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The grip strength test is used to measure the neuromuscular function as maximal muscle strength of forelimbs and combined forelimbs and hind limbs. These are assessed by the grasping applied by the mouse on a grid that is connected to a sensor. Three trials are carried out in succession measuring forelimb-strength only, followed by three successive trials measuring the combined forelimb/hindlimb grip strength. All grip strength values obtained are normalized against mouse body weight.Ontological description: MP:0001515 - abnormal grip strength.
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Indirect calorimetry provides detailed information on the energy metabolism of mutant mice. Energy expenditure is evaluated through indirect calorimetry by measuring oxygen consumption with an open flow respirometric system. CO2 and O2 sensors measure the difference in CO2 and O2 concentrations in air volumes flowing through control or animal cages. The amount of oxygen consumed over a given period of time can thus be calculated, as far as the air flow through the cage is known. Data are expressed as ml O2 h-1animal-1. The system also monitors CO2 production, therefore, the respiratory exchange ratio (RER) and heat production can be calculated. An activity and food and water intake monitoring system can also be integrated into the set up in order to investigate circadian pattern and behaviour.Ontological description: MP:0005266 - abnormal metabolism.
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The glucose tolerance test measures the clearance of an intraperitoneally injected glucose load from the body. It is used to detect disturbances in glucose metabolism that can be linked to human conditions such as diabetes or metabolic syndrome. Animals are fasted for approximately 16 hours, fasted blood glucose levels are determined before a solution of glucose is administered by intra-peritoneal (IP) injection. Subsequently, the blood glucose level is measured at different time points during the following 2 hours.Ontological description: MP:0005559 - increased circulating glucose level, MP:0005560 - decreased circulating glucose level, MP:0005293 - impaired glucose tolerance, MP:0005292 - improved glucose tolerance, MP:0005291 abnormal glucose tolerance, MP:0000188 - abnormal circulating glucose level.
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The acoustic startle response is characterized by an exaggerated flinching response to an unexpected strong auditory stimulus (pre-pulse). This response can be attenuated when it is preceded by a weaker stimulus (pre-pulse) and is the principle underlying pre-pulse inhibition (PPI). PPI has been described in numerous species, including mice and humans and provides an operational measure of sensorimotor gating reflecting the ability of an animal to successfully integrate and inhibit sensory information. Several clinical studies have shown that a number of human disorders have impaired PPI including: schizophrenia, Huntington’s disease, fragile X syndrome, and autism. The acoustic startle and PPI paradigm is therefore largely used to assess sensorimotor gating and the effects of a number of treatment modalities such as putative anti-psychotics, and to explore genetic and neurobiological mechanisms underlying behaviors of relevance to psychosis (Geyer, 1999; Ouagazzal et al., 2001).Ontological description: MP:0002067 - abnormal sensory capabilities/reflexes/nociception.
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Clinical chemistry determines biochemical parameters in plasma including enzymatic activity, specific substrates and electrolytes. Ontological description: MP:0001545 – blood physiology abnormalities.
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The Open Field test is used to assess anxiety and exploratory behaviors. It is based on the natural tendency of an animal to explore and to protect itself using avoidance which translates to a normal animal spending more time in the periphery of the Open Field arena than in the center (the most anxiogenic area).DescriptionMP Term or Commentabnormal anxiety-related reponseMP:0001362abnormal locomoter activityMP:0001392abnormal response to novel environmentMP:0001449
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Hematological assessment of blood determines blood cell counts (white blood cells, red blood cells, hemoglobin, and platelets) and additional hematological parameters (hematocrit, mean cell volume, mean corpuscular hemoglobin, mean cell hemoglobin concentration) can be derived using these indices. These tests will indicate abnormalities in the production of blood and its components (blood cells and hemoglobin) as well as in the associated blood-forming organs. Ontological description: MP:0002429 - abnormal blood cell morphology/development.
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The characterisation of of main leukocyte subsets in peripheral blood cells from mice by flow cytometry.Reliable values of frequencies of leukocyte clusters are very much dependent on the appropriate preparation, acquisition and gating of leukocytes. This methodinvolves ammonium chloride erythrocyte lyses, thus preventing interference by larger amounts of erythrocytes. For all stainings, a monoclonal antibody to thecell surface glycoprotein CD45 is used to be able to create a CD45+ gate, allowing discrimination of leukocytes from debris, erythrocytes and thrombocytes. Furthermore, staining with propidium iodide (PI) gates out dead cells. Samples are acquired until a number of 30000 living CD45+ cells is reached for each sample (ref Adler and Busch 2007, submitted). Data are compensated analysed and using FlowJo software.This protocol is part of the EMPReSSslim pipeline and is carried out on a cohort of 10 males and 10 females (age matched) at a time.
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To perform a complete necropsy to detect and record abnormal external findings and macroscopic alterations in internal and external organs, record body and heart weights (see IMPC Heart Weight SOP), and collect a standardized list of tissues for fixation with or without further processing (see non-mandatory IMPC Tissue Embedding & Block Banking SOP).
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The insulin concentration in the blood is an important indicator of diabetes.Ontological description: abnormal circulating insulin level [MP:0001560]; increased circulating insulin level [MP:0002079]; decreased circulating insulin level [MP:0002727].
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