Name: olfactory assays for mouse models of neurodegenerative disease
Uploaded: 2014-08-25T14:00:00
Duration: 7 min 27 s
Description: Watch this Scientific Journal Video about Olfactory Assays for Mouse Models of Neurodegenerative Disease at JoVE.com

This protocol follows any and all animal care and use guidelines and regulations set by IACUC at the University of Cincinnati. This work is funded by NIH/NINDS NS07722 and the Gardner Family Center for Parkinson&#8217;s Disease and Movement Disorders.

所有步骤的协议遵循动物护理和使用由辛辛那提大学的IACUC设定的准则和法规。 1，一般注意事项<ol><li>如果可能的话，在此期间在黑暗周期的活跃期测试小鼠。通常情况下，进行低光照下，并在至少1小时，检测到黑暗的循环。然而，如果测试在暗周期是不可行的，光周期期间执行所有三个测试。请记住，当老鼠在他们的睡眠周期被嗅探测试时间可能会减少。注意：当测试可以在睡眠周期中执行的老鼠可能会显示在埋颗粒，块，或香味的墨盒不感兴趣。 </li><li>为了减少外部嗅觉暗示的任何效果，穿碱基对手套在任何时候都贯穿每个试验，然后将清洁的一对手套在碱基对每只小鼠和每个试验。一般来说，如果有疑问，换上了一套手套的清洁。有一个盒子手套读随手可用，因为它们会经常发生变化。注：我们用丁腈手套所有动物处理。 </li><li>清洁测试材料和受试者用消毒之间笼/消毒剂也正是在这种电池的试验的一个重要步骤，以减少潜在的疾病传播，以及除去不需要的嗅觉暗示。 </li></ol> 2，埋颗粒测试<ol><li>在测试前至少两天，记录每只小鼠的体重，然后食物限制到90％的体重。在测试之前和期间，食物限制，给每只小鼠1-2件粒料的测试过程中使用（ 例如，一块甜化谷物的）。不要跳过这一步，因为老鼠是恐新食品和测试过程中不得搜查的颗粒，如果他们都没有吃小球之前。 </li><li>在所有的测试天，通过将笼子的考场，没有一瓶水或器B的，观察者小鼠1小时前测试英寸房子他们在笼子只有一个过滤器顶盖。 </li><li>在习惯化，填补了清洁鼠笼〜3个厘米高有干净的被褥使某些被褥均匀地分布在整个笼子。 5分钟设置一个计时器。 </li><li>继1小时的习惯，埋在下面的床上用品1加糖谷物颗粒0.5厘米所以它是不可见的。 </li><li>从家笼中取出测试鼠标，将其放置在试验网箱的中心，将过滤器顶盖上的笼子里，开始计时。 </li><li>停止定时器，当鼠标揭示沉淀，并开始吃它。注意，露出埋丸所需的时间。 </li><li>如果鼠标没有发现在5分钟内颗粒，停止审判，并记下该鼠标的得分为300秒。将沉淀物以这样的鼠标可以访问它的寝具的顶部，并允许小鼠吃的粒料。 </li><li>下面的试验中，返回鼠标的家笼。 </li><li>清空从测试笼的床上用品和清洁的C年龄与动物房清洗溶液。 </li><li>重复步骤2.4至2.9为每个滑鼠，在每次鼠标后更换手套。 </li><li>毕竟小鼠进行试验，给他们足够的食物来维持在90％的体重。 </li><li>执行第2-5天以同样的方式测试作为测试第1天有一个例外;埋葬沉淀在笼子里的每个试验不同的位置。 </li><li>在测试的第6天，进行表面颗粒审判。 </li><li>对于表面颗粒试验，遵循相同的程序测试第1天但不是在床上用品将它放在被褥上面埋葬的沉淀。记录时间为鼠标找到并开始进食沉淀。 </li></ol> 3，测试座<ol><li>个别家鼠在清洁笼子里，并放置5块笼（标记为AE），每个网箱设置改变之前的手套。 </li><li>随后进行测试24小时。移动测试小鼠的试验区，取出一瓶水和馈线箱，然后将人升从笼子里有5个街区还有少数床上用品装入塑料袋标有动物的身份。放置在密封的塑料袋上鼠标的笼子的顶部。 </li><li>的，观察者动物在其没有水瓶，给料机仓笼，并阻止1小时前测试。 </li><li>更改每笼之间的手套，这样的气味是不是动物/块之间交换。 </li><li>排队对方旁边的笼子鼠标在桌子上，每个网箱之间至少有10厘米。放置一台摄像机，使笼（不笼的长边）的前面是明确的说法。 </li><li>设置定时器30秒，标记有一个重要的实验细节（日期，鼠标的ID，审判等 ）的记忆卡。录像带的标签为2-3秒。 </li><li>从塑料袋中与鼠标的标识删除块广告并将其放置在笼子的中间，使他们清楚地摄像机上可见。确保有〜1厘米Ø每个块的F之间的空间。将过滤器顶部回到笼子里。启动定时器和录像带，持续30秒。 </li><li> 30秒后，停​​止录像并从笼子块，并把它们放回塑料袋。 </li><li>更换手套，然后转移到下一个鼠标，重复用于第一试验同样地进行。做到这一点，共计6项研究对每只小鼠确保有中〜5分钟的试验间隔。 </li><li>在第 7 次试验，遵循相同的程序为试验1-6除了没有把块D从鼠标本身的笼子从另一个老鼠的笼子里添加E座让鼠标从接触到块A，B和C的从另一个老鼠的笼子自己的家笼及E座。注意，在实验室中书的方框E各小鼠暴露于和备用其中方框E被放置在每只小鼠的笼子（ 例如 ABCE或AEBC）。以E座从鼠标同性而不是从原始cagemate。六deotape 30秒。 </li><li>继7项试验，动物回归到一个新的干净的笼子与原cagemates。用于测试用中性清洁剂和水的清洁块，晾干在后面的测试中使用。 </li></ol> 4，习惯化/ Dishabituation <ol><li>以类似的方式，以块测试执行习惯/ dishabituation测试，但代替使用具有鼠标的气味块，使用保持棉花香味的不同对提取物的组织盒。 </li><li>按照步骤3.1-3.4使用一个单一的，无味的组织盒，而不是块。 </li><li> ，观察者的小鼠在试验室。 </li><li>在一个单独的房间准备香味组织墨盒。将小棉球放入干净的墨盒，然后加入5微升提取物（ 如杏仁萃取物）成绵。捕捉墨盒顶部在棉花和发生在专用的盒子或袋子标记与香味。 </li><li>做同样的程序F或第二的气味（ 如八角提取物）。改变处理不同气味的手套。确保棉花球，在组织盒完全包封。注：裸露的棉花球纤维可导致动物在棉花球嵌套的目的，而不是气味本身的兴趣。 </li><li>然后，按照步骤3.6-3.10使用一种带香味的组织盒的第6项试验，第二香味组织墨盒第七审判。 </li></ol>录像带的五分析<ol><li>对于该块和习惯/ dishabituation测试，有一个评估者盲目的实验条件下测量了嗅探每个块，用嗅探定义为与该块鼻腔接触的时间。如果需要的话，测量的其他行为，如时间到试验7和总活性各试验期间（沿长度和宽度的家笼所作运动的次数）在接近新的嵌段（E）。分数嗅花的时间ð时间从用秒表录像带接近。 </li><li>同样，对于习惯/ dishabituation测试，有一个评估者盲目的实验条件下测嗅花的墨盒，时间接近墨盒，自主​​活动的时间。 </li></ol> 6，统计<ol><li>分析采用非参数统计，因为等待时间，找到小球得分往往表现出差异的异质埋颗粒测试。平均等待时间以找到在整个3-5天的沉淀，然后用曼 - 惠特尼U检验的小鼠组之间的比较。类似地，对于测试的表面沉淀部分，分析延迟使用曼 - 惠特尼U检验比较基因型发现沉淀。 </li><li>对于该块试验，分析试验从7（引进新的香味块）中的数据。通常情况下，采用非参数的分析来解释方差的异质性。为了分析在时间差使用了嗅探日ê小说块以及时间接近块不同实验组，用曼 - 惠特尼U检验。使用Wilcoxon符号秩检验比较花时间嗅探小说块花费的时间嗅着熟悉的块。 </li><li>对于习惯/ dishabituation试验，进行参数分析了试验1,6和7，类似于以前的研究20。如果小鼠表现出几乎没有嗅探，使用非参数的分析。 </li></ol>

<ol>
	<li>Hawkes, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfaction+in+neurodegenerative+disorder.">Olfaction in neurodegenerative disorder.</a> Adv. Otorhinolaryngol. 63, 133-151 (2006).</li><li>Ward, C., Hess, W., Calne, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+impairment+in+Parkinson's+disease.">Olfactory impairment in Parkinson's disease.</a> Neurology. 33, 943-946 (1983).</li><li>Doty, R., Stern, M., Pfeiffer, C., Gollomp, S., Hurtig, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+olfactory+dysfunction+in+early+stage+treated+and+untreated+idiopathic+Parkinson's+disease.">Bilateral olfactory dysfunction in early stage treated and untreated idiopathic Parkinson's disease.</a> J. Neurol. Neurosurg. Psychiatry. 55, 138-142 (1992).</li><li>Doty, R., Deems, D., Stellar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+dysfunction+in+parkinsonism:+a+general+deficit+unrelated+to+neurologic+signs,+disease+stage,+or+disease+duration.">Olfactory dysfunction in parkinsonism: a general deficit unrelated to neurologic signs, disease stage, or disease duration.</a> Neurology. 38, 1237-1244 (1988).</li><li>Tissingh, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+olfaction+in+de+novo+and+treated+Parkinson's+disease:+possible+implications+for+early+diagnosis.">Loss of olfaction in de novo and treated Parkinson's disease: possible implications for early diagnosis.</a> Movement Disorders. 16 (1), 41-46 (2001).</li><li>Montgomery, E., Baker, K., Lyons, K., Koller, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abnormal+performance+on+the+PD+test+battery+by+asymptomatic+first-degree+relatives.">Abnormal performance on the PD test battery by asymptomatic first-degree relatives.</a> Neurology. 52, 757-762 (1999).</li><li>Berendse, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subclinical+dopaminergic+dysfunction+in+asymptomatic+Parkinson's+disease+patients'+relatives+with+a+decreased+sense+of+smell.">Subclinical dopaminergic dysfunction in asymptomatic Parkinson's disease patients' relatives with a decreased sense of smell.</a> Ann. Neurol. 50 (1), 34-41 (2001).</li><li>Ponsen, M., Stoffers, D., Booij, J., van Eck-Smit, B., Wolters, E., Berendse, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Idiopathic+hyposmia+as+a+preclinical+sign+of+Parkinson's+disease.">Idiopathic hyposmia as a preclinical sign of Parkinson's disease.</a> Ann. Neurol. 56, 173-181 (2004).</li><li>Langston, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Parkinson's+complex:+parkinsonism+is+just+the+tip+of+the+iceberg.">The Parkinson's complex: parkinsonism is just the tip of the iceberg.</a> Ann. Neurol. 59 (4), 591-596 (2006).</li><li>Ross, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+olfactory+dysfunction+with+risk+for+future+Parkinson's+disease.">Association of olfactory dysfunction with risk for future Parkinson's disease.</a> Ann. Neurol. 63, 167-173 (2008).</li><li>Fleming, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+and+progressive+sensorimotor+anomalies+in+mice+overexpressing+wild-type+human+alpha-synuclein.">Early and progressive sensorimotor anomalies in mice overexpressing wild-type human alpha-synuclein.</a> J. Neurosci. 24, 9434-9440 (2004).</li><li>Fleming, S., Ekhator, O., Ghisays, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+sensorimotor+function+in+mouse+models+of+Parkinson's+disease.">Assessment of sensorimotor function in mouse models of Parkinson's disease.</a> J. Vis. Exp. , e50303 (2013).</li><li>Schallert, T., Tillerson, 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=Intervention+strategies+for+degeneration+of+DA+neurons+in+parkinsonism:+Optimizing+behavioral+assessment+of+outcome.">Intervention strategies for degeneration of DA neurons in parkinsonism: Optimizing behavioral assessment of outcome.</a> Central Nervous System Diseases. , 131-151 (2000).</li><li>Whishaw, I., O'Connor, W., Dunnett, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+contributions+of+motor+cortex,+nigrostriatal+dopamine+and+caudate-putamen+to+skilled+forelimb+use+in+the+rat.">The contributions of motor cortex, nigrostriatal dopamine and caudate-putamen to skilled forelimb use in the rat.</a> Brain. 109 (5), 805-843 (1986).</li><li>Schwarting, R., Huston, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+unilateral+6-hydroxydopamine+lesion+model+in+behavioral+brain+research.+Analysis+of+functional+deficits,+recovery+and+treatments.">The unilateral 6-hydroxydopamine lesion model in behavioral brain research. Analysis of functional deficits, recovery and treatments.</a> Prog Neurobiol. 49 (3), 215-266 (1996).</li><li>Glasl, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pink1-deficiency+in+mice+impairs+gait,+olfaction+and+serotonergic+innervation+of+the+olfactory+bulb.">Pink1-deficiency in mice impairs gait, olfaction and serotonergic innervation of the olfactory bulb.</a> Experimental Neurology. 235, 214-227 (2012).</li><li>Kim, Y., Lussier, S., Rane, A., Choi, S., Andersen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducible+dopaminergic+glutathione+depletion+in+an+&#945;-synuclein+transgenic+mouse+model+results+in+age-related+olfactory+dysfunction.">Inducible dopaminergic glutathione depletion in an &#945;-synuclein transgenic mouse model results in age-related olfactory dysfunction.</a> Neuroscience. 172, 379-386 (2011).</li><li>Fleming, S., Tetreault, N., Mulligan, C., Huston, C., Masliah, E., Chesselet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+deficits+in+mice+overexpressing+human+wildtype+alpha-synuclein.">Olfactory deficits in mice overexpressing human wildtype alpha-synuclein.</a> The European Journal of Neuroscience. 28 (2), 247-256 (2008).</li><li>Nuber, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+progressive+dopaminergic+phenotype+associated+with+neurotoxic+conversion+of+&#945;-synuclein+in+BAC-transgenic+rats.">A progressive dopaminergic phenotype associated with neurotoxic conversion of &#945;-synuclein in BAC-transgenic rats.</a> Brain. 136 (2), 412-432 (2013).</li><li>Macknin, J., Higuchi, M., Lee, V., Trojanowski, J., Doty, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+dysfunction+occurs+in+transgenic+mice+overexpressing+human+tau+protein.">Olfactory dysfunction occurs in transgenic mice overexpressing human tau protein.</a> Brain Res. 1000, 174-178 (2004).</li><li>Lazic, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+abnormalities+in+Huntington's+disease:+decreased+plasticity+in+the+primary+olfactory+cortex+of+R6/1+transgenic+mice+and+reduced+olfactory+discrimination+in+patients.">Olfactory abnormalities in Huntington's disease: decreased plasticity in the primary olfactory cortex of R6/1 transgenic mice and reduced olfactory discrimination in patients.</a> Brain Res. 1151, 219-226 (2007).</li><li>Wesson, D., Wilson, D., Nixon, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Should+olfactory+dysfunction+be+used+as+a+biomarker+of+Alzheimer's+disease?.">Should olfactory dysfunction be used as a biomarker of Alzheimer's disease?.</a> Expert Review of Neurotherapeutics. 10 (5), 633-635 (2010).</li><li>Taylor, T., Caudle, W., Miller, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VMAT2-deficient+mice+display+nigral+and+extranigral+pathology+and+motor+and+nonmotor+symptoms+of+Parkinson's+disease.">VMAT2-deficient mice display nigral and extranigral pathology and motor and nonmotor symptoms of Parkinson's disease.</a> Parkinsons Dis. 2011, 124-165 (2011).</li><li>Tillerson, J., Caudle, W., Parent, J., Gong, C., Schallert, T., Miller, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+discrimination+deficits+in+mice+lacking+the+dopamine+transporter+or+the+D2+dopamine+receptor.">Olfactory discrimination deficits in mice lacking the dopamine transporter or the D2 dopamine receptor.</a> Behav. Brain Res. 172, 97-105 (2006).</li><li>Fleming, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pilot+trial+of+the+microtubule-interacting+peptide+(NAP)+in+mice+overexpressing+alpha-synuclein+shows+improvement+in+motor+function+and+reduction+of+alpha-synuclein+inclusions.">A pilot trial of the microtubule-interacting peptide (NAP) in mice overexpressing alpha-synuclein shows improvement in motor function and reduction of alpha-synuclein inclusions.</a> Mol. Cell. Neurosci. 4, 597-606 (2011).</li><li>Crawley, 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> What's Wrong with My Mouse? Behavioral Phenotyping of Transgenic and Knockout Mice. , (2000).</li><li>Nathan, B., Yost, J., Litherland, M., Struble, R., Switzer, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+function+in+apoE+knockout+mice.">Olfactory function in apoE knockout mice.</a> Behav. Brain Res. 150, 1-7 (2004).</li><li>Spinetta, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alcohol-induced+retrograde+memory+impairment+in+rats:+prevention+by+caffeine.">Alcohol-induced retrograde memory impairment in rats: prevention by caffeine.</a> Psychopharmacology. 201 (3), 361-371 (2008).</li><li>Bielsky, I., Hu, S., Szegda, K., Westphal, H., Young, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profound+impairment+in+social+recognition+and+reduction+in+anxiety-like+behavior+in+vasopressin+V1a+receptor+knockout+mice.">Profound impairment in social recognition and reduction in anxiety-like behavior in vasopressin V1a receptor knockout mice.</a> Neuropsychopharmacology. 29, 483-493 (2004).</li></ol>

We have no conflicts of interest.

每个在此研究中所描述的测试测量嗅觉功能的小鼠的不同方面。埋颗粒测试测量嗅觉的粮食动机方面，测试的饥饿（食物的限制，而不是禁食）来检测可口一块甜麦片埋在被褥能力的小鼠。块试验测量更嗅觉功能的社交方面，检测小鼠自身气味和的同种之间进行区分的能力。习惯化/ dishabituation试验评估了小鼠的熟悉和新颖的，无害的气味来区分的能力。在鉴定一个新的模型，因为食物中的动机或恐惧异常有助于在突变小鼠中观察到与对照组相比差异使用多个测试是非常重要的。例如，如果仅在块执行测试，并降低嗅探已另一个鼠标的气味被观察到的块，也不清楚是否有olfact储器缺陷或增强的恐惧反应，可能导致回避该块的同种。当多个测试执行和缺陷在所有试验中观察到，它强烈支持的嗅觉障碍的解释。但是，如果差别只在一个测试，但没有其他人看到那么有可能是一个更微妙的嗅觉障碍，但是这将是必不可少的，以排除其他的解释（ 即增强恐惧或减少食物的动机）。测试嗅觉功能的小鼠时，要记住其他重要因素，包括小鼠的背景和应变性。不同遗传背景的品系（ 即，C57BL / 6，DBA， 等等 ）可以具有对小鼠行为产生深远的影响，因此，建议在进行整个实验前，以适应在同一背景的野生型小鼠的每个协议。此外，还建议在雄性和雌性小鼠分别测试彼此因为男性的嗅觉系统是高LY到女性发情和嗅觉检测的发情雌性敏感可能干扰在所有三个测试中的表现。 有些步骤是绝对关键的小鼠嗅觉测试时要遵循的。要弄清实验者被引入到动物的气味是重要的。戴手套的程序是动物之间手套必不可少的，频繁的更换是必要的。它始终是为实验者不穿古龙水或香水的嗅觉测试天一个很好的做法。而该过程的某些部分是不灵活的，也有可以进行修改和改编，而不降低测试的有效性或灵敏度等部位。例如，习惯化试验的数目可以增加或减少取决于小鼠的菌株进行测试，以及如何快速地它们，观察者对刺激的反应。这些测试的主要限制是，它们都被驱动通过某种形式的动力，无论是食品或社会，使得难以完全排除异常动机作为解释当改变的响应变化。这可以通过测试和时间接近的刺激中分析的其他参数，如活性被最小化。 一旦掌握了这些测试可以很容易地用于研究的神经退行性疾病的新的小鼠模型的表型。此外，具有最高功率的测试可以包括在临床前研究中测试的潜在疗法（对于一个例子见参考文献25）。

嗅觉障碍是与一些神经变性疾病，包括帕金森氏病（PD），阿尔茨海默氏病和亨廷顿氏病1。在PD，嗅觉障碍包括在气味识别，检测和识别缺陷和被发现在高达的患者2-5 70-95％。这些缺陷可以通过长达4年之前PD的大是大非运动症状，表明嗅觉障碍可能预示6-10帕金森的早期阶段。在帕金森病早期出现嗅觉赤字已经导致嗅觉障碍所涉及的基本机制产生了浓厚的兴趣和。在啮齿类动物的临床前研究，嗅觉障碍可能是一个敏感的测量结果来预测新的治疗策略的治疗潜力。 许多测试已经设计并广泛用于帕金森病的啮齿动物模型和TES表征感觉障碍吨的新的治疗方法11-15的治疗潜力。尽管嗅觉赤字是有据可查的帕金森病，嗅觉功能尚未经常在很多车型进行测量。这种观点正在发生变化，虽然与遗传形式的帕金森病的发现和比较公认的观点，即帕金森病是一种全身性疾病影响的不仅仅是感觉功能。目前，有在帕金森病和其他神经变性疾病的遗传小鼠模型大量的研究，现在包括嗅觉的分析中表征16-24。给定的嗅觉障碍在神经退行性疾病的越来越大的兴趣，我们试图组装的，可用于临床前研究以表征神经变性的新模型，以及测试潜在的疾病修饰治疗嗅觉测试的电池。在本研究中所描述的试验中采用了两个特征和临床前研究18,25。 测试极力推ighted在这项研究中已被证明是在帕金森病（18）的频繁使用的α-突触核蛋白过表达的小鼠模型中检测嗅觉障碍敏感。它们包括掩埋沉淀试验26,27，块试验23,24,28,29的改编版本，并且习惯/ dishabituation测试30。需要注意的是有位在小鼠嗅觉功能的敏感措施在本研究中所描述的测试的几个适应是非常重要的，那些在本研究强调是测试我们的实验室拥有最经验和常规使用的。

<table border="0" cellpadding="0" cellspacing="0" width="352">
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:88px;">Cap&#39;n Crunch</td>
			<td style="width:88px;">Quaker Oats</td>
			<td align="right" style="width:88px;">3E+10</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:88px;">Wooden Blocks</td>
			<td style="width:88px;">Lara&#8217;s Crafts</td>
			<td align="right" style="width:88px;">10144</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="63">
			<td height="379" rowspan="6" style="height:379px;width:88px;">Flavor Extracts</td>
			<td rowspan="6" style="width:88px;">Kroger</td>
			<td style="width:88px;">Almond: 011110664716</td>
			<td rowspan="6" style="width:88px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:88px;">Anise: 011110615619</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:88px;">Banana: 011110669919</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:88px;">Coconut: 011110669889</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:88px;">Lemon: 011110669957</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:88px;">Orange: 0011110669964</td>
		</tr>
		<tr height="41">
			<td height="59" rowspan="2" style="height:59px;width:88px;">Tissue Cartridges</td>
			<td rowspan="2" style="width:88px;">Sigma-Aldrich</td>
			<td rowspan="2" style="width:88px;">Z672122-500EA</td>
			<td rowspan="2" style="width:88px;">Manufactured by Simport no: M490-2</td>
		</tr>
		<tr height="18">
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:88px;">Cotton Balls</td>
			<td style="width:88px;">Kroger</td>
			<td align="right" style="width:88px;">1.1111E+10</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:88px;">Mouse Cages</td>
			<td style="width:88px;">Ancare</td>
			<td style="width:88px;"></td>
			<td style="width:88px;">19 x 29 x 12.7 cm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:88px;">Camcorder</td>
			<td style="width:88px;">Sony</td>
			<td style="width:88px;">&#160;HDR-HC9</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:88px;">MiniDV Tapes</td>
			<td style="width:88px;">Sony DVC premium</td>
			<td align="right" style="width:88px;">2.7243E+10</td>
			<td style="width:88px;">60 minute long play&#160;</td>
		</tr>
	</tbody>
</table>

olfactory assays for mouse models of neurodegenerative disease

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process and may be a useful behavioral marker for early detection. Earlier detection in neurodegenerative disease is a major goal in the field because this is when neuroprotective therapies have the best potential to be effective. Therefore, in preclinical studies testing novel neuroprotective strategies in rodent models of neurodegenerative disease, olfactory assessment could be highly useful in determining therapeutic potential of compounds and translation to the clinic. In the present study we describe a battery of olfactory assays that are useful in measuring olfactory function in mice. The tests presented in this study were chosen because they measure olfaction abilities in mice related to food odors, social odors, and non-social odors. These tests have proven useful in characterizing novel genetic mouse models of Parkinson&#8217;s disease as well as in testing potential disease-modifying therapies.

掩埋丸，块，并习惯/ dishabituation测试都是嗅觉小鼠的高度有利的评价。使用这些试验中，我们发现在嗅觉在帕金森氏病的多个基因小鼠模型，包括α-突触核蛋白过表达（大鼠Thy1-ASYN）18和Atp13a2敲除小鼠显著变化。这两种大鼠Thy1-ASYN和Atp13a2基因敲除小鼠需要较长时间才能找到埋藏颗粒比对照组18。 图从野生型和Atp13a2基因敲除小鼠收集1-3显示的数据。在埋入沉淀试验，Atp13a2敲除小鼠显示增加的延迟，以找到掩埋粒料相比野生型对照小鼠（ 图1）。在块试验都野生型和Atp13a2敲除小鼠嗅探从另一小鼠的笼子的新颖块相比，从自己家笼（ 图2）块。在习惯/ dishabituation测试野生型小鼠表现出习惯一个气味和<html>然后，当增加了一种新的气味引入（ 图3）嗅。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51804/51804fig1highres.jpg" width="500" /> 图1。埋颗粒测试。延迟发现沉淀在野生型（N = 10）和Atp13a2 KO（N = 14），在20-27米以上。 * P &lt;0.05，曼 - 惠特尼U。 <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51804/51804fig2highres.jpg" width="500" /> 图2块测试时间嗅探小说块（五）在该块试验中野生型组（n = 10）和Atp13a2 KO（N = 12）的雌性小鼠20 - 27米岁。 ΔΔ表示P &lt;0.01相对于块A，B和C从同一基因型。采用Mann-Whitney U。 “SRC =”/文件/ ftp_upload / 51804 / 51804fig3highres.jpg“WIDTH =”500“/&gt; 图3：习惯化/ dishabituation测试，平均时间在嗅野生型小鼠香味盒（9米的年龄，N = 7），在6个试验，并引进墨盒与一种新型香味审判7。

Watch this Scientific Journal Video about Olfactory Assays for Mouse Models of Neurodegenerative Disease at JoVE.com

Olfactory Assays for Mouse Models of Neurodegenerative Disease

Impairment in olfactory function is a common feature in many neurodegenerative disorders including Parkinson, Alzheimer, and Huntington diseases. In the present article, we describe a set of tests for assessing olfaction discrimination and detection in mice that can be used to measure olfactory abilities in mouse models of neurodegenerative diseases.

嗅觉测定的神经退行性疾病的小鼠模型

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process ...

Research

Education

JoVE Core

JoVE Journal

Anatomy and Physiology

Neuroscience

Unit 1

The Special Senses

SCIENTISTS IN ACTION

Mouse Models of Periventricular Leukomalacia

脑室周围白质软化的小鼠模型

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature ...

科研

神经科学

Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence

检测内源NADH的荧光双光子成像的小鼠大脑皮层Microregional缺氧

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. ...

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

评估神经退行性表型<em&gt;果蝇</em&gt;爬坡试验和全脑多巴胺能神经元免疫组化染色

Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the ...

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

一个鼠标，两种文化：隔离和成人神经的培养干细胞从单个小鼠的两个神经源性区

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

嗅觉受体神经元在全山免疫标记<em&gt;果蝇</em&gt;天线

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction

新生小鼠脑侧脑室注射病毒的持续和广泛的神经元传导

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene ...

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

分析突触调制<em&gt;果蝇</em&gt;曝光时间延长后，光光感受器

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at ...

Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease

Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay ...

In Vivo Single-Molecule Tracking at the Drosophila Presynaptic Motor Nerve Terminal

In Vivo Single-Molecule Tracking at the Drosophila Presynaptic Motor Nerve Terminal

在体内果蝇前运动神经末端的单分子跟踪

An increasing number of super-resolution microscopy techniques are helping to uncover the mechanisms that govern the nanoscale cellular world. ...

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

嗜德维人神经元树突状树突状复杂度的定量分析

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. ...