Name: A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS (Video) | JoVE Journal
Uploaded: 2015-10-06T15:00:00.000+00:00
Duration: 6 min 49 s
Description: Learn a quick phenotypic neurological scoring system to evaluate neuromuscular function in SOD1-G93A mice. This standardized method assesses hind limb function through daily observations of gait and reflexes.

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5 minutes

We would like to acknowledge Beth Levine for critically reviewing the manuscript, Valerie Tassinari for developing the genotyping protocol, and Matt Ferola and Carlos Maya for their exceptional animal care.

All experiments are conducted in accordance with the protocols described by the National Institutes of Health Guide for the Care and Use of Animals and were approved by ALS TDI&#8217;s institutional animal care and use committee (IACUC).
1. Animals, Housing, and Study Design
Note: The SOD1-G93A mouse colony was derived from the high copy B6SJLTgN(SOD1G93A)1Gur strain originally produced by Gurney et al. 7. The colony is currently being maintained by crossing transgenic C57BL/6-SJL males with wild-type C57BL/6-SJL F1 females. The F1 animals are generated by crossing SJL males with B6 females. Mice are shipped to ALS TDI weekly at 35&#8211;45 days of age.
<ol>
	<li>Allow mice at least one week to acclimate to the animal facility (12-hr light/dark cycle at a temperature of 18-23&#160;&#176;C&#160;and 40-60% humidity) before assigning to a study.</li>
	<li>House male SOD1-G93A mice one mouse per cage, since group housed males tend to fight. House female SOD1-G93A mice up to two&#160;per cage. Provide environmental enrichment, in the form of plastic huts. Provide food and water ad libitum, unless the mice are part of a study requiring drug administration in drinking water/food. In these cases, ration and record food and water.</li>
	<li>Begin a drug-efficacy study around age day 50 and include 64 litter-matched gender-balanced mice (32 per cohort). Finally, ensure that the researchers conducting the studies are blinded to the experimental conditions. 
	Note: It is recommended that a typical drug-efficacy study starts at age day 50 however based on the type of therapeutic intervention it might be preferable to start the study earlier or later. 
	Note: A study with 32 mice per cohort is powered to detect gender-specific drug-effects. If no gender-specific comparisons are required, 24 mice per cohort should suffice to detect overall drug-effects.</li>
</ol>
2. Body Weight
<ol>
	<li>Record body weights for all mice in studies daily with a precision of one tenth of a gram. Note: Body weight is a sensitive indicator of disease progression and of any malaise that might result from chronic drug treatment.</li>
	<li>Mouse body weights vary considerably during the course of a day, being higher in the mornings, thus record them around the same time every day.</li>
</ol>
3. Neurological Scoring System (NeuroScore)
<ol>
	<li>Assess neurological scores (NS) for each mouse daily by observing the mouse under the following three conditions performed sequentially: a) the mouse is suspended by its tail, b) the mouse is allowed to walk, and after the onset of paresis c) the mouse is placed on its side.
	<ol>
		<li>For the tail suspension test, hold the mouse approximately 1.5&#8221; from the base of the tail over the wire top of their home cage for 1-2 sec, away from the food basin, while observing the hindlimbs. Repeat the suspension test 3 times and record the most consistent outcome.</li>
		<li>For the walking test, place the mouse on a clean surface that provides some traction and a walking distance of 25&#160;cm (e.g., a paper towel taped down to prevent slipping). Allow the mouse to walk a total of 75&#160;cm (3X length of the paper towel) while observing its gait.</li>
		<li>For the &#8220;righting reflex&#8221; test, place the mouse on its left or right side and using a stop-watch measure the time it takes to right itself up unassisted from both sides. Note: Only one trial of this test is required.</li>
	</ol>
	</li>
	<li>Determine the NS of each hindlimb (left or right) independently on a scale from 0 to 4. Note: For instance a score of 0, 1 indicates that the left hindlimb NS is 0, while the right hindlimb NS is 1.</li>
	<li>Assign neurological scores (0-4) corresponding to the following set of observations (summary provided in Table 1):
	<ol>
		<li>Assign NS 0 (Pre-symptomatic) if the following is observed: When the mouse is suspended by the tail, the hindlimb presents a normal splay i.e., it is fully extended away from the lateral midline and it stays in this position for 2 sec&#160;or longer. When the mouse is allowed to walk, normal gait is observed.</li>
		<li>Assign NS 1 (First symptoms) if the following is observed: When the mouse is suspended by the tail, the hindlimb presents an abnormal splay, i.e., it is collapsed or partially collapsed towards lateral midline OR it trembles during tail suspension OR it is retracted/clasped. When the mouse is allowed to walk, normal OR slightly slow gait is observed.</li>
		<li>Assign NS 2 (Onset of paresis) if the following is observed: When the mouse is suspended by the tail, the hindlimb is partially OR completely collapsed, not extending much. (There might still be joint movement). When the mouse is allowed to walk, the hindlimb is used for forward motion however the toes curl downwards at least twice during a 90 cm walk OR any part of the foot is dragging along cage bottom/table. When the mouse is placed on its left AND right side, it is able to right itself within 10 sec&#160;from BOTH sides.</li>
		<li>Assign NS 3 (Paralysis) if the following is observed: When the mouse is suspended by the tail, there is rigid paralysis in the hindlimb OR minimal joint movement. When the mouse is allowed to walk there is forward motion however the hindlimb is NOT being used for forward motion. When the mouse is placed on its left AND right side, it is able to right itself within 10 sec&#160;from BOTH sides.&#160;Note: Rarely, after the onset of paralysis, urine moisture might appear on the hindlimbs.&#160; Left untreated urine moisture might result to urine &#8220;burn&#8221; and skin lesions.&#160; Treat urine moisture by clipping the hair, apply warm water soaks to remove the urine, and gently blot dry. If skin lesions are present apply antibiotic ointment.</li>
		<li>Assign NS 4 (Humane end-point) if the following is observed: When the mouse is suspended by the tail, there is rigid paralysis in the hindlimbs. When the mouse is allowed to walk, there is no forward motion. When the mouse is placed on its left AND right side it is NOT able to right itself within 10 sec&#160;from EITHER side. i.e., absence of righting reflex.</li>
	</ol>
	</li>
</ol>
4. Data Entry and Analysis
<ol>
	<li>During the course of a drug efficacy study, enter data collected for each mouse into a Laboratory Information Management System (LIMS) or into a commercially available spreadsheet software. 
	Note: The ALS TDI-developed LIMS software is a central repository for all data collected from internal studies, making these data conveniently accessible over time for ALS TDI investigators. LIMS tracks any individual mouse, with its unique ID, back to its lineage, body weight or neurological score on all study days, and includes any other observations noted during its lifetime. LIMS ultimately generates a spreadsheet which is used for subsequent analyses.</li>
	<li>Upon the completion of a drug efficacy study, i.e., when all mice reach the humane end-point, use spreadsheet software (see Table of Materials) to compute the combined NS for each mouse, for each day by averaging the left and right hindlimb NS. Next, compute the median age at each NS for each mouse and the number of days each mouse spends at a particular NS.</li>
	<li>Analyze overall changes in NS over time by importing the data from the spreadsheet software to the statistical software (see Table of Materials). Fit these data using a model of ordinal logistic regression which estimates the expected NS, with median age at NS and drug-treatment as effects in the regression. Using the log-rank chi square test estimate the effect likelihood probability and determine whether the drug-treatment effect on NS progression is statistically significant (see Supplemental Code File for step-by-step instructions). 
	Note: A p-value &#60;0.05 is considered statistically significant. 
	Note: This method of statistical analysis respects the ordinal nature of the NS data and the variable nature of the NS trajectories for individual mice. It is preferred because individual NS trajectories can be markedly different resulting in NS time-associated group averages that are not sufficiently reflective of NS responses. In addition, in some cases NS can fluctuate, particularly when in the zero-to-one range.</li>
	<li>Finally, use graphing software (see Table of Materials) to generate &#8220;Ordinal Expected NS vs Median Age at NS&#8221; graphs for drug-treated and control animals (Figure 2; see Supplemental Code File for step-by-step instructions). Use these graphs to conveniently estimate shifts between drug-treated and control groups by interpolating from the y-axis the median age of disease onset (i.e., NS 2) on the x-axis. 
	Note: The difference in median age between the two groups represents a drug-induced shift and is treated as a practical measure of the magnitude of effect. 
	Note: Rightward shifts in the expected NS vs. median age curve indicate slowed symptomatic disease progression. Conversely, leftward shifts in the expected NS vs. median age curve indicate accelerated symptomatic disease progression.</li>
</ol>

<ol><li>Gurney, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Motor+neuron+degeneration+in+mice+that+express+a+human+Cu%2CZn+superoxide+dismutase+mutation%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation.</a> Science. 264 (5166), 1772-1775 (1994).</li>
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<li>Hegedus, J., Putman, C. T., Gordon, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Time+course+of+preferential+motor+unit+loss+in+the+SOD1+G93A+mouse+model+of+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22Neurobiol.+Dis%22%5BJournal%5D)">Time course of preferential motor unit loss in the SOD1 G93A mouse model of amyotrophic lateral sclerosis.</a> Neurobiol. Dis. 28 (2), 154-164 (2007).</li>
<li>Gordon, T., Ly, V., Hegedus, J., Tyreman, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Early+detection+of+denervated+muscle+fibers+in+hindlimb+muscles+after+sciatic+nerve+transection+in+wild+type+mice+and+in+the+G93A+mouse+model+of+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22Neurol.+Res%22%5BJournal%5D)">Early detection of denervated muscle fibers in hindlimb muscles after sciatic nerve transection in wild type mice and in the G93A mouse model of amyotrophic lateral sclerosis.</a> Neurol. Res. 31 (1), 28-42 (2009).</li>
<li>Gurney, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Motor+neuron+degeneration+in+mice+that+express+a+human+Cu%2CZn+superoxide+dismutase+mutation%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation.</a> Science. 264 (5166), 1772-1775 (1994).</li>
<li>Kim, K., Moore, D. H., Makriyannis, A., Abood, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((AM1241%2C+a+cannabinoid+CB2+receptor+selective+compound%2C+delays+disease+progression+in+a+mouse+model+of+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22Eur.+J.+Pharmacol%22%5BJournal%5D)">AM1241, a cannabinoid CB2 receptor selective compound, delays disease progression in a mouse model of amyotrophic lateral sclerosis.</a> Eur. J. Pharmacol. 542 (1-3), 1-3 (2006).</li>
<li>Weydt, P., Hong, S. Y., Kliot, M., Moller, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Assessing+disease+onset+and+progression+in+the+SOD1+mouse+model+of+ALS%5BTitle%5D)+AND+%22Neuroreport%22%5BJournal%5D)">Assessing disease onset and progression in the SOD1 mouse model of ALS.</a> Neuroreport. 14 (7), 1051-1054 (2003).</li>
<li>Azari, M. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Behavioural+and+anatomical+effects+of+systemically+administered+leukemia+inhibitory+factor+in+the+SOD1%28G93A+G1H%29+mouse+model+of+familial+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22Brain+Res%22%5BJournal%5D)">Behavioural and anatomical effects of systemically administered leukemia inhibitory factor in the SOD1(G93A G1H) mouse model of familial amyotrophic lateral sclerosis.</a> Brain Res. 982 (1), 92-97 (2003).</li>
<li>Guan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Thiazolidinediones+expand+body+fluid+volume+through+PPARgamma+stimulation+of+ENaC-mediated+renal+salt+absorption%5BTitle%5D)+AND+%22Nat.+Med%22%5BJournal%5D)">Thiazolidinediones expand body fluid volume through PPARgamma stimulation of ENaC-mediated renal salt absorption.</a> Nat. Med. 11 (8), 861-866 (2005).</li>
<li>Savcheniuk, O. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effect+of+probiotic+therapy+on+development+of+experimental+obesity+in+rats+caused+by+monosodium+glutamate%5BTitle%5D)+AND+%22Fiziol.+Zh%22%5BJournal%5D)">The effect of probiotic therapy on development of experimental obesity in rats caused by monosodium glutamate.</a> Fiziol. Zh. 60 (2), 63-69 (2014).</li>
<li>Smittkamp, S. E., Brown, J. W., Stanford, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Time-course+and+characterization+of+orolingual+motor+deficits+in+B6SJL-Tg%28SOD1-G93A%291Gur%2FJ+mice%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Time-course and characterization of orolingual motor deficits in B6SJL-Tg(SOD1-G93A)1Gur/J mice.</a> Neuroscience. 151 (2), 613-621 (2008).</li>
<li>C, M., Gurney, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+low+expressor+line+of+transgenic+mice+carrying+a+mutant+human+Cu%2CZn+superoxide+dismutase+%28SOD1%29+gene+develops+pathological+changes+that+most+closely+resemble+those+in+human+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22Acta+Neuropathol%22%5BJournal%5D)">A low expressor line of transgenic mice carrying a mutant human Cu,Zn superoxide dismutase (SOD1) gene develops pathological changes that most closely resemble those in human amyotrophic lateral sclerosis.</a> Acta Neuropathol. 93 (6), 537-550 (1997).</li>
<li>Wegorzewska, I., Baloh, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((TDP-43-based+animal+models+of+neurodegeneration%3A+new+insights+into+ALS+pathology+and+pathophysiology%5BTitle%5D)+AND+%22Neurodegener.+Dis%22%5BJournal%5D)">TDP-43-based animal models of neurodegeneration: new insights into ALS pathology and pathophysiology.</a> Neurodegener. Dis. 8 (4), 262-274 (2011).</li>
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</ol>

The authors have nothing to disclose.

In this report, we describe a quick and simple video protocol which, if properly applied, is capable of reliably assessing the disease progression in SOD1-G93A mice and identifying drug-induced changes. While different groups have developed phenotypic scoring systems for SOD1-G93A mice 8-10, they often do not provide sufficient details on the procedure and are inadequate for replication. As a result, scoring systems are rarely used outside the specific group that develops them. In this report, the combination ...

Ever since its development in the mid 1990&#8217;s the SOD1-G93A transgenic mouse model has been the most widely used animal model of amyotrophic lateral sclerosis (ALS) 1. This transgenic mouse model was genetically engineered to overexpress a mutant form of the human Cu/Zn superoxide dismutase 1(SOD1) gene harboring the ALS-associated glycine to alanine mutation at amino acid 93 (G93A). The G93A mutation is one of many mutations in the SOD1 gene that collectively make-up approximately one fifth of familial ALS cases 2.
The SOD1-G93A model has become a workhorse of ALS drug development research because in addition to its obvious genetic link to ALS, it recapitulates many of the pathological features of ALS in humans such as motor neuron loss, progressive muscle weakness and atrophy, with eventual paralysis and death 3. Though, as is often the case with transgenic animal models, there is an inherent biological variability associated with SOD1-G93A mice. Scott et al. have determined that cohorts of at least 24 litter-matched gender-balanced mice are needed in drug-efficacy study designs in order to overcome the noise created by the biological variability 4. The large number of animals required in these studies in combination with the aggressive disease progression and the need for daily monitoring prohibits the use of elaborate, time-consuming and potentially stressful techniques for measuring neuromuscular strength and function (e.g., electromyography, grip strength, rotorod, etc.). Instead, it highlights the need for an in vivo screening tool that reliably assesses the neuromuscular function of SOD1-G93A mice in a quick manner.
At ALS TDI a protocol was developed that allows for the reliable assessment of the neuromuscular function of SOD1-G93A mice in less than 30 sec&#160;per mouse on average. This protocol encompasses daily assessment of hindlimb function using a simple neurological scoring system (NeuroScore) which is described in this report in press and in video. NeuroScore is focused on hindlimb function because hindlimb deficits are the earliest reported neurological sign of disease in SOD1-G93A mice 5,6. Further, the protocol developed by ALS TDI provides an unbiased assessment of onset of paresis (slight or partial paralysis), progression and severity of paralysis and it is sensitive enough to identify drug-induced changes in disease progression.

<table><tbody><tr><td>B6SJLTgN(SOD1G93A)1Gur strain</td><td>The Jackson Laboratory, Bar Harbor, ME</td><td>002726</td><td>Strain of mice used in this study</td></tr><tr><td>Breeders</td><td>Biomedical Research Models, Inc., Worcester, MA</td><td></td><td>Colony maintainance</td></tr><tr><td>Scale</td><td>Navigator OHAUS, Parsippany, NJ</td><td>Model N14120</td><td>Used to weigh mice</td></tr><tr><td>Nutra-GelH</td><td>Bio-Serv, Flemington, NJ</td><td>#S4798</td><td>Wet food provided to sick mice</td></tr><tr><td>Irradiated Lab Animal Diet</td><td>Harlan, Indianapolis, IN</td><td>2918-111914M</td><td>Chow diet</td></tr><tr><td>Lab-grade Sani-chips</td><td>Harlan Teklad, Indianapolis, IN</td><td>7090</td><td>Bedding</td></tr><tr><td>JMPH&amp;nbsp;</td><td>SAS Institute, Inc., SAS Campus Drive, Cary, NC</td><td>&amp;nbsp;v10.0.2</td><td>Statistical software</td></tr><tr><td>Microsoft Excel</td><td>Microsoft, One Microsoft Way, Redmond, WA</td><td>Excel 2013 (v 15.0)</td><td>Spreadsheet software</td></tr><tr><td>GraphPad Prism 5</td><td>GraphPad software Inc., La Jolla, CA</td><td>v5.4</td><td>Graphing software</td></tr><tr><td>Mouse Huts</td><td>Bio-Serv, Flemington, NJ</td><td>K3272</td><td>For environmental enrichment</td></tr></tbody></table>

a quick phenotypic neurological scoring system for evaluating disease progression in the sod1-g93a mouse model of als

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening protocol, demonstrated in video herein, which reliably assesses the neuromuscular function of SOD1-G93A mice in a quick manner. This protocol encompasses a simple neurological scoring system (NeuroScore) designed to assess hindlimb function. NeuroScore is focused on hindlimb function because hindlimb deficits are the earliest reported neurological sign of disease in SOD1-G93A mice. The protocol developed by ALS TDI provides an unbiased assessment of onset of paresis (slight or partial paralysis), progression and severity of paralysis and it is sensitive enough to identify drug-induced changes in disease progression. In this report, the combination of a detailed manuscript with video minimizes scoring ambiguities and inter-experimenter variability thus allowing for the protocol to be adopted by other laboratories and enabling comparisons between studies taking place at different settings. We believe that this video protocol can serve as an excellent training tool for present and future ALS researchers.

The neurological score data for 90 male and 94 female non-treated SOD1-G93A mice studied at ALS TDI during 2014 were evaluated. The results show that male mice typically have a more aggressive disease progression than female mice, as evidenced by a greater proportion of their lifespan at NS 1 compared to females. NS 2 and NS 3 occur with virtually equal frequency across genders. NS 0 was excluded from the analysis because it is considered a “normal” phenotype. NS 4 was also excluded because by definition it can only occu...

This video protocol describes a sensitive, reliable, and quick method for evaluating the neuromuscular deficits in a transgenic mouse model of amyotrophic lateral sclerosis.

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

The overall goal of this procedure is to quickly and reliably evaluate the neuromuscular function of SOD one G 93 a mice by daily assessment of their hind limb function using a simple phenotypic neurological scoring system. This is accomplished by first observing the hind limbs of the mice while suspending them by the tail. The second step is to allow the mice to walk while observing their gait.Next, the mice are placed on their sides to test for the presence or absence of the writing reflex. The final step is to perform the data analysis by fitting neurological score data of mouse cohorts into an ordinal logistic regression model. Ultimately, our phenotypic neurological scoring system is used to assess disease progression in the SOD one G 93, a mouse model of a LS for the purpose of discerning the efficacy of therapeutic interventions.The main advantage of this method over all other existing techniques is that it is quick, simple, reliable, and minimally stressful for the mice. The implications of this method extend to a therapy of neotropic lateral sclerosis or a LS because it allows for the quick assessment of therapeutic interventions in the most common mouse model of the disease. Visual demonstration is critical as the neurological scoring method is difficult to learn because the neurological differences can be subtle.Begin by housing male SOD one G 93 A mice so that there is one mouse per cage house, female SOD one G 93 A mice up to two per cage. Provide environmental enrichment for the mice in the form of plastic huts. Provide the mice with food and water at Lido record.Body weights for all mice and studies at the same time. Daily body weight is a sensitive indicator of disease progression and of any malaise that might result from chronic drug treatment. During the study, assess the neurologic score of each mouse daily through a regimen of three tests performed sequentially.Watch carefully as delineating scores at phenotypic transition points is very difficult In the first test. The tail suspension test, hold the mouse approximately 1.5 inches from the base of the tail over the wire top of the home cage. Observe the function of the hind limbs for a period of one to two seconds and repeat the test three times.For the walking test. Place the mouse on a clean surface that provides some traction and has a walking distance of at least 25 centimeters. Allow the mouse to walk up to 75 centimeters while observing its ga.For the writing reflex test, place the mouse on its side and use a stopwatch to measure the time it takes to write itself unassisted from both sides. Only one trial of this test is required. Score each hind limb left or right independently on a scale from zero to four based on the collective outcome of the three phenotypic observation tests.A sinus score of zero indicating the mouse is presymptomatic. When during the suspension test, the hind limbs are fully extended away from the lateral midline and stay in this position for two seconds or longer. Also, during the walking test, a normal gait is observed.Assign a score of one indicating the appearance of first symptoms. If during the suspension test, the hind limb is either collapsed towards the lateral midline, partially collapsed towards the lateral midline trembling or abnormally splayed in some other fashion. In addition to the suspension test symptoms just shown, the gait of a mouse that should receive a score of one should still appear normal or slightly lower.Assign a score of two indicating the onset of Parsis. If during the suspension test, the hind limb is partially or completely collapsed, and in addition during the walking test, the hind limbs are used for forward motion. However, the toes either curl downwards at least twice during a 75 centimeter walk or any part of the foot drags to get a score of two.The mouse must also still be able to write itself after being placed on both of its sides within 10 seconds. A sinus score of three indicating paralysis. If during the suspension test the mouse shows rigid paralysis in the hind limb or has minimal joint movement, the mouse may still show forward motion during the walking test, however, not by using its hind limbs.In addition to a sinus score of three, the mouse must still be able to write itself after being placed on both of its sides within 10 seconds. Finally, a of score of four, defining the humane endpoint whenever the mouse shows rigid paralysis during the suspension. Test shows no forward motion during the walking test using the hind limbs, and is also unable to write itself within 10 seconds after being placed on either side.Male mice typically have a more aggressive disease progression than female mice, as evidenced by a greater proportion of their lifespan displaying symptoms that can be classified with a neurological score of one compared to females. As symptoms increase into the range of a neurological score of two or three, the number of days spent at each score is equal in frequency across genders. Shown here are ordinal logistics, statistical analysis graphs generated using the neuroco protocol that was just described.These data are from a drug efficacy pharmacology study in which a cohort of SOD one G 93 a mice receiving the test compound TDI 1 73 is compared to a cohort of mice receiving vehicle. No statistically significant differences were observed when genders were separately analyzed, but statistical significance was reached when genders were merged. Though showing similar trends, Kaplan Meyer analysis of survival was not sensitive enough to reveal significant differences even when genders were merged.Once mastered, the scoring system can be applied in less than a minute per mouse if it is properly performed. After watching this video, you should have a good understanding of how to assess the disease progression in the SOD one G 93, a mouse model of a LS by using the Neuroco system.

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Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of β-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

This paper introduces a method for repeated measurements of ventilation and respiratory muscle activity in a freely behaving amyotrophic lateral sclerosis (ALS) mouse model throughout disease progression with whole-body plethysmography and electromyography via an implanted telemetry device.

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Experimental autoimmune encephalomyelitis (EAE) serves as an animal model of multiple sclerosis. This article describes an approach for scoring spinal cord inflammation, demyelination, and axonal injury in EAE. Additionally, a method to quantify soluble neurofilament light levels in the mice serum is presented, facilitating the assessment of axonal injury in live mice.

Scoring Central Nervous System Inflammation, Demyelination, and Axon Injury in Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis is one of the most widely used murine models of multiple sclerosis. In the current protocol, C57BL/6J mice of both sexes are immunized with myelin oligodendrocyte glycoprotein peptide, resulting mainly in ascending paresis of the tail and limbs. Here we discuss the protocol of EAE induction and evaluation.

Modeling Multiple Sclerosis in the Two Sexes: MOG35-55-Induced Experimental Autoimmune Encephalomyelitis

This article presents a protocol for executing the highly translational and independently proven mouse Stroke Unit (mSU) concept in large-stroke mice. It also provides a standardized protocol for executing the focal experimental stroke scale in mice, which evaluates stroke focal deficits detected even long-term.

The Mouse Stroke Unit Protocol with Standardized Neurological Scoring for Translational Mouse Stroke Studies

This article details intrathecal delivery methods for CNS-targeted therapeutics in adult mice, focusing on protocol precision and enabling repeated injection. It includes findings from a validation study where antisense oligonucleotides reduced mutant SOD1 expression in SOD1-ALS model mice, supporting the utility of the approach in preclinical research on neurodegenerative diseases.

Lumbar Intrathecal Injection of SOD1-ASOs for Precise CNS Targeting and Predictive Efficacy in Human SOD1-G93A ALS Mice

Learn a quick phenotypic neurological scoring system to evaluate neuromuscular function in SOD1-G93A mice. This standardized method assesses hind limb function through daily observations of gait and reflexes.

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

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