Experimental Animals
Online ISSN : 1881-7122
Print ISSN : 1341-1357
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Volume 60 , Issue 1
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Review
  • Takashi KURAMOTO
    Type: Review
    Volume 60 (2011) Issue 1 Pages 1-6
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    During the 18th century, raising the "nezumi" rodent became so prevalent in Japan that two guidebooks were published on the topic. The first guidebook was entitled Yoso-tama-no-kakehashi (1775) and the second was entitled Chinganso-date-gusa (1787). It remains unclear in these texts whether the term nezumi was used to refer to the rat (Rattus norvegicus) or the mouse (Mus musculus). In this review, I explore Yoso-tama-no-kakehashi (English translation: A bridge to obtaining novel jewel-like nezumi). It was written by the owner of "Shunpo-do" and comprises two volumes; the first is 34 pages in length and the second is 14 pages. It introduces the nezumi and then provides details on novel varieties and the methods that were used to raise them. The nezumi dwells in peoples' homes. It is noteworthy that the "norako" species is classified in the same group as the nezumi. The norako is smaller than the nezumi. Its alias is "hatsuka-nezumi", a term which is still used in Japan today when referring to the mouse. This indicates that when the guidebook was written people distinguished the rat from the mouse by identifying the rat using the word nezumi and the mouse using the word norako. Moreover, I recently confirmed that the rat varieties which are introduced in Yoso-tama-no-kakehashi, such as "white", "spotted", "black bear-like", "deer-spotted", and "cracked-mark", can be found in modern laboratory rats. Taken together, it is very likely that the term nezumi is used to refer to the rat in Yoso-tama-no-kakehashi and that this was indeed a guidebook on the rat.
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  • Makoto MURAKAMI
    Type: Review Review Series: Frontiers of Model Animals for Human Diseases
    Volume 60 (2011) Issue 1 Pages 7-20
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    "Lipid mediators" represent a class of bioactive lipids that are produced locally through specific biosynthetic pathways in response to extracellular stimuli. They are exported extracellularly, bind to their cognate G protein-coupled receptors (GPCRs) to transmit signals to target cells, and are then sequestered rapidly through specific enzymatic or non-enzymatic processes. Because of these properties, lipid mediators can be regarded as local hormones or autacoids. Unlike proteins, whose information can be readily obtained from the genome, we cannot directly read out the information of lipids from the genome since they are not genome-encoded. However, we can indirectly follow up the dynamics and functions of lipid mediators by manipulating the genes encoding a particular set of proteins that are essential for their biosynthesis (enzymes), transport (transporters), and signal transduction (receptors). Lipid mediators are involved in many physiological processes, and their dysregulations have been often linked to various diseases such as inflammation, infertility, atherosclerosis, ischemia, metabolic syndrome, and cancer. In this article, I will give an overview of the basic knowledge of various lipid mediators, and then provide an example of how research using mice, gene-manipulated for a lipid mediator-biosynthetic enzyme, contributes to life science and clinical applications.
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Review
  • Haruo HASHIMOTO
    Type: Review
    Volume 60 (2011) Issue 1 Pages 21-32
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    In research into type 2 diabetes, diet-based approaches, i.e., nutritional intake, are important approaches for therapeutic research. We would like to make the following two proposals from the standpoint of laboratory animal science for reproducible animal studies using type 2 diabetes mouse models. These include congenic strains of diabetes mouse models and improvement of diets used in daily care and management. In this research, the Irs2homo-knockout mouse with both impaired glucose tolerance and insulin resistance, and thus type 2 diabetes, was established as a congenic strain. The effect of the genetic background on the onset of diabetes was examined. Next, we discussed which diets are appropriate for general care and management of mouse models in which the pathophysiology is controlled by nutritional conditions. Therefore, we prepared diets by converting the current Japanese and US diets to mice and adjusting the diet contents accordingly. We compared the insulin signals such as those of the liver, pancreas and white fat. We were thus able to establish an evaluation system closer to diabetes in the current population. Using this data as an example, we should consider the quality and ordinary diet of animals as important factors in animal experiments.
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Original
  • Kazuo KATAO, Reiko HONMA, Satoko KATO, Shinya WATANABE, Jun-ichi IMAI
    Type: Original
    Volume 60 (2011) Issue 1 Pages 33-45
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    Platinum is recognized as a harmless metal and is widely used in many industrial products. Recent studies have proposed that platinum in the form of nanoparticles has antioxidant properties, suggesting potential uses for platinum nanoparticles as additives in foods and cosmetics, with direct exposure consequences for humans. However, the influence of platinum nanoparticles on humans has not been sufficiently evaluated, thus far. Therefore, to investigate the influence of platinum nanoparticles on a living body, we comprehensively examined the expression profiles of genes obtained from 25 organs and tissues of rats after oral administration of platinum nanoparticles by gavage. Comparative analysis revealed that the expression levels of 18 genes were altered in 12 organs and tissues after the administration (approximately 0.17% of all the genes examined). Of the tissues examined, those of the glandular stomach, whichi were most directly exposed to the orally administered platinum nanoparticles, showed altered expression levels of genes associated with inflammation. In subcutaneous adipose tissue, the expression levels of genes whose products exhibited ATPase activity were altered. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) analysis confirmed the alteration in the expression levels of these genes in these 2 different tissues. Our findings indicate that orally administered platinum nanoparticles do not have a marked effect on systemic gene expression levels, except on a small number of genes expressed in rat tissues, including peripheral tissues indirectly exposed to the orally administered nanoparticles.
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  • Longquan REN, Qiang WENG, Miori KISHIMOTO, Gen WATANABE, Sukanya JAROE ...
    Type: Original
    Volume 60 (2011) Issue 1 Pages 47-56
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    The present study was undertaken to clarify changes in secretions of FSH, LH, inhibin and testosterone, and sperm motility after bilateral vasectomy in adult male rats. Bilateral vasectomy was created surgically (treated group) and intact rats were used as control (control group). On days 3, 5, 7, 14, 30, 60, and 90 after surgery, plasma concentrations of FSH, LH, inhibin, and testosterone were measured by radioimmunoassay, and sperm motility characteristics were measured by computer-assisted sperm analysis (CASA). The results show that weights of epididymides significantly increased in vasectomized rats as compared to control rats. Histologically, damage to spermatogenesis was observed in vasectomized rats. Multinucleated giant cells were observed in the lumen of some seminiferous tubules, and there were degenerative spermatids in the epididymides of vasectomized rats. Plasma levels of LH, FSH, and testosterone only decreased on day 3 after vasectomy; however, plasma levels of ir-inhibin significantly increased on day 3 after vasectomy. In addition, the sperm motility parameters, straight-line velocity, curvilinear velocity, deviation of the sperm head from the mean trajectory and the maximum amplitude of lateral head displacement were decreased from day 60 after vasectomy. These results suggest that vasectomy reduces sperm motility starting from day 60 after vasectomy, and early bilateral vasectomy does not strongly affect the endocrine function of the testis, though it may result in damage to spermatogenesis in vasectomized rats.
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  • Takashi KURAMOTO, Mitsuru KUWAMURA, Fumi TAGAMI, Tomoji MASHIMO, Masat ...
    Type: Original
    Volume 60 (2011) Issue 1 Pages 57-63
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    N-ethyl-N-nitrosourea (ENU) mutagenesis is an important tool for studying gene function and establishing human disease models. Here, we report the characterization of a novel hairless mutant rat strain that carries a recessive mutation called Kyoto rhino (krh), which was created by ENU-mutagenesis. We produced a F344-krh strain through inbreeding without backcrossing to F344 rats. The krh/krh rats lost their coat hair by eight weeks of age. They also developed wrinkled skin, cystic hair canals and long curved nails by four months of age. Markedly dilated hair follicles that contained keratin debris were observed during histological analysis of the skin. The krh locus was mapped near the hairless (Hr) gene on chromosome 15. Sequence analysis revealed a nonsense mutation (c. 1238 C>A, p. S413X) in the Hr gene. The truncated HR protein was deduced to lack a zinc-finger domain and repression domains. In aged Hrkrh/Hrkrh rats, focal glomerulosclerosis (FGS) was observed in which collapsed glomeruli contained protein exudates in Bowman's capsule. Mesangial matrices that had proliferated in segments and foot processes that were fused in podocytes were also observed. The Hrkrh/Hrkrh rats also suffered from significant proteinuria. Given its breeding history, the F344-Hrkrh strain may harbor ENU-induced mutation(s) that underlie FGS in addition to having the Hrkrh mutation. The F344-Hrkrh rat is a useful model of skin disease and may provide a new model system for the examination of the pathogenesis of FGS.
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  • Eui-Suk JEONG, Kyoung-Sun LEE, Seung-Ho HEO, Jin-Hee SEO, Yang-Kyu CHO ...
    Type: Original
    Volume 60 (2011) Issue 1 Pages 65-70
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    The accurate and economical diagnosis of pathogenic bacteria is necessary for the microbiological control of laboratory animals. In this study, we developed a triplex PCR method for the direct detection of three common gastroenteric bacteria, Pseudomonas aeruginosa, Helicobacter hepaticus, and Salmonella typhimurium. Targets were specifically amplified by conventional PCR assay using a genomic fragment from P. aeruginosa, 16S ribosomal RNA from H. hepaticus, and the invA gene from S. typhimurium. To investigate the specificity of our primers, they were tested against purified DNA from many other bacterial species. There were no amplification products from other bacteria. Under optimized conditions, the triplex assay simultaneously yielded a 726-bp product from P. aeruginosa, a 417-bp product from H. hepaticus, and a 246-bp product from S. typhimurium. The detection limits of this assay in pure culture were 10 pg for P. aeruginosa, and 0.1 pg for H. hepaticus and S. typhimurium. All three bacteria were successfully detected in the liver, cecum, and feces of experimentally infected mice. This method is a useful and convenient assay that allows the simultaneous identification of bacterial pathogens in mice. Our triplex method will be used to improve quality control in the detection of pathogenic bacterial infections in laboratory animal facilities.
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  • Lin XU, Zixing WANG, Xiwen XIONG, Xingxing GU, Xiang GAO, Xia GAO
    Type: Original
    Volume 60 (2011) Issue 1 Pages 71-78
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    N-ethyl-N-nitrosourea (ENU)-induced mutagenesis is an important approach in the study of gene function and the establishment of human disease models. Here we report an ENU-induced mutation, Elfin, as a mouse model with hearing loss. Homozygous mutants were deaf and displayed severe ataxia, while heterozygous mice had a significant hearing loss. Histological analysis of the inner ear revealed that Elfin had progressive degeneration of the organ of Corti, spiral ganglion cells and an absence of otoconia in the vestibular system. The new mutation was mapped to chromosome 6 between microsatellite markers D6Mit39 and D6Mit254, where the Ca2+-ATPase type 2 (Atp2b2) gene resides. Sequence analysis revealed a unique T-to-A transition mutation at amino acid 655 resulting in Ile-to-Asn substitution. These results for the Elfin mutant confirm the role of ATP2B2 in balance, hearing and formation of otoconia and suggest it may serve as a new model of human hereditary hearing loss.
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  • Masaki UENO, Ken URABE, Kouji NARUSE, Kentaroo UCHIDA, Hiroaki MINEHAR ...
    Type: Original
    Volume 60 (2011) Issue 1 Pages 79-87
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    This study aimed to clarify the relationship between the mechanical environment at the fracture site and endogenous fibroblast growth factor-2 (FGF-2). We compared two types of fracture healing with different callus formations and cellular events using MouseFixTM plate fixation systems for murine fracture models. Left femoral fractures were induced in 72 ten-week-old mice and then fixed with a flexible (Group F) or rigid (Group R) Mouse FixTM plate. Mice were sacrificed on days 3, 5, 7, 10, 14, and 21. The callus volumes were measured by 3D micro-CT and tissues were histologically stained with hematoxylin & eosin or safranin-O. Sections from days 3, 5, and 7 were immunostained for FGF-2 and Proliferating Cell Nuclear Antigen (PCNA). The callus in Group F was significantly larger than that in Group R. The rigid plate allowed bone union without a marked external callus or chondrogenesis. The flexible plate formed a large external callus as a result of endochondral ossification. Fibroblastic cells in the granulation tissue on days 5 and 7 in Group F showed marked FGF-2 expression compared with Group R. Fibroblastic cells showed ongoing proliferation in granulation tissue in group F, as indicated by PCNA expression, which explained the relative granulation tissue increase in group F. There were major differences in early phase endogenous FGF-2 expression between these two fracture healing processes, due to different mechanical environments.
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Note
  • Hideharu OCHIAI, Ken ONDA, Takuya MARUO, Nobuyuki KANEMAKI
    Type: Note
    Volume 60 (2011) Issue 1 Pages 89-91
    Released: February 15, 2011
    JOURNALS FREE ACCESS
    To evaluate the relationship between aquaporin-1 (AQP1) expression and the cell volume of red blood cells (RBCs), canine peripheral RBCs were separated according to specific gravity, and expression of the AQP1 protein on the membrane of RBCs was compared using anti-dog AQP1 polypeptide serum. Western blot analysis indicated that there was no significant difference in AQP1 expression between large and small cell fractions. In addition, the AQP1 expression of inherited high K/low Na RBCs which are known to be 20% larger than normal RBCs, was comparable to that of normal RBCs. These results suggest that AQP1, the major water channel in RBCs, does not determine the cell volume of peripheral canine RBCs.
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