本研究では、助成金NIHM R01 MH091816とTDGのR21 MH084043によってサポートされています。

<ol>
	<li>Hasco&#233;t, M., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In Mood and Anxiety Related Phenotypes in Mice. 42, 85-118 (2009).</li><li>Sunal, R., G&#252;m&#252;&#231;el, a. B., Kayaalp, S. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+changes+in+swimming+area+on+results+of+"behavioral+despair+test".">Effect of changes in swimming area on results of "behavioral despair test".</a> Pharmacology Biochemistry and Behavior. 49, 891-896 (1994).</li><li>Petit-Demouliere, B., Chenu, F., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forced+swimming+test+in+mice:+a+review+of+antidepressant+activity.">Forced swimming test in mice: a review of antidepressant activity.</a> Psychopharmacology. (Berl). 177, 245-255 (2005).</li><li>Cryan, J. F., Markou, A., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+antidepressant+activity+in+rodents:+recent+developments+and+future+needs.">Assessing antidepressant activity in rodents: recent developments and future needs.</a> Trends in Pharmacological Sciences. 23, 238-245 (2002).</li><li>Can, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant-like+responses+to+lithium+in+genetically+diverse+mouse+strains.">Antidepressant-like responses to lithium in genetically diverse mouse strains.</a> Genes, Brain and Behavior. 10, 434-443 (2011).</li><li>Lucki, I., Dalvi, A., Mayorga, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitivity+to+the+effects+of+pharmacologically+selective+antidepressants+in+different+strains+of+mice.">Sensitivity to the effects of pharmacologically selective antidepressants in different strains of mice.</a> Psychopharmacology. (Berl). 155, 315-322 (2001).</li><li>David, D. J., Renard, C. E., Jolliet, P., Hascoet, M., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant-like+effects+in+various+mice+strains+in+the+forced+swimming+test.">Antidepressant-like effects in various mice strains in the forced swimming test.</a> Psychopharmacology (Berl). 166, 373-382 (2003).</li><li>Bai, F., Li, X., Clay, M., Lindstrom, T., Skolnick, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-+and+interstrain+differences+in+models+of+"behavioral+despair".">Intra- and interstrain differences in models of "behavioral despair".</a> Pharmacol. Biochem. Behav. 70, 187-192 (2001).</li><li>Guzzetti, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain+differences+in+paroxetine-induced+reduction+of+immobility+time+in+the+forced+swimming+test+in+mice:+Role+of+serotonin.">Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: Role of serotonin.</a> European Journal of Pharmacology. 594, 117-124 (2008).</li><li>Cervo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotype-dependent+activity+of+tryptophan+hydroxylase-2+determines+the+response+to+citalopram+in+a+mouse+model+of+depression.">Genotype-dependent activity of tryptophan hydroxylase-2 determines the response to citalopram in a mouse model of depression.</a> J. Neurosci. 25, 8165-8172 (2005).</li><li>Jiao, J., Nitzke, A., Doukas, D., Seiglie, M., Dulawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant+response+to+chronic+citalopram+treatment+in+eight+inbred+mouse+strains.">Antidepressant response to chronic citalopram treatment in eight inbred mouse strains.</a> Psychopharmacology. 213, 509-520 (2011).</li><li>Porsolt, R. D., Pichon, M. L. e., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression:+a+new+animal+model+sensitive+to+antidepressant+treatments.">Depression: a new animal model sensitive to antidepressant treatments.</a> Nature. 266, 730-732 (1977).</li><li>Porsolt, R. D., Bertin, A., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+despair+in+mice:+a+primary+screening+test+for+antidepressants.">Behavioral despair in mice: a primary screening test for antidepressants.</a> Arch. Int. Pharmacodyn. Ther. 229, 327-336 (1977).</li><li>Porsolt, R. D., Bertin, A., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term="Behavioural+despair"+in+rats+and+mice:+strain+differences+and+the+effects+of+imipramine.">"Behavioural despair" in rats and mice: strain differences and the effects of imipramine.</a> Eur. J. Pharmacol. 51, 291-294 (1978).</li><li>Mineur, Y. S., Belzung, C., Crusio, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+unpredictable+chronic+mild+stress+on+anxiety+and+depression-like+behavior+in+mice.">Effects of unpredictable chronic mild stress on anxiety and depression-like behavior in mice.</a> Behav. Brain. Res. 175, 43-50 (2006).</li><li>Millstein, R. A., Holmes, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+repeated+maternal+separation+on+anxiety-+and+depression-related+phenotypes+in+different+mouse+strains.">Effects of repeated maternal separation on anxiety- and depression-related phenotypes in different mouse strains.</a> Neuroscience &#38; Biobehavioral Reviews. 31, 3-17 (2007).</li><li>Cryan, J. F., Valentino, R. J., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+substrates+underlying+the+behavioral+effects+of+antidepressants+using+the+modified+rat+forced+swimming+test.">Assessing substrates underlying the behavioral effects of antidepressants using the modified rat forced swimming test.</a> Neuroscience &#38; Biobehavioral Reviews. 29, 547-569 (2005).</li><li>Crowley, J. J., Jones, O. '. L. e. a. r. y., F, O., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+tests+for+measuring+the+effects+of+antidepressants+in+mice.">Automated tests for measuring the effects of antidepressants in mice.</a> Pharmacology Biochemistry and Behavior. 78, 269-274 (2004).</li><li>Kurtuncu, M., Luka, L. J., Dimitrijevic, N., Uz, T., Manev, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliability+assessment+of+an+automated+forced+swim+test+device+using+two+mouse+strains.">Reliability assessment of an automated forced swim test device using two mouse strains.</a> Journal of Neuroscience Methods. 149, 26-30 (2005).</li><li>Hayashi, E., Shimamura, M., Kuratani, K., Kinoshita, M., Hara, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+experimental+system+capturing+three+behavioral+components+during+murine+forced+swim+test.">Automated experimental system capturing three behavioral components during murine forced swim test.</a> Life Sciences. 88, 411-417 (2011).</li><li>Cryan, J., Page, M., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+behavioral+effects+of+the+antidepressants+reboxetine,+fluoxetine,+and+moclobemide+in+a+modified+forced+swim+test+following+chronic+treatment.">Differential behavioral effects of the antidepressants reboxetine, fluoxetine, and moclobemide in a modified forced swim test following chronic treatment.</a> Psychopharmacology. 182, 335-344 (2005).</li><li>Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta-catenin+overexpression+in+the+mouse+brain+phenocopies+lithium-sensitive+behaviors.">Beta-catenin overexpression in the mouse brain phenocopies lithium-sensitive behaviors.</a> Neuropsychopharmacology. 32, 2173-2183 (2007).</li><li>Can, A., Grahame, N. J., Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Affect-related+related+behaviors+in+mice+selectively+bred+for+high+and+low+voluntary+alcohol+consumption.">Affect-related related behaviors in mice selectively bred for high and low voluntary alcohol consumption.</a> Behav. Genet. , (2011).</li><li>McKinney, W. T., Bunney, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Model+of+Depression:+I+Review+of+Evidence:+Implications+for+Research..">Animal Model of Depression: I Review of Evidence: Implications for Research..</a> Arch. Gen. Psychiatry. 21, 240-248 (1969).</li><li>Willner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+validity+of+animal+models+of+depression.">The validity of animal models of depression.</a> Psychopharmacology.(Berl). 83, 1-16 (1984).</li><li>Detke, M. J., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+serotonergic+and+noradrenergic+antidepressants+in+the+rat+forced+swimming+test:+the+effects+of+water+depth.">Detection of serotonergic and noradrenergic antidepressants in the rat forced swimming test: the effects of water depth.</a> Behav. Brain Res. 73, 43-46 (1995).</li><li>Gould, T. D., Dao, D. T., Kovacsics, C. E., Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In Mood and Anxiety Related Phenotypes in mice: characterization using behavioral tests. 42, (2009).</li><li>Bourin, M., Fiocco, A. J., Clenet, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+valuable+are+animal+models+in+defining+antidepressant+activity.">How valuable are animal models in defining antidepressant activity.</a> Human Psychopharmacology: Clinical and Experimental. 16, 9-21 (2001).</li></ol>

著者は、利害の衝突を宣言しません。

FST（と呼ばれることもありますPorsoltの水泳試験）は、ラットで最初に開発し、Porsoltと同僚12,13によってマウスのために変更されました。我々の研究室で成功し、上記のプロトコルに加えて、主に微妙なテストの修正の数は、（完全な見直し1 HascoëtとBourinを参照してください）発表されています。それは抗うつ薬と基礎および臨床研究3,14-16で様々な行動と神経生物学...

<ul><li>水タンク</li><li>温度計</li><li>タイマー</li><li>ビデオカメラ</li><li>ホワイトノイズ発生器（オプション） </li><li>乾燥紙</li></ul>

the mouse forced swim test

強制水泳試験は抗うつ薬、新規化合物の抗うつ効果、および状態のようなうつ病をレンダリングまたは予防を目的としている実験操作の評価に使用されるげっ歯類の行動試験です。マウスは、水で満たされており、彼らの脱出に関連するモビリティの挙動が測定され避けられない透明なタンクに配置されます。強制水泳試験では、確実に実施するのは簡単であり、それは最小限の特殊な装置を必要とします。強制水泳試験の実装を成功させるには、一定の手続きの詳細およびマウスへの不当なストレスの最小化に従う必要があります。プロトコルの説明および添付のビデオでは、我々はどのようにそれらを避けるために、結果の解釈とに有害かもしれない潜在的な落とし穴に重点を置いてこの試験のマウスのバージョンを実施する方法について説明します。さらに、我々はテストで明らかに動作が評価される方法について説明します。

Watch this Scientific Journal Video about The Mouse Forced Swim Test at JoVE.com

The Mouse Forced Swim Test

強制水泳試験は、げっ歯類の潜在的な抗うつ効果を評価するための実験的なアプローチとして検証されます。実験動物は、水のタンクに配置され、エスケープ関連のモビリティの挙動が定量化される。この試験のマウスのバージョンに共通の手順が説明されています。

マウス強制水泳試験

The forced swim test is a rodent behavioral test used for evaluation of antidepressant drugs, antidepressant efficacy of new compounds, and experimental ...

Research

JoVE Journal

Neuroscience

104.7K ビュー.  University of Maryland School of Medicine. 強制水泳試験は、げっ歯類の潜在的な抗うつ効果を評価するための実験的なアプローチとして検証されます。実験動物は、水のタンクに配置され、エスケープ関連のモビリティの挙動が定量化される。この試験のマウスのバージョンに共通の手順が説明されています。

ビデオ: The Mouse Forced Swim Test

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

複数のマウスの神経解剖学的磁気共鳴イメージング

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

神経科学

Detection of Neuritic Plaques in Alzheimer's Disease Mouse Model

Alzheimer's disease (AD) is the most common neurodegenerative disorder leading to dementia. Neuritic plaque formation is one of the pathological hallmarks of Alzheimer's disease. The central component of neuritic plaques is a small filamentous protein called amyloid &beta; protein (A&beta;)1, which is derived from sequential proteolytic cleavage of the beta-amyloid precursor protein (APP) by &beta;-secretase and &gamma;-secretase. The amyloid hypothesis entails that A&gamma;-containing plaques as the underlying toxic mechanism in AD pathology2. The postmortem analysis of the presence of neuritic plaque confirms the diagnosis of AD. To further our understanding of A&gamma; neurobiology in AD pathogenesis, various mouse strains expressing AD-related mutations in the human APP genes were generated. Depending on the severity of the disease, these mice will develop neuritic plaques at different ages. These mice serve as invaluable tools for studying the pathogenesis and drug development that could affect the APP processing pathway and neuritic plaque formation. In this protocol, we employ an immunohistochemical method for specific detection of neuritic plaques in AD model mice. We will specifically discuss the preparation from extracting the half brain, paraformaldehyde fixation, cryosectioning, and two methods to detect neurotic plaques in AD transgenic mice: immunohistochemical detection using the ABC and DAB method and fluorescent detection using thiofalvin S staining method.

One of the pathological characteristics of AD is the formation of Amyloid &#x03B2; protein positive neuritic plaques. In this protocol we describe two methods to detect neuritic plaques in transgenic AD model mice: immunohistochemical detection using the ABC and DAB method and fluorescent detection using thioflavin S staining method.

アルツハイマー病のマウスモデルにおける神経突起斑の検出

Alzheimer's disease (AD) is the most common neurodegenerative disorder leading to dementia. Neuritic plaque formation is one of the pathological hallmarks ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

マウスで作業していて、メモリの参照を評価するためのT-迷路強制交替と左右弁別課題

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Tissue Preparation and Immunostaining of Mouse Sensory Nerve Fibers Innervating Skin and Limb Bones

Detection and primary processing of physical, chemical and thermal sensory stimuli by peripheral sensory nerve fibers is key to sensory perception in animals and humans. These peripheral sensory nerve fibers express a plethora of receptors and ion channel proteins which detect and initiate specific sensory stimuli. Methods are available to characterize the electrical properties of peripheral sensory nerve fibers innervating the skin, which can also be utilized to identify the functional expression of specific ion channel proteins in these fibers. However, similar electrophysiological methods are not available (and are also difficult to develop) for the detection of the functional expression of receptors and ion channel proteins in peripheral sensory nerve fibers innervating other visceral organs, including the most challenging tissues such as bone. Moreover, such electrophysiological methods cannot be utilized to determine the expression of non-excitable proteins in peripheral sensory nerve fibers. Therefore, immunostaining of peripheral/visceral tissue samples for sensory nerve fivers provides the best possible way to determine the expression of specific proteins of interest in these nerve fibers. So far, most of the protein expression studies in sensory neurons have utilized immunostaining procedures in sensory ganglia, where the information is limited to the expression of specific proteins in the cell body of specific types or subsets of sensory neurons. Here we report detailed methods/protocols for the preparation of peripheral/visceral tissue samples for immunostaining of peripheral sensory nerve fibers. We specifically detail methods for the preparation of skin or plantar punch biopsy and bone (femur) sections from mice for immunostaining of peripheral sensory nerve fibers. These methods are not only key to the qualitative determination of protein expression in peripheral sensory neurons, but also provide a quantitative assay method for determining changes in protein expression levels in specific types or subsets of sensory fibers, as well as for determining the morphological and/or anatomical changes in the number and density of sensory fibers during various pathological states. Further, these methods are not confined to the staining of only sensory nerve fibers, but can also be used for staining any types of nerve fibers in the skin, bones and other visceral tissue.

Immunocytochemical identification of peripheral sensory nerve fiber subtypes (and detection of protein expression therein) are key to the understanding of molecular mechanisms underlying peripheral sensation. Here we describe methods for preparation of peripheral/visceral tissue samples, such as skin and limb bones, for specific immunostaining of peripheral sensory nerve fibers.

スキンと四肢骨の神経支配マウス感覚神経線維の組織の調製と免疫染色

Detection and primary processing of physical, chemical and thermal sensory stimuli by peripheral sensory nerve fibers is key to sensory perception in ...

The Tail Suspension Test

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that are expected to affect depression related behaviors. Mice are suspended by their tails with tape, in such a position that it cannot escape or hold on to nearby surfaces. During this test, typically six minutes in duration, the resulting escape oriented behaviors are quantified. The tail-suspension test is a valuable tool in drug discovery for high-throughput screening of prospective antidepressant compounds. Here, we describe the details required for implementation of this test with additional emphasis on potential problems that may occur and how to avoid them. We also offer a solution to the tail climbing behavior, a common problem that renders this test useless in some mouse strains, such as the widely used C57BL/6. Specifically, we prevent tail climbing behaviors by passing mouse tails through a small plastic cylinder prior to suspension. Finally, we detail how to manually score the behaviors that are manifested in this test.

The tail-suspension test is validated as an experimental procedure to assess antidepressant efficacy of drug treatments in mice. Mice are suspended by their tails for six minutes and escape-related behaviors are assessed. We describe procedures used in conducting the tail suspension test.

尾懸垂テスト

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that ...

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and lifespan. In addition, telomerase protects the mitochondria from oxidative stress and confers resistance to apoptosis, suggesting its possible importance for the surviving of non-mitotic, highly active cells such as neurons. We previously demonstrated the ability of novel telomerase activators to increase telomerase activity and expression in the various mouse brain regions and to protect motor neurons cells from oxidative stress. These results strengthen the notion that telomerase is involved in the protection of neurons from various lesions. To underline the role of telomerase in the brain, we here compare the activity of telomerase in male and female mouse brain and its dependence on age. TRAP assay is a standard method for detecting telomerase activity in various tissues or cell lines. Here we demonstrate the analysis of telomerase activity in three regions of the mouse brain by non-denaturing protein extraction using CHAPS lysis buffer followed by modification of the standard TRAP assay.
In this 2-step assay, endogenous telomerase elongates a specific telomerase substrate (TS primer) by adding TTAGGG 6 bp repeats (telomerase reaction). The telomerase reaction products are amplified by PCR reaction creating a DNA ladder of 6 bp increments. The analysis of the DNA ladder is made by 4.5% high resolution agarose gel electrophoresis followed by staining with highly sensitive nucleic acid stain.
Compared to the traditional TRAP assay that utilize 32P labeled radioactive dCTP&#39;s for DNA detection and polyacrylamide gel electrophoresis for resolving the DNA ladder, this protocol offers a non-toxic time saving TRAP assay for evaluating telomerase activity in the mouse brain, demonstrating the ability to detect differences in telomerase activity in the various female and male mouse brain region.

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

マウス脳のさまざまな領域におけるテロメラーゼ活性：非放射性テロメラーゼリピート増幅プロトコル（TRAP）アッセイ

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and ...

Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the over-production of amyloid-beta and the deposition of amyloid-beta plaques in brain regions of the afflicted individuals. Throughout the years scientists have generated numerous Alzheimer&#8217;s disease mouse models that attempt to replicate the amyloid-beta pathology. Unfortunately, the mouse models only selectively mimic the disease features. Neuronal death, a prominent effect in the brains of Alzheimer&#8217;s disease patients, is noticeably lacking in these mice. Hence, we and others have employed a method of directly infusing soluble oligomeric species of amyloid-beta - forms of amyloid-beta that have been proven to be most toxic to neurons - stereotaxically into the brain. In this report we utilize male C57BL/6J mice to document this surgical technique of increasing amyloid-beta levels in a select brain region. The infusion target is the dentate gyrus of the hippocampus because this brain structure, along with the basal forebrain that is connected by the cholinergic circuit, represents one of the areas of degeneration in the disease. The results of elevating amyloid-beta in the dentate gyrus via stereotaxic infusion reveal increases in neuron loss in the dentate gyrus within 1 week, while there is a concomitant increase in cell death and cholinergic neuron loss in the vertical limb of the diagonal band of Broca of the basal forebrain. These effects are observed up to 2 weeks. Our data suggests that the current amyloid-beta infusion model provides an alternative mouse model to address region specific neuron death in a short-term basis. The advantage of this model is that amyloid-beta can be elevated in a spatial and temporal manner.

Here, we present a protocol for direct stereotaxic brain infusion of amyloid-beta. This methodology provides an alternative in vivo mouse model to address the short-term effects of amyloid-beta on brain neurons.

マウス海馬にオリゴマーアミロイドベータの定位注入

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the ...

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has proven to be an invaluable model organism in helping to tackle this challenge, from inspiring technological approaches,&#160;building the human brain connectome, to actually shedding light on the specific molecular drivers of basic functional patterns. The bulk of the behavioral studies in C. elegans have been performed on solid substrates. In liquid, animals exhibit behavioral patterns that include movement at a range of speeds in 3D, as well as partial body movements, such as a posterior curl without anterior shape change, which introduce new challenges for quantitation. The steps of a simple procedure, and use of a software that enables high-resolution analysis of C. elegans swim behavior, are presented here. The software, named CeleST, uses a specialized computer program that tracks multiple animals simultaneously and provides novel measures of C. elegans locomotion in liquid (swimming). The measures are mostly grounded in animal posture and based on mathematics used in computer vision and pattern recognition, without computational requirements for threshold cut-offs. The software tool can be used to both assess overall swimming prowess in hundreds of animals from combined small batch trials and to reveal novel phenotypes even in well-characterized genetic mutants. The preparation of specimens for analysis with CeleST is simple and low-tech, enabling wide adaptation by the scientific community. Use of the computational approach described here should therefore contribute to the greater understanding of behavior and behavioral circuits in the C. elegans model.

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

An efficient and simple methodology for computer-based analysis of nematode swimming behavior in liquid is described. The method requires little to no investment for C. elegans laboratories. The hardware used is standard, and the computer software for the behavioral analysis (CeleST) is an open source one.

の自動解析<em&gt; C。エレガンス</emセレストソフトウェアを使用&gt;スイム挙動

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb

The capillary refill test was introduced in 1947 to help estimate circulatory status in critically ill patients. Guidelines commonly state that refill should occur within 2 s after releasing 5 s of firm pressure (e.g., by the physician&#39;s finger) in the normal healthy supine patient. A slower refill time indicates poor skin perfusion, which can be caused by conditions including sepsis, blood loss, hypoperfusion, and hypothermia. Since its introduction, the clinical usefulness of the test has been debated. Advocates point out its feasibility and simplicity and claim that it can indicate changes in vascular status earlier than changes in vital signs such as heart rate. Critics, on the other hand, stress that the lack of standardization in how the test is performed and the highly subjective nature of the naked eye assessment, as well as the test&#39;s susceptibility to ambient factors, markedly lowers the clinical value. The aim of the present work is to describe in detail the course of the refill event and to suggest potentially more objective and exact endpoint values for the capillary refill test using diffuse polarization spectroscopy.

This protocol describes how the use of diffuse polarization spectroscopy can improve the clinical usefulness of the capillary refill test. We suggest a more detailed analysis of the course of the capillary refill in healthy volunteers using diffuse reflectance spectroscopy videos and new informatic endpoints.

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィル テスト

The capillary refill test was introduced in 1947 to help estimate circulatory status in critically ill patients. Guidelines commonly state that refill ...

Localization of the Locus Coeruleus in the Mouse Brain

The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional abnormalities of LC impact several brain regions including cortex, hippocampus, and cerebellum and may contribute to depression, bipolar disorder, anxiety, as well as Parkinson disease and Alzheimer disease. These disorders are often associated with metal misbalance, but the role of metals in LC is only partially understood. Morphologic and functional studies of LC are needed to better understand the human pathologies and contribution of metals. Mice are a widely used experimental model, but the mouse LC is small (~0.3 mm diameter) and hard to identify for a non-expert. Here, we describe a step-by-step immunohistochemistry-based protocol to localize the LC in the mouse brain. Dopamine-&#946;-hydroxylase (DBH), and alternatively, tyrosine hydroxylase (TH), both enzymes highly expressed in the LC, are used as immunohistochemical markers in brain slices. Sections adjacent to LC-containing sections can be used for further analysis, including histology for morphological studies, metabolic testing, as well as metal imaging by X-ray fluorescence microscopy (XFM).

The locus coeruleus is a small cluster of neurons involved in a variety of physiological processes. Here, we describe a protocol to prepare mouse brain sections for studies of proteins and metals in this nucleus.

マウスの脳、青斑核の局在

The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional ...

マウス強制水泳試験

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この動画の章

複数のマウスの神経解剖学的磁気共鳴イメージング

アルツハイマー病のマウスモデルにおける神経突起斑の検出

マウスで作業していて、メモリの参照を評価するためのT-迷路強制交替と左右弁別課題

スキンと四肢骨の神経支配マウス感覚神経線維の組織の調製と免疫染色

尾懸垂テスト

マウス脳のさまざまな領域におけるテロメラーゼ活性：非放射性テロメラーゼリピート増幅プロトコル（TRAP）アッセイ

マウス海馬にオリゴマーアミロイドベータの定位注入

の自動解析

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィルテスト

マウスの脳、青斑核の局在

マウス強制水泳試験

要約

さらに動画を探す

この動画の章

複数のマウスの神経解剖学的磁気共鳴イメージング

アルツハイマー病のマウスモデルにおける神経突起斑の検出

マウスで作業していて、メモリの参照を評価するためのT-迷路強制交替と左右弁別課題

スキンと四肢骨の神経支配マウス感覚神経線維の組織の調製と免疫染色

尾懸垂テスト

マウス脳のさまざまな領域におけるテロメラーゼ活性：非放射性テロメラーゼリピート増幅プロトコル（TRAP）アッセイ

マウス海馬にオリゴマーアミロイドベータの定位注入

の自動解析

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィル テスト

マウスの脳、青斑核の局在

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィルテスト