Phenotyping Protocols for Pipeline: MGP Select Pipeline MGP_001

Unrestricted

Welfare Observations IMPC_WEL_001 (v.1.0)

The Welfare Observations procedure is used for the recording of welfare issues as and when they occur.

Body Weight IMPC_BWT_001 (v.1.3)

The body weight test measures the weight of the mouse in a time series, allowing monitoring of its evolution; also, it is required in many other procedures.

Fertility of Homozygous Knock-out Mice IMPC_FER_001 (v.1.1)

To assess the fertility of homozygous knockout mice.

Week 9

Grip Strength IMPC_GRS_001 (v.1.1)

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.

Week 12

Indirect Calorimetry IMPC_CAL_003 (v.3.5)

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.

Previous Versions:(v.2.7)(v.1.8)

Week 13

Intraperitoneal glucose tolerance test (IPGTT) IMPC_IPG_001 (v.1.1)

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.

Previous Versions:(v.1.0)

Week 14

Auditory Brain Stem Response IMPC_ABR_002 (v.2.5)

Auditory brainstem response test determines hearing sensitivity and other physiological parameters using evoked potential recordings in anesthetized mice.Ontological description: MP:0004738 - abnormal brainstem auditory evoked potential.

Previous Versions:(v.1.7)

X-ray MGP_XRY_001 (v.1.1)

Construct and analyse digital X-ray images in immobilised mice using a Faxitron X-Ray system or NTB digital X-ray scanner.

Body Composition (DEXA lean/fat) IMPC_DXA_001 (v.1.8)

Measure bone mineral content and density as well as body composition in mice using the DEXA (Dual Energy X-ray Absorptiometry) analyser.

Week 15

Eye Morphology IMPC_EYE_002 (v.2.8)

To detect abnormalities in eye morphology.

Previous Versions:(v.1.9)

Week 16

Tissue Embedding and Block Banking IMPC_BLK_001 (v.1.9)

Collect and fix a standard list of tissues from the complete necropsy (see IMPC Gross Pathology & Tissue Collection SOP)Trim the fixed tissues into cassettes for processingEmbed the tissues in paraffin in a standard orientation

Hematology IMPC_HEM_002 (v.2.9)

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.

Previous Versions:(v.1.11)

Mesenteric Lymph Node Immunophenotyping MGP_MLN_001 (v.1.0)

3i Mesenteric Lymph Node Immunophenotyping

Immunophenotyping IMPC_IMM_001 (v.1.8)

Immunophenotyping of immune system cells

Clinical Chemistry IMPC_CBC_003 (v.3.9)

Clinical chemistry determines biochemical parameters in plasma including enzymatic activity, specific substrates and electrolytes. Ontological description: MP:0001545 – blood physiology abnormalities.

Previous Versions:(v.2.1)(v.1.12)

Gross Pathology and Tissue Collection IMPC_PAT_002 (v.2.9)

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).

Previous Versions:(v.1.9)

Heart Weight IMPC_HWT_001 (v.1.9)

To evaluate cardiac size using heart weight and body weight.

Insulin Blood Level IMPC_INS_003 (v.3.2)

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].

Previous Versions:(v.2.3)(v.1.3)