Amber Southwell cihazın kurmak ve kullanımı yazarlar yetiştirdi. Finansman NINDS Paul Patterson hibe sağlanmıştır.

1. Bu Protokol olanlar Southwell et dayanarak. arkadaşları (2009) ve Carter ve ark. arkadaşları (2001). <ol><li> Işını cihaz 12 mm düz bir yüzey veya 6 mm genişliğinde, 1 metre kirişler oluşur) istirahat iki kutup masa üstünden 50 cm. Bir kara kutu kiriş sonunda bitiş noktası olarak yerleştirilir. (Kontrplak ve ahşap kirişler, direkleri, ve kutu diğer türleri çoğu donanım mağazalarında bulunabilir). Ev kafeslerden Yuvalama malzeme bitiş noktasına fare çekmek için kara kutu yerleştirilir. Bir lambası (60 watt ampul ile) başlangıç ​​noktası üzerinde ışık tutmak için kullanılan ve caydırıcı bir uyarıcı olarak hizmet vermektedir. Bir zamanlayıcı ve bir zamanlayıcı durur 80 cm başlar 0 cm merkezinde 80 cm çapraz zaman iki hareket dedektörleri (Visopia tarafından uydurma) ile ölçülür. Bir naylon hamak (yerel bir kumaş mağazasından temin edilebilir), kiriş altında gerilir, herhangi bir düşme minderi masa üstünden ~ 7.5 cm. Bir video kamera performans kaydetmek için bir tripod üzerinde ayarlanır. Bu gösteride kullanılan aparat yerinde inşa edildi. </li><li> Bu test için, her durum için en az 10 farelerin bir seride tavsiye edilir. Fareler 12 saat ışık grupları, mümkün (07:00-19:00) / karanlık döngüsü gibi, ev sahipliği yapmaktadır ve davranış tesis test başlamadan önce en az 2 hafta boyunca kafeslerde acclimated. Eğitim / test öncesinde yaklaşık 10 dakika, farelerin ışın cihazı bulunduğu oda taşınabilmektedir. </li><li> Eğitim günü, her bir fare 12 mm ışın 3 kez ve daha sonra 6 mm ışın 3 kez geçer. Geniş genişlikleri (yani 28 mm) kirişler dar olanlar geçmek mümkün değildir fareler için veya daha büyük fareler için kullanılabilir. Fare bir ışının bir ucunda yer ve zaman, (80 cm uzaklıkta) hareket dedektörleri tarafından ölçülür diğer ucunda kaçış kutusuna geçmek için gerekli. Zamanlayıcı 80 cm merkezine giren fare burun başladı ve 80 cm hayvan sonuna geldiğinde durdu. Fareler evlerine kafesleri iki ışın eğitim oturumları arasında 10 dakika bekletin. Bu intertrial dönemde, başka bir 3-4 fareler zaman verimli kullanımını sağlamak için eğitilmiş olabilir. C57BL / 6 farelerde, durdurma ya da düşme az miktarda ışın genellikle çapraz. Ancak, ahır, sniff veya ileri geçmeden olmadan etrafına bakmak, araştırmacı fare dürtüsüyle, alay ya da eldivenli parmakları ile arkasından iterek ilerlemeye devam etmek için teşvik etmelidir. Fareler kasa sonra, bir sonraki duruşma öncesi dinlenmek için bir süre (~ 15 saniye) izin verilir. Her antrenmandan sonra, farelerin evlerine kafesleri doğrudan iade edilir ve onların konut odasına geri alınır. Onlar üzerinde eğitimli ve görev ve aparatları ile çok aşina, farelerin daha fazla durak olabilir. Eğitim 2 gün sonra fareler hala başarıyla tüm ışın hareket yok eğer Öte yandan, eğitim ek gün süreyle devam edilebilir. Farelerin tedaviler uygulanır, performansı ölçmek için temel tedavi öncesinde elde edilmelidir. </li><li> Test gününde, her kiriş çapraz kez kaydedilmektedir. İki başarılı çalışmalarda fare kiriş üzerinde durak değildi ortalaması alınır. Video kayıtlarının kayma ince analiz ve diğer gözlemlenebilir motor açıkları için kullanılabilir. &quot;1&quot; puan olarak Kiriş yetersizlik ve ışın yüzeyde hem pençeleri ile normal ışın hareket yeteneği ve daha az oranları nörolojik skorlama sistemi 1,2, ışın yürüme değerlendirmek için alternatif bir yol. olarak iki ayak fişleri &quot;7&quot; Kiriş üstüne çıkıyorsa ayak sürçmesi olarak tanımlanır. Bu endeksleme sistemi, &quot;3&quot; puan bacaklarda sürükleyerek ışın kapısına karşılık gelmektedir. 1 - iki çalışmanın puanları yeniden test olmadığı bir hayvan. Diğer puanları ile ilgili kiriş üzerinde performans Açıklama Feeney ve ark, (1982) ve Carter ve ark, (2001) bulunabilir. </li><li> Kirişler ve kutu fare pisliği temizlenir ve sonraki ışını, cihazın yerleştirilmeden önce% 70 etanol ve sonra su ile ıslatılmış havlu ile silinir. </li></ol> 2. Temsilcisi Sonuçlar: <img src="/files/ftp_upload/2376/2376fig1.jpg" alt="Şekil 1" /> Şekil 1 Tipik çapraz kez 6mm ve 12mm kirişler ve erkek C57Bl6. 9 ila 11 hafta eski C57BL/6-NCRL WT farelerin bir kohort eğitim 2 gün sonra iki kirişler üzerinde test edildi. 12 mm ışın geçiş için ortalama 4.6 ± erkekler için 0.4 sn (n = 14) ve 3.3 ± 0.3 sn (n = 11) kadınlar için. 6 mm kiriş üzerinde Times 6.8 ± 0.7, erkeklerde sn (n = 13) ve 5.9 ± kadınlarda 0.5 sn (n = 10) (hatalar SEM olarak bildirilmiştir).

<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>

Caltech Kurumsal Hayvan Bakım ve Kullanım Kurulu (IACUC) tarafından belirlenen kurallara ve yönetmeliklere uygun olarak hayvanlar üzerinde deneyler yapıldı.

Ince koordinasyon ve denge denge ışın Performans yararlı bir ölçüsüdür ve önceki çalışma tarafından onaylandı. Işın testi, genç ve yaşlı kemirgenler 2,3,4 yaş nedeniyle motor açıkları, merkezi sinir Sist `lezyonlar, farmakolojik ve genetik manipülasyonlar algılayabilir . Fare, kortikal etkisi lezyonlar genellikle ışın 5,6 kaymasını kontralateral sergi . Işın testi de zamanla yabani tip motor becerileri (WT) ve Huntington hastalığı (HD) fare modelleri ayırt etmek...

<ul><li> Kirişler: 1-m, küçük, orta ve büyük kare ile düzgün odun (yani, 6, 12, ve 28 mm genişliğinde) kesit şeritler </li><li> Destek başlangıç ​​bölümünde yükseltilmiş ışın (~ 1.5 cm, 50 cm kesit yüksek) kadar tutun standı </li><li> Amaç kutusu (her iki tarafta 20 cm) bir destek standı üzerine monte edilmiş (3 cm kesit, 50 cm yüksekliğinde) </li><li> Masa seviyesinin Hamak-~ 7.5 cm </li><li> 60-watt&#39;lık (W) masa lambası </li><li> Video kamera, tripod ve boş video kasetler </li><li> Etanol, kağıt havlu, temizlik malzemeleri </li></ul>

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

Beyin hasarı, genetik manipülasyonlar ve farmakolojik tedaviler farelerde motor becerileri değişiklikler neden olabilir. İnce motor koordinasyon ve denge ışın yürüme testi ile tespit edilebilir. Bu testin amacı, ayakta kalmak ve güvenli bir platforma yükseltilmiş bir dar bir ışın üzerinde yürümek fare için. 2 gün eğitim ve 1 gün test: Bu test, birbirini takip eden 3 gün içinde gerçekleşir. Kiriş, kiriş ve bu süreç içinde ortaya çıkan pençe fişleri numara çapraz fare için gereken zamanı ölçmek için Performans miktarı. Burada bizim laboratuvarda kullanılan protokol ve C57BL / 6 farelerin bir kohort temsilcisi sonuçları rapor. Bu görev, ince motor becerileri ve denge Rotarod gibi diğer motor testleri ile tespit olmayabilir açıkları tespit etmek için özellikle yararlıdır.

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

Ince motor koordinasyonu Açıkları denge ışın testi ile tespit edilebilir. Performans kiriş, kiriş geçiş hızını ve kiriş üzerinde fare kayar sayısını miktarı.

Denge kullanarak Fare Motor Denge ve Koordinasyon Değerlendirme

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

46.0K Görüntüleme.  California Institute of Technology. Ince motor koordinasyonu Açıkları denge ışın testi ile tespit edilebilir. Performans kiriş, kiriş geçiş hızını ve kiriş üzerinde fare kayar sayısını miktarı.

Video: 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.

Farelerde Motor Koordinasyon Ölçüm

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

Nörobilim

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.

Termal ve Mekanik nosiseptik Usuller kullanma Farelerde Morfin kaynaklı Hiperaljezi ve Analjezik Hoşgörü Değerlendirilmesi

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.

Nöromusküler Kavşak: Ölçme Synapse Boyut, Konfokal Floresan Mikroskobu kullanılarak sinaptik Protein Yoğunluk Parçalanma ve Değişiklikler

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.

Farelerde Hipokampal Bölgesi Dentate Gyrus dendritik arborization Değerlendirilmesi

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.

İnme sonrası Denge Rehabilitasyon bir Nöromüsküler Elektriksel Stimülasyon Sistemi ile düşük maliyetli Sensörler entegre bir İnsan-makine arayüzü

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.

Frenik Motor Nöronlar tarafından Diyafram İNNERVASYONU Fonksiyonel ve Morfolojik Değerlendirmesi

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.

Yaşlı Bir Farede Hipokampal Dendritik Karmaşıklığın Golgi-Cox Yöntemi ile Değerlendirilmesi

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.

Farelerde frenik Motor nöronların retrograd nöroanatomik izleme

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.

Siyatik Sinir Ezilme Hasarının Sıçan Modelinde Fonksiyonel Değerlendirme için 3D Kinematik Analiz

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.

Farelerin Motor Korteksinde LazerE Bağlı Beyin Hasarı

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