We thank Anniwaer Yilifate for proofreading our manuscript. This study was supported by the National Science Foundation under Grant No. 81902281 and No. 82072544, the General Guidance Project of Guangzhou Health and Family Planning Commission under Grant No. 20191A011091 and No. 20211A011106, the Guangzhou Key Laboratory Fund under Grant No. 201905010004 and Guangdong Basic and Applied Basic Research Foundation under Grant No.2020A1515010578.

NOTE: The clinical study was approved by the Medical Ethics Association of the Fifth Affiliated Hospital of Guangzhou Medical University (NO. KY01-2019-02-27) and has been registered at the China Clinical Trial Registration Center (No. ChiCTR1800017487 and entitled, &#34;The multiple modal tasks on gait control and motor cognition after stroke&#34;).
1. Recruitment
<ol>
	<li>Recruit stroke patients with the following inclusion criteria: patients meeting the diagnostic criteria for cerebrovascular disease of the Neurological Branch of the Chinese Medical Association (2005); cerebral infarction confirmed by computed tomography or magnetic resonance imaging; damage to the unilateral cortex or with a subcortical lesion; ability to walk independently, Brunnstrom stage &#8805; 4 stages; Modified Ashworth Scale11&#160;&#8804; 2 points; meeting the requirements of three-dimensional (3D) gait analysis and the ability to tolerate the whole process; and the ability to give informed consent.</li>
	<li>Ensure the following exclusion criteria are met: congestive heart failure, deep vein thrombosis of the lower extremities, malignant progressive hypertension, respiratory failure or other diseases, and serious risk of falling.</li>
	<li>Obtain written informed consent from all patients before beginning the study.</li>
</ol>
2. Clinical evaluation
<ol>
	<li>Record the demographic characteristics of the patient including the name, gender, date of birth, level of education, chief complaint, current medical history, past history, medical treatment, and current medications.</li>
	<li>Cognitive function assessment
	<ol>
		<li>Ask the patient to complete the Mini-Mental State Examination (MMSE)12 record the patient&#39;s responses to a 30-question scale with a total score of 30 points for cognition evaluation, which involves the following seven aspects: time orientation, position orientation, instant memory, attention and computing power, delayed memory, language, and visual space. 
		&#8203;NOTE: The scores of MMSE are closely related to the level of education. The normal cognitive standard is illiteracy &#62; 17 points, primary school &#62; 20 points, and junior high school &#62; 24 points13.</li>
		<li>Ask the patient to complete the Montreal Cognitive Assessment (MoCA)14 record the patient&#39;s responses to an 11-question scale with a total score of 30 points for cognition evaluation, which involves the following eight aspects: attention and concentration, executive function, memory, language, visual structure skills, abstract thinking, calculation, and orientation. 
		&#8203;NOTE: The normal cognitive standard is &#8805; 26 points. If the subject has been educated for less than 12 years, they should add 1 point to the score15.</li>
	</ol>
	</li>
	<li>Walking ability assessment
	<ol>
		<li>Conduct the 10-m walk test (10 MWT)16. Ask the patient to perform three consecutive trials at a self-selected pace for safety, comfort, and higher speed, respectively. Record the time taken to walk to the middle 6 m in each trial (to exclude acceleration and deceleration effects).</li>
		<li>Conduct the timed up and go test (TUGT)17. Ask the patient to perform three consecutive TUG trials (stand up, walk 3 m, turn, walk back, and sit down) at a self-selected pace for safety and comfort18.</li>
	</ol>
	</li>
</ol>
3. 3D gait analysis
<ol>
	<li>Patient preparation
	<ol>
		<li>Inform the patient about the precautions and the purpose of the experiment.</li>
		<li>Ask the patient to wear tight underwear to fully expose the neck, shoulders, waist, and lower limbs.</li>
		<li>Record the values of various anthropometric indicators including height, weight, bilateral width of the ankle joints, bilateral knee diameter, pelvic width, bilateral pelvic depth, and bilateral leg length.</li>
		<li>Place 22 markers on key points of the patient based on the Davis protocol19: three markers on the trunk (7th cervical vertebrae, shoulders on both sides); three markers on the pelvis (both sides of the anterior superior iliac spine&#160;and ankle joint); six markers on the thigh (bilateral femoral greater trochanter, femoral condyle, and middle point of femoral greater trochanter and femoral condyle on the same side); six markers on the calf (bilateral humeral head, lateral ankle joint, and middle point of humeral head and lateral ankle joint on the same side); four markers on the foot (the fifth metatarsal head and the heel on both sides) (Figure 1).</li>
		<li>Click on the Start button of the 3D gait analysis system, and make a new profile for the patient.</li>
		<li>Enter basic patient information and previously measured parameters.</li>
	</ol>
	</li>
	<li>Standing data acquisition
	<ol>
		<li>Instruct the patient to maintain an upright position on the force plate for at least 3-5 s to gather the baseline data.</li>
		<li>Click on the Proc_Davis_Standing button to quickly check the position of the marker.</li>
	</ol>
	</li>
	<li>Walking task data acquisition
	<ol>
		<li>Determine the random order of three walking tasks by drawing lots.</li>
		<li>Ask the patient to walk on the walking pass for five trials at a self-selected comfortable speed, which is marked as Task 0 (consider the single walking task as the Baseline task).</li>
		<li>Ask the patient to walk while holding a bottle of water on the walking pass for five trials at a self-selected comfortable speed, which is marked as Task 1 (simple dual-motor task). 
		NOTE: Ask the patient to hold a 550 mL bottle of water in the unaffected hand while holding the arm position of the shoulder joint at 0&#176; and elbow flexion at 90&#176;.</li>
		<li>Ask the patient to walk across the line in the middle of the walking pass for five trials at a self-selected comfortable speed, which is marked as Task 2 (complex dual-motor task). 
		&#8203;NOTE: Place a soft ruler in the middle of the walking pass before Task 2 data acquisition.</li>
	</ol>
	</li>
</ol>
4. Data processing and analysis
<ol>
	<li>Select the middle three trials of each walking task to be processed to ensure the patient is stable.</li>
	<li>Identify each gait cycle with two consecutive heel stride points on the same side.</li>
	<li>Mark the toe-off point in each gait cycle20.</li>
	<li>Click on the Proc_DavisHeel+GI_AE button to compute the kinematic parameters of gait, as well as the computation of the Gait Performance Score (GPS) index.</li>
</ol>
5. Data extraction and statistical analysis of interest
<ol>
	<li>Select region of interest parameters from the processed data, which include special-temporary parameters (stance phase, swing phase, single stance, double stance, cadence), joint angle parameters (trunk obliquity (frontal plane), trunk tilt (sagittal plane), trunk rotation (transversal plane), pelvic obliquity (frontal plane), pelvic tilt (sagittal plane), pelvic rotation (transversal plane), hip flex-extension, hip ab-adduction, hip rotation, knee flex-extension, ankle dorsi-plantarflexion, and GPS index.</li>
	<li>Calculate DTC values based on the following formula[10]: 
	([single-task gait velocity - dual-task gait velocity]/ single-task gait velocity) &#215; 100 (1)</li>
	<li>Perform the statistical analysis (see the Table of Materials) using the methodology described previously20,21.
	<ol>
		<li>Present parametric data as means and standard deviation if normally distributed or as medians if not.</li>
		<li>Use the paired t-test to compare the differences in kinematic parameters between patients in Task 1 and Task 2 conditions.</li>
		<li>Use one-way analysis of variance to compare three different tasks (Task 0, Task 1, and Task 2) of the kinematic parameters. Set statistical significance at P &#60; 0.05.</li>
	</ol>
	</li>
</ol>

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The authors have nothing to disclose.

This study describes a protocol for the clinical assessment of dual motor task gait analysis in stroke patients with motor control deficits. The design of this protocol was based on two main points. First, most previous studies used a single walking task to assess the gait function of stroke patients, and the related discussions on motor control were inadequate, especially because the principles of complex motor movements were rarely involved22,23. Therefore, in ...

The restoration of independent walking function is one of the requisites for the participation of post-stroke patients in community life1. The recovery of walking ability requires not only the interaction of the perception and cognitive systems, but also motor control2,3,4. Furthermore, in real community life, people require higher abilities such as performing two or more tasks at the same time (e.g., walking while holding objects or crossing obstacles). Therefore, studies have begun to focus on the interference of dual-tasks in gait performance5,6. Previous dual-task studies were mostly targeted to elderly and cognitively impaired patients owing to the difficulty in motor performance and heterogeneity in stroke patients; the gait function in stroke patients was mostly evaluated by a single walking task7,8,9. However, further research on dual-task gait analysis, especially motor dual-tasks related to motor control, is required.
This study introduces a methodology for dual motor task gait analysis and evaluation. This protocol not only includes clinical assessment of the walking ability in stroke patients, but also focuses on two dual-motor tasks: the holding-water-and-walking task (a simple dual motor task) and the crossing-obstacle walking task (a complex dual motor task). The aim of this study was to explore the effects of dual motor tasks on the gait of stroke patients and to employ the dual-task gait cost (DTC) values10 of dual-task parameters (the difference between a single task and dual-task) to exclude the heterogeneity among stroke patients. The design of the experimental tasks facilitated an in-depth discussion of the motor control function of stroke patients, which provided new ideas for the clinical diagnosis and evaluation of the gait function of stroke patients.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>BTS Smart DX system</td>
			<td>Bioengineering Technology System, Milan, Italy</td>
			<td>1</td>
			<td>Temporospatial data collection</td>
		</tr>
		<tr>
			<td>BTS SMART-Clinic software</td>
			<td>Bioengineering Technology System, Milan, Italy</td>
			<td>2</td>
			<td>Data processing</td>
		</tr>
		<tr>
			<td>SPSS software (version 25.0)</td>
			<td>IBM Crop., Armonk, NY, USA</td>
			<td></td>
			<td>Statistical analysis</td>
		</tr>
	</tbody>
</table>

motor dual-tasks for gait analysis and evaluation in post-stroke patients

Eighteen stroke patients were recruited for this study involving the evaluation of cognition and walking ability and multitask gait analysis. Multitask gait analysis consisted of a single walking task (Task 0), a simple motor dual-task (water-holding, Task 1), and a complex motor dual-task (crossing obstacles, Task 2). The task of crossing obstacles was considered to be equivalent to the combination of a simple walking task and a complex motor task as it involved more nervous system, skeletal movement, and cognitive resources. To eliminate heterogeneity in the results of the gait analysis of the stroke patients, the dual-task gait cost values were calculated for various kinematic parameters. The major differences were observed in the proximal joint angles, especially in the angles of the trunk, pelvis, and hip joints, which were significantly larger in the dual motor tasks than in the single walking task. This research protocol aims to provide a basis for the clinical diagnosis of gait function and an in-depth study of motor control in stroke patients with motor control deficits through the analyses of dual-motor walking tasks.

Eighteen patients with hemiplegia after stroke were recruited in this study. The average age of the participants was 51.61 &#177; 12.97 years; all were males. The proportion of left and right hemiplegia was 10/8; the average Brunnstrom stage was 4.50 &#177; 0.76. The average of MMSE and MoCA were 26.56 &#177; 1.67 and 20.06 &#177; 2.27, respectively. Other demographic characteristics (including stroke type and time of onset) are shown in Table 1. For the original data of gait dual-tasks (Task 1 and Task ...

Watch this Scientific Journal Video about Motor Dual-Tasks for Gait Analysis and Evaluation in Post-Stroke Patients at JoVE.com

Motor Dual-Tasks for Gait Analysis and Evaluation in Post-Stroke Patients

This paper presents a protocol specifically for dual motor task gait analysis in stroke patients with motor control deficits.

Eighteen stroke patients were recruited for this study involving the evaluation of cognition and walking ability and multitask gait analysis. Multitask ...

motor-dual-tasks-for-gait-analysis-evaluation-post-stroke

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2.3K Views.  The Fifth Affiliated Hospital of Guangzhou Medical University. This paper presents a protocol specifically for dual motor task gait analysis in stroke patients with motor control deficits. 

Video: Motor Dual-Tasks for Gait Analysis and Evaluation in Post-Stroke Patients

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.
This extracellular technique has advantages. First, extracellular electrodes can stimulate and record neurons through the sheath5, so it does not need to be removed. Thus, neurons will be healthier in extracellular experiments than in intracellular ones. Second, if ganglia are rotated by appropriate pinning of the sheath, extracellular electrodes can access neurons on both sides of the ganglion, which makes it easier and more efficient to identify multiple neurons in the same preparation. Third, extracellular electrodes do not need to penetrate cells, and thus can be easily moved back and forth among neurons, causing less damage to them. This is especially useful when one tries to record multiple neurons during repeating motor patterns that may only persist for minutes. Fourth, extracellular electrodes are more flexible than intracellular ones during muscle movements. Intracellular electrodes may pull out and damage neurons during muscle contractions. In contrast, since extracellular electrodes are gently pressed onto the sheath above neurons, they usually stay above the same neuron during muscle contractions, and thus can be used in more intact preparations.
To uniquely identify motor neurons for a motor pool (in particular, the I1/I3 muscle in Aplysia) using extracellular electrodes, one can use features that do not require intracellular measurements as criteria: soma size and location, axonal projection, and muscle innervation4,6,7. For the particular motor pool used to illustrate the technique, we recorded from buccal nerves 2 and 3 to measure axonal projections, and measured the contraction forces of the I1/I3 muscle to determine the pattern of muscle innervation for the individual motor neurons.
We demonstrate the complete process of first identifying motor neurons using muscle innervation, then characterizing their timing during motor patterns, creating a simplified diagnostic method for rapid identification. The simplified and more rapid diagnostic method is superior for more intact preparations, e.g. in the suspended buccal mass preparation8 or in vivo9. This process can also be applied in other motor pools10,11,12 in Aplysia or in other animal systems2,3,13,14.

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.

In animals with large  identified neurons (e.g. mollusks), analysis of motor pools is done using  intracellular techniques1,2,3,4. Recently,  we developed ...

A Human-machine-interface Integrating Low-cost Sensors with a Neuromuscular Electrical Stimulation System for Post-stroke Balance Rehabilitation

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing delivery of oxygen and nutrients. About half of the stroke survivors are left with some degree of disability. Innovative methodologies for restorative neurorehabilitation are urgently required to reduce long-term disability. The ability of the nervous system to reorganize its structure, function and connections as a response to intrinsic or extrinsic stimuli is called neuroplasticity. Neuroplasticity is involved in post-stroke functional disturbances, but also in rehabilitation. Beneficial neuroplastic changes may be facilitated with non-invasive electrotherapy, such as neuromuscular electrical stimulation (NMES) and sensory electrical stimulation (SES). NMES involves coordinated electrical stimulation of motor nerves and muscles to activate them with continuous short pulses of electrical current while SES involves stimulation of sensory nerves with electrical current resulting in sensations that vary from barely perceivable to highly unpleasant. Here, active cortical participation in rehabilitation procedures may be facilitated by driving the non-invasive electrotherapy with biosignals (electromyogram (EMG), electroencephalogram (EEG), electrooculogram (EOG)) that represent simultaneous active perception and volitional effort. To achieve this in a resource-poor setting, e.g., in low- and middle-income countries, we present a low-cost human-machine-interface (HMI) by leveraging recent advances in off-the-shelf video game sensor technology. In this paper, we discuss the open-source software interface that integrates low-cost off-the-shelf sensors for visual-auditory biofeedback with non-invasive electrotherapy to assist postural control during balance rehabilitation. We demonstrate the proof-of-concept on healthy volunteers.

A novel low-cost human-machine interface for interactive post-stroke balance rehabilitation system is presented in this article. The system integrates off-the-shelf low-cost sensors towards volitionally driven electrotherapy paradigm. The proof-of-concept software interface is demonstrated on healthy volunteers.

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing ...

Automated Gait Analysis in Mice with Chronic Constriction Injury

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some disadvantages such as subjective data and the requirement of a skilled, experienced experimenter. To date, a variety of modifications have improved the von Frey method, but it still has a few limitations. Recent reports have suggested that gait analysis produces more accurate and objective data from the neuropathic animals. This protocol demonstrates how to perform the automated gait analysis to determine the degree of neuropathic pain in mice. After several days of acclimation, the mice were allowed to walk freely on the glass floor to illuminate footprints. Then, quantification of the footprints and gait were performed through video clips with automatic analysis of various walking parameters, such as area of paw print, swing time, angle of paw, etc.
The main purpose of this study is to describe the methodology of automated gait analysis and briefly compare it with data from the classical sensory test using von Frey filament.

The precise assessment of pain response in a neuropathic animal model is critical to investigate the pathophysiology of pain diseases and develop new analgesics. We present a sensitive and objective method to determine the sensory function of the rodent hind paw by an automated gait analysis system.

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some ...

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

With the rising prevalence of neurodegenerative diseases, it is increasingly important to understand the underlying pathophysiology that leads to neuronal dysfunction and loss. Fluorescence-based imaging tools and technologies enable unprecedented analysis of subcellular neurobiological processes, yet there is still a need for unbiased, reproducible, and accessible approaches for extracting quantifiable data from imaging studies. We have developed a simple and adaptable workflow to extract quantitative data from fluorescence-based imaging studies using Drosophila models of neurodegeneration. Specifically, we describe an easy-to-follow, semi-automated approach using Fiji/ImageJ to analyze two cellular processes: first, we quantify protein aggregate content and profile in the Drosophila optic lobe using fluorescent-tagged mutant huntingtin proteins; and second, we assess autophagy-lysosome flux in the Drosophila visual system with ratiometric-based quantification of a tandem fluorescent reporter of autophagy. Importantly, the protocol outlined here includes a semi-automated segmentation step to ensure all fluorescent structures are analyzed to minimize selection bias and to increase resolution of subtle comparisons. This approach can be extended for the analysis of other cell biological structures and processes implicated in neurodegeneration, such as proteinaceous puncta (stress granules and synaptic complexes), as well as membrane-bound compartments (mitochondria and membrane trafficking vesicles). This method provides a standardized, yet adaptable reference point for image analysis and quantification, and could facilitate reliability and reproducibility across the field, and ultimately enhance mechanistic understanding of neurodegeneration.

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

We have developed a simple and adaptable workflow to extract quantitative data from fluorescence-imaging-based cell biological studies of protein aggregation and autophagic flux in the central nervous system of Drosophila models of neurodegeneration.

With the rising prevalence of neurodegenerative diseases, it is increasingly important to understand the underlying pathophysiology that leads to neuronal ...

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of sciatic nerve injury rodent models. In this protocol, we describe a novel kinematic analysis method that uses a three-dimensional (3D) motion capture apparatus for functional evaluations using a rat sciatic nerve crush injury model. First, the rat is familiarized with treadmill walking. Markers are then attached to the designated bone landmarks and the rat is made to walk on the treadmill at the desired speed. Meanwhile, the posterior limb movements of the rat are recorded using four cameras. Depending on the software used, marker tracings are created using both automatic and manual modes and the desired data are produced after subtle adjustments. This method of kinematic analysis, which uses a 3D motion capture apparatus, offers numerous advantages, including superior precision and accuracy. Many more parameters can be investigated during the comprehensive functional evaluations. This method has several shortcomings that require consideration: The system is expensive, can be complicated to operate, and may produce data deviations due to skin shifting. Nevertheless, kinematic analysis using a 3D motion capture apparatus is useful for performing functional anterior and posterior limb evaluations. In the future, this method may become increasingly useful for generating accurate assessments of various traumas and diseases.

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...

Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke involves middle cerebral artery occlusion (MCAO). Infarct zone, brain edema and blood-brain barrier (BBB) breakdown are measured as parameters that reflect the extent of brain injury after MCAO. A significant limitation to this method is that these measurements are normally obtained in different rat brain samples, leading to ethical and financial burdens due to the large number of rats that need to be euthanized for an appropriate sample size. Here we present a method to accurately assess brain injury following MCAO by measuring infarct zone, brain edema and BBB permeability in the same set of rat brains. This novel technique provides a more efficient way to evaluate the pathophysiology of stroke.

This protocol describes a novel technique of measuring the three most important parameters of ischemic brain injury on the same set of rodent brain samples. Using only one brain sample is highly advantageous in terms of ethical and economic costs.

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke ...

Automated Gait Analysis to Assess Functional Recovery in Rodents with Peripheral Nerve or Spinal Cord Contusion Injury

Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and a lot of comparative data has been published in the literature. This includes a multitude of assessment methods to study functional recovery following nerve injury and repair. Besides evaluation of nerve regeneration by means of histology, electrophysiology, and other in vivo and in vitro assessment techniques, functional recovery is the most important criterion to determine the degree of neural regeneration. Automated gait analysis allows recording of a vast quantity of gait-related parameters such as Paw Print Area and Paw Swing Speed as well as measures of inter-limb coordination. Additionally, the method provides digital data of the rats&#8217; paws after neuronal damage and during nerve regeneration, adding to our understanding of how peripheral and central nervous injuries affect their locomotor behavior. Besides the predominantly used sciatic nerve injury model, other models of peripheral nerve injury such as the femoral nerve can be studied by means of this method. In addition to injuries of the peripheral nervous systems, lesions of the central nervous system, e.g., spinal cord contusion can be evaluated. Valid and reproducible data assessment is strongly dependent on meticulous adjustment of the hard- and software settings prior to data acquisition. Additionally, proper training of the experimental animals is of crucial importance. This work aims to illustrate the use of computerized automated gait analysis to assess functional recovery in different animal models of peripheral nerve injury as well as spinal cord contusion injury. It also emphasizes the method&#8217;s limitations, e.g., evaluation of nerve regeneration in rats with sciatic nerve neurotmesis due to limited functional recovery. Therefore, this protocol is thought to help researchers interested in peripheral and central nervous injuries to assess functional recovery in rodent models.

Automated gait analysis is a feasible tool to evaluate functional recovery in rodent models of peripheral nerve injury and spinal cord contusion injury. While it requires only one setup to assess locomotor function in various experimental models, meticulous hard- and soft-ware adjustment and training of the animals is highly important.

Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and ...

Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars compacta. Gait abnormalities, including decreased arm swing, slower walking speed, and shorter steps are common in PD patients and appear early in the course of disease. Thus, the quantification of motor patterns in animal models of PD will be important for phenotypic characterization during disease course and upon therapeutic treatment. Most cases of PD are idiopathic; however, the identification of hereditary forms of PD uncovered gene mutations and variants, such as loss-of-function mutations in Pink1 and Parkin, two proteins involved in mitochondrial quality control that could be harnessed to create animal models. While mice are resistant to neurodegeneration upon loss of Pink1 and Parkin (single and combined deletion), in rats, Pink1 but not Parkin deficiency leads to nigral DA neuron loss and motor impairment. Here, we report the utility of FTIR imaging to uncover gait changes in freely walking young (2 months of age) male rats with combined loss of Pink1 and Parkin prior to the development of gross visually apparent motor abnormality as these rats age (observed at 4-6 months), characterized by hindlimb dragging as previously reported in Pink1 knockout (KO) rats.

Here we describe the RatWalker system, built by redesigning the MouseWalker apparatus to accommodate for the increased size and weight of rats. This system uses frustrated total internal reflection (FTIR), high-speed video capture, and open-access analysis software to track and quantify gait parameters.

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars ...

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor impairment that worsens with time. While considerable advances have been made in determining genetic drivers of ALS for a subset of patients, the majority of cases have an unknown etiology. Further, the mechanisms underlying motor neuron dysfunction and degeneration are not well understood; therefore, there is an ongoing need to develop and characterize representative models to study these processes. Caenorhabditis elegans can adapt their movement to the physical constraints of their surroundings, with two primary movement paradigms studied in a laboratory environment- crawling on a solid surface and swimming in liquid. These represent a complex interplay between sensation, motor neurons, and muscles. C. elegans models of ALS can exhibit impairment in one or both of these movement paradigms. This protocol describes two sensitive assays for evaluating motility in C. elegans: an optimized radial locomotion assay measuring crawling on a solid surface and an automated method for tracking and analyzing swimming in liquid (thrashing). In addition to the characterization of baseline motor impairment of ALS models, these assays can detect suppression or enhancement of the phenotypes from genetic or small molecule interventions. Thus, these methods have utility for studying ALS&#160;models and any C. elegans strain that exhibits altered motility.

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

This protocol describes two sensitive assays for discriminating among mild, moderate, and severe motor impairment in C. elegans models of amyotrophic lateral sclerosis, with general utility for C. elegans strains, with altered motility.

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor ...