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      Test-retest reproducibility of cardiac magnetic resonance imaging in healthy mice at 7-Tesla: effect of anesthetic procedures

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          Abstract

          Cardiac magnetic resonance (CMR) has emerged as a powerful tool for in vivo assessments of cardiac parameters in experimental animal models of cardiovascular diseases, but its reproducibility in this setting remains poorly explored. To address this issue, we investigated the test-retest reproducibility of preclinical cardiac magnetic resonance imaging (CMR) at 7 Tesla in healthy C57BL/6 mice, including an analysis of the impact of different anesthetic procedures (isoflurane or pentobarbital). We also analyzed the intra-study reproducibility and the intra- and inter-observer post-processing reproducibility of CMR images. Test-retest reproducibility was high for left ventricular parameters, especially with the isoflurane anesthetic procedure, whereas right ventricular parameters and deformation measurements were less reproducible, mainly due to physiological variability. Post-processing reproducibility of CMR images was high both within and between observers. These results highlight that anesthetic procedures might influence CMR test-retest reproducibility, an important ethical consideration for longitudinal studies in rodent models of cardiomyopathy to limit the number of animals used.

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          Effects of anesthetics on systemic hemodynamics in mice.

          The aim of this study was to compare the systemic hemodynamic effects of four commonly used anesthetic regimens in mice that were chronically instrumented for direct and continuous measurements of cardiac output (CO). Mice (CD-1, Swiss, and C57BL6 strains) were instrumented with a transit-time flow probe placed around the ascending aorta for CO measurement. An arterial catheter was inserted into the aorta 4 or 5 days later for blood pressure measurements. After full recovery, hemodynamic parameters including stroke volume, heart rate, CO, mean arterial pressure (MAP), and total peripheral resistance were measured with animals in the conscious state. General anesthesia was then induced in these mice using isoflurane (Iso), urethane, pentobarbital sodium, or ketamine-xylazine (K-X). The doses and routes of administration of these agents were given as required for general surgical procedures in these animals. Compared with the values obtained for animals in the conscious resting state, MAP and CO decreased during all anesthetic interventions, and hemodynamic effects were smallest for Iso (MAP, -24 +/- 3%; CO, -5 +/- 7%; n = 15 mice) and greatest for K-X (MAP, -51 +/- 6%; CO, -37 +/- 9%; n = 8 mice), respectively. The hemodynamic effects of K-X were fully antagonized by administration of the alpha(2)-receptor antagonist atipamezole (n = 8 mice). These results indicate that the anesthetic Iso has fewer systemic hemodynamic effects in mice than the nonvolatile anesthetics.
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            Anesthesia and other considerations for in vivo imaging of small animals.

            The use of small animal imaging is increasing in biomedical research thanks to its ability to localize altered biochemical and physiological processes in the living animal and to follow these processes longitudinally and noninvasively. In contrast to human studies, however, imaging of small animals generally requires anesthesia, and anesthetic agents can have unintended effects on animal physiology that may confound the results of the imaging studies. In addition, repeated anesthesia, animal preparation for imaging, exposure to ionizing radiation, and the administration of contrast agents may affect the processes under study. We discuss this interplay of factors for small animal imaging in the context of four common imaging modalities for small animals: positron emission tomography (PET) and single photon emission computed tomography (SPECT), computed tomography (CT), magnetic resonance imaging (MRI), and optical imaging. We discuss animal preparation for imaging, including choice of animal strain and gender, the role of fasting and diet, and the circadian cycle. We review common anesthesias used in small animal imaging, such as pentobarbital, ketamine/xylazine, and isoflurane, and describe techniques for monitoring the respiration and circulation of anesthetized animals that are being imaged as well as developments for imaging conscious animals. We present current imaging literature exemplifying how anesthesia and animal handling can influence the biodistribution of PET tracers. Finally, we discuss how longitudinal imaging studies may affect animals due to repeated injections of radioactivity or other substrates and the general effect of stress on the animals. In conclusion, there are many animal handling issues to consider when designing an imaging experiment. Reproducible experimental conditions require clear, consistent reporting, in the study design and throughout the experiment, of the animal strain and gender, fasting, anesthesia, and how often individual animals were imaged.
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              Effects of anesthesia on echocardiographic assessment of left ventricular structure and function in rats.

              Echocardiography is an essential diagnostic tool for accurate noninvasive assessment of cardiac structure and function in vivo. However, the use of anesthetic agents during echocardiographic studies is associated with alterations in cardiac anatomical and functional parameters. We sought to systematically compare the effects of three commonly used anesthetic agents on echocardiographic measurements of left ventricular (LV) systolic and diastolic function, LV dimensions, and LV mass in rats. Adult male Fischer 344 rats underwent echocardiographic studies under pentobarbital (PB, 25 mg/kg i.p.) (group I, n = 25), inhaled isoflurane (ISF, 1.5%) (group II, n = 25),or ketamine/xylazine (K/X, 37 mg/kg ketamine and 7 mg/kg xylazine i.p.) (group III, n = 25) anesthesia in a cross-over design. Echocardiography was also performed in an additional group of unanesthetized conscious rats (group IV, n = 5). Postmortem studies were performed to validate echocardiographic assessment of LV dimension and mass. Rats in group I exhibited significantly higher LV ejection fraction, fractional shortening, fractional area change, velocity of circumferential fiber shortening corrected for heart rate, and heart rate as compared with groups II and III. LV end-diastolic volume, end-diastolic diameter, and cross-sectional area in diastole were significantly smaller in group I compared with groups II and III. Cardiac output was significantly lower in group III compared with groups I and II. Postmortem LV mass measurements correlated well with echocardiographic estimation of LV mass for all anesthetic agents, and the correlation was best with PB anesthesia. Limited echocardiographic data obtained in conscious rats were similar to those obtained under PB anesthesia. We conclude that compared with ISF and K/X anesthesia, PB anesthesia at a lower dose yields echocardiographic LV structural and functional data similar to those obtained in conscious rats. In addition, PB anesthesia also facilitates more accurate estimation of LV mass.
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                Author and article information

                Contributors
                joubert-m@chu-caen.fr
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                27 July 2017
                27 July 2017
                2017
                : 7
                : 6698
                Affiliations
                [1 ]ISNI 0000 0001 2186 4076, GRID grid.412043.0, EA4650, , Université de Caen Normandie, ; 14000 Caen, France
                [2 ]ISNI 0000 0004 0472 0160, GRID grid.411149.8, Nuclear Medicine, , CHU de Caen, ; 14000 Caen, France
                [3 ]ISNI 0000 0004 0472 0160, GRID grid.411149.8, , Cardiology, CHU de Caen, ; 14000 Caen, France
                [4 ]ISNI 0000 0001 2294 713X, GRID grid.7942.8, , Cardiology, Université Catholique de Louvain, ; B-1348 Louvain-la-Neuve Brussels, Belgium
                [5 ]ISNI 0000 0004 0472 0160, GRID grid.411149.8, , Biostatistic, CHU de Caen, ; 14000 Caen, France
                [6 ]ISNI 0000 0004 0472 0160, GRID grid.411149.8, , Diabetes Care Unit, CHU de Caen, ; 14000 Caen, France
                Article
                7083
                10.1038/s41598-017-07083-1
                5532227
                28751730
                1984b3a8-2a03-431d-8001-3b8910175c66
                © The Author(s) 2017

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 6 March 2017
                : 5 July 2017
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