Bernstein Southwell Einrichten der Geräte und geschultes die Autoren in ihrer Nutzung. Die Finanzierung wurde durch einen Zuschuss aus dem NINDS zu Paul Patterson zur Verfügung gestellt.

1. Dieses Protokoll ist am Diejenigen von Southwell et. al. (2009) und Carter et. al. (2001). <ol><li> Der Strahl Apparat besteht aus 1 Meter Balken mit einer ebenen Fläche von 12 mm oder 6 mm Breite) ruhen 50 cm über der Tischplatte auf zwei Pole. Ein Black-Box befindet sich am Ende des Balkens als der Endpunkt gesetzt. (Sperrholz und andere Arten von Holz für die Balken, Pfosten, und Box kann in den meisten Baumärkten zu finden). Nestmaterial von zu Hause aus Käfigen ist in der Black Box platziert, um die Maus, um den Zielpunkt zu gewinnen. Eine Lampe (mit 60-Watt-Glühbirne) wird verwendet, um Licht über dem Startpunkt Glanz und dient als aversiven Reiz. Nacheinander 0 cm, die einen Timer und eine auf 80 cm, dass der Timer stoppt beginnt: Die Zeit der Mitte 80 cm Kreuz wird von zwei Bewegungsmeldern (hergestellt von Visopia) gemessen. Ein Nylon-Hängematte (die von einem lokalen Stoffgeschäft erhältlich) unter dem Strahl gestreckt, ~ 7,5 cm über der Tischplatte, jede fällt Kissen. Eine Videokamera auf einem Stativ, um die Leistung Rekord aufgestellt. Der Apparat in dieser Demonstration diente vor Ort gebaut. </li><li> Eine Kohorte von mindestens 10 Mäuse für jede Bedingung ist für diesen Test empfohlen. Mäuse sind als Gruppen, wenn möglich, in einem 12 Stunden Licht (07.00 bis 19.00) / Dunkel-Zyklus untergebracht und eingewöhnt in ihre Käfige in den Verhaltenswissenschaften Anlage für mindestens 2 Wochen vor dem Beginn der Prüfung. Etwa 10 Minuten vor dem Training / Prüfung können die Mäuse in den Raum mit dem Strahl Apparat transportiert werden. </li><li> An trainingsfreien Tagen kreuzt jeder Maus die 12 mm Breite 3 mal und dann die 6 mm Breite 3 mal. Beams von größeren Breiten (dh 28 mm) kann für Mäuse, die nicht in der Lage, die schmalere Kreuz oder für größere Mäuse verwendet werden. Mäuse sind an einem Ende eines Trägers gelegt und die benötigte Zeit, um die Flucht-Box am anderen Ende überqueren (80 cm entfernt) ist durch die Bewegung Detektoren gemessen. Der Timer wird durch die Nase der Maus in die Mitte 80 cm begonnen und beendet, wenn das Tier das Ende des 80 cm. Die Mäuse Rest für 10 min in ihre Käfige zwischen den Trainingseinheiten auf dem zwei Strahlen. Während dieser intertrial Zeit kann noch 3-4 Mäusen geschult, um eine effiziente Nutzung von Zeit zu machen. C57BL / 6 Mäusen in der Regel über die Balken mit einer minimalen Menge absetzen oder Abwürgen. Als sie jedoch Stall tun, schnüffeln oder umzusehen, ohne Verfahren nach vorne, sollte der Prüfer fördern die Maus, um vorwärts durch Drängen, Stoßen, Schieben oder ihn von hinten mit behandschuhten Fingern. Nachdem die Mäuse im Safe sind, werden sie einige Zeit (~ 15 Sekunden), um dort Ruhe vor dem nächsten Versuch erlaubt. Nach jedem Training werden die Mäuse direkt auf ihre Käfige zurück und wieder in ihr Gehäuse Zimmer gebracht. Zu beachten ist, können die Mäuse Stall mehr, wenn sie über ausgebildete und sind zu sehr mit dieser Aufgabe vertraut und Apparate. Auf der anderen Seite, können Schulungen für weitere Tage fortgesetzt werden, wenn die Mäuse noch nicht durchqueren den gesamten Strahl erfolgreich nach 2 Tagen Training. Wenn Behandlungen, um die Mäuse aufgetragen werden, sollte eine Baseline-Messung der Leistung vor der Behandlung erreicht werden. </li><li> Am Testtag werden die Zeiten für jeden Strahlquerschnitt aufgezeichnet. Zwei erfolgreiche Versuche, bei denen die Maus nicht auf dem Balken hatten Stall gemittelt werden. Video-Aufnahmen können zur feineren Analyse der Rutschen und anderen beobachtbaren motorischen Defiziten eingesetzt werden. Eine alternative Möglichkeit zum Strahl zu Fuß zu beurteilen ist durch eine neurologische Scoring-System 1,2, die die Unfähigkeit, um den Strahl wie eine Partitur von &quot;1&quot; Kreuz und die Fähigkeit, den Strahl normalerweise Traverse mit beiden Pfoten auf dem Balken Oberfläche und weniger als Preise zwei Fuß rutscht als &quot;7&quot;. Ein Slip ist wie der Fuß kommen von der Spitze des Strahls definiert. In diesem Indexierung, würde einen Wert von &quot;3&quot; entsprechen Kreuzung der Balken durch Ziehen Gliedmaßen. Ein Tier, das zählt 1 auf zwei Studien nicht getestet. Beschreibung der Leistung auf dem Balken im Zusammenhang mit anderen Noten können in Feeney et al., (1982) und Carter et al., (2001) gefunden werden. </li><li> Die Balken und Kasten sind Mäusedreck gereinigt und wischte sich mit Handtüchern mit 70% Ethanol und dann mit Wasser eingeweicht, bevor der nächste Strahl auf das Gerät gestellt wird. </li></ol> 2. Repräsentative Ergebnisse: <img src="/files/ftp_upload/2376/2376fig1.jpg" alt="Abbildung 1" /> Abbildung 1. Typische Kreuz mal von männlichen und weiblichen C57BL6 auf 6mm und 12mm Balken. Eine Kohorte von 9 bis 11 Wochen alte C57BL/6-NCRL WT-Mäusen wurden auf beiden Balken nach 2 Tagen Training getestet. Durchschnittliche Zeit für das Überschreiten der 12 mm Balken wurden 4,6 ± 0,4 s für Männer (n = 14) und 3,3 ± 0,3 sec für Frauen (n = 11). Times über die 6 mm Balken waren 6,8 ± 0,7 sec bei den Männern (n = 13) und 5,9 ± 0,5 sec für Frauen (n = 10) (Fehler werden als SEM berichtete).

<ol>
	<li>Feeney, D. M., Gonzalez, A., Law, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphetamine,+haloperidol,+and+experience+interact+to+affect+rate+of+recovery+after+motor+cortex+injury.">Amphetamine, haloperidol, and experience interact to affect rate of recovery after motor cortex injury.</a> Science. 217, 855-857 (1982).</li><li>Carter, R. J., Morton, J., 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=Motor+coordination+and+balance+in+rodents.">Motor coordination and balance in rodents.</a> Curr Protoc Neurosci. Chapter 8, (2001).</li><li>Wallace, J. E., Krauter, E. E., Campbell, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+and+reflexive+behavior+in+the+aging+rat.">Motor and reflexive behavior in the aging rat.</a> J Gerontol. 35, 364-370 (1980).</li><li>Brooks, S. P., 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=Tests+to+assess+motor+phenotype+in+mice:+a+user's+guide.">Tests to assess motor phenotype in mice: a user's guide.</a> Nat Rev Neurosci. 10, 519-529 (2009).</li><li>Fox, G. B., Fan, L., Levasseur, R. A., Faden, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+sensory/motor+and+cognitive+deficits+with+neuronal+apoptosis+following+controlled+cortical+impact+brain+injury+in+the+mouse.">Sustained sensory/motor and cognitive deficits with neuronal apoptosis following controlled cortical impact brain injury in the mouse.</a> J Neurotrauma. 15, 599-614 (1998).</li><li>Buccafusco, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods of behavioral analysis in neuroscience. , (2009).</li><li>Southwell, A. L., Ko, J., Patterson, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrabody+gene+therapy+ameliorates+motor,+cognitive,+and+neuropathological+symptoms+in+multiple+mouse+models+of+Huntington's+disease.">Intrabody gene therapy ameliorates motor, cognitive, and neuropathological symptoms in multiple mouse models of Huntington's disease.</a> J Neurosci. 29, 13589-13602 (2009).</li><li>Carter, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+progressive+motor+deficits+in+mice+transgenic+for+the+human+Huntington's+disease+mutation.">Characterization of progressive motor deficits in mice transgenic for the human Huntington's disease mutation.</a> J Neurosci. 19, 3248-3257 (1999).</li><li>Gulinello, M., Chen, F., Dobrenis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+deficits+in+motor+coordination+and+cognitive+dysfunction+in+a+mouse+model+of+the+neurodegenerative+lysosomal+storage+disorder,+Sandhoff+disease.">Early deficits in motor coordination and cognitive dysfunction in a mouse model of the neurodegenerative lysosomal storage disorder, Sandhoff disease.</a> Behav Brain Res. 193, 315-319 (2008).</li><li>Stanley, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mouse+beam+walking+assay+offers+improved+sensitivity+over+the+mouse+rotarod+in+determining+motor+coordination+deficits+induced+by+benzodiazepines.">The mouse beam walking assay offers improved sensitivity over the mouse rotarod in determining motor coordination deficits induced by benzodiazepines.</a> J Psychopharmacol. 19, 221-227 (2005).</li></ol>

Tierversuche wurden in Übereinstimmung mit den Richtlinien und Vorschriften her von der Caltech Institutional Animal Care und Verwenden Committee (IACUC) gesetzt durchgeführt.

Leistung auf dem Schwebebalken ist ein nützliches Maß für feine Koordination und das Gleichgewicht und wurde durch frühere Arbeiten bestätigt worden. Der Strahl testen können motorische Defizite aufgrund von Alter, Zentralnervensystem syst `Läsionen und genetische und pharmakologische Manipulationen bei jungen und älteren Nagetiere 2,3,4 erkennen. Mäuse mit kortikalen Auswirkungen Läsionen weisen oft Gegenseite rutscht auf den Balken 5,6. Der Strahl-Test wurde auch verwendet, um die motor...

<ul><li> Beams: 1-m-Streifen aus glattem Holz mit kleinen, mittleren und großen Platz (dh, 6, 12 und 28 mm breit) Querschnitte </li><li> Stativ zu halten Beginn Teil der erhöhten Strahl (~ 1,5-cm Querschnitt, 50 cm hoch) </li><li> Ziel Box (20 cm auf jeder Seite) montiert auf einem Stativ (3 cm im Querschnitt, 50 cm hoch) </li><li> Hängematten-~ 7,5 cm über Tischplatte </li><li> 60-Watt (W) Schreibtischlampe </li><li> Video-Kamera, Stativ und leere Videokassetten </li><li> Ethanol, Papierhandtücher, Reinigungsmittel </li></ul>

assessment of motor balance and coordination in mice using the balance beam

Hirn-Trauma, genetische Manipulationen oder pharmakologische Behandlungen können in Veränderungen der motorischen Fähigkeiten bei Mäusen führen. Feinmotorische Koordination und Gleichgewicht können durch den Strahl zu Fuß Assay beurteilt werden. Das Ziel dieses Tests ist für die Maus, um aufrecht zu bleiben und zu Fuß über einen erhöhten schmalen Lichtstrahl, um eine sichere Plattform. Dieser Test erfolgt über 3 aufeinander folgende Tage: 2 Tage Training und 1 Tag der Prüfung. Leistung auf dem Balken wird durch die Messung der Zeit, die für die Maus, um den Strahl und die Anzahl der Pfote gleitet, die im Prozess auftreten durchqueren quantifiziert. Hier berichten wir über das Protokoll in unserem Labor verwendet werden, und repräsentative Ergebnisse aus einer Kohorte von C57BL / 6 Mäusen. Diese Aufgabe ist besonders nützlich für die Erkennung subtiler Defizite in Motorik und Gleichgewichtssinn, die nicht durch andere motorische Tests, wie die Rotarod erkannt werden kann.

Watch this Scientific Journal Video about Assessment of Motor Balance and Coordination in Mice using the Balance Beam at JoVE.com

Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Defizite in der Feinmotorik können mit dem Schwebebalken Test festgestellt werden. Leistung auf dem Balken wird durch die Geschwindigkeit, mit der der Strahl durchquert und die Anzahl, wie oft die Maus gleitet auf dem Balken quantifiziert.

Beurteilung der Motor Balance und Koordination in Mäuse mit dem Schwebebalken

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice.  Fine motor coordination and ...

assessment-motor-balance-coordination-mice-using-balance

Research

JoVE Journal

Neuroscience

46.2K Aufrufe.  California Institute of Technology. Defizite in der Feinmotorik können mit dem Schwebebalken Test festgestellt werden. Leistung auf dem Balken wird durch die Geschwindigkeit, mit der der Strahl durchquert und die Anzahl, wie oft die Maus gleitet auf dem Balken quantifiziert.

Serie: Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

Mess-motorische Koordination in Mäuse

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

Forschung

Neurowissenschaften

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular mechanisms underlying both phenomena are not well understood and their elucidation should benefit from the study of animal models and from the design of appropriate experimental protocols. 
We describe here a methodological approach for inducing, recording and quantifying morphine-induced hyperalgesia as well as for evidencing analgesic tolerance, using the tail-immersion and tail pressure tests in wild-type mice. As shown in the video, the protocol is divided into five sequential steps. Handling and habituation phases allow a safe determination of the basal nociceptive response of the animals. Chronic morphine administration induces significant hyperalgesia as shown by an increase in both thermal and mechanical sensitivity, whereas the comparison of analgesia time-courses after acute or repeated morphine treatment clearly indicates the development of tolerance manifested by a decline in analgesic response amplitude. This protocol may be similarly adapted to genetically modified mice in order to evaluate the role of individual genes in the modulation of nociception and morphine analgesia. It also provides a model system to investigate the effectiveness of potential therapeutic agents to improve opiate analgesic efficacy.

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of na&#239;ve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

Bewertung von Morphin-induzierte Hyperalgesie und Schmerztoleranz in Mäuse mit Thermal-und Maschinen Nozizeptive Modalitäten

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. Structural changes at the NMJ can result in neurotransmission failure, resulting in weakness, atrophy and even death of the muscle fiber. Many studies have investigated how genetic modifications or disease can alter the structure of the mouse NMJ. Unfortunately, it can be difficult to directly compare findings from these studies because they often employed different parameters and analytical methods. Three protocols are described here. The first uses maximum intensity projection confocal images to measure the area of acetylcholine receptor (AChR)-rich postsynaptic membrane domains at the endplate and the area of synaptic vesicle staining in the overlying presynaptic nerve terminal. The second protocol compares the relative intensities of immunostaining for synaptic proteins in the postsynaptic membrane. The third protocol uses Fluorescence Resonance Energy Transfer (FRET) to detect changes in the packing of postsynaptic AChRs at the endplate. The protocols have been developed and refined over a series of studies. Factors that influence the quality and consistency of results are discussed and normative data are provided for NMJs in healthy young adult mice.

The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

Der neuromuskulären Synapse: Mess Synapse Größe, Fragmentierung und Veränderungen der synaptischen Protein Dichte Mit der konfokalen Fluoreszenzmikroskopie

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

Bewertung der dendritischen Verzweigung im Gyrus dentatus des Hippocampus-Region an der Maus

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

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

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

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

Eine Mensch-Maschine-Schnittstelle Integration von Low-Cost-Sensoren mit einem Neuromuskuläre Elektrostimulation System zur Post-Stroke Balance Rehabilitation

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

Functional and Morphological Assessment of Diaphragm Innervation by Phrenic Motor Neurons

This protocol specifically focuses on tools for assessing phrenic motor neuron (PhMN) innervation of the diaphragm at both the electrophysiological and morphological levels. Compound muscle action potential (CMAP) recording following phrenic nerve stimulation can be used to quantitatively assess functional diaphragm innervation by PhMNs of the cervical spinal cord in vivo in anesthetized rats and mice. Because CMAPs represent simultaneous recording of all myofibers of the whole hemi-diaphragm, it is useful to also examine the phenotypes of individual motor axons and myofibers at the diaphragm NMJ in order to track disease- and therapy-relevant morphological changes such as partial and complete denervation, regenerative sprouting and reinnervation. This can be accomplished via whole-mount immunohistochemistry (IHC) of the diaphragm, followed by detailed morphological assessment of individual NMJs throughout the muscle. Combining CMAPs and NMJ analysis provides a powerful approach for quantitatively studying diaphragmatic innervation in rodent models of CNS and PNS disease.

Compound muscle action potential recording quantitatively assesses functional diaphragm innervation by phrenic motor neurons. Whole-mount diaphragm immunohistochemistry assesses morphological innervation at individual neuromuscular junctions. The goal of this protocol is to demonstrate how these two powerful methodologies can be used in various rodent models of spinal cord disease.

Funktionellen und morphologischen Bewertung der Membran Innervation durch phrenicus Motoneuronen

This protocol specifically focuses on tools for assessing phrenic motor neuron (PhMN) innervation of the diaphragm at both the electrophysiological and ...

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching variations of the neuronal dendrites within the hippocampus are implicated in cognition and memory formation. There are several approaches to Golgi staining, all of which have been useful for determining the morphological characteristics of dendritic arbors and produce a clear background. The present Golgi-Cox method, (a slight variation of the protocol that is provided with a commercial Golgi staining kit), was designed to assess how a relatively low dose of the chemotherapeutic drug 5-flurouracil (5-Fu) would affect dendritic morphology, the number of spines, and the complexity of arborization within the hippocampus. The 5-Fu significantly modulated the dendritic complexity and decreased the spine density throughout the hippocampus in a region-specific manner. The data presented show that the Golgi staining method effectively stained the mature neurons in the CA1, the CA3, and the dentate gyrus (DG) of the hippocampus. This protocol reports the details for each step so that other researchers can reliably stain tissue throughout the brain with high quality results and minimal troubleshooting.

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

Bewertung der Hippocampalen Dendritischen Komplexität bei gealterten Mäusen unter Verwendung der Golgi-Cox-Methode

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching ...

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic nerves innervating the respiratory diaphragm. In spinal cord slices, phrenic motor neurons cannot be identified from other motor neurons on morphological or biochemical criteria. We provide the description of procedures for visualizing phrenic motor neuron cell bodies in mice, following intrapleural injections of cholera toxin subunit beta (CTB) conjugated to a fluorophore. This fluorescent neuroanatomical tracer has the ability to be caught up at the diaphragm neuromuscular junction, be carried retrogradely along the phrenic axons and reach the phrenic cell bodies. Two methodological approaches of intrapleural CTB delivery are compared: transdiaphragmatic versus transthoracic injections. Both approaches are successful and result in similar number of CTB-labeled phrenic motor neurons. In conclusion, these techniques can be applied to visualize or quantify the phrenic motor neurons in various experimental studies such as those focused on the diaphragm-phrenic circuitry.

Here, we describe a protocol for identifying phrenic motor neurons in mice after intrapleural delivery of fluorophore conjugated cholera toxin subunit beta. Two techniques are compared to inject the pleural cavity: transdiaphragmatic versus transthoracic approaches.

Retrograde neuroanatomischen Tracing der Phrenicus Motoneuronen bei Mäusen

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...

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.

3D Kinematische Analyse zur funktionellen Bewertung im Rattenmodell von Ischias-Nerven-Crush-Verletzungen

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

Laser-Induced Brain Injury in the Motor Cortex of Rats

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) occlusion of the middle cerebral artery (MCA) using a catheter. This generally accepted technique, however, has some limitations, thereby limiting its extensive use. Stroke induction by this method is often characterized by high variability in the localization and size of the ischemic area, periodical occurrences of hemorrhage, and high death rates. Also, the successful completion of any of the transient or permanent procedures requires expertise and often lasts for about 30 minutes. In this protocol, a laser irradiation technique is presented that can serve as an alternative method for inducing and studying brain injury in rodent models.
When compared to rats in the control and MCAO groups, the brain injury by laser induction showed reduced variability in body temperature, infarct volume, brain edema, intracranial hemorrhage, and mortality. Furthermore, the use of a laser-induced injury caused damage to the brain tissues only in the motor cortex unlike in the MCAO experiments where destruction of both the motor cortex and striatal tissues is observed.
Findings from this investigation suggest that laser irradiation could serve as an alternative and effective technique for inducing brain injury in the motor cortex. The method also shortens the time for completing the procedure and does not require expert handlers.

The protocol presented here shows a technique to create a rodent model of brain injury. The method described here uses laser irradiation and targets motor cortex.

Laserinduzierte Hirnverletzung im Motor Cortex von Ratten

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) ...