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Experimental Animals

Atsuo Ogura
Reproductive engineering techniques are essential for assisted reproduction of animals and generation of genetically modified animals. They may also provide invaluable research models for understanding the mechanisms involved in the developmental and reproductive processes. At the RIKEN BioResource Center (BRC), I have sought to develop new reproductive engineering techniques, especially those related to cryopreservation, microinsemination (sperm injection), nuclear transfer, and generation of new stem cell lines and animals, hoping that they will support the present and future projects at BRC...
October 19, 2016: Experimental Animals
Akiyoshi Ishikawa, Keita Sakai, Takehiro Maki, Yuri Mizuno, Kimie Niimi, Yasuhiro Oda, Eiki Takahashi
To understand sleep mechanisms and develop treatments for sleep disorders, investigations using animal models are essential. The sleep architecture of rodents differs from that of diurnal mammals including humans and non-human primates. Sleep studies have been conducted in non-human primates; however, these sleep assessments were performed on animals placed in a restraint chair connected via the umbilical area to the recording apparatus. To avoid restraints, cables, and other stressful apparatuses and manipulations, telemetry systems have been developed...
October 18, 2016: Experimental Animals
Minoru Kato, Yi-Ying Huang, Mina Matsuo, Yoko Takashina, Kazuyo Sasaki, Yasushi Horai, Aya Juni, Shin-Ichi Kamijo, Kaoru Saigo, Kumiko Ui-Tei, Hajime Tei
RNA interference (RNAi) is a powerful tool for the study of gene function in mammalian systems, including transgenic mice. Here, we report a gene knockdown system based on the human mir-187 precursor. We introduced small interfering RNA (siRNA) sequences against the mouse melanocortin-4 receptor (mMc4r) to alter the targeting of miR-187. The siRNA-expressing cassette was placed under the control of the cytomegalovirus (CMV) early enhancer/chicken β-actin promoter. In vitro, the construct efficiently knocked down the gene expression of a co-transfected mMc4r-expression vector in cultured mammalian cells...
September 30, 2016: Experimental Animals
Jaeman Bae, Hyeonhae Choi, Yuri Choi, Jaesook Roh
We previously showed that prepubertal chronic caffeine exposure adversely affected the development of the testes in male rats. Here we investigated dose- and time-related effects of caffeine consumption on the testis throughout sexual maturation in prepubertal rats. A total of 80 male SD rats were randomly divided into four groups: controls and rats fed 20, 60, or 120 mg caffeine/kg/day, respectively, via gavage for 10, 20, 30, or 40 days. Preputial separation was monitored daily before the rats were sacrificed...
September 21, 2016: Experimental Animals
Dejanovic Bratislav, Lavrnja Irena, Ninkovic Milica, Stojanovic Ivana, Djuric Ana, Dilber Sanda, Stevanovic Ivana
Chlorpromazine (CPZ) is a member of a widely used class of antipsychotic agents. The metabolic pathways of CPZ toxicity were examined by monitoring oxidative/nitrosative stress markers. The aim of the study was to investigate the hypothesis that agmatine (AGM) prevents oxidative stress in the liver of Wistar rats 48 hours after administration of CPZ. All tested compounds were administered intraperitoneally (i.p.) in one single dose. The animals were divided into control (C, 0.9% saline solution), CPZ (CPZ, 38...
August 11, 2016: Experimental Animals
Yasuyo Ito-Fujishiro, Hiroshi Koie, Hiroaki Shibata, Sachi Okabayashi, Yuko Katakai, Chieko Ohno, Kiichi Kanayama, Yasuhiro Yasutomi, Naohide Ageyama
Regenerative therapy with stem cell transplantation is used to treat various diseases such as coronary syndrome and Buerger's disease. For instance, stem-cell transplantation into the infarcted myocardium is an innovative and promising strategy for treating heart failure due to ischemic heart disease. Basic studies using small animals have shown that transplanted cells improve blood flow in the infarcted region. Magnetic resonance imaging (MRI) can noninvasively identify and track transplanted cells labeled with superparamagnetic iron oxide (SPIO)...
July 29, 2016: Experimental Animals
Yoshikazu Hasegawa, Yoshikazu Hoshino, Abdelaziz E Ibrahim, Kanako Kato, Yoko Daitoku, Yoko Tanimoto, Yoshihisa Ikeda, Hisashi Oishi, Satoru Takahashi, Atsushi Yoshiki, Ken-Ichi Yagami, Hiroyoshi Iseki, Seiya Mizuno, Fumihiro Sugiyama
In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1(em1 (cre) Utr) strain was produced from an oocyte injected with pX330 containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying 2A-cre. (R26GRR x C57BL/6J-Ins1(em1 (cre) Utr)) F1 mice were histologically characterized for cre-loxP recombination in the embryonic and adult stages; cre-loxP recombination was observed in all pancreatic islets examined in which almost all insulin-positive cells showed tdsRed fluorescence, suggesting β cell-specific recombination...
July 29, 2016: Experimental Animals
Chi Hai-Ying, Kiori Nagano, Sayeh Ezzikouri, Chiho Yamaguchi, Mohammad Enamul Hoque Kayesh, Khadija Rebbani, Bouchra Kitab, Hirohumi Nakano, Hiroyuki Kouji, Michinori Kohara, Kyoko Tsukiyama-Kohara
Previous studies have shown that intermittent cold stress (ICS) induces depression-like behaviors in mammals. Tupaia belangeri (the tree shrew) is the only experimental animal other than the chimpanzee that has been shown to be susceptible to infection by hepatitis B and C viruses. Moreover, full genome sequence analysis has revealed strong homology between host proteins in Tupaia and in humans and other primates. Tupaia neuromodulator receptor proteins are also known to have a high degree of homology with their corresponding primate proteins...
July 29, 2016: Experimental Animals
Yasufumi Shirasaki, Ayumi Matsuura, Masashi Uekusa, Yoshihiro Ito, Toshiaki Hayashi
Chlorine dioxide (ClO2) is a strong oxidant that possesses an antimicrobial activity. We demonstrated here that ClO2 gas is easily generated by mixing 3.35% sodium chlorite solution (Purogene) and 85% phosphoric acid at a 10:1 volume ratio without using an expensive machine. In a test room (87 m(3)), experiments were carried out using various amounts of sodium chlorite solution (0.25 ml/m(3) to 20.0 ml/m(3)). The gas concentration increased in a sodium chlorite volume-dependent manner and reached peak values of from 0...
July 29, 2016: Experimental Animals
Ai Nishitani, Miyuu Tanaka, Saki Shimizu, Naofumi Kunisawa, Mayuko Yokoe, Yusaku Yoshida, Toshiro Suzuki, Tetsushi Sakuma, Takashi Yamamoto, Mitsuru Kuwamura, Shigeo Takenaka, Yukihiro Ohno, Takashi Kuramoto
Essential tremor (ET) is a common movement disorder with a poorly understood etiology. The TRM/Kyo mutant rat, showing spontaneous tremor, is an animal model of ET. Recently, we demonstrated that tremors in these rats emerge when two mutant loci, a missense mutation in the hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (Hcn1) and the tremor (tm) deletion, are present simultaneously. However, we did not identify which gene within the tm deletion causes tremor expression in TRM/Kyo rats...
July 29, 2016: Experimental Animals
Miho Terao, Moe Tamano, Satoshi Hara, Tomoko Kato, Masato Kinoshita, Shuji Takada
The CRISPR/Cas9 system is a powerful genome editing tool for the production of genetically modified animals. To produce mutant mice, chimeric single-guide RNA (sgRNA) is cloned in a plasmid vector and a mixture of sgRNA and Cas9 are microinjected into the fertilized eggs. An issue associated with gene manipulation using the CRISPR/Cas9 system is that there can be off-target effects. To simplify the production of mutant mice with low risks of off-target effects caused by the CRISPR/Cas9 system, we demonstrated that genetically modified mice can be efficiently obtained using chemically synthesized CRISPR RNA (crRNA), trans-activating crRNA (tracrRNA), and modified Cas9s, such as the nickase version and FokI-fused catalytically inactive Cas9, by microinjection into fertilized eggs...
July 29, 2016: Experimental Animals
Hossam H Shawki, Takumi Kigoshi, Yuki Katoh, Manabu Matsuda, Chioma M Ugboma, Satoru Takahashi, Hisashi Oishi, Akihiro Kawashima
Previously, we have identified a calcium-binding protein that is specifically expressed in spermatids and localized to the flagella of the mature sperm in mouse, so-called mCABS1. However, the physiological roles of CABS1 in the male reproductive system have not been fully elucidated yet. In the current study, we aimed to localize and clarify the role of CABS1 in porcine (pCABS1). We determined for the first time the full nucleotides sequence of pCABS1 mRNA. pCABS1 protein was detected on SDS-PAGE gel as two bands at 75 kDa and 70 kDa in adult porcine testis, whereas one band at 70 kDa in epididymal sperm...
July 29, 2016: Experimental Animals
Hidemi Yasuda, Akiko Harauma, Maki Kato, Yuki Ootomo, Erisa Hatanaka, Toru Moriguchi
It is important to establish experimental animal techniques that are applicable to the newborn and infant phases for nutrition and pharmacological studies. Breeding technology using the artificial suckling method without breast milk is very effective for the study of newborn nutrition. Using this method, we separated newborn mice from dams within 48 h of birth and provided them with artificial milk. We evaluated mouse anxiety levels after early postnatal maternal separation. Artificially reared mice were subjected to elevated plus-maze tests to assess emotional behavior at 9 weeks of age...
July 29, 2016: Experimental Animals
Hirotake Ichise, Akiko Hori, Seiji Shiozawa, Saki Kondo, Yumi Kanegae, Izumu Saito, Taeko Ichise, Nobuaki Yoshida
Temporal genetic modification of mice using the ligand-inducible Cre/loxP system is an important technique that allows the bypass of embryonic lethal phenotypes and access to adult phenotypes. In this study, we generated a tamoxifen-inducible Cre-driver mouse strain for the purpose of widespread and temporal Cre recombination. The new line, named CM32, expresses the GFPneo-fusion gene in a wide variety of tissues before FLP recombination and tamoxifen-inducible Cre after FLP recombination. Using FLP-recombined CM32 mice (CM32Δ mice) and Cre reporter mouse lines, we evaluated the efficiency of Cre recombination with and without tamoxifen administration to adult mice, and found tamoxifen-dependent induction of Cre recombination in a variety of adult tissues...
July 29, 2016: Experimental Animals
Bing Chen, Hui-Ling Ouyang, Wen-Hua Wang, Yi-Heng Yin, Lin-Na Yan, Bin Yang, Zheng-Feng Xue
Hirschsprung disease (HSCR), or colonic aganglionosis, is a congenital disorder characterized by the absence of intramural ganglia along variable lengths of the colon, resulting in intestinal obstruction. It is the most common cause of congenital intestinal obstruction, with an incidence of 1 in 5,000 live births. N-ethyl-N-nitrosourea (ENU)-induced mutagenesis is a powerful tool for the study of gene function and the generation of human disease models. In the current study, a novel mutant mouse with aganglionic megacolon and coat color spotting was generated by ENU-induced mutagenesis...
July 29, 2016: Experimental Animals
Tae Yeon Kim, Kimie Niimi, Eiki Takahashi
Neuronal voltage-gated Cav2.1 channel controls a broad array of functions, including neurotransmitter release, neuronal excitability, activity-dependent gene expression, and neuronal survival. The Cav2.1 channel is molecular complexes consisting of several subunits: α1, α2/δ, β, and γ. The pore-forming subunit, α1, is encoded by the Cacna1a gene. Tottering-6j mice, generated by the Neuroscience Mutagenesis Facility at The Jackson Laboratory, are a recessive mutant strain in which the mutation has been chemically induced by ethylnitrosourea...
July 29, 2016: Experimental Animals
Masayoshi Kato, Masaya Marumo, Jun Nakayama, Misaki Matsumoto, Chihiro Yabe-Nishimura, Tohru Kamata
Accumulating evidence suggests that reactive oxygen species (ROS) generated by endogenous metabolic enzymes are involved in a variety of intracellular mechanisms. In particular, superoxide-generating NADPH oxidase (Nox) 1 is highly expressed in the colon and has been implicated in physiological and pathophysiological states of colon tissues. However, its role in tissue repair following colitis has not been fully elucidated. Our study using experimental colitis in mice showed that repair of the mucosal layer did not occur in Nox1-deficient mice following dextran sulfate sodium-induced colitis...
July 29, 2016: Experimental Animals
Hideyuki H Motohashi, Hidetoshi Ishibashi
The ovary of neonatal nonhuman primates contains the highest number of immature oocytes, but its cryopreservation has not yet been sufficiently investigated in all life stages. In the current study, we investigated cryodamage after vitrification/warming of neonatal ovaries from a nonhuman primate, the common marmoset (Callithrix jacchus). A Cryotop was used for cryopreservation of whole ovaries. The morphology of the vitrified/warmed ovaries was found to be equivalent to that of fresh ovaries. No significant difference in the number of oocytes retaining normal morphology per unit area in histological sections was found between the two groups...
July 29, 2016: Experimental Animals
Atsushi Tsukamoto, Kaho Uchida, Shizuka Maesato, Reiichiro Sato, Eiichi Kanai, Tomo Inomata
Representative inhalant anesthetic agent, isoflurane is commonly used during surgery in rats. However, isoflurane mediates relatively strong respiratory depression. In human and veterinary medicine, sedatives and analgesics are co-administered to complement the anesthetic action of inhalant anesthesia. The present study aimed to establish the novel balanced anesthesia that combines midazolam and butorphanol with isoflurane (MBI) in rats. Male Sprague Dawley rats were divided into 2 groups, and administered either isoflurane monoanesthesia or isoflurane with midazolam (2...
July 29, 2016: Experimental Animals
Rieko Yoshiyuki, Ryo Tanaka, Ryuji Fukushima, Noboru Machida
The present study aimed to evaluate the preventive effect of sildenafil treatment on pulmonary hypertension (PH) induced by monocrotaline (MCT) in rats. Fifty-four 12-week-old male Sprague-Dawley rats were injected with MCT or saline solution (MCT-injected rats: n=36; saline: n=18). Serial echocardiography and right ventricular systolic pressure (RVSP) measurements via a cardiac catheter were performed at 2, 4 and 6 weeks after the injection. After injection of MCT, rats received oral sildenafil (MCT/sildenafil group: n=18) or no treatment (MCT group: n=18) until undergoing echocardiography and cardiac catheterization...
July 29, 2016: Experimental Animals
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