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      Effects of a mixture of medetomidine, midazolam and butorphanol on anesthesia and blood biochemistry and the antagonizing action of atipamezole in hamsters

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          Abstract

          Syrian golden hamsters ( Mesocricetus auratus) are useful laboratory rodents for studying human infectious diseases, metabolic diseases and cancer. In other rodents, such as mice and rats, a mixture of medetomidine, midazolam and butorphanol functions as a useful anesthetic, although it alters some blood biochemical parameters. In this study, we examined the effects of this mixture on anesthesia and blood biochemical parameters, and the action of atipamezole, a medetomidine antagonist, in hamsters. Intramuscular injection of a mixture of medetomidine, midazolam and butorphanol at doses of 0.15, 2.0 and 2.5 mg/kg, respectively, had a short induction time (within 5 min) and produced an anesthetic duration of approximately 100 min in hamsters. We also demonstrated that 0.15 mg/kg of atipamezole, corresponding to the same dose as medetomidine, made hamsters recover quickly from anesthesia. The anesthetic agent markedly altered metabolic parameters, such as plasma glucose and insulin; however, 0.15 mg/kg of atipamezole returned these levels to normal range within approximately 10 min after the injection. The anesthetic also slightly altered mineral levels, such as plasma inorganic phosphorus, calcium and sodium; the latter two were also improved by atipamezole. Our results indicated that the mixture of medetomidine, midazolam, and butorphanol at doses of 0.15, 2.0 and 2.5 mg/kg, respectively, functioned as an effective anesthetic, and atipamezole was useful for antagonizing both anesthesia and biochemical alteration in hamsters.

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          Effect of three types of mixed anesthetic agents alternate to ketamine in mice.

          Ketamine is usually used for murine anesthesia in animal experiments with other anesthetics for its sedation and analgesic effects. However, ketamine was categorized as a narcotic drug in Japan on January 1, 2007. After this act came into effect, a narcotic handling license became necessary for using and possessing ketamine. Pentobarbital sodium, which is also used for laboratory animal experiments as Nembutal, is no longer being manufactured. For these reasons, other anesthetic agents that can be used without a license are needed. In this paper, we examined the use of anesthetics other than ketamine and pentobarbital sodium. A combination anesthetic (M/M/B: 0.3/4/5) was prepared with 0.3 mg/kg of medetomidine, 4.0 mg/kg of midazolam, and 5.0 mg/kg of butorphanol. The anesthetics were administered to male ICR mice by intraperitoneal injection. In order to assess anesthetic depth and duration, we stimulated the mice directly after loss of righting reflexes to recovery of these same reflexes and then recorded four parameters--a tail pinch reflex, a pedal withdrawal reflex in the forelimbs, a pedal withdrawal reflex in the hindlimbs, and corneal reflex. Each parameter was scored, and the anesthetic depth, expressed by the total score, was summed. The surgical anesthesia duration of M/M/B: 0.3/4/5 mg/kg was almost identical to the surgical anesthetic duration with a ketamine and xylazine mixture (80-8 mg/kg). These data suggested that mice can be anesthetized by M/M/B: 0.3/4/5 as an alternate to ketamine. We thus can recommend M/M/B: 0.3/4/5 for murine surgical anesthesia.
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            Anesthetic Effects of a Mixture of Medetomidine, Midazolam and Butorphanol in Two Strains of Mice

            The combination of ketamine and xylazine is a widely used anesthetic for laboratory animals. However, due to an abuse problem in Japan, ketamine has been specified as a narcotic since 2007. Instead of using ketamine, Kawai et al. reported an injectable formula with an equivalent effect to the mixture of ketamine and xylazine [11]. The mixture of 0.3 mg/kg body weight (b.w.) medetomidine (Med.), 4.0 mg/kg b.w. midazoram (Mid.), and 5.0 mg/kg b.w. butorphanol (But.) produced an anesthetic duration of around 40 min in outbred ICR mice. However, the anesthetic effect of the mixture for inbred mice strains remains unknown. Therefore, we examined anesthetic effects of the mixture of Med., Mid., and But. in the BALB/c and C57BL/6J strains. After intraperitoneal injection into mice, right front paw, left hind paw, and tail pinch reflexes as well as corneal and righting reflexes were observed. Every 5 min, we scored each reflex category as 0 for reaction or 1 for no reaction. As long as the total score was at least 4 out of 5, we considered the mixture as putting a mouse in a surgical anesthetic state. The mixture produced an anesthetic duration of more than 45 min in both strains of mice. These results indicate that the mixture of Med., Mid., and But. can be a useful and effective anesthesia for the BALB/c and C57BL/6J strains of inbred mice as well as outbred ICR mice.
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              Efficient Gene Targeting in Golden Syrian Hamsters by the CRISPR/Cas9 System

              The golden Syrian hamster is the model of choice or the only rodent model for studying many human diseases. However, the lack of gene targeting tools in hamsters severely limits their use in biomedical research. Here, we report the first successful application of the CRISPR/Cas9 system to efficiently conduct gene targeting in hamsters. We designed five synthetic single-guide RNAs (sgRNAs)—three for targeting the coding sequences for different functional domains of the hamster STAT2 protein, one for KCNQ1, and one for PPP1R12C—and demonstrated that the CRISPR/Cas9 system is highly efficient in introducing site-specific mutations in hamster somatic cells. We then developed unique pronuclear (PN) and cytoplasmic injection protocols in hamsters and produced STAT2 knockout (KO) hamsters by injecting the sgRNA/Cas9, either in the form of plasmid or mRNA, targeting exon 4 of hamster STAT2. Among the produced hamsters, 14.3% and 88.9% harbored germline-transmitted STAT2 mutations from plasmid and mRNA injection, respectively. Notably, 10.4% of the animals produced from mRNA injection were biallelically targeted. This is the first success in conducting site-specific gene targeting in hamsters and can serve as the foundation for developing other genetically engineered hamster models for human disease.
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                Author and article information

                Journal
                J Vet Med Sci
                J. Vet. Med. Sci
                JVMS
                The Journal of Veterinary Medical Science
                The Japanese Society of Veterinary Science
                0916-7250
                1347-7439
                11 June 2017
                July 2017
                : 79
                : 7
                : 1230-1235
                Affiliations
                [1) ]Section of Biological Safety Research, Chitose Laboratory, Japan Food Research Laboratories, Chitose, Hokkaido 066-0052, Japan
                [2) ]Laboratory of Anatomy, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan
                [3) ]Department of Histology and Cytology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44519, Egypt
                [4) ]Section of Biological Safety Research, Tama Laboratory, Japan Food Research Laboratories, Tama, Tokyo 206-0025, Japan
                Author notes
                [* ]Correspondence to: Nakamura, T., Section of Biological Safety Research, Chitose Laboratory, Japan Food Research Laboratories, 2-3 Bunkyo, Chitose, Hokkaido 066-0052, Japan. e-mail: nakamurate@ 123456jfrl.or.jp
                Article
                17-0210
                10.1292/jvms.17-0210
                5559369
                28603217
                01a7ec95-1707-4a0d-a122-587599eccce2
                ©2017 The Japanese Society of Veterinary Science

                This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)

                History
                : 17 April 2017
                : 30 May 2017
                Categories
                Laboratory Animal Science
                Full Paper

                atipamezole,butorphanol,hamster,medetomidine,midazolam
                atipamezole, butorphanol, hamster, medetomidine, midazolam

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