The Wellcome Trust for providing Open Access funding to Oxford University. Robert Deacon is a member of Oxford OXION group, funded by Wellcome Trust grant WT084655MA.

1. Apparatus
1.1 Kondziela's inverted screen test
The inverted screen is a 43 cm square of wire mesh consisting of 12 mm squares of 1 mm diameter wire (Figure 1). It is surrounded by a 4 cm deep wooden beading (which prevents the occasional mouse which attempts to from climbing on to the other side) (Figure 2).
1.2 Weights test
Seven weights constitute the apparatus. Each consists of a ball of tangled fine gauge stainless steel wire, a "scale collector" as used to prevent limestone scale formation in domestic kettles. The weight of this is small (7 g) but each scale collector is attached to a series of steel chain links, each weighing approximately 13 g. The number of links ranges from one to seven. Their weights are therefore: 20, 33, 46, 59, 72, 85 and 98 g (Figures 3 and 4).
2. Procedure
For all tests, bring the mice to the experimental room 5-20 min before testing to ensure they are properly awake. As a general rule, to allow recovery of muscular strength and a return to normal levels of arousal, rest the mice by a return to the home cage after each motor test.
<ol>
	<li>Kondziela's inverted screen test</li>
	<li>Procedure: Place the mouse in the center of the wire mesh screen, start a stopclock, rotate the screen to an inverted position over 2 sec, with the mouse's head declining first. Hold the screen steadily 40-50 cm above a padded surface. Note the time when the mouse falls off, or remove it when the criterion time of 60 sec is reached. Longer criterion times may be useful for some experiments.</li>
	<li>Scoring the inverted screen:</li>
</ol>
Falling between 1-10 sec = 1 
Falling between 11-25 sec = 2 
Falling between 26-60 sec = 3 
Falling after 60 sec = 4
Or, e.g. for 2 min: 
Falling between 1-10 sec = 1 
Falling between 11-25 sec = 2 
Falling between 26-60 sec = 3 
Falling between 61-90 sec = 4 
Falling after 90 sec = 5
3. Weights Test
Hold a mouse by the middle/base of the tail and lower it to allow it to grasp the first weight (20 g) which is lying on the laboratory bench. As it grasps the wire scale collector with its forepaws, start a stop clock and raise the mouse until the link is clear of the bench. A hold of three seconds is the criterion. If the mouse drops the weight in less than 3 sec, note the time it held the weight. Rest the mouse for about 10 sec and try it on the weight once again. If it fails three times, that terminates the trial, and the mouse is assigned the maximum time/weight achieved. If it holds it for 3 sec then try it on the next heaviest weight. If it lifts this for 3 sec, test it on the next heavier weight, after you have tested all its cagemates on the first weight. Again, it is given three chances to hold the weight for 3 sec. A final total score is calculated as the product of the number of links in the heaviest chain held for the full 3 sec, multiplied by the time (sec) it is held. If the heaviest weight is dropped before 3 sec an appropriate intermediate value is calculated. Thus a mouse holding a 5-link weight for 3 seconds, but unable to lift a 6-link weight, is assigned a score of (5 x 3) = 15. If it holds the 6-link weight for 1 second, it scores (5 x 3) + (1) =16.
Sometimes mice may not be adequately motivated to grip the apparatus. This generally occurs in older animals. This problem can be minimized by holding the mice by the tail for a longer time before bringing it within gripping distance of the wire ball.

<ol>
	<li>Bulfield, G., Siller, W. G., Wight, P. A., Moore, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=K.J.X+chromosome-linked+muscular+dystrophy+(mdx)+in+the+mouse.">K.J.X chromosome-linked muscular dystrophy (mdx) in the mouse.</a> Proc. Natl. Acad. Sci. U.S.A. 81, 1189-1192 (1984).</li><li>Contet, C., Rawlins, J. N. P., Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+129S2/SvHsd+and+C57BL/6JOlaHsd+mice+on+a+test+battery+assessing+sensorimotor,+affective+and+cognitive+behaviours:+implications+for+the+study+of+genetically+modified+mice.">A comparison of 129S2/SvHsd and C57BL/6JOlaHsd mice on a test battery assessing sensorimotor, affective and cognitive behaviours: implications for the study of genetically modified mice.</a> Behavioural Brain Research. 124, 33-46 (2001).</li><li>Deacon, R. M. J., Brook, R. C., Meyer, D., Haeckel, O., Ashcroft, F. M., Miki, T., Seino, S., Liss, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+mice+lacking+the+KATP+channel+subunit+Kir6.2.">Behavioral phenotyping of mice lacking the KATP channel subunit Kir6.2.</a> Physiol. Behav. 87, 723-733 (2006).</li><li>Deacon, R. M. J., Croucher, A., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+cytotoxic+lesion+effects+on+species-typical+behaviors+in+mice.">Hippocampal cytotoxic lesion effects on species-typical behaviors in mice.</a> Behav. Brain Res. 132, 203-213 (2002).</li><li>Rawlins, J. N. P., Morley, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+the+behavior+of+C57BL/6+and+C57BL/10+mice.">A comparison of the behavior of C57BL/6 and C57BL/10 mice.</a> Behav. Brain Res. 179, 239-247 (2007).</li><li>Guenther, K., Deacon, R. M. J., Perry, V. H., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+behavioural+changes+in+scrapie-affected+mice+and+the+influence+of+dapsone.">Early behavioural changes in scrapie-affected mice and the influence of dapsone.</a> Eur. J. Neurosci. 14, 401-409 (2001).</li><li>Kondziela, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eine+neue+method+zur+messung+der+muskularen+relaxation+bei+weissen+mausen.">Eine neue method zur messung der muskularen relaxation bei weissen mausen.</a> Arch. Int. Pharmacodyn. 152, 277-284 (1964).</li></ol>

No conflicts of interest declared.

Mouse models of the many diseases that affect the human motor systems are continually being developed, therefore good assays of strength in mice are essential. Examples include the many forms of muscular dystrophy, Parkinson's disease and Huntington's chorea. Strength assays also find use in models of nerve regeneration after spinal or peripheral nerve damage. Industrial and automobile accidents continue to cause considerable disability to humans, but much preclinical work is presently investigating possible avenues of treatment based on nerve growth factors.
Muscles never work individually. They are activated by the brain, local or higher spinal reflexes, via the neuromuscular junctions and work within the constraints of other muscles and the skeletal system. Therefore, the complexity of the system means in vivo assays of muscular force in the awake animal are essential in the evaluation of its present condition and the effects of any putative treatments. Mice instinctively hold on to materials with a strong grip, thus Mus musculus is an ideal model animal in which to assess disorders of muscular strength and possible treatments.

To a lay person, the idea of measuring strength in mice might appear absurd, even something of an oxymoron. However, it is not without reason that house/lab mice are named Mus musculus. They actually have a very high strength/weight ratio, greater than that of rats. For instance, adult mice can easily support their own body weight, even using only the fore or hind paws; very few adult lab rats can do this.
Mice are playing an ever greater role in biomedical research. They can be used to model many human motor disorders, both somatic and central nervous system in origin. The former include the more than thirty forms of inherited muscular dystrophy1 and myasthenia gravis, while examples of the latter are multiple sclerosis, spinal muscular atrophy, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. For all of these models, a full assessment of their motor deficits must include specific tests of strength.

measuring the strength of mice

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score maximum on this task; it is a quick but insensitive gross screen, and the weights test described in this article will provide a finer measure of muscular strength.
There are also several strain gauge-based pieces of apparatus available commercially that will provide more graded data than the inverted screen test, but their cost may put them beyond the reach of many laboratories which do not specialize in strength testing. Hence in 2000 a cheap and simple apparatus was devised by the author. It consists of a series of chain links of increasing length, attached to a "fur collector" a ball of fine wire mesh sold for preventing limescale build up in hard water areas. An accidental observation revealed that mice could grip these very tightly, so they proved ideal as a grip point for a weight-lifting apparatus. A common fault with commercial strength meters is that the bar or other grip feature is not thin enough for mice to exert a maximum grip. As a general rule, the thinner the wire or bar, the better a mouse can grip with its small claws.
This is a pure test of strength, although as for any test motivational factors could potentially play a role. The use of scale collectors, however, seems to minimize motivational problems as the motivation appears to be very high for most normal young adult mice.

Expected Results
Contet et al.2 demonstrated that 129S2/sv mice performed similarly on the weights test to C57BL/6 mice. Female C57BL/6 mice scored approximately 15, 129 mice 17, which means they could lift approximately 5 chain links, around 70 g. This is a high value; normally 2-3 chain links (score 6-9, 33-46 g) are lifted by C57BL/6 mice5. These differences are presumably related to factors such as how much handling the mice have received and the details of the experimenter's technique.
Hippocampal lesions did not affect strength as measured either by the weights test or the inverted screen4. Prion (scrapie) diseased mice were impaired on the inverted screen by 18 weeks after infection6. Although somewhat impaired on the horizontal bar and rotarod relative to C57BL/6 mice, C57BL/10 mice were not impaired on weightlifting5. Knockout mice for the KATP channel subunit Kir6.2 were not impaired on the inverted screen or weightlifting tests, although the females scored higher than males on the latter3. This was unusual; normally the heavier males display greater strength than the females.
<img alt="Figure 1" src="/files/ftp_upload/2610/2610fig1.jpg" /> 
 Figure 1. The inverted screen.
<img alt="Figure 2" src="/files/ftp_upload/2610/2610fig2.jpg" /> 
 Figure 2. Close view of a mouse on the inverted screen.
<img alt="Figure 3" src="/files/ftp_upload/2610/2610fig3.jpg" /> 
 Figure 3. Weights, ranging from 20 g to 98 g.
<img alt="Figure 4" src="/files/ftp_upload/2610/2610fig4.jpg" /> 
 Figure 4. A mouse weightlifting.

Watch this Scientific Journal Video about Measuring the Strength of Mice at JoVE.com

Measuring the Strength of Mice

Deficits in muscular strength occur in many clinical conditions such as motor neuron disease. The inverted screen and weight lifting tests described here measure strength in mice almost exclusively, with minimal influence of factors such as coordination.

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score ...

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53.2K ビュー.  University of Oxford. 筋力における赤字は、このような運動ニューロン疾患などの多くの臨床条件で発生します。反転画面と重量挙げのテストはここでこのような調整などの要因の影響が最小限で、ほぼ独占的にマウスでの強さを測定する説明。

ビデオ: Measuring the Strength of Mice

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In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

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Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, and persistent nerve and muscle impairments after hospital discharge.1-6 Using an explicit protocol with a structured approach to training and quality assurance of research staff, manual muscle testing (MMT) is a highly reliable method for assessing strength, using a standardized clinical examination, for patients following ARDS, and can be completed with mechanically ventilated patients who can tolerate sitting upright in bed and are able to follow two-step commands. 7, 8 
This video demonstrates a protocol for MMT, which has been taught to &ge;43 research staff who have performed &gt;800 assessments on &gt;280 ARDS survivors. Modifications for the bedridden patient are included. Each muscle is tested with specific techniques for positioning, stabilization, resistance, and palpation for each score of the 6-point ordinal Medical Research Council scale.7,9-11 Three upper and three lower extremity muscles are graded in this protocol: shoulder abduction, elbow flexion, wrist extension, hip flexion, knee extension, and ankle dorsiflexion. These muscles were chosen based on the standard approach for evaluating patients for ICU-acquired weakness used in prior publications. 1,2.

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We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

We present a protocol for measuring in vivo aortic stiffness in mice using high-resolution ultrasound imaging. Aortic diameter is measured by ultrasound and aortic blood pressure is measured invasively with a solid-state pressure catheter. Blood pressure is raised then lowered incrementally by intravenous infusion of vasoactive drugs phenylephrine and sodium nitroprusside. Aortic diameter is measured for each pressure step to characterize the pressure-diameter relationship of the ascending aorta. Stiffness indices derived from the pressure-diameter relationship can be calculated from the data collected. Calculation of arterial compliance is described in this protocol.
This technique can be used to investigate mechanisms underlying increased aortic stiffness associated with cardiovascular disease and aging. The technique produces a physiologically relevant measure of stiffness compared to ex vivo approaches because physiological influences on aortic stiffness are incorporated in the measurement. The primary limitation of this technique is the measurement error introduced from the movement of the aorta during the cardiac cycle. This motion can be compensated by adjusting the location of the probe with the aortic movement as well as making multiple measurements of the aortic pressure-diameter relationship and expanding the experimental group size.

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Hip weakness is a common symptom affecting walking ability in people with multiple sclerosis (MS). It is known that resistance strength training (RST) can improve strength in individuals with MS, however; it remains unclear the duration of RST that is needed to make strength gains and how to adapt hip strengthening exercises for individuals of varying strength using only resistance bands. This paper describes the methodology to set up and implement an adapted resistance strength training program, using resistance bands, for individuals with MS. Directions for pre- and post-strength tests to evaluate efficacy of the strength-training program are included. Safety features and detailed instructions outline the weekly program content and progression. Current evidence is presented showing that significant strength gains can be made within 8 weeks of starting a RST program. Evidence is also presented showing that resistance strength training can be successfully adapted for individuals with MS of varying strength with little equipment.

Hip weakness is a common symptom affecting walking ability in people with multiple sclerosis. Isolated muscle strengthening is a useful method to target specific weaknesses. This protocol describes a progressive resistance-training program using exercise bands to increase hip muscle strength.

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This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in elderly people performing short-term resistance training (RET). Supervised progressive resistance training for 1 h three times a week over 8 weeks was performed by RET participants (71&#177;1 years, range 65-80). Compared to a control group without training, the RET showed improvements on the measures used to indicate strength, power, glucose tolerance, and several parameters of muscle aerobic capacity. Strength training was performed in a gym with only robust fitness equipment. An isokinetic dynamometer for knee extensor strength permitted the measurement of concentric, eccentric, and static strength, which increased for the RET group (8-12% post- versus pre-test). The power (rate of force development, RFD) at the initial 0-30 ms also showed an increase for the RET group (52%). A glucose tolerance test with frequent blood glucose measurements showed improvements only for the RET group in terms of blood glucose values after 2 h (14%) and the area under the curve (21%). The blood lipid profile also improved (8%). From muscle biopsy samples prepared using histochemistry, the amount of fiber type IIa increased, and a trend towards a decrease in IIx in the RET group reflected a change to a more oxidative profile in terms of fiber composition. Western blot (to determine the protein content related to the signaling for muscle protein synthesis) showed a rise of 69% in both Akt and mTOR in the RET group; this also showed an increase in mitochondrial proteins for OXPHOS complex II and citrate synthase (both ~30%) and for complex IV (90%), in only the RET group. We demonstrate that this type of progressive resistance training offers various improvements (e.g., strength, power, aerobic capacity, glucose tolerance, and plasma lipid profile).

The effect of short-term resistance training on elderly people was investigated through the simultaneous use of several methods. Compared to a control group, many improvements were seen, including on muscle aerobic capacity, glucose tolerance, strength, power, and muscle quality (i.e., protein involved in cell signaling and muscle fiber type composition).

This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in ...

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

Toxicological evaluation is crucial for understanding the effects of chemicals on living organisms in basic and applied biological science fields. A non-mammalian soil round worm, Caenorhabditis elegans, is a valuable model organism for toxicology studies due to its convenience and lack of animal ethics issues compared with mammalian animal systems. In this protocol, a detailed procedure of toxicological evaluation of chemicals in C. elegans is described. A clinical anticancer drug, etoposide, which targets human topoisomerase II and inhibits DNA replication of human cancer cells, was selected as a model testing chemical. Age-synchronized C. elegans eggs were exposed to either dimethyl sulfoxide (DMSO) or etoposide, and then the growth of C. elegans was monitored every day for 4 days by the stereo microscope observation. The total number of eggs laid from C. elegans treated with DMSO or etoposide was also counted by using the stereo microscope. Etoposide treatment significantly affected the growth and reproduction of C. elegans. By comparison of the total number of eggs laid from worms with different treatment periods of chemicals, it can be decided that the reproductive toxicity of chemicals on C. elegans reproduction is reversible or irreversible. These protocols may be helpful for both the development of various drugs and risk assessment of environmental toxicants.

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

A basic protocol to evaluate the toxicity of chemicals in a model animal, Caenorhabditis elegans, is described. The method is convenient and useful for the development of pharmaceuticals as well as for the risk assessment of various environmental pollutants.

Toxicological evaluation is crucial for understanding the effects of chemicals on living organisms in basic and applied biological science fields. A ...

Blood Collection Through Subclavian Vein Puncture in Mice

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in research work. Tail blood collection is a popular method when a small amount of blood sample is needed. Orbital artery could be considered if a large amount of blood is needed but this blood collection method has ethical issues. Formerly, we demonstrated the feasibility and safety of blood sample collection through subclavian vein puncture in rats, and here we investigate whether this method could be used in mice. We report that this method is safe and practical for blood collection in mice. Blood collection through the subclavian vein puncture in mice can be a convenient method in daily research works.

Here, we present a protocol to take blood samples from the subclavian vein of mice.

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in ...

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a nonmammalian animal model that is widely used as an assay system due to its short lifespan, transparency, cost-effectiveness, and lack of animal ethics issues. In this study, a method was developed to investigate the effects of different bacteria and 3,3'-diindolylmethane (DIM) on the intestinal permeability of C. elegans with a high-throughput image analysis system. The worms were infected with different gut bacteria or cotreated with DIM for 48 h and fed with fluorescein isothiocyanate (FITC)-dextran overnight. Then, the intestinal permeability was examined by comparing the fluorescence images and the fluorescence intensity inside the worm bodies. This method may also have the potential to identify probiotic and pathogenic intestinal bacteria that affect intestinal permeability in the animal model and is effective for examining the effects of harmful or health-promoting chemicals on intestinal permeability and intestinal health. However, this protocol also has some considerable limitations at the genetic level, especially for determining which genes are altered to control illness, because this method is mostly used for phenotypic determination. In addition, this method is limited to determining exactly which pathogenic substrates cause inflammation or increase the permeability of the worms' intestines during infection. Therefore, further in-depth studies, including investigation of the molecular genetic mechanism using mutant bacteria and nematodes as well as chemical component analysis of bacteria, are required to fully evaluate the function of bacteria and chemicals in determining intestinal permeability.

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

This protocol describes how to measure intestinal permeability of Caenorhabditis elegans. This method is helpful for basic biological research on intestinal health related to the interaction between intestinal bacteria and their host and for screening to identify probiotic and chemical agents to cure leaky gut syndrome and inflammatory bowel diseases.

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...