Subventions du NIH NS038079 (MSB &amp; JCB), des subventions du NIH AG032518 (MSB &amp; JCB), État de New York CoRE 19772 (MSB &amp; JCB), NIH NS069537 (ARF) et le NIH NS053059 (ARF).

<ol>
	<li>Anderson, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+recovery:+priorities+of+the+spinal+cord-injured+population.">Targeting recovery: priorities of the spinal cord-injured population.</a> J Neurotrauma. 21 (10), 1371-1383 (2004).</li><li>Anderson, K. D., Sharp, K. G., Steward, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+cervical+contusion+spinal+cord+injury+in+rats.">Bilateral cervical contusion spinal cord injury in rats.</a> Exp. Neurol. 220 (1), 9-22 (2009).</li><li>Ballermann, M., Metz, G. A., McKenna, J. E., Klassen, F., Whishaw, Q. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pasta+matrix+reaching+task:+a+simple+test+for+measuring+skilled+reaching+distance,+direction,+and+dexterity+in+rats.">The pasta matrix reaching task: a simple test for measuring skilled reaching distance, direction, and dexterity in rats.</a> J Neurosci Methods. 106, 39-45 (2001).</li><li>Bertelli, J. A., Mira, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+evaluating+methods+in+the+objective+clinical+assessment+of+motor+function+after+experimental+brachial+plexus+reconstruction+in+the+rat.">Behavioral evaluating methods in the objective clinical assessment of motor function after experimental brachial plexus reconstruction in the rat.</a> J Neurosci Methods. 46, (1993).</li><li>Gensel, J. C., Tovar, C. A., Hamers, F. P., Deibert, R. J., Beattie, M. S., Bresnahan, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+histological+characterization+of+unilateral+cervical+spinal+cord+contusion+injury+in+rats.">Behavioral and histological characterization of unilateral cervical spinal cord contusion injury in rats.</a> J Neurotrauma. 23, 36-54 (2006).</li><li>Girgis, J., Merrett, D., Kirkland, S., Metz, G. A., Verge, V., Fouad, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reaching+training+in+rats+with+spinal+cord+injury+promotes+plasticity+and+task+specific+recovery.">Reaching training in rats with spinal cord injury promotes plasticity and task specific recovery.</a> Brain. 130 (PT 11), 2993-3003 (2007).</li><li>Metz, G. A., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skilled+reaching+an+action+pattern:+stability+in+rat+(Rattus+norvegicus)+grasping+movements+as+a+function+of+changing+food+pellet+size.">Skilled reaching an action pattern: stability in rat (Rattus norvegicus) grasping movements as a function of changing food pellet size.</a> Behav Brain Res. 116 (2), 111-122 (2000).</li><li>Montoya, C. P., Campbell-Hope, L. J., Pemberton, K. D., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+"staircase+test":+a+measure+of+independent+forelimb+reaching+and+grasping+abilities+in+rats.">The "staircase test": a measure of independent forelimb reaching and grasping abilities in rats.</a> J Neurosci Methods. 36 (2-3), 219-228 (1991).</li><li>Pearse, D. D., Lo, T. P. J. r., Cho, K. S., Lynch, M. P., Garg, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histopathological+and+behavioral+characterization+of+a+novel+cervical+spinal+cord+displacement+contusion+injury+in+the+rat.">Histopathological and behavioral characterization of a novel cervical spinal cord displacement contusion injury in the rat.</a> J Neurotrauma. 22 (6), 680-702 (2005).</li><li>Whishaw, I. Q., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+structure+of+skilled+forelimb+reaching+in+the+rat:+a+proximally+driven+movement+with+a+single+distal+rotatory+component.">The structure of skilled forelimb reaching in the rat: a proximally driven movement with a single distal rotatory component.</a> Behav. Brain Res. 41 (1), 49-59 (1990).</li></ol>

L&#39;échelle de Irvine, Beatties et Bresnahan forelimb (IBB) est une échelle de 10 points de récupération ordinale de la progression des membres antérieurs. L&#39;IBB analyses de nombreuses caractéristiques de l&#39;utilisation des membres antérieurs à la fois proximale et distale lors de la consommation de deux morceaux entiers de céréales mis à la tâche, une forme sphérique et une céréale en forme de beignet. Vidéos des animaux qui mangent des céréales sont évalués pour les comportements les plus communs des membres antérieurs (y compris la position commune, le soutien objet, le mouvement et la technique de chiffres préhension) utilisés pour les deux formes, puis un score est attribué IBB basée sur une combinaison de ces comportements choisis pour représenter les plus grande quantité de la récupération des membres antérieurs. Les évaluations doivent être faites avec des céréales qui sont d&#39;une taille uniforme et de la forme: ainsi, les animaux ne devraient pas être marqué en mangeant les morceaux brisés de céréales. Les animaux doivent remplir l&#39;ensemble des comportements qui décrivent chaque point ou l&#39;animal est marqué le point le plus proche de déficit. L&#39;IBB a été créé pour l&#39;évaluation de la récupération des membres antérieurs suivants induite expérimentalement cervicale SCI. Les premières études dans notre laboratoire a constaté que, comme avec d&#39;autres tests comportementaux, la récupération évaluée à l&#39;aide de l&#39;IBB score est sensible à la gravité des blessures. L&#39;utilisation de l&#39;IBB ne peut pas être limitée aux cervicales SCI, mais peut aussi être adapté à d&#39;autres formes de lésion nerveuse centrale et périphérique qui affectent la fonction des membres antérieurs appropriée.

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<td>Froot Loops</td>
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<td>Doughnut-shaped cereal Available in: India, Australia, Canada, USA, Germany, Middle east and South America. 
Diameter: ~ 81 mm</td>
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<td>Reese's Pieces</td>
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<td>Herseys Co.</td>
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<td>Spherical-shaped cereal Available in: USA 
Diameter: ~ 63 mm</td>
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a novel method for assessing proximal and distal forelimb function in the rat: the irvine, beatties and bresnahan (ibb) forelimb scale

Plusieurs modèles expérimentaux de lésions de la moelle épinière cervicale (SCI) ont été développés récemment pour évaluer les conséquences des dommages à ce niveau de la moelle épinière (Pearse et al., 2005, Gensel et al., 2006, Anderson et al., 2009) , comme la majorité des humains se produisent ici SCI (Young, 2010; www.sci-info-pages.com). Déficits comportementaux comprennent la perte de fonction des membres antérieurs en raison de dommages à la matière blanche affectant à la fois moteur de descendants et ascendants systèmes sensoriels, et de la matière grise contenant les circuits segmentaire pour le traitement des informations sensorielles et de sortie du moteur pour le membre antérieur. En outre, une priorité clé pour des patients humains atteints du col SCI est la restauration des mains / bras de fonction (Anderson, 2004). Ainsi, les mesures des résultats qui permettent d&#39;évaluer la fonction des membres antérieurs à la fois proximale et distale sont nécessaires. Bien qu&#39;il existe plusieurs tests de comportement qui sont sensibles aux différents aspects de la récupération des membres antérieurs dans des modèles expérimentaux de cancer du col SCI (Girgis et coll., 2007, Gensel et al., 2006, Ballerman et al., 2001, Metz et Whishaw, 2000, Bertelli et Mira, 1993, Montoya et al., 1991, Whishaw et Pellis, 1990), quelques techniques de fournir des informations détaillées sur la récupération de la motricité fine et les mouvements chiffres. La technique de mesure actuelle, Irvine, Beatties et Bresnahan forelimb échelle (IBB), peut détecter récupération de la fonction des membres antérieurs à la fois proximale et distale, y compris les mouvements de chiffres lors d&#39;un comportement naturel qui ne nécessite pas une formation poussée ou une privation pour renforcer la motivation. L&#39;IBB a été générée par l&#39;observation de la récupération après une SCI unilatérale C6, et implique l&#39;enregistrement vidéo d&#39;animaux de manger deux céréales de forme différente (sphérique et beignets) d&#39;une taille uniforme. Ces vidéos ont ensuite été utilisés pour évaluer les caractéristiques d&#39;utilisation des membres antérieurs, tels que la position commune, support de l&#39;objet, le mouvement chiffres et la technique de préhension. L&#39;IBB, comme d&#39;autres membres antérieurs des tâches comportementales, montre un modèle cohérent de récupération qui est sensible à la gravité des blessures. Par ailleurs, l&#39;échelle IBB pourrait être utilisée pour évaluer la récupération après d&#39;autres types de blessures que la fonction des membres antérieurs d&#39;impact normale.

Watch this Scientific Journal Video about A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale at JoVE.com

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale

Ici, nous allons décrire un test de comportement chez les rongeurs qui peuvent détecter récupération de la fonction des membres antérieurs à la fois proximale et distale, y compris les mouvements de chiffres lors d&#39;un comportement naturel qui ne nécessite pas une formation poussée ou une privation pour renforcer la motivation.

Une nouvelle méthode pour évaluer la fonction forelimb proximales et distales chez le rat: l&#39;Irvine, Beatties et Bresnahan (IBB) Echelle forelimb

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the ...

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Neuroscience

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan (IBB) Forelimb Scale

19.7K vues.  University of California, San Francisco. Ici, nous allons décrire un test de comportement chez les rongeurs qui peuvent détecter récupération de la fonction des membres antérieurs à la fois proximale et distale, y compris les mouvements de chiffres lors d'un comportement naturel qui ne nécessite pas une formation poussée ou une privation pour renforcer la motivation.

Vidéo: A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

Appareil et méthode pour Novel Drug Renforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

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Neurosciences

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended upon this work to develop a simple quantitative measure of forepaw dexterity that is sensitive to lateralized impairments and age-dependent changes. Rodents learn skillful forepaw and digit movements to manage thin pasta pieces, which they eagerly consume. We have previously described methods for quantifying vermicelli handling in rats and showed that the measures are very sensitive to forelimb impairments resulting from unilateral ischemic lesions, middle cerebral artery occlusions and unilateral striatal dopamine depletion [Allred, R.P., Adkins, D.L., Woodlee, M.T., Husbands, L.C., Maldonado M.A., Kane, J.R., Schallert, T. &amp; Jones, T.A. The Vermicelli Handling Test: a simple quantitative measure of dexterous forepaw function in rats. J. Neurosci. Methods 170, 229-244 (2008)]. Here we present a more detailed protocol for this test in rats and compare it with a newly developed version for mice, the Capellini Handling Test. Rats and mice are videotaped while handling short lengths of uncooked vermicelli or capellini pasta, respectively, with a camera positioned to optimize the view of paw movements. Slow motion video playback allows for the identification of forepaw adjustments, defined as any distinct removal and replacement of the paw, or of any number of digits, on the pasta piece after eating commences. Forepaw adjustments per piece are averaged over trials per each testing session. Repeated testing permits sensitive quantitative analysis of changes in forepaw dexterity over time. Protocols for pre-testing habituation and handling practice, as well as procedures for characterizing atypical handling patterns, are described. Because rats and mice perform the pasta handling tests slightly differently, species-specific differences in administration and scoring of these tests are highlighted. All animal use was in accordance with protocols approved by the University of Texas at Austin Animal Care and Use Committee.

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

Les vermicelles et tests de manipulation Capellini: Simple mesures quantitatives de la fonction patte habile dans les rats et les souris

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended ...

Cryopreservation of Cortical Tissue Blocks for the Generation of Highly Enriched Neuronal Cultures

In this study, we outline a standardized protocol for the successful cryopreservation and thawing of cortical brain tissue blocks to generate highly enriched neuronal cultures. For this protocol the freezing medium used is 10% dimethyl sulfoxide (DMSO) diluted in Hank's Buffered Salt Solution (HBSS). Blocks of cortical tissue are transferred to cryovials containing the freezing medium and slowly frozen at -1&deg;C/min in a rate-controlled freezing container. Post-thaw processing and dissociation of frozen tissue blocks consistently produced neuronal-enriched cultures which exhibited rapid neuritic growth during the first 5 days in culture and significant expansion of the neuronal network within 10 days. Immunocytochemical staining with the astrocytic marker glial fibrillary acidic protein (GFAP) and the neuronal marker beta-tubulin class III, revealed high numbers of neurons and astrocytes in the cultures. Generation of neural precursor cell cultures after tissue block dissociation resulted in rapidly expanding neurospheres, which produced large numbers of neurons and astrocytes under differentiating conditions. This simple cryopreservation protocol allows for the rapid, efficient, and inexpensive preservation of cortical brain tissue blocks, which grants increased flexibility for later generation of neuronal, astrocyte, and neuronal precursor cell cultures.

Here, we describe a method for efficient cryopreservation and thawing of cortical brain tissue blocks to generate highly enriched neuronal cultures. This simple protocol provides flexibility for later generation of neuronal, astrocyte, and neuronal precursor cell cultures.

La cryoconservation de tissu cortical Blocs pour la génération des cultures de neurones hautement enrichi

In this study, we outline a standardized protocol for the successful cryopreservation and thawing of cortical brain tissue blocks to generate highly ...

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of behaviors from the spiking activity of its neurons. One of the most effective methods to monitor spiking activity from a large number of neurons in multiple local neuronal circuits simultaneously is by using silicon electrode arrays1-3.
Action potentials produce large transmembrane voltage changes in the vicinity of cell somata. These output signals can be measured by placing a conductor in close proximity of a neuron. If there are many active (spiking) neurons in the vicinity of the tip, the electrode records combined signal from all of them, where contribution of a single neuron is weighted by its 'electrical distance'. Silicon probes are ideal recording electrodes to monitor multiple neurons because of a large number of recording sites (+64) and a small volume. Furthermore, multiple sites can be arranged over a distance of millimeters, thus allowing for the simultaneous recordings of neuronal activity in the various cortical layers or in multiple cortical columns (Fig. 1). Importantly, the geometrically precise distribution of the recording sites also allows for the determination of the spatial relationship of the isolated single neurons4. Here, we describe an acute, large-scale neuronal recording from the left and right forelimb somatosensory cortex simultaneously in an anesthetized rat with silicon probes (Fig. 2).

Extracellular recordings of neuronal activity using silicon probes in the anesthetized rat will be described. This technique allows information to be obtained across multiple brain areas from more than 100 neurons simultaneously. It provides information with single cell resolution about neuronal ensembles dynamics in multiple local circuits.

Enregistrement à grande échelle ensembles neuronaux avec des sondes en silicone chez le rat anesthésié

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

Co-analyse de la structure du cerveau et son fonctionnement en utilisant l&#39;IRMf et imagerie de diffusion

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

A Visual Description of the Dissection of the Cerebral Surface Vasculature and Associated Meninges and the Choroid Plexus from Rat Brain

This video presentation was created to show a method of harvesting the two most important highly vascular structures, not residing within the brain proper, that support forebrain function. They are the cerebral surface (superficial) vasculature along with associated meninges (MAV) and the choroid plexus which are necessary for cerebral blood flow and cerebrospinal fluid (CSF) homeostasis. The tissue harvested is suitable for biochemical and physiological analysis, and the MAV has been shown to be sensitive to damage produced by amphetamine and hyperthermia 1,2. As well, the major and minor cerebral vasculatures harvested in MAV are of potentially high interest when investigating concussive types of head trauma. The MAV dissected in this presentation consists of the pial and some of the arachnoid membrane (less dura) of the meninges and the major and minor cerebral surface vasculature. The choroid plexus dissected is the structure that resides in the lateral ventricles as described by Oldfield and McKinley3,4,5,6. The methods used for harvesting these two tissues also facilitate the harvesting of regional cortical tissue devoid of meninges and larger cerebral surface vasculature, and is compatible with harvesting other brain tissues such as striatum, hypothalamus, hippocampus, etc. The dissection of the two tissues takes from 5 to 10 min total. The gene expression levels for the dissected MAV and choroid plexus, as shown and described in this presentation can be found at GSE23093 (MAV) and GSE29733 (choroid plexus) at the NCBI GEO repository. This data has been, and is being, used to help further understand the functioning of the MAV and choroid plexus and how neurotoxic events such as severe hyperthermia and AMPH adversely affect their function.

This video presentation shows a method of harvesting the two most important highly vascular structures that support forebrain function. They are the cerebral surface (superficial) vasculature along with associated meninges (MAV) and the choroid plexus which are necessary for cerebral blood flow and cerebrospinal fluid (CSF) homeostasis.

Une description visuelle de la dissection de la vascularisation cérébrale et de surface associées Méninges et le plexus choroïde du cerveau de rat

This video presentation was created to show a method of harvesting the two most important highly vascular structures, not residing within the brain ...

Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal cord of the majority of these fiber systems have not been thoroughly investigated in any species. Serotonergic fibers, which project to the spinal cord, have been studied in rats and opossums on histological sections and their functional significance has been deduced based on their fiber termination pattern in the spinal cord. With the development of CLARITY and CUBIC techniques, it is possible to investigate this fiber system and its distribution in the spinal cord, which is likely to reveal previously unknown features of serotonergic supraspinal pathways. Here, we provide a detailed protocol for imaging the serotonergic fibers in the mouse spinal cord using the combined CLARITY and CUBIC techniques. The method involves perfusion of a mouse with a hydrogel solution and clarification of the tissue with a combination of clearing reagents. Spinal cord tissue was cleared in just under two weeks, and the subsequent immunofluorescent staining against serotonin was completed in less than ten days. With a multi-photon fluorescent microscope, the tissue was scanned and a 3D image was reconstructed using Osirix software.

Supraspinal projections are important for pain perception and other behaviors, and serotonergic fibers are one of these fiber systems. The present study focused on the application of the combined CLARITY/CUBIC protocol to the mouse spinal cord in order to investigate the termination of these serotonergic fibers.

Imagerie fibres sérotoninergiques dans la moelle épinière de souris Utilisation du CLARITY / Technique CUBIC

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal ...

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.

Analyse cinématique 3D pour l’évaluation fonctionnelle dans le modèle de rat de la blessure d’écrasement de nerf sciatique

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

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

Une méthode de nettoyage des tissus hydrophobes pour le tissu cérébral de rat

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...

Establishment of a Rat Model of Superior Sagittal-Sinus Occlusion via a Thread-Embolism Method

The mechanisms contributing to the natural onset of cerebral venous sinus thrombosis (CVST) are mostly unknown, and a variety of uncontrollable factors are involved in the course of the disease, resulting in great limitations in clinical research. Therefore, the establishment of stable CVST animal models that can standardize a variety of uncontrollable confounding factors have helped to circumvent shortcomings in clinical research. In recent decades, a variety of CVST animal models have been constructed, but the results based on these models have been inconsistent and incomplete. Hence, in order to further explore the pathophysiological mechanisms of CVST, it is necessary to establish a novel and highly compatible animal model, which has important practical value and scientific significance for the diagnosis and treatment of CVST. In the present study, a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis was established via a thread-embolization method, and the stability and reliability of the model were verified. Additionally, we evaluated changes in cerebral venous blood flow in rats after the formation of CVST. Collectively, the SD-rat SSS-thrombosis model represents a novel CVST animal model that is easily established, minimizes trauma, yields good stability, and allows for accurately controlling ischemic timing and location.

Here, we establish a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis via a thread-embolization method, and the stability and reliability of the model were verified.

Établissement d’un modèle de rat d’occlusion sagittale-sinus supérieure par l’intermédiaire d’une méthode d’embolie filimatique

The mechanisms contributing to the natural onset of cerebral venous sinus thrombosis (CVST) are mostly unknown, and a variety of uncontrollable factors ...