Experimental Animals
Online ISSN : 1881-7122
Print ISSN : 1341-1357
ISSN-L : 0007-5124
59 巻 , 2 号
選択された号の論文の18件中1~18を表示しています
Review
  • Kazunari K. YOKOYAMA, Takehide MURATA, Jianzhi PAN, Koji NAKADE, Shota ...
    原稿種別: Review Review Series: Animal Bioresource in Japan
    2010 年 59 巻 2 号 p. 115-124
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Genetic materials are one of the most important and fundamental research resources for studying biological phenomena. Scientific need for genetic materials has been increasing and will never cease. Ever since it was established as RIKEN DNA Bank in 1987, the Gene Engineering Division of RIKEN BioResource Center (BRC) has been engaged in the collection, maintenance, storage, propagation, quality control, and distribution of genetic resources developed mainly by the Japanese research community. When RIKEN BRC was inaugurated in 2001, RIKEN DNA Bank was incorporated as one of its six Divisions, the Gene Engineering Division. The Gene Engineering Division was selected as a core facility for the genetic resources of mammalian and microbe origin by the National BioResource Project (NBRP) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan in 2002. With support from the scientific community, the Division now holds over 3 million clones of genetic materials for distribution. The genetic resources include cloned DNAs, gene libraries (e.g., cDNA and genomic DNA cloned into phage, cosmid, BAC, phosmid, and YAC), vectors, hosts, recombinant viruses, and ordered library sets derived from animal cells, including human and mouse cells, microorganisms, and viruses. Recently genetic materials produced by a few MEXT national research projects were transferred to the Gene Engineering Division for further dissemination. The Gene Engineering Division performs rigorous quality control of reproducibility, restriction enzyme mapping and nucleotide sequences of clones to ensure the reproducibility of in vivo and in vitro experiments. Users can easily access our genetic materials through the internet and obtain the DNA resources for a minimal fee. Not only the materials, but also information of features and technology related to the materials are provided via the web site of RIKEN BRC. Training courses are also given to transfer the technology for handling viral vectors. RIKEN BRC supports scientists around the world in the use of valuable genetic materials.
  • Masa-Toshi YAMAMOTO
    原稿種別: Review Review Series: Animal Bioresource in Japan
    2010 年 59 巻 2 号 p. 125-138
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    The fruit fly, Drosophila melanogaster, is not categorized as a laboratory animal, but it is recognised as one of the most important model organisms for basic biology, life science, and biomedical research. This tiny fly continues to occupy a core place in genetics and genomic approaches to studies of biology and medicine. The basic principles of genetics, including the variations of phenotypes, mutations, genetic linkage, meiotic chromosome segregation, chromosome aberrations, recombination, and precise mapping of genes by genetic as well as cytological means, were all derived from studies of Drosophila. Recombinant DNA technology was developed in the 1970s and Drosophila DNA was the first among multicellular organisms to be cloned. It provided a detailed characterization of genes in combination of classical cytogenetic data. Drosophila thus became the pioneering model organism for various fields of life science research into multicellular organisms. Here, I briefly describe the history of Drosophila research and provide a few examples of the application of the abundant genetic resources of Drosophila to basic biology and medical investigations. A Japanese national project, the National BioResource Project (NBRP) for collection, maintainance, and provision of Drosophila resources, that is well known and admired by researchers in other countries as an important project, is also briefly described.
  • Yutaka BANNO, Toru SHIMADA, Zenta KAJIURA, Hideki SEZUTSU
    原稿種別: Review Review Series: Animal Bioresource in Japan
    2010 年 59 巻 2 号 p. 139-146
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Silkworms have played an important agricultural role in supporting Japan’s modernization, and traditionally, Japan has led the world as a repository of silkworm bioresources. The silkworm is a small and highly domesticated insect, which is ideal as a laboratory tool, although it is a bioresource that is relatively infrequently used in experiments at present. In this review, we describe the potential for silkworm resources to contribute to life sciences.
Original
  • Takashi KURAMOTO, Mayuko YOKOE, Kayoko YAGASAKI, Tatsuya KAWAGUCHI, Ke ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 147-155
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    To collect rat mutations and increase the value of the rat model system, we introduced fancy-derived mutations to the laboratory and carried out genetic analyses. Six fancy rats were shipped from a fancy rat colony in the USA and used as founders. After initial crosses with a laboratory strain, TM/Kyo or PVG/Seac, inbreeding started and 6 partially inbred lines, including 2 sublines, were produced as Kyoto Fancy Rat Stock (KFRS) strains. During inbreeding, we isolated 9 mutations: 5 coat colors, American mink (am), Black eye (Be), grey (g), Pearl (Pel), siamese (sia); 1 coat pattern, head spot (hs); 2 coat textures, Rex (Re), satin (sat); and an ear pinnae malformation, dumbo (dmbo). Genetic analyses mapped 7 mutations to particular regions of the rat chromosomes (Chr): am to Chr 1, sia to Chr 1, sat to Chr 3, Re to Chr 7, g to Chr 8, dmbo to Chr 14, and hs to Chr 15. Candidate gene analysis revealed that a missense mutation in the tyrosinase gene, Ser79Pro, was responsible for sia. From mutant phenotypes and mapping positions, it is likely that all mutations isolated in this study were unique to the fancy rat. These findings suggest that fancy rat colonies are a good source for collecting rat mutations. The fancy-derived mutations, made available to biomedical research in the current study, will increase the scientific value of laboratory rats.
  • Seiji NISHIBAYASHI, Katsuji HATTORI, Takahiro HIRANO, Kenji UEHARA, Yo ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 157-170
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    The aim of this study was to develop and characterize a rat glomerulonephritis model, which progresses to renal fibrosis and renal failure. A single immunization of female WKY rats with more than 10 μg of recombinant α3(IV)NC1 protein caused severe proteinuria followed by progressive increases in plasma creatinine and blood urea nitrogen (BUN) level within 42 days. Sequential histopathological evaluation revealed crescent formation in glomeruli followed by tubular dilation and interstitial fibrosis. Hydroxyproline content and expression of type I collagen and smooth muscle actin genes in the renal cortex increased as renal dysfunction progressed. Furthermore, the TGF-β1 level in the renal cortex also increased. In the evaluation of antinephritic agents in this model, prednisolone and mycophenolate mofetil (MMF) treatment significantly decreased plasma creatinine and BUN, and suppressed renal fibrosis and histological changes involving crescent formation, compared with the vehicle-treated nephritic rats, whereas lisinopril treatment failed to improve renal function and histology. We demonstrated that immunization of female WKY rats with a sufficient dose of recombinant α3(IV)NC1 induces end-stage kidney disease accompanied by renal fibrosis. The relatively short period needed to induce the disease and the high incidence of functional and structural changes were considered a great advantage of this model for clarifying the mechanisms of progressive glomerulonephritis and for evaluating agents used to treat renal failure.
  • Haru OKAMOTO, Shinsuke UMEDA, Takehiro NOZAWA, Michihiro T. SUZUKI, Ya ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 171-182
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    The central region of the primate retina is called the macula. The fovea is located at the center of the macula, where the photoreceptors are concentrated to create a neural network adapted for high visual acuity. Damage to the fovea, e.g., by macular dystrophies and age-related macular degeneration, can reduce central visual acuity. The molecular mechanisms leading to these diseases are most likely dependent on the proteins in the macula which differ from those in the peripheral retina in expression level. To investigate whether the distribution of proteins in the macula is different from the peripheral retina, proteomic analyses of tissues from these two regions of cynomolgus monkeys were compared. Two-dimensional gel electrophoresis and mass spectrometry identified 26 proteins that were present only in the macular gel spots. The expression levels of five proteins, cone photoreceptor specific arrestin-C, γ-synuclein, epidermal fatty acid binding protein, tropomyosin 1α chain, and heterogeneous nuclear ribonucleoproteins A2/B1, were significantly higher in the macula than in the peripheral retina. Immunostaining of macula sections by antibodies to each identified protein revealed unique localization in the retina, retinal pigment epithelial cells and the choroidal layer. Some of these proteins were located in cells with higher densities in the macula. We suggest that it will be important to study these proteins to determine their contribution to the pathogenesis and progression of macula diseases.
  • Hideki KATOH, Tetsu NISHIKAWA, Jiro KIMURA, Yumika YAMAUCHI, Shuji TAK ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 183-190
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    We attempted to detect natural mutations existing in the Jcl:ICR closed colony of mice which is maintained by random mating. We used ordinary genetic backcrosses to efficiently detect recessive mutations carried by individual mice in the colony. Crosses of DBA/2 females and ICR males were performed to obtain F1 mice. Four F1 females randomly selected from each cross were backcrossed to the male parent. More than thirty backcross progeny were obtained from each F1 female by several deliveries. Phenotypes of the backcross progeny were observed macroscopically at about one month of age. As a result, 18 (26.1%) of 69 Jcl:ICR males carried 11 recessive mutation(s). Based on the phenotypes, the tentative names were abnormal kidney, aplasia of eyelids/hind limb digits, circling, dwarfism, heterotaxy, hind limb paralysis, hydrocephalus, rigidity (or rigor), testicular hypoplasia, tremor, and wobbling. The genes responsible for aplasia of eyelids/hind limb digits and dwarfism were each carried by two males, the genes responsible for hydrocephalus and testicular hypoplasia were each carried by three males and the gene responsible for wobbling by four males. It was strongly suggested that the genes shared by several males originated from an identical mutated gene. Surprisingly, male No. 43 had the responsible genes of abnormal kidneys and testicular hypoplasia, and No. 79 had those of dwarfism and tremor. The results obtained in this study suggest that breeders need to be aware of the presence of natural mutations in their colonies.
  • Nobutaka DOE, Tsutomu TAKAHASHI, Mika KIRIYAMA
    原稿種別: Original
    2010 年 59 巻 2 号 p. 191-197
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    In the case of mice, when the difficulty of a water maze learning task is increased, some animals gradually cease to swim, abandon adaptive learning, and become immobile. We trained 99 male C57BL/6N mice in a pool containing a hidden platform. The pool was surrounded by white featureless walls, and almost all external cues were removed. On the eighth day of escape training, 36 inferior-learners exhibited behavioral despair. The predictive validity of the inferior-learners as a depression model was verified by testing their sensitivity to clinically efficacious antidepressants. The inferior-learners treated with a selective serotonin reuptake inhibitor (SSRI), fluvoxamine, or a serotonin noradrenaline reuptake inhibitor (SNRI), milnacipran, resumed swimming and adaptive learning. Because of facial similarities between inferior-learners and depressive patients and their sensitivity to antidepressant drugs, our experimental method is expected to be an effective tool in basic research on depression.
  • Hideki SANO, Matsuyuki DOI, Soichiro MIMURO, Shuchun YU, Tadayoshi KUR ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 199-205
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    This study examined the sedative effect of, and hemodynamic response to dexmedetomidine administration in propofol-sedated swine. Sixteen swine were subjects. After anesthetic induction and preparation, the propofol infusion rate was adjusted to maintain a bispectral index (BIS) value between 55 and 65 (i.e., baseline). With the propofol infusion rate fixed at the baseline rate, dexmedetomidine was infused continuously at a rate of 0.2, 0.4, and 0.7 μg·kg –1·h–1 for one hour at each rate. The BIS value and hemodynamic parameters were recorded at each step. Dexmedetomidine decreased the BIS value, mean arterial blood pressure, heart rate, cardiac output, and mixed venous oxygen saturation in a dose-dependent manner. The systemic vascular resistance (SVR) did not change, but the pulmonary vascular resistance (PVR) increased. Oxygen delivery (DO2) and oxygen consumption (VO2) decreased. A small dose of dexmedetomidine (0.2 μg·kg–1·h–1) greatly enhanced the sedative effects of propofol with only small changes in hemodynamics and systemic oxygen balance, suggesting it may be useful in reducing the propofol dose requirement. However, dexmedetomidine 0.4 μg·kg–1·h –1 suppressed cardiac contractility, and 0.7 μg·kg–1·h–1 induced hemodynamic instability and further systemic oxygen imbalance while the additional sedative effect was limited. A lower dose of dexmedetomidine may be recommended when using it in combination with propofol.
  • Xue-Ya ZHANG, Xi HUANG, Feng QIN, Ping REN
    原稿種別: Original
    2010 年 59 巻 2 号 p. 207-214
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    A traditional Chinese medicine, Guan-Xin-Er-Hao (GXEH), is a famous multiple target therapeutic polypharmaceutical. Our aim was to evaluate whether or not oral administration of GXEH has an anti-inflammatory effect associated with blockade of nuclear factor-kappa B (NF-κB), and to investigate the NF-κB-mediated pro-inflammatory cytokines expression pathway during acute myocardial infarction (AMI) in rats. Sprague-Dawley rats were randomly assigned to four groups: oral GXEH administered at 15 or 5 g/kg, the vehicle control and sham-operated groups. Thirty minutes after giving GXEH or 0.9% NaCl (p.o.) once, coronary arteries were occluded except for the sham-operated rats. We measured 24-h infarct size, 3-h expression of NF-κB protein in the myocardial left ventricular tissues and serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and C-reactive protein (CRP). Compared with rats receiving vehicle, rats administered 15 g/kg GXEH had significantly reduced 24-h infarct size, expression of NF-κB protein and serum concentrations of TNF-α, IL-6, and CRP. GXEH at 5 g/kg did not have a significant effect on these parameters. In conclusion, GXEH exhibited an anti-inflammatory effect through inhibition of the NF-κB-mediated signaling pathway leading to downregulation of pro-inflammatory cytokine expression. These findings provide new evidence of the cardioprotective effect of GXEH through reduction of infarct size by mediating lots of endogenous materials via multiple pathways to act on myocardial cells in the treatment of cardiovascular disease.
  • Giuseppe PICCIONE, Claudia GIANNETTO, Francesco FAZIO, Elisabetta GIUD ...
    原稿種別: Original
    2010 年 59 巻 2 号 p. 215-223
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    The present study was undertaken to determine the temporal variation in intraocular pressure (IOP) and if this variation is under circadian clock control. The authors exposed five female and five male Beagles to four different artificial lighting regimes: 12/12 light/dark (L/D) period, 12/12 D/L period, constant light, and constant darkness. IOP was measured at 3 h intervals over a 48-h period. Statistical analysis of the data was performed by multivariate ANOVA, one-way repeated measure ANOVA and by the single cosinor method. Results showed no statistical effect of gender, eye and photoperiod on IOP values. A significant effect of time for each gender and each eye during all lighting regimes was seen, except during constant light, and also robust daily rhythmicity of IOP values in all L/D periods, except during constant light. In conclusion IOP values in the dog show a circadian rhythm and this rhythm is driven by a central pacemaker.
  • Kazuki NAKAO, Kenichi INOUE
    原稿種別: Original
    2010 年 59 巻 2 号 p. 225-229
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Mutant mice are indispensable to biological and medical research, and transgenic mouse production by DNA injection into zygotes has been an important method for producing these mice which are used to examine the over-expression of genes and to analyze gene transcriptional regulatory sequences. Recently, cryopreservation of zygotes by a simple vitrification method has become popular, saving the labor of isolating zygotes following each injection. However, the DNA injection technique requires training in the use of special equipment, and the injection cannot be accomplished routinely in every laboratory. The exchange of live mice also risks the propagation of common murine pathogens and possible escape of the animals; therefore the transfer of mice in frozen zygotes or embryos is recommended. Here we propose injecting DNA into frozen and thawed zygotes, refreezing them before transportation to any destination, where they can be thawed and developed into pups. The rate of transgenic mouse production using this method does not decrease significantly, even if zygotes are frozen and thawed before and after their DNA injection, and will make transgenic studies more popular on a worldwide scale.
  • Xiao Hui LV, Da Zhao SHI
    原稿種別: Original
    2010 年 59 巻 2 号 p. 231-237
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Serum estradiol (E2) and progesterone (P4) levels were measured by chemiluminescence immunoassay (CLIA) during consecutive reproductive states in Mongolian gerbils. The results show that E2 and P4 levels throughout the estrous cycle in Mongolian gerbils peaked at proestrus and estrus, respectively. During gestation in primiparous and multiparous gerbils, E2 levels reached a small peak on day 6 and a maximum peak on day 21, followed by a slight decline until parturition. Thereafter, they dropped sharply during the first 3 days of lactation and remained stable until lactation ended. P4 levels reached a peak on day 12 during gestation in primiparous gerbils, while they peaked on day 6 in multiparous gerbils. The levels of P4 then decreased until parturition. During lactation, P4 levels peaked on day 9 in primiparous gerbils, while they peaked on day 6 in multiparous gerbils, and then the levels declined gradually until lactation ended. The findings suggest that the variations of E2 levels during the estrous cycle, gestation and lactation in Mongolian gerbils are similar to those observed in rats and mice. Changes in E2 levels during different reproductive states were the same in the primiparous and multiparous gerbils, but the variations in P4 levels did not display this tendency. Changing patterns of E2 and P4 levels are suitable for the reproductive stages of Mongolian gerbils.
Note
  • Maria Shirley HERBAS, Hiroshi SUZUKI
    原稿種別: Note
    2010 年 59 巻 2 号 p. 239-243
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    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.
  • Takayuki ANZAI, Ludwig G. ULLMANN, Daisuke HAYASHI, Tetsuo SATOH, Take ...
    原稿種別: Note
    2010 年 59 巻 2 号 p. 245-249
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    The Local Lymph Node Assay (LLNA) is now regarded as the worldwide standard. The analysis of accumulated LLNA data reveals that the animal strains and vehicles employed are likely to affect LLNA results. Here we show that an obvious strain difference in the local lymph node response was observed between DMSO-treated CBA/CaOlaHsd and CBA/CaHsdRcc mice. We also show that a vehicle difference in the response was observed when CBA/CaHsdRcc mice were exposed to 6 vehicles; 4:1 v/v acetone/olive oil (AOO), ethanol/water (70% EtOH), N,N-dimethylformamide (DMF), 2-butanone (BN), propylene glycol (PG), and dimethylsulfoxide (DMSO). The dpm/LN level was lowest in the 70% EtOH group and highest in the DMSO group. When alpha-hexylcinnamaldehyde (HCA) was used as a sensitizer for the LLNA, HCA was a weak sensitizer when AOO or DMSO was used as a vehicle, but a moderate sensitizer when the other 4 vehicles were used. This study showed that there are vehicle differences in the local lymph node response (dpm/LN level) in the LLNA and that the sensitization potency of HCA may be classified in different categories when using different vehicles. This suggests that careful consideration should be exercised in selecting a vehicle for the LLNA. A further comprehensive study will be needed to investigate why vehicle differences are observed in the LLNA.
  • Takashi YOSHIDA, Koji HANARI, Koji FUJIMOTO, Tadashi SANKAI
    原稿種別: Note
    2010 年 59 巻 2 号 p. 251-254
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    We studied two mating systems for a large-scale breeding colony of cynomolgus monkeys: (i) a 3-day timed system that keeps one female and one male together in a cage for 3 days beginning 11 days after menstruation, and (ii) a 7-day timed system beginning 9 days after menstruation. Regardless of the mating system used, we propose a practical definition of an infertile female as one that does not become pregnant in up to six pairings. If we eliminate these animals from the breeding colony, the pregnancy rates would be 3.6 times higher. Eliminating infertile females from a breeding colony is important in order to save labor, time and cost and to maintain a healthy breeding colony.
  • Fusako MITSUNAGA, Miyuki UEIWA, Yoshirou KAMANAKA, Mayumi MORIMOTO, Sh ...
    原稿種別: Note
    2010 年 59 巻 2 号 p. 255-260
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Non-invasive fetal sex determination is required for biomedical studies, in which some sexual difference would be expected in fetal events, in order to make a choice of male or female fetus. To detect male fetal DNA of the sex-determining region Y gene (SRY) in maternal macaque plasma, nested real-time PCR using the SYBR Green system was developed. In all cases of pregnant macaques with male fetuses, a nested PCR product of SRY was amplified from the mother’s plasma, while no amplicon was detected in any case of pregnancy with a female fetus. Interestingly, fetal SRY DNA appeared to be cleared rapidly from the maternal blood after parturition. The current method is sensitive and can be performed with a regular PCR machine.
  • Kaori YABUUCHI, Masaru TAJIMA, Yuko KOTANI, Katsumi AIHARA, Aiko KAWAS ...
    原稿種別: Note
    2010 年 59 巻 2 号 p. 261-267
    発行日: 2010年
    公開日: 2010/05/19
    ジャーナル フリー
    Immunodeficient animals are in demand in current biomedical research, and they contribute to medical progress. In Pneumocystis infections, a specific histological diagnostic tool may be immunochemistry (IC). However, it was recently reported that the antibody (3F6) was not suitable for detecting Pneumocystis in rats. We purchased another antibody [PNC007] from a commercial source for IC. We could detect positive signals at identical locations with IC and Toluidine blue O in lungs of infected rats. These results corresponded to the results obtained with PCR. We should study the relationship between unexpected positive signals seen in IC and trophic forms in lungs of infected rats. We could clinically diagnose pneumonia caused by Pneumocystis carinii with the diagnostic method we developed.
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