This work has been funded by an American Heart Association Scientist Development Grant (#12SDG12200001) to E. Vasudevan. R. Hamzey's current affiliation is Department of Mechanical Engineering, Boston University, Boston, MA, USA. E. Kirk's current affiliation is the MGH Institute of Health Professions Department of Physical Therapy.

All procedures have been approved by the Institutional Review Board at Stony Brook University.
1. Experimental Set-up
Note: Refer to Supplementary File 1-Definitions for definitions of common terms used in split-belt treadmill experiments.
<ol>
	<li>Screen all participants for prior experience with the split-belt treadmill. 
	NOTE: People have been shown to readapt faster to the split-belt treadmill following a prior exposure to it29,30. The timescale over which people &#34;forget&#34; the split-belt treadmill is not presently known; thus, prior experience with the split-belt treadmill may be a confounding variable if it is not controlled.</li>
	<li>Conduct all testing in a quiet environment, and minimize activity in the testing room.</li>
	<li>Set up a motion tracking system (according to system instructions) to record movement on a split-belt treadmill and on an over ground walkway. 
	NOTE: For example, the current protocol used a motion tracking system with active LED markers. Four tripod-mounted sensor units detected the three-dimensional position of the active markers, with two units placed on either side (right and left) of the treadmill and two on either side of a 7 m over ground walkway.</li>
	<li>Outfit the participant with motion tracking markers, electromyography, etc.</li>
	<li>Consider including a partition between the two belts of the split-belt treadmill to prevent the legs from crossing over to the contralateral belt. This partition is not strictly necessary for neurologically-intact adults but may be helpful for testing children or clinical populations. Note that the presence of a partition likely increases step width; however, the extent to which this affects split-belt adaptation is unknown.</li>
	<li>Set up a safety harness over the treadmill to protect the participant from falling during treadmill walking. 
	NOTE: The harness should not support body weight, unless this is part of the research question. Although falling during treadmill walking is exceedingly rare, many research ethics boards require safety harness use.</li>
	<li>Maintain consistency in arm movement across the experimental paradigm and across participants. When deciding on the type of arm movement (e.g., holding handrails, swinging arms naturally), consider what will be comfortable for the subject group and whether typical arm swing will obscure the visibility of critical markers used for motion capture (e.g., for markers placed on the hips).
	<ol>
		<li>Regardless of arm movement, instruct all participants to hold handrails while starting and stopping the treadmill for safety.</li>
	</ol>
	</li>
	<li>Maintain consistency in incline across the experimental paradigm. 
	NOTE: To our knowledge, all published split-belt treadmill protocols, including the current one, have used zero incline for treadmill and over ground walking.</li>
</ol>
2. Baseline Period
Note: The purpose of the baseline period is to establish what normal walking coordination is for each person. Baseline coordination should be tested in all the conditions in which after-effects are tested. For example, in the current protocol, after-effects were tested during treadmill and over ground walking at different speeds (0.7 and 1.4 m/s). Therefore, baseline over ground and treadmill trials at 0.7 and 1.4 m/s were included. This allows a direct comparison of after-effects to baseline walking coordination at the same speed and context. Over ground walking baseline trials can be eliminated when the experimental objectives do not include generalization to over ground walking.
<ol>
	<li>For over ground baseline trials, instruct the participant to walk over the ground on a walkway where motion capture data can be collected. Collect a minimum of 10 stride cycles to establish the baseline over ground walking.
	<ol>
		<li>If the motion capture system only allows for motion capture within a limited space, have the participant perform several passes (e.g., trials) through the motion capture space. At the end of each trial, instruct the participant to stop, turn in place, and prepare for the researcher&#39;s cue to begin the next trial.</li>
		<li>For each trial, ensure that at least two stride cycles are performed within the motion capture space, not including the first and last stride cycles. 
		NOTE: These initial and final stride cycles will be discarded from analysis as they are acceleration/deceleration strides, not steady-state walking.</li>
		<li>Have the participants perform several (typically 10) over ground walking trials.
		<ol>
			<li>If a specific speed is desired, have the participant walk at that speed on the treadmill (on tied-belts) to familiarize him/her with the task. Then, move back to the walkway, instruct the participant to walk at the same speed as he/she did on the treadmill, and time the participant during each trial of over ground walking. Give verbal feedback in between each trial to speed up or slow down, if needed25.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>For treadmill baseline trials, instruct the participant to walk on tied-belts for 1 - 5 min. 
	NOTE: This constitutes a single baseline trial. If the participant is unfamiliar with treadmill walking, this period may be lengthened to allow the person to become comfortable with the task.
	<ol>
		<li>Match the speed(s) of baseline trials to the speed(s) at which the after-effects will be tested, to allow for comparison of pre- and post-adaptation gait coordination at equivalent speeds. 
		NOTE: Multiple baseline trials (i.e., 1 - 5 min blocks) at different tied-belt speeds may be required; for example, in the current protocol, baseline trials at tied-belt speeds of 0.7 m/s and 1.4 m/s were collected because those were the speeds used to evaluate after-effects.</li>
	</ol>
	</li>
</ol>
3. Adaptation Period
Note: Participants do not need to be instructed that they are about to walk on split-belts. In many experiments, including the current one, participants are not told whether belts will be tied- or split-; they are simply told when the treadmill will be starting or stopping. This allows the experimenter to measure the effects of an unanticipated change in the walking environment.
<ol>
	<li>While the participant is standing on the stationary treadmill belts, start the split-belt treadmill with one belt running faster than the other and allow the participant to walk for at least 7 min (10 - 15 min is more common).
	<ol>
		<li>Instruct the participant to look straight ahead, not down at their feet.</li>
		<li>Set one belt speed faster than the other (e.g.,2 - 3 fold differences between belt speeds). 
		NOTE: Higher speed ratios have been used in the past8,31. The current protocol uses 0.7:1.4 m/s for a 2:1 ratio.
		<ol>
			<li>Either randomize which leg is driven by the slower belt or consistently choose one leg (either dominant or non-dominant) as the leg that is driven by the slower belt.</li>
			<li>The belt speed differential may be introduced gradually (fast belt speed is incrementally increased and/or slow belt speed incrementally decreased over several min) or abruptly (from stopped position, belts accelerate to target speed within seconds). 
			NOTE: The way that split-belts are introduced can affect how individuals adapt, how well they transfer the adapted pattern to different walking environments, and how well they re-adapt to split-belts 24 h later27,32. Presently, most split-belt walking protocols (including the current one) introduce the split-belts abruptly.</li>
			<li>If it is anticipated that breaks will be needed (e.g., for young children, older adults, or individuals with limited mobility), add predetermined rest breaks to the protocol for all participants. Ensure that the length of these breaks is consistent; unanticipated breaks should be recorded and timed, as this may be a factor to consider in analysis33.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>
4. Catch Trial
Note: Catch trials are performed on the treadmill (tied-belts) and are used to briefly test the participant&#39;s after-effects thus far in the protocol, indicating how much they have adapted. A catch trial is a short (usually &#60; 20 s) period of tied-belt walking to quickly evaluate the development of after-effects during the split-belt adaptation period.
<ol>
	<li>Once the participant has fully adapted to split-belts (minimum of 7 min split-belt walking), briefly stop the belts and restart the treadmill with both belts running at the same speed. Perform the catch trial by starting the treadmill at the same speed as the slower belt during split-belt adaptation28 as after-effects will be largest here.
	<ol>
		<li>To maximize after-effect amplitude following split-belt adaptation at 0.7:1.4 m/s, perform the catch trial at 0.7 m/s.</li>
	</ol>
	</li>
	<li>To mitigate de-adaptation, end the catch trial (i.e., stop the treadmill) once the participant has taken about five strides at the desired catch trial speed (~ 10-15 s).</li>
	<li>To evaluate after-effects in catch trials performed at multiple different walking speeds (or other changes in walking contexts, e.g., forwards and backwards walking24), re-adapt the participant for at least 2 min on split-belts between each catch trial. 
	NOTE: The order of catch trials should be randomized25 and/or the first catch trial should be re-tested near the end of the adaptation period to determine if there was a systematic decrease in after-effect size with repeated switching between tied-belts (catch trials) and split-belts (re-adaptation)28.</li>
	<li>Following the last catch trial, stop the treadmill and restart it with split-belts (same configuration as adaptation - see step 3.1.2) for 2-5 min to allow the participant to re-adapt.</li>
</ol>
5. Post-adaptation - Testing After-effects During Over Ground Walking
Note: This step is optional and depends on the objectives of the experiment. In the present protocol, the objectives included assessment of generalization to over ground walking, thus a post-adaptation over ground testing period was included.
<ol>
	<li>Stop the treadmill and transfer the participant to the over ground walkway using a wheelchair, to prevent participants from taking unrecorded steps prior to reaching the recording area.</li>
	<li>Instruct the participant to walk along the over-ground walkway, as in step 2.1.
	<ol>
		<li>If a specific walking speed is desired, instruct individuals to replicate the baseline walking speed25.</li>
		<li>To completely wash-out over ground walking after-effects so that people return to their baseline coordination, have the participants perform 10-15 walking passes on a 6 m over ground walkway 
		NOTE: This has been shown to be sufficient26,27 and amounts to roughly 30 strides27. If over ground walking is not continuously recorded (e.g., several passes are taken through the recording area), there will be several steps that are not analyzed in between each over ground walking trial, as the participant slows down, turns in place, and starts walking in the other direction. The rate of de-adaptation in over ground post-adaptation (OG PA) trials should be cautiously interpreted unless the experimental set-up allows for continuous recording of over ground walking.</li>
	</ol>
	</li>
</ol>
6. Post-adaptation - Testing After-effects During Treadmill Walking
NOTE: As in step 5, this step is optional and depends upon the study objectives. If an OG PA period was included, the subsequent treadmill post-adaptation period tests for the presence of treadmill after-effects after over ground after-effects have been washed-out23,26,27. If there was no OG PA period, the treadmill post-adaptation period can be used to evaluate treadmill after-effects (first 1 - 5 strides of post-adaptation) and/or treadmill de-adaptation rates22,29,34.
<ol>
	<li>If there was no OG PA, at the end of the adaptation period, stop the treadmill briefly and re-start with tied-belts. If there was an over ground walking period, use the wheelchair to transport the participant back to the stationary treadmill and re-start with tied-belts; the wheelchair is important to minimize the number of steps that are not recorded.
	<ol>
		<li>To simply measure after-effect size, record tied-belt walking for a short period (e.g., 30 s). In order to assess rates of de-adaptation, record continuous tied-belt walking for a minimum of 10 min to ensure complete wash-out of after-effects.</li>
		<li>Set the speed of tied-belts during the post-adaptation period as per the hypotheses posed, as the largest treadmill after-effects occur when the tied-belt speed matches that of the slower belt during split-belt adaptation25,28. If adaptation is performed at split-belt speeds of 0.7 and 1.4 m/s, set the tied belt speed at 0.7 m/s to observe the largest after-effects.</li>
	</ol>
	</li>
</ol>

The authors have nothing to disclose.

Numerous studies have now shown that people adapt gait coordination on a split-belt treadmill in order to restore symmetry in interlimb coordination parameters like step length and double support duration. When natural walking conditions are restored following split-belt walking, participants continue using the adapted gait pattern, leading to after-effects that have to be unlearned in order to return to normal walking coordination. Researchers primarily use adaptation rate and after-effect amplitude to quantify the abil...

A split-belt treadmill has two belts that can drive each leg at a different speed or in a different direction. This device was first used over 45 years ago as a tool to study coordination between the legs (i.e., interlimb coordination) during walking1. This, and other early studies primarily used cats as an experimental model1,2,3, but insects were also studied4. The first investigations of split-belt locomotion in human infants and adults were published in 1987 and 1994, respectively5,6. These initial studies in both human and non-human animals mostly investigated short-term (i.e., immediate) adjustments in interlimb coordination to preserve stability and forward progression when the legs are driven at different speeds. A 1995 study noted that longer periods (several minutes) of split-belt walking impaired the ability of human adults to accurately perceive treadmill belt speed and make adjustments to equalize speeds on each side. This suggests that the sensorimotor mapping of walking had been recalibrated7. However, it was not until 2005 that the first detailed kinematic report of human motor adaptation over 10 minutes of split-belt treadmill walking was published8.
Motor adaptation refers to an error-driven process during which sensorimotor mappings of well-learned movements are adjusted in response to a new, predictable demand9. It is a form of motor learning that occurs over an extended practice period (minutes to hours) and results in after-effects, which are changes in the movement pattern when the demand is removed and/or conditions return to normal. For example, walking on split-belts initially causes people to walk with asymmetric interlimb coordination, resembling a limp. Over several minutes of split-belt walking, people adapt their walking coordination so that their gait becomes more symmetric. When the two belts subsequently return to the same speed (i.e. tied-belts), thus restoring normal walking conditions, people demonstrate after-effects by walking with asymmetric coordination. These after-effects must be actively de-adapted or unlearned over several minutes of tied-belt walking before normal walking coordination is restored8.
Following the 2005 Reisman et al.8 kinematic analysis of split belt walking in humans, use of the split-belt treadmill in published research has increased approximately ten-fold compared to the previous decade. Why is the split-belt treadmill becoming more popular as an experimental tool? Split-belt ambulation is clearly a laboratory task &#8211; the closest real-world analog is turning or walking in a tight circle, but the split-belt treadmill induces a much more extreme version of turning, with one leg being driven two- to four-times faster than the other. The fact that the split-belt treadmill is a highly unusual walking task offers several advantages for studying locomotor learning. First, it is novel for most people regardless of age and independent of walking experience; it is easy to screen experimental participants for novelty of split-belt walking. Second, the split-belt treadmill induces sizeable changes in interlimb coordination that are not quickly resolved. The relatively slow rates of adaptation and de-adaptation permit us to study how different training interventions can alter these rates without approaching a ceiling. Third, the kinematic8,10, kinetic11,12,13,14, electromyographic6,15,16, and perceptual7,17,18,19 modifications that occur with split-belt treadmill adaptation have been well-studied, as has the neural control of this task20,21,22. In other words, adaptations to the split-belt treadmill have been documented and replicated by several different groups, making this a well-characterized locomotor learning task.
Over the past ten years, several studies have demonstrated the task- and context-specific nature of split-belt adaptation. After-effects following split-belt adaptation are significantly reduced in amplitude if they are tested under different conditions from the training condition. For example, after-effects are smaller if the person is moved to a different environment (e.g., over ground walking23), performs a different locomotor task (e.g., backward walking or running13,24), or even walks at a speed that differs from the speed of the slower belt during adaptation25. Efforts to establish parameters governing the generalization of locomotor adaptation are ongoing.
The objective of this paper is to describe a protocol for using the split-belt treadmill to investigate human locomotor adaptation and generalization of the adapted pattern to other walking contexts (i.e., different walking speeds and environments). While the protocol described here is most directly derived from that used in Hamzey et al.25 (Figure 1a), it should be noted that this protocol was informed by a number of studies that preceded it8,23,24,26,27,28. The method was originally developed to test the hypothesis that maintaining constancy in walking speed between the treadmill and over ground environments would improve generalization of split-belt walking across these different environments25. In the protocol section below, we give instructions on how to replicate this version of the split-belt treadmill method, with notes that indicate how certain protocol steps may be modified to for different method purposes.

<table border="0" cellpadding="0" cellspacing="0" width="997">
	<tbody>
		<tr height="220">
			<td height="220" style="height:220px;width:216px;">Split-belt treadmill</td>
			<td style="width:71px;">Woodway</td>
			<td style="width:119px;"></td>
			<td style="width:591px;">The WOODWAY SPLIT-BELT is an advanced gate measurement and analysis tool used for synchronous or asynchronous running/walking. With its unique and innovative dual belt system, the &#34;SPLIT-BELT,&#34; provides infinitely variable speed control of each leg independently. Used for gait rehab, the gas-assisted, fully adjusted handrail options provide more room for therapists and patients.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Codamotion CX1</td>
			<td>Charmwood Dynamics, Ltd, Leicestershire, UK</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

using a split-belt treadmill to evaluate generalization of human locomotor adaptation

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. However, studying human locomotor learning can be challenging. During infancy and childhood, the neuromuscular system is quite immature, and it is unlikely that locomotor learning during early stages of development is governed by the same mechanisms as in adulthood. By the time humans reach maturity, they are so proficient at walking that it is difficult to come up with a sufficiently novel task to study de novo locomotor learning. The split-belt treadmill, which has two belts that can drive each leg at a different speed, enables the study of both short- (i.e., immediate) and long-term (i.e., over minutes-days; a form of motor learning) gait modifications in response to a novel change in the walking environment. Individuals can easily be screened for previous exposure to the split-belt treadmill, thus ensuring that all experimental participants have no (or equivalent) prior experience. This paper describes a typical split-belt treadmill adaptation protocol that incorporates testing methods to quantify locomotor learning and generalization of this learning to other walking contexts. A discussion of important considerations for designing split-belt treadmill experiments follows, including factors like treadmill belt speeds, rest breaks, and distractors. Additionally, potential but understudied confounding variables (e.g., arm movements, prior experience) are considered in the discussion.

Walking on a split-belt treadmill initially causes large asymmetries in interlimb coordination. Over a period of 10 - 15 min, symmetry in many of these measures is gradually restored. Detailed descriptions of how kinematic walking parameters change over the course of split-belt treadmill adaptation have been published elsewhere8,10. This paper focuses on two measures of interlimb coordination: ste...

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Using a Split-belt Treadmill to Evaluate Generalization of Human Locomotor Adaptation

We describe a protocol for investigating human locomotor adaptation using the split-belt treadmill, which has two belts that can drive each leg at a different speed. We specifically focus on a paradigm designed to test the generalization of adapted locomotor patterns to different walking contexts (e.g., gait speeds, walking environments).

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. ...

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Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

8.2K Views.  Stony Brook University. We describe a protocol for investigating human locomotor adaptation using the split-belt treadmill, which has two belts that can drive each leg at a different speed. We specifically focus on a paradigm designed to test the generalization of adapted locomotor patterns to different walking contexts (e.g., gait speeds, walking environments). 

Video: Using a Split-belt Treadmill to Evaluate Generalization of Human Locomotor Adaptation

Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice and other rodents. Improvements in video tracking software (VTS) have allowed it to be coupled to LMA testing, dramatically improving specificity and sensitivity when compared to the line crossings method with manual scoring. In addition, VTS enables high-throughput experimentation. While similar to automated video tracking used for the open field test (OFT), LMA testing is unique in that it allows mice to remain in their home cage and does not utilize the anxiogenic stimulus of bright lighting during the active phase of the light-dark cycle. Traditionally, LMA has been used for short periods of time (mins), while longer movement studies (hrs-days) have often used implanted transmitters and biotelemetry. With the option of real-time tracking, long-, like short-term LMA testing, can now be conducted using videography. Long-term LMA testing requires a specialized, but easily constructed, cage so that food and water (which is usually positioned on the cage top) does not obstruct videography. Importantly, videography and VTS allows for the quantification of parameters, such as path of mouse movement, that are difficult or unfeasible to measure with line crossing and/or biotelemetry. In sum, LMA testing coupled to VTS affords a more complete description of mouse movement and the ability to examine locomotion over an extended period of time.

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice. Coupling of video tracking software (VTS) and LMA allows for the improvement of specificity and sensitivity, especially when compared with the manual, line crossing method of LMA analysis. Additionally VTS allows long-term tracking of mouse LMA.

Locomotor  activity (LMA) is a simple and easily performed measurement of behavior in mice  and other rodents. Improvements in video tracking software ...

Using Eye Movements to Evaluate the Cognitive Processes Involved in Text Comprehension

The present article describes how to use eye tracking methodologies to study the cognitive processes involved in text comprehension. Measuring eye movements during reading is one of the most precise methods for measuring moment-by-moment (online) processing demands during text comprehension. Cognitive processing demands are reflected by several aspects of eye movement behavior, such as fixation duration, number of fixations, and number of regressions (returning to prior parts of a text). Important properties of eye tracking equipment that researchers need to consider are described, including how frequently the eye position is measured (sampling rate), accuracy of determining eye position, how much head movement is allowed, and ease of use. Also described are properties of stimuli that influence eye movements that need to be controlled in studies of text comprehension, such as the position, frequency, and length of target words. Procedural recommendations related to preparing the participant, setting up and calibrating the equipment, and running a study are given. Representative results are presented to illustrate how data can be evaluated. Although the methodology is described in terms of reading comprehension, much of the information presented can be applied to any study in which participants read verbal stimuli.

The present article describes how to use eye tracking methodologies to study the cognitive processes involved in text comprehension. Descriptions of eye tracking equipment, how to develop experimental stimuli, and procedural recommendations are included. The information presented can be applied to most any study using verbal stimuli.

The present article describes how to use eye  tracking methodologies to study the cognitive processes involved in text  comprehension. Measuring eye ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing locomotive impairment in animal models of neuromuscular disease and efficacy of therapeutic drugs that may improve locomotion and/or muscle function. The open field activity measurement provides a different measure than muscle strength, which is commonly assessed by grip strength measurements. It can also show how drugs may affect other body systems as well when used with additional outcome measures. In addition, measures such as total distance traveled mirror the 6 min walk test, a clinical trial outcome measure. However, open field activity monitoring is also associated with significant challenges: Open field activity measurements vary according to animal strain, age, sex, and circadian rhythm. In addition, room temperature, humidity, lighting, noise, and even odor can affect assessment outcomes. Overall, this manuscript provides a well-tested and standardized open field activity SOP for preclinical trials in animal models of neuromuscular diseases. We provide a discussion of important considerations, typical results, data analysis, and detail the strengths and weaknesses of open field testing. In addition, we provide recommendations for optimal study design when using open field activity in a preclinical trial.

Open field activity levels are used to assess locomotive and behavioral activity levels. This protocol provides a well-designed, standardized protocol to use in preclinical trials for neuromuscular disorders.

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies for modeling the genetic bases of vocal communication deficits. Characterizing the vocal behavior of mouse models carrying mutations in genes associated with neuropsychiatric disorders such as autism spectrum disorders will help to understand the mechanisms leading to social communication deficits. We provide here protocols to reliably elicit ultrasonic vocalizations in pups and in adult mice. This standardization will help reduce inter-study variability due to the experimental settings. Pup isolation calls are recorded throughout development from individual pups isolated from dam and littermates. In adulthood, vocalizations are recorded during same-sex interactions (without a sexual component) by exposing socially motivated males or females to an unknown same-sex conspecific. We also provide a protocol to record vocalizations from adult males exposed to an estrus female. In this context, there is a sexual component in the interaction. These protocols are established to elicit a large amount of ultrasonic vocalizations in laboratory mice. However, we point out the important inter-individual variability in the vocal behavior of mice, which should be taken into account by recording a minimal number of individuals (at least 12 in each condition). These recordings of ultrasonic vocalizations are used to evaluate the call rate, the vocal repertoire and the acoustic structure of the calls. Data are combined with the analysis of synchronous video recordings to provide a more complete view on social communication in mice. These protocols are used to characterize the vocal communication deficits in mice lacking ProSAP1/Shank2, a gene associated with autism spectrum disorders. More ultrasonic vocalizations recordings can also be found on the mouseTube database, developed to favor the exchange of such data.

Mouse ultrasonic vocalizations are used as proxies to model the genetic bases of vocal communication deficits in mouse models for neuropsychiatric disorders. The present protocol describes three experimental contexts that reliably elicit ultrasonic vocalizations from pups (throughout development) and adult mice (same-sex interactions, male-estrus female interactions).

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies ...

Visualizing Visual Adaptation

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical or age differences, or a color deficiency or disease. This protocol describes a technique for incorporating sensory adaptation into the simulations. The protocol is illustrated with the example of color vision, but is generally applicable to any form of visual adaptation. The protocol uses a simple model of human color vision based on standard and plausible assumptions about the retinal and cortical mechanisms encoding color and how these adjust their sensitivity to both the average color and range of color in the prevailing stimulus. The gains of the mechanisms are adapted so that their mean response under one context is equated for a different context. The simulations help reveal the theoretical limits of adaptation and generate &#34;adapted images&#34; that are optimally matched to a specific environment or observer. They also provide a common metric for exploring the effects of adaptation within different observers or different environments. Characterizing visual perception and performance with these images provides a novel tool for studying the functions and consequences of long-term adaptation in vision or other sensory systems.

This article describes a novel method for simulating and studying adaptation in the visual system.

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

Sit-to-stand-and-walk from 120% Knee Height: A Novel Approach to Assess Dynamic Postural Control Independent of Lead-limb

Individuals with sensorimotor pathology e.g., stroke have difficulty executing the common task of rising from sitting and initiating gait (sit-to-walk: STW). Thus, in clinical rehabilitation separation of sit-to-stand and gait initiation - termed sit-to-stand-and-walk (STSW) - is usual. However, a standardized STSW protocol with a clearly defined analytical approach suitable for pathological assessment has yet to be defined.
Hence, a goal-orientated protocol is defined that is suitable for healthy and compromised individuals by requiring the rising phase to be initiated from 120% knee height with a wide base of support independent of lead limb. Optical capture of three-dimensional (3D) segmental movement trajectories, and force platforms to yield two-dimensional (2D) center-of-pressure (COP) trajectories permit tracking of the horizontal distance between COP and whole-body-center-of-mass (BCOM), the decrease of which increases positional stability but is proposed to represent poor dynamic postural control.
BCOM-COP distance is expressed with and without normalization to subjects&#39; leg length. Whilst COP-BCOM distances vary through STSW, normalized data at the key movement events of seat-off and initial toe-off (TO1) during steps 1 and 2 have low intra and inter subject variability in 5 repeated trials performed by 10 young healthy individuals. Thus, comparing COP-BCOM distance at key events during performance of an STSW paradigm between patients with upper motor neuron injury, or other compromised patient groups, and normative data in young healthy individuals is a novel methodology for evaluation of dynamic postural stability.

Here, we present a novel protocol to measure positional stability at key events during the sit-to-stand-to-walk using the center-of-pressure to the whole-body-center-of-mass distance. This was derived from the force platform and three-dimensional motion-capture technology. The paradigm is reliable and can be utilized for the assessment of neurologically compromised individuals.

Individuals with sensorimotor pathology e.g., stroke have difficulty executing the common task of rising from sitting and initiating gait (sit-to-walk: ...

A Method to Study Adaptation to Left-Right Reversed Audition

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the previous studies have used special spectacles with prisms to achieve unusual spaces in the visual domain, a methodology for studying the adaptation to unusual auditory spaces has yet to be fully established. This study proposes a new protocol to set-up, validate, and use a left-right reversed stereophonic system using only wearable devices, and to study the adaptation to left-right reversed audition with the help of neuroimaging. Although individual acoustic characteristics are not yet implemented, and slight spillover of unreversed sounds is relatively uncontrollable, the constructed apparatus shows high performance in a 360&#176; sound source localization coupled with hearing characteristics with little delay. Moreover, it looks like a mobile music player and enables a participant to focus on daily life without arousing curiosity or drawing attention of other individuals. Since the effects of adaptation were successfully detected at the perceptual, behavioral, and neural levels, it is concluded that this protocol provides a promising methodology for studying adaptation to left-right reversed audition, and is an effective tool for uncovering the adaptability of humans to a novel environments in the auditory domain.

The present study proposes a protocol to investigate the adaptation to left-right reversed audition achieved only by wearable devices, using neuroimaging, which can be an effective tool for uncovering the adaptability of humans to a novel environment in the auditory domain.

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the ...

Using a Classroom-Based Deese Roediger McDermott Paradigm to Assess the Effects of Imagery on False Memories

Associated word list procedures can elicit false memories in predictable ways by inducing associative processing, thus making it harder to monitor the accuracy of memories. The purpose of the method presented here was to induce false memories using lists of either semantically or phonologically related words and to assess the effects of imagery instructions on the recall and recognition of those false memories. To do this, we used a modified version of the Deese Roediger McDermott (DRM) paradigm. We adapted word lists from previous DRM studies to suit imagery procedures and created an automated presentation to present the word lists in classroom settings. We then recruited undergraduate classes and instructed some of the classes to create mental images of the list words as they were being presented, while instructing the other classes to simply remember the words. The automated presentation presented word lists to participants, one word at a time, alternating between phonologically and semantically related lists. Participants used paper-pencil recall packets to immediately recall list items, complete a distractor activity, and take a subsequent final recognition test. Often, participants immediately recalled and later recognized words that were related to the list items but were not actually presented; these are known as critical lures and indicate a false memory. The protocol detailed here describes a four-step procedure - list presentation, immediate recall, distractor phase, and final recognition - that can assess the effects of list type and imagery instruction within the DRM paradigm on memory. The automated nature of the list presentation provides the ability to systematically vary variables of interest, and the paper and pencil method of data collection affords an easily accessible method for collecting data in classroom settings. The protocol also offers options to modify the procedure to a more traditional DRM paradigm without imagery and/or list type manipulations. The use of this protocol can provide results relevant to both classroom learning and cognitive science principles.

The method presented here induced false memories using lists of related words and also assessed the effects of imagery instructions on the recall and recognition of those false memories. This protocol details a modified version of the Deese Roediger McDermott (DRM) paradigm.

Associated word list procedures can elicit false memories in predictable ways by inducing associative processing, thus making it harder to monitor the ...