Name: postural organization of gait initiation for biomechanical analysis using force platform recordings
Uploaded: 2022-07-26T14:30:00
Description: Watch this Scientific Journal Video about Postural Organization of Gait Initiation for Biomechanical Analysis Using Force Platform Recordings at JoVE.com

The authors would like to thank the ANRT and the LADAPT.

The protocol described below follows the guideline of the human research ethics committee of the Universit&#233; Paris-Saclay. Participants approved and signed a consent form.
1. Participants
<ol>
	<li>Include at least 15 healthy young adult participants in the experiment (aged 20 to 40 years old). 
	NOTE: This recommended number of subjects corresponds to what is classically considered in the literature on GI.</li>
	<li>Exclude participants with walking aids, visual, h...

The goal of this paper was to provide scholars, clinicians, and higher education students information on the method (the &#34;global&#34; method) used in our laboratory to investigate the biomechanical organization of gait initiation (GI). Critical steps of the protocol, limitations of the method, and alternative methods and applications are discussed below.
A critical step in the protocol is the detection of the timing events of GI (i.e., APA onset, swing heel-off and toe-off, and rear foot-o...

Gait initiation (GI), the transient phase between orthograde posture and steady-state locomotion, is a functional task and an experimental paradigm that is classically used in the literature to investigate postural control during a complex motor task requiring simultaneous whole-body propulsion and stability1. Patients with neurological conditions, such as Parkinson&#39;s disease2, stroke3, progressive supranuclear palsy4, and &#34;higher level gait disorders&#34;5, are known to have difficulty initiating gait, which exposes them to an increased risk of falling. It is therefore important for both basic and clinical sciences to develop concepts and methods to gain insight into the postural control mechanisms in play during gait initiation, to gain scientific knowledge and a better understanding of the pathophysiology of gait and balance disorders and be able to remediate them through adequate interventions.
The concept of biomechanical organization of gait initiation is described below, and the classical method designed to investigate this organization is detailed in the protocol section. GI can be subdivided into three successive phases: the &#34;anticipatory postural adjustments&#34; (APA) phase corresponding to the dynamic phenomena occurring in the whole body before swing heel-off, the &#34;unloading&#34; phase (between swing heel-off and toe-off), and the &#34;swing&#34; phase that ends at the time of swing foot contacting the support surface. This classical subdivision of the GI process originates from the pioneering studies of Belenkii et al.6 and others7,8, focusing on the coordination between posture and movement during voluntary arm raising to horizontal in the erect posture. In this paradigm, the body segments that are directly involved in the arm raising correspond to the &#34;focal&#34; chain, while the body segments that are interposed between the proximal part of the focal chain and the support surface correspond to the &#34;postural&#34; chain9. These authors reported that raising the arm was systematically preceded by dynamic and electromyographical phenomena in the postural chain, which they called &#34;anticipatory postural adjustments&#34;. For GI, swing heel-off (or swing toe-off, depending on the authors) is considered as the onset of gait movement10. Consequently, the dynamic phenomena occurring before this instant correspond to APA, and the swing limb is considered to be a component of the focal chain11. This statement is in agreement with the classical conception of movement biomechanical organization, according to which any motor act must involve a focal and a postural component12,13.
From a biomechanical point of view, APA associated with GI manifests as a backward and mediolateral (swing leg side-oriented) displacement of the center of pressure, which acts to propel the center of gravity in the opposite direction - forward and toward the stance leg side. The larger the anticipatory backward center of pressure displacement, the higher the motor performance in terms of the forward center of gravity velocity at foot contact10,14. In addition, by propelling the center of gravity toward the stance leg side, APA contribute to maintain mediolateral stability during the swing phase of GI1,15,16,17. The current literature stresses that alteration in this anticipatory control of stability is a major source of falls in the elderly1. Stability during GI has been quantified in the literature with an adaptation of the &#34;margin of stability&#34;18, a quantity that takes into account both the velocity and the position of the center of gravity within the base of support. In addition to the development of APA, the fall of the center of gravity during the swing phase of GI under the effect of gravity has been reported to be actively braked by the triceps surae of the stance leg. This active braking facilitates stability maintenance after foot contact, allowing a smooth foot landing on the support surface4.
The goal of this paper is to provide scholars, clinicians, and higher education students information on the material and method developed in our laboratory to investigate the postural organization of GI via a biomechanical approach. This &#34;global&#34; method (which can also be assimilated to a &#34;kinetic&#34; method for the reasons detailed below) was initiated by Breni&#232;re and collaborators10,19. It is based on the direct principle of mechanics to calculate both the acceleration of the center of gravity, as well as the instantaneous positions of the center of pressure. Each of these points is a global expression specific to the movement.
One is the instantaneous expression of the movements of all body segments related to the purpose of the movement (the center of gravity; e.g., the progression velocity of the body during GI); the other (the center of pressure) is the expression of the support conditions necessary to reach this objective. The instantaneous positions of these two points reflect the posturo-dynamic conditions to be satisfied for gait initiation. The force platform is the appropriate instrument for this model because it allows the direct measurement of the external forces and moments acting at the supporting surface during movement. It also allows the performance of natural movements and requires no special preparation.
Many factors are known to influence the postural organization of GI, including biomechanical, (neuro)physiological, psychological, environmental, and cognitive factors1,20. This paper focuses on the influence of two factors - velocity of GI and temporal pressure - and provides typical values obtained in healthy young adults.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Force platform(s)</td>
			<td>AMTI</td>
			<td></td>
			<td>One large [120 cm x 60 cm] or two small [60 cm x 40 cm] force platform(s)</td>
		</tr>
		<tr>
			<td>Python or Matlab</td>
			<td>Python or MathWorks</td>
			<td></td>
			<td>Programming language for the computation of experimental variables</td>
		</tr>
		<tr>
			<td>Qualisys track manage</td>
			<td>Qualisys</td>
			<td></td>
			<td>Software for the synchronization of the force platform(s), the recording and the on-line visualization of raw biomechanical traces (3D forces and moments)</td>
		</tr>
		<tr>
			<td>Visual3D</td>
			<td>C-Motion Inc</td>
			<td></td>
			<td>Software for the processing of raw biomechanical traces (low-pass filtering)</td>
		</tr>
	</tbody>
</table>

postural organization of gait initiation for biomechanical analysis using force platform recordings

Gait initiation (GI), the transient phase between orthograde posture and steady-state locomotion, is a functional task and an experimental paradigm that is classically used in the literature to obtain insight into the basic postural mechanisms underlying body motion and balance control. Investigating GI has also contributed to a better understanding of the physiopathology of postural disorders in elderly and neurological participants (e.g., patients with Parkinson&#39;s disease). As such, it is recognized to have important clinical implications, especially in terms of fall prevention.
This paper aims to provide scholars, clinicians, and higher education students information on the material and method developed to investigate GI postural organization via a biomechanical approach. The method is based on force platform recordings and the direct principle of mechanics to compute the kinematics of the center of gravity and center of pressure. The interaction between these two virtual points is a key element in this method since it determines the conditions of stability and whole-body progression. The protocol involves the participant initially standing immobile in an upright posture and starting to walk until the end of an at least 5 m track.
It is recommended to vary the GI velocity (slow, spontaneous, fast) and the level of temporal pressure - gait may be initiated as soon as possible after the deliverance of a departure signal (high level of temporal pressure) or when the participant feels ready (low level of temporal pressure). Biomechanical parameters obtained with this method (e.g., duration and amplitude of anticipatory postural adjustments, step length/width, performance, and stability) are defined, and their computation method is detailed. In addition, typical values obtained in healthy young adults are provided. Finally, critical steps, limitations, and significance of the method with respect to the alternative method (motion capture system) are discussed.

Description of representative biomechanical time plots obtained from the force platform during gait initiation 
Whatever the level of temporal pressure or the instruction on GI velocity, swing heel-off is systematically preceded by APA. These APA can be characterized by a backward and swing leg side shift of the center of pressure (Figure 2). This anticipatory center of pressure shift promotes the acceleration of the center of gravity in the opposite direction (i.e., fo...

Watch this Scientific Journal Video about Postural Organization of Gait Initiation for Biomechanical Analysis Using Force Platform Recordings at JoVE.com

Postural Organization of Gait Initiation for Biomechanical Analysis Using Force Platform Recordings

This paper describes the material and method developed to investigate the postural organization of gait initiation. The method is based on force platform recordings and on the direct principle of mechanics to compute center of gravity and center of pressure kinematics.

Gait initiation (GI), the transient phase between orthograde posture and steady-state locomotion, is a functional task and an experimental paradigm that ...

This method make it possible to obtain anticipatory postural adjustment, or APA. APA reflects the functioning of the central level system. It therefore make it possible to insert the question of what the APA, the MOS, and the BI look like.The main advantage of this technique is precision. Moreover, it is a technique that has the possibility of being adapted, and less tiring for patient who have motor fixation, for example. The implication of this technique can be extended to all working pathology.For Parkinson disease, it is especially interesting because it could help to find the mechanical marker for providing an earlier diagnosis or detecting subclinical balance and gait disorder. This method is applicable in certain sport, such as running start or sport with gesture with a still position, like shooting basketball or baseball. I think a person who have never practiced this method can also perform well.You just have to be rigorous. To begin, ask the participants to stand barefoot and immobile on a force platform in their natural upright posture. With the arms hanging loosely against their sides and their gaze directed to a target at eye level at least five meters away.Determine the participant's preferential starting leg by pushing lightly against their back while in the initial posture with their eyes closed to provoke a step forward. Explain to the participants that the tasks they are to perform is to initiate gait from the standing posture with the preferred leg. To continue walking to the end of the track and then return quietly to the initial standing posture.Explain that gait is to be initiated following two successive signals, a preparatory signal and a departure signal. Then explain the instructions on velocity and temporal pressure. Trigger data acquisition from the force platform and deliver the first or preparatory signal to the participants.Instruct them to stand immobile and avoid anticipating gait initiation at this first signal. Ensure that the participants are visually immobile. Check the immobility online with the time plots of the antero-posterior or medial lateral center of pressure displacement.Then deliver the second or departure signal. Vary the conditions of temporal pressure imposed on gait initiation, and then vary the conditions of gait initiation velocity. Instruct the participants to perform a series of 10 successive trials in each experimental condition and impose a rest of at least two minutes between successive conditions to avoid the effects of fatigue.In each condition, allow the participants to perform two familiarization trials before the recordings. Finally, stop the data acquisition once the participant has left the force platform. The biomechanical time plots obtained from the force platform during gait initiation and selected spacial temporal variables are shown here.Gait was initiated quickly in a reaction time condition. The acceleration of the center of gravity and the velocity of the center of gravity along the antero-posterior, medial lateral and vertical directions are depicted here. The displacement of the center of pressure along the antero-posterior and medial lateral directions is also depicted in this figure.The onset of anticipatory postural adjustments, or APA, along the media lateral and antero-posterior directions, time of swing heel off, time of swing toe off, time of swing foot contact and time of rear foot off are shown here. The figure also depicts the representative values of different temporal variables like time windows for APA, unloading phase and swing phase of gait initiation. The representative values of different spatial variables like antero-posterior velocity of the center of gravity at foot off and foot contact.The maximal anticipatory center of pressure displacement, step length, step width, peak downward center of gravity velocity and vertical center of gravity velocity at swing foot contact time are shown here. The most important part of the foot occur is to ensure that the participant are visibly immobile in order to have a good baseline. Beside it is also important that the participant perform two formalization to have before the recording.This will allow us to have a really better trial during the passages. So kinematic method can be used to add data during stationary working. I think might interest some researchers and think how they can use this technique in the field, from engineering to clinical replication.

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High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex

Computation in the brain relies on neurons responding appropriately to their synaptic inputs. Neurons differ in their complement and distribution of membrane ion channels that determine how they respond to synaptic inputs. However, the relationship between these cellular properties and neuronal function in behaving animals is not well understood. One approach to this problem is to investigate topographically organized neural circuits in which the position of individual neurons maps onto information they encode or computations they carry out1. Experiments using this approach suggest principles for tuning of synaptic responses underlying information encoding in sensory and cognitive circuits2,3.
The topographical organization of spatial representations along the dorsal-ventral axis of the medial entorhinal cortex (MEC) provides an opportunity to establish relationships between cellular mechanisms and computations important for spatial cognition. Neurons in layer II of the rodent MEC encode location using grid-like firing fields4-6. For neurons found at dorsal positions in the MEC the distance between the individual firing fields that form a grid is on the order of 30 cm, whereas for neurons at progressively more ventral positions this distance increases to greater than 1 m. Several studies have revealed cellular properties of neurons in layer II of the MEC that, like the spacing between grid firing fields, also differ according to their dorsal-ventral position, suggesting that these cellular properties are important for spatial computation2,7-10.
Here we describe procedures for preparation and electrophysiological recording from brain slices that maintain the dorsal-ventral extent of the MEC enabling investigation of the topographical organization of biophysical and anatomical properties of MEC neurons. The dorsal-ventral position of identified neurons relative to anatomical landmarks is difficult to establish accurately with protocols that use horizontal slices of MEC7,8,11,12, as it is difficult to establish reference points for the exact dorsal-ventral location of the slice. The procedures we describe enable accurate and consistent measurement of location of recorded cells along the dorsal-ventral axis of the MEC as well as visualization of molecular gradients2,10. The procedures have been developed for use with adult mice (&gt; 28 days) and have been successfully employed with mice up to 1.5 years old. With adjustments they could be used with younger mice or other rodent species. A standardized system of preparation and measurement will aid systematic investigation of the cellular and microcircuit properties of this area.

We describe procedures for preparation and electrophysiological recording from brain slices that maintain the dorsal-ventral axis of the medial entorhinal cortex (MEC). Because neural encoding of location follows a dorsal-ventral organization within the MEC, these procedures facilitate investigation of cellular mechanisms important for navigation and memory.

Computation in the brain relies on neurons responding appropriately to their synaptic inputs. Neurons differ in their complement and distribution of ...

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date this tool has been widely used to study the regulation of cellular proliferation, differentiation and neuronal migration especially in the developing cerebral cortex 6-8. Here we detail our protocol to electroporate in utero the cerebral cortex and the hippocampus and provide evidence that this approach can be used to study dendrites and spines in these two cerebral regions.
Visualization and manipulation of neurons in primary cultures have contributed to a better understanding of the processes involved in dendrite, spine and synapse development. However neurons growing in vitro are not exposed to all the physiological cues that can affect dendrite and/or spine formation and maintenance during normal development. Our knowledge of dendrite and spine structures in vivo in wild-type or mutant mice comes mostly from observations using the Golgi-Cox method 9. However, Golgi staining is considered to be unpredictable. Indeed, groups of nerve cells and fiber tracts are labeled randomly, with particular areas often appearing completely stained while adjacent areas are devoid of staining. Recent studies have shown that IUE of fluorescent constructs represents an attractive alternative method to study dendrites, spines as well as synapses in mutant / wild-type mice 10-11 (Figure 1A). Moreover in comparison to the generation of mouse knockouts, IUE represents a rapid approach to perform gain and loss of function studies in specific population of cells during a specific time window. In addition, IUE has been successfully used with inducible gene expression or inducible RNAi approaches to refine the temporal control over the expression of a gene or shRNA 12. These advantages of IUE have thus opened new dimensions to study the effect of gene expression/suppression on dendrites and spines not only in specific cerebral structures (Figure 1B) but also at a specific time point of development (Figure 1C).
Finally, IUE provides a useful tool to identify functional interactions between genes involved in dendrite, spine and/or synapse development. Indeed, in contrast to other gene transfer methods such as virus, it is straightforward to combine multiple RNAi or transgenes in the same population of cells. 
In summary, IUE is a powerful method that has already contributed to the characterization of molecular mechanisms underlying brain function and disease and it should also be useful in the study of dendrites and spines.

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

This article describes in detail a protocol to electroporate in utero the cerebral cortex and the hippocampus at E14.5 in mice. We also show that this is a valuable method to study dendrites and spines in these two cerebral regions.

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron. When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased. This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue. A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation. Paired whole cell recordings are often perceived as too challenging to perform. While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed. The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings. While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation. In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology.

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct ...

Ex Vivo Imaging of Postnatal Cerebellar Granule Cell Migration Using Confocal Macroscopy

During postnatal development, immature granule cells (excitatory interneurons) exhibit tangential migration in the external granular layer, and then radial migration in the molecular layer and the Purkinje cell layer to reach the internal granular layer of the cerebellar cortex. Default in migratory processes induces either cell death or misplacement of the neurons, leading to deficits in diverse cerebellar functions. Centripetal granule cell migration involves several mechanisms, such as chemotaxis and extracellular matrix degradation, to guide the cells towards their final position, but the factors that regulate cell migration in each cortical layer are only partially known. In our method, acute cerebellar slices are prepared from P10 rats, granule cells are labeled with a fluorescent cytoplasmic marker and tissues are cultured on membrane inserts from 4 to 10 hr before starting real-time monitoring of cell migration by confocal macroscopy at 37 &#176;C in the presence of CO2. During their migration in the different cortical layers of the cerebellum, granule cells can be exposed to neuropeptide agonists or antagonists, protease inhibitors, blockers of intracellular effectors or even toxic substances such as alcohol or methylmercury to investigate their possible role in the regulation of neuronal migration.

Ex Vivo Imaging of Postnatal Cerebellar Granule Cell Migration Using Confocal Macroscopy

During postnatal cerebellum development, immature granule cells originating from the germinal zone exhibit distinct modalities of migration to reach their final destination and to establish neuronal networks. This protocol describes the preparation of cerebellar slices and the confocal macroscopic approach used to investigate the factors that regulate neuronal migration.

During postnatal development, immature granule cells (excitatory interneurons) exhibit tangential migration in the external granular layer, and then ...

Peering at Brain Polysomes with Atomic Force Microscopy

The translational machinery, i.e., the polysome or polyribosome, is one of the biggest and most complex cytoplasmic machineries in cells. Polysomes, formed by ribosomes, mRNAs, several proteins and non-coding RNAs, represent integrated platforms where translational controls take place. However, while the ribosome has been widely studied, the organization of polysomes is still lacking comprehensive understanding. Thus much effort is required in order to elucidate polysome organization and any novel mechanism of translational control that may be embedded. Atomic force microscopy (AFM) is a type of scanning probe microscopy that allows the acquisition of 3D images at nanoscale resolution. Compared to electron microscopy (EM) techniques, one of the main advantages of AFM is that it can acquire thousands of images both in air and in solution, enabling the sample to be maintained under near physiological conditions without any need for staining and fixing procedures. Here, a detailed protocol for the accurate purification of polysomes from mouse brain and their deposition on mica substrates is described. This protocol enables polysome imaging in air and liquid with AFM and their reconstruction as three-dimensional objects. Complementary to cryo-electron microscopy (cryo-EM), the proposed method can be conveniently used for systematically analyzing polysomes and studying their organization.

While ribosome structure has been extensively characterized, the organization of polysomes is still understudied. To overcome this lack of knowledge, we present here a detailed preparation protocol for accurate imaging of mammalian polysomes by atomic force microscopy (AFM) in air and liquid.

The translational machinery, i.e., the polysome or polyribosome, is one of the biggest and most complex cytoplasmic machineries in cells. Polysomes, ...

A Visual Guide to Sorting Electrophysiological Recordings Using 'SpikeSorter'

Few stand-alone software applications are available for sorting spikes from recordings made with multi-electrode arrays. Ideally, an application should be user friendly with a graphical user interface, able to read data files in a variety of formats, and provide users with a flexible set of tools giving them the ability to detect and sort extracellular voltage waveforms from different units with some degree of reliability. Previously published spike sorting methods are now available in a software program, SpikeSorter, intended to provide electrophysiologists with a complete set of tools for sorting, starting from raw recorded data file and ending with the export of sorted spikes times. Procedures are automated to the extent this is currently possible. The article explains and illustrates the use of the program. A representative data file is opened, extracellular traces are filtered, events are detected and then clustered. A number of problems that commonly occur during sorting are illustrated, including the artefactual over-splitting of units due to the tendency of some units to fire spikes in pairs where the second spike is significantly smaller than the first, and over-splitting caused by slow variation in spike height over time encountered in some units. The accuracy of SpikeSorter's performance has been tested with surrogate ground truth data and found to be comparable to that of other algorithms in current development.

The article shows how to use the program SpikeSorter to detect and sort spikes in extracellular recordings made with multi-electrode arrays.

Few stand-alone software applications are available for sorting spikes from recordings made with multi-electrode arrays. Ideally, an application should be ...

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 ...

Preparation of Acute Human Hippocampal Slices for Electrophysiological Recordings

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for sudden death. Despite an abundance of available treatment options, about 30% of patients are drug-resistant. Several novel therapeutics have been developed using animal models, though the rate of drug-resistant patients remains unaltered. One of probable reasons is the lack of translation between rodent models and humans, such as a weak representation of human pharmacoresistance in animal models. Resected human brain tissue as a preclinical evaluation tool has the advantage to bridge this translational gap. Described here is a method for high quality preparation of human hippocampal brain slices and subsequent stable induction of epileptiform activity. The protocol describes the induction of burst activity during application of 8 mM KCl and 4-aminopyridin. This activity is sensitive to established AED lacosamide or novel antiepileptic candidates, such as dimethylethanolamine (DMEA). In addition, the method describes induction of seizure-like events in CA1 of human hippocampal brain slices by reduction of extracellular Mg2+ and application of bicuculline, a GABAA receptor blocker. The experimental set-up can be used to screen potential antiepileptic substances for their effects on epileptiform activity. Furthermore, mechanisms of action postulated for specific compounds can be validated using this approach in human tissue (e.g., using patch-clamp recordings). To conclude, investigation of vital human brain tissue ex vivo (here, resected hippocampus from patients suffering from temporal lobe epilepsy) will improve the current knowledge of physiological and pathological mechanisms in the human brain.

The presented protocol describes the transport and preparation of resected human hippocampal tissue with the ultimate goal to use vital brain slices as a preclinical evaluation tool for potential antiepileptic substances.

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for ...

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 ...