Phenotyping Protocols for Pipeline: JAX Pipeline JAX_001


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.

Viability Primary Screen IMPC_VIA_002 (v.2.0)

Assess the viability of mutant mice of each sex and zygosity.

Previous Versions:(v.1.0)

Week 8

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.

Open Field IMPC_OFD_001 (v.1.1)

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).

Adult LacZ IMPC_ALZ_001 (v.1.9)

To stain adult mutant  and control tissues for bacterial beta-galactosidase (LacZ) activity in order to assess the adult organ, tissue, substructure and cell type of gene expression of IKMC alleles utilizing a LacZ reporter.  Description: Adult mouse tissues are scored for presence of LacZ staining which is distinct from either nonspecific staining observed in wildtype control mice, or is too faint to score as present.  Intensity of staining is not required but can be reported.   Anatomical terms to localize staining to the organ, structure, substructure, or cell type will use standard mouse anatomy ontology terms.

Open Field - centre start JAX_OFD_001 (v.1.1)

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).

Combined SHIRPA and Dysmorphology IMPC_CSD_003 (v.3.9)

SHIRPA and dysmorphology were originally always separate assessments.  However they have recently been combined as assessments, so that they take place at the same time.The purpose of the assessments is to examine mice for obvious physical characteristics, behaviors and morphological abnormalities.Descriptions include abnormal locomotion/appearance/behavior/reflex reactions.

Previous Versions:(v.2.7)(v.1.11)

Week 9

Hole-board Exploration JAX_HBD_001 (v.1.1)

To monitor the activity of mice in hole-board apparatus over a defined period of time in order to assess the exploratory and attentional phenotype.

Light-Dark Test JAX_LDT_001 (v.1.2)

To monitor the anxiety-related behavior of mice in a two-chambered light-dark arena over a defined period of time in order to assess the anxiety phenotype.

Previous Versions:(v.1.1)

Week 10

Acoustic Startle and Pre-pulse Inhibition (PPI) IMPC_ACS_003 (v.3.4)

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.

Previous Versions:(v.2.4)(v.1.5)

Y-maze IMPC_YMZ_001 (v.1.1)

Y-maze assesses learning and memory; it can be used instead/in parallel of Fear Conditioning

Week 11

Rotarod JAX_ROT_001 (v.1.1)

To assess motor co-ordination, motor learning, and balance phenotype in rodents.

Electrocardiogram (ECG) IMPC_ECG_003 (v.3.0)

To provide a high throughput method to obtain Electrocardiograms in a conscious mouse or an anesthetized mouse.

Previous Versions:(v.2.6)(v.1.9)

Tail Suspension JAX_TLS_001 (v.1.1)

To monitor the immobility response to tail suspension to test behavioral despair in mice, an induced depression model.

Week 12

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.

Week 13

Urinalysis JAX_URI_001 (v.1.0)

Urinalysis is performed to monitor renal function and will be performed as part of the adult phenotyping pipeline for KOMP2 mice. Samples will be analyzed using the Beckman AU680 autoanalyzer. Levels of urinary albumin, creatinine, magnesium and glucose are measured. The albumin to creatinine ratio (ACR) will be derived. A standard curve is run for microalbumin with each set of samples and used to derive an adjusted albumin value for use in the ACR.

Plasma Chemistry JAX_PLC_001 (v.1.1)

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

Week 14

X-ray IMPC_XRY_001 (v.1.9)

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_003 (v.3.0)

To detect abnormalities in eye morphology.

Previous Versions:(v.2.8)(v.1.9)

Sleep Wake JAX_SLW_001 (v.1.1)

To quantitate sleep behavior in mice using automatic detection of movement with a piezoelectric floor, and threshold/statistical analysis software. Sleep-Wake Monitoring Quad Cage and Sensor System (Signal Solutions, Lexington KY) is used to monitor mouse activity throughout a given time period. It records the amplitude of the electrical signal generated by force on the piezoelectric sensors, then uses software to set signal thresholds and statistically analyse and classify the activity as sleep or wake. Data are recorded as a confidence classification of sleep/wake over time, and include amplitude.

Week 16

Electroretinography JAX_ERG_001 (v.1.2)

Full-field electroretinogram (ERG) is a mass electrical response of the retina to a light stimulus. The ERG contains two components, reflecting the response of the photoreceptors (a-wave) and ON bipolar cells (b-wave; Kofujj et al., 2000 J Neurosci 20: 5733-5740, 2000)

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)

Immunophenotyping IMPC_IMM_002 (v.2.0)

Immunophenotyping of immune system cells

Previous Versions:(v.1.8)

Week 17

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

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)

Edema JAX_EDM_001 (v.1.0)

Complete Freund's Adjuvant (CFA) induced paw edema is used to measure local inflammatory response.

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)

Von Frey Test JAX_VFR_001 (v.1.0)

To investigate mechanical sensitivity in wild type and genetically altered mice.

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)

Organ Weight IMPC_OWT_001 (v.1.4)

Organ weights from different tissues collected for IMPC

Heart Weight IMPC_HWT_001 (v.1.9)

To evaluate cardiac size using heart weight and body weight.

Hargreaves Test JAX_HRG_001 (v.1.0)

To assess thermal hyperalgesia in the mouse paw.

Electroconvulsive Threshold Testing JAX_ECT_001 (v.1.3)

Electroconvulsive threshold testing is used to understand mechanisms that underlie susceptibility to epilepsy. An electrical current is used to induce minimal clonic (forebrain), maximal tonic (hindbrain/brainstem) and psychomotor (limbic) seizures.

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)