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A Study on Freeze-Drying as a Method of Preserving Mouse Sperm OAK
KAWASE, Yosuke; SUZUKI, Hiroshi; 鈴木, 宏志.
SRD Young Investigator Award
Palavras-chave: Comet assay; Freeze-dried sperm; ICSI; SCSA.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3032
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Artificial Insemination with Canine Spermatozoa Frozen in a Skim Milk/Glucose-Based Extender OAK
ABE, Yasuyuki; LEE, Dong-Soo; SANO, Hikaru; AKIYAMA, Koji; YANAGIMOTO-UETA, Yoshiko; ASANO, Tomoyoshi; SUWA, Yoshinori; SUZUKI, Hiroshi; 鈴木, 宏志.
Due to the recent outbreak of avian influenza, transportation of frozen canine semen with egg yolk has been sharply restricted. Thus, there is urgent need to develop a novel egg yolk-free extender for freezing canine spermatozoa. In the present study, the effect of using skim milk/glucose (SG)-based extender without egg yolk on the motility and fertilizing capacity of canine spermatozoa frozen-thawed in the presence of glycerol was examined. There was a tendency for the proportion of motile spermatozoa exposed to SG-based extender for 3 h to be higher than that exposed for 1 h, but the difference was not significant. The motility and other viability parameters of canine spermatozoa after thawing were similar to those obtained with an egg yolk-based...
Palavras-chave: Canine; Cryopreservation; Skim milk/glucose-based extender; Spermatozoa.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3030
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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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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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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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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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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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Expression of Amphiregulin During the Pre- and Post-implantation Period in the Mouse Reproductive Tract OAK
LEE, Dong-Soo; YANAGIMOTO UETA, Yoshiko; SUZUKI, Hiroshi; 鈴木, 宏志.
In mammals, embryo implantation is an essential step in reproduction. Implantation is a phenomenon that involves crosstalk between the blastocyst and the maternal endometrium. However, the molecular basis of the connections between the blastocyst and endometrium is not yet fully understood. Amphiregulin is a member of the epidermal growth factor family and is known to be expressed in the luminal epithelium of the mouse uterus on 3.5 days post coitum (dpc). Thus, to clarify the mechanism of amphiregulin at fetomaternal interface, we analyzed the expression pattern of amphiregulin mRNA in the oviducts and uteri of pregnant and psuedopregnant mice by means of real-time PCR. Amphiregulin expression in the pregnant uterus dramatically increased on 2.5 dpc,...
Palavras-chave: Mice; Implantation; Embryo transfer; Amphiregulin; MRNA.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3026
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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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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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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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Freeze-drying in Mouse Spermatozoa OAK
川瀬, 洋介; 鈴木, 宏志; SUZUKI, Hiroshi.
現在,液体窒素を用いた精子の凍結保存法が積極的に利用されている.しかしながら,凍結精子の保存・輸送には様々な問題が存在し,これらの問題点を解決すべく,液体窒素を利用しない次世代の精子の保存・輸送方法として,凍結乾燥が注目されている.運動能をもたない凍結乾燥された精子においても,卵細胞質内精子注入法(ICSI)を利用することで凍結乾燥精子由来の産子を得ることが可能であることから,遺伝子資源の保存に凍結乾燥精子を利用することへの期待が高まっている.従って,凍結乾燥精子の数十年,数百年といった長期保存の可能性の保証や凍結乾燥精子を比較的高い温度で長期保存可能な凍結乾燥条件を見出すことが重要な課題である.最近,これらの課題を克服するために,医薬品の安定性を速度論的に考察するのに利用されている「アレニウスプロット」を用いた加速試験を応用することによって,凍結乾燥精子の長期保存性の予測が実現化された.
Palavras-chave: マウス; 凍結乾燥精子; ICSI; 加速試験; 真空度.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4188
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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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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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Transportation of Freeze-Dried Mouse Spermatozoa Under Different Preservation Conditions OAK
KAWASE, Yosuke; TACHIBE, Takanori; JISHAGE, Kou-ichi; SUZUKI, Hiroshi; 鈴木, 宏志.
Freeze-dried mouse spermatozoa can be used for normal embryonic development after injection into oocytes, thus indicating that freeze-drying is a useful method for the storage and transportation of genetic materials from animals. We recently reported that storage of freeze-dried mouse spermatozoa requires maintenance at temperatures lower than -80 C for long-term preservation and a pressure of 0.37 mbar at primary drying and that these conditions significantly improve the developmental rate to the blastocyst stage. In this study, we examined the influence of transportation and preservation conditions on freeze-dried spermatozoa. Freeze-dried spermatozoa stored for 2 or 2.5 years at 4 or -80 C were transported round trip overland between Shizuoka and...
Palavras-chave: Freeze-dried spermatozoa; Intra cytoplasmic sperm injection (ICSI); Mouse; Transportation.
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3027
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Vitamin C Deficiency Fails to Protect Mice from Malaria OAK
HERBAS, Maria Shirley; SUZUKI, Hiroshi; 鈴木, 宏志.
Nutritional deficiencies are frequent in malaria-endemic areas. It seems that micronutrient antioxidants play an important role in malaria parasite’s proliferation. Thus, the effect of vitamin C deficiency on malaria infection was examined in mice. When vitamin C deficient mice, L-gulono-γ-lactone oxidase gene knockout mice which are unable to synthesize ascorbic acid, were infected with a lethal dose of Plasmodium berghei NK65-infected red blood cells, the knockout mice showed similar parasitemia kinetics and survival rates as wild-type mice. The results indicate that deficiency of vitamin C might not affect the development of the malaria parasite in mice.
Palavras-chave: Free radical; Malaria; Vitamin C.
Ano: 2010 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3016
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