUKI, Hiroshi; 鈴木, 宏志.
Zona incision using a piezo-micromanipulator (ZIP) has been demonstrated to be effective for in vitro fertilization (IVF) using cryopreserved C57BL/6 spermatozoa. In this study, ZIP oocytes inseminated with frozen-thawed genetically modified C57BL/6J or FVB mice spermatozoa (21 lines) showed fertilization rates of 22-75% and live fetus rates of 8-49%. In 6 of the lines, the fertilization rates for oocytes compared with ZIP (42-75%) were significantly higher than that of nontreated oocytes (0-50%). Using only 90 oocytes for IVF with ZIP, 5 breeding pairs were produced from cryopreserved genetically modified mice spermatozoa. Our results indicate that application of the ZIP technique is effective for IVF using cryopreserved genetically modified mouse...
Palavras-chave: Frozen-thawed spermatozoa; IVF; ZIP.
Ano: 2009 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3017
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CLONING AND EXPRESSION OF AN ANTIGEN OF BABESIA GIBSONI IN ESCHERICHIA COLI AND ITS USE FOR THE IMMUNODIAGNOSIS OF CANINE BABESIOSIS OAK
Fukumoto, Shinya; Sekine, Yukiko; Kimbita, Elikira; Huang, Xiaohong; Xuan, Xuenan; Inoue, Noboru; Yokoyama, Naoaki; Igarashi, Ikuo; Fujisaki, Kozo; Sugimoto, Chihiro; Nagasawa, Hideyuki; Mikami, Takeshi; Suzuki, Hiroshi; 福本, 晋也; 玄, 学南; 井上, 昇; 横山, 直明; 五十嵐, 郁男; 鈴木, 宏志.
Palavras-chave: Babesia gibsoni; Babesia canis; CDNA library; ELISA; P30 gene.
Ano: 2002 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/624
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動物の原虫感染に対する抵抗性遺伝子の発生工学的研究 OAK
鈴木, 宏志; Suzuki, Hiroshi.
発行年は推定
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3505
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Cryopreservation of Canine Ovaries by Vitrification OAK
ISHIJIMA, Takako; KOBAYASHI, Yoshiyasu; LEE, Dong-Soo; UETA, Yoshiko Yanagimoto; MATSUI, Motozumi; LEE, Jung-Youn; SUWA, Yoshinori; MIYAHARA, Kazuro; SUZUKI, Hiroshi; 松井, 基純; 宮原, 和郎; 鈴木, 宏志; 古林, 与志安.
The cryopreservation of ovarian tissues is a technology with significant potential for the preservation of the genetic resource materials of working dogs, including guide dogs for the blind. However, no attempt has been reported on cryopreservation of the canine ovary. Thus, we evaluated a vitrification method for cryopreservation of canine ovaries and determined the potential functionality of vitrified-warmed canine ovaries by means of transplantation into non-obese diabetic-severe combined immunodeficiency (NOD-SCID) mice. All ovarian tissues cryopreserved by vitrification were morphologically normal in terms of histology. Cryopreserved ovaries were transplanted into the ovarian bursa of the NOD-SCID mice, and the xenografts were recovered from 23 of 23...
Palavras-chave: Canine; Ovary transfer; Transplantation; Vitrification; Xenograft.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3023
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Expression Patterns of the Implantation-associated Genes in the Uterus during the Estrous Cycle in Mice OAK
LEE, Dong-Soo; YANAGIMOTO UETA, Yoshiko; XUAN, Xuenan; IGARASHI, Ikuo; FUJISAKI, Kozo; SUGIMOTO, Chihiro; TOYODA, Yutaka; SUZUKI, Hiroshi; 玄, 学南; 五十嵐, 郁男; 鈴木, 宏志.
The mRNA expression patterns of EGF, HB-EGF, Amphiregulin, EGF receptor, IGF-1, CSF-1, IL-1 alpha, IL-1 beta, IL-1 receptor type 1, IL-1 receptor antagonist, LIF, COX-1, COX-2, Mucin-1, calcitonin, and rat USAG-1 mouse homologue, all of which are involved in the process of conceptus implantation to the endometrium, were examined during the estrous cycle by means of real-time quantitative PCR. COX-2, HB-EGF, LIF, Mucin-1, CSF-1, IL-1 alpha, IL-1 beta, and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage. In the case of COX-1, EGF, IGF-1, and EGF receptor, the highest mRNA expression was during the diestrous stage. In contrast, the rat USAG-1 mouse homologue mRNA expression did not...
Palavras-chave: Estrous cycle; Implantation; Mice; Ovarian hormone.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3022
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Reproductive bio-technology for the guide dog for the blind OAK
鈴木, 宏志.
Although guide dogs make a remarkable contribution to the quality of life of the blind people, many countries suffer from an acute shortage of guide dogs such that not all those who need them are able to be supplied. One of the important challenges from the point of view of animal and veterinary science as well as social welfare is to establish a stable and effective breeding and reproduction system for guide dogs for the blind. Although the development and extension of assisted reproductive techniques such as artificial insemination, embryo transfer and in vitro fertilization has been remarkable, in canines, it is not a successful as other mammalian spices such as mice, pigs, cows and humans. In this review, recent advances of cryopreservation of sperm,...
Palavras-chave: Guide dog; Artifical insemination; Embryo transfer; Recipient; Cryopreservation; イヌ; 人工授精; 移植; 受容雌; 凍結保存.
Ano: 2009 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4511
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Effect of Intraperitoneal Administration of Desialylated Erythropoietin on the Follicular Survival in Cryopreserved Canine Ovaries after Xenotransplantation OAK
Hirayama, Yukari; Inoue, Kou-suke; Suzuki, Hiroshi; 鈴木, 宏志.
The effect of intraperitoneal administration of desialylated erythropoietin (asialo EPO) on the follicular reserve of cryopreserved canine ovarian tissues was examined using non obese diabetic-severe combined immunodeficient (NOD-SCID) mice was examined. Vitrified-warmed canine ovarian tissues were placed under the dorsal skin of mice, and were then treated with saline or 400 or 4,000 U/kg/day of asialo EPO by intraperitoneal injection from one day before transplantation for 5 days. Transplanted tissues were removed and subjected to a histological examination 4 weeks after the transplantation. Without asialo EPO treatment, the mean number of primordial follicles in the cryopreserved ovarian tissues had decreased from 4.3/mm2 to 0.2/mm2 at 4 weeks after...
Palavras-chave: Canine; Cryopreservation; Ovary; Transplantation.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3856
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The Role of Scavenger Receptor Type A (SR-A) during Infection with Babesia microti OAK
Kamada, Takenori; Igarashi, Ikuo; Inoue, Noboru; Nagasawa, Hideyuki; Suzuki, Hiroshi; Kodama, Tatsuhiko; Suzuki, Naoyoshi; Toyoda, Yutaka; 五十嵐, 郁男; 井上, 昇; 長澤, 秀行; 鈴木, 宏志.
Palavras-chave: Babesiosis; Protective immune response; Knock-out mouse; SRKO.
Ano: 1997 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/265
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Babesia gibsoni internal transcribed spacer 1 region is highly conserved amongst isolates from dogs across Japan OAK
LIU, Mingming; CAO, Shinuo; VUDRIKO, Patrick; SUZUKI, Hiroshi; SOMA, Takehisa; XUAN, Xuenan; 鈴木, 宏志; 玄, 学南.
Babesia gibsoni is a tick-borne apicomplexan parasite of dogs that often causes fever and hemolytic anemia with highly variable clinical outcome. In this study, we sequenced the 254bp Internal Transcribed Spacer 1 region (ITS1) of 54 B. gibsoni isolates from 14 different geographical regions of Japan. The 54 isolates shared high sequence identity with each other and with B. gibsoni isolates reported in GenBank database (97.2–100%). Consistent with previous reports, phylogenetic analysis showed that B. gibsoni isolates from Japan formed the same clade with those from U.S.A., Australia, India and Taiwan. Our finding indicates that B. gibsoni ITS1 region is highly conserved among isolates from dogs in Japan, making it a useful genetic marker for molecular...
Palavras-chave: Babesia gibsoni; Conserved gene; Internal transcribed spacer 1 (ITS1); Japan.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4302
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Preimplantation Development of Embryos in Labrador Retrievers OAK
ABE, Yasuyuki; SUWA, Yoshinori; YANAGIMOTO-UETA, Yoshiko; SUZUKI, Hiroshi; 鈴木, 宏志.
Preimplantation development of canine embryos is not well understood. To understand the timing of preattachment embryogenesis relative to the luteinizing hormone (LH) surge, early embryonic development was examined in Labrador Retrievers after artificial insemination. The embryos migrated from the oviduct to the uterus beginning on day 11 after the LH surge. This transport must be completed within 24 h. By day 13 after the LH surge, all of the embryos had moved and were localized in the uterus. The embryos developed to the morula stage within 11-13 days and to the blastocyst stage within 14 days after the LH surge, respectively. These findings add to the current understanding concerning the physiology of preimplantation development and should help further...
Palavras-chave: Dog; Embryo; Preimplantation development.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3029
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Exposure of C57BL/6J Male Mice to an Electric Field Improves Copulation Rates with Superovulated Females OAK
HORI, Takuya; YAMSAARD, Thicomeporn; UETA, Yoshiko Yanagimoto; HARAKAWA, Shinji; KANEKO, Etsushi; MIYAMOTO, Akio; XUAN, Xuenan; TOYODA, Yutaka; SUZUKI, Hiroshi; 宮本, 明夫; 玄, 学南; 鈴木, 宏志.
It is well-known that there are considerable strain differences in the relative copulation rates between male and superovulated female mice. In particular, the C57BL/6J strain of mice has a lower rate of successful copulation. We examined the effect of exposure to an electric field on sexual behavior in C57BL/6J male mice. When C57BL/6J males were exposed to a 50 Hz, 45 kV/m electric field for 30 min per day for 11 days and placed in a cage with a superovulated female of the same strain, the successful copulation rates of males was significantly improved compared with unexposed males (P<0.05). These results suggest that the exposure of C57BL/6J male mice to an electric field improves their sub-fertility activity in mating with superovulated females.
Palavras-chave: Mice; Male; Electric field; Copulation; Superovulation; Sexual behavior.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3021
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