This work is supported by a Baylor University Research Council grant and by a research grant from the Epilepsy Foundation.

在以下实验中使用的成年雄性小鼠产生并容纳在贝勒大学在22℃的环境温度下，用14小时光照和10小时黑暗（20:00至6:00小时）昼夜周期。给予小鼠随意获得食物和水。所有的程序给小鼠均符合的护理和使用实验动物和动物卫生协议指引全国学院是经贝勒大学动物护理和使用委员会。小鼠在上午晚些时候/下午早些时候进行测试。对于研究者下的所有程序，应戴手套，以消除异味，防止污染。 1.准备非社会的气味<ol><li>测试中的每一天准备新鲜的非社会的气味。 注:研究者应使用手套，准备气味。 <ol><li>填3离心管，用10毫升的水每。 </li><li>吸取100μl的香蕉提取物的入15ml＆＃160;锥形管和100μl的杏仁提取物的成第二的15毫升锥形管中以1:100稀释。 注意:第三15ml试管将仅含有水。 </li></ol></li></ol> 2.制备社会的气味<ol><li>准备在上午的测试会出现新的社会气味。从还没有被清理为至少三天，并包含小鼠相等数目两个保持得到社会的气味。获得从性别相同的测试受试者的小鼠社会气味。 注意:这些是要被存储在RT，并应在4-6小时用于维持强烈的和一致的气味。 <ol><li>确定有多少社会气味棉签需要创建:每只小鼠被测试将看到来自社会的气味1，和三个三拭子从社会的气味2。 </li><li>通过在一个肮脏的笼子底部擦拭棉签以Z字形方式编制社会的气味。抖掉多余的被褥抱住一角。 </li><li>商店棉签在大玻璃罐子的盖子或盖上封口膜烧杯。指定一个容器社会气味1和第二社会的气味2。 </li></ol></li><li>保持气味源网箱实验测试室外面。 </li></ol> 3.驯化测试鼠标<ol><li>戴上手套并称重小鼠进行测试并单独放置在一个干净的笼子，顶部的线盖和水瓶除去。 </li><li>让鼠标适应新的笼子45分钟在一个房间以外的检测室。确保有没有在房间里不寻常的气味或噪音。 </li><li>由于测试需要约45分钟，通过将鼠标旁边的错开时间驯化要测试到一个干净的笼子测试当前鼠标前适应的权利。 </li></ol> 4.编写的测试室<ol><li>将新鲜的棉签，实验室带，包含社会气味拭子瓶，n的管上社会的气味，微量，两个定时器和一个废液瓶在试验室的盖子。 </li><li>确保考场是合理的自由气味，大声喧哗，明亮的光线。不要穿带香味的古龙水或洗液。 注意:观察者不应该在测试期间口香糖或糖果，并应尽量限制其暴露在强烈气味的食物。 </li></ol> 5.测试程序<ol><li>把鼠标驯化它在笼中带入考场，并放置在柜台上笼的高度舒适的观看，同时就位。 </li><li>呈现每个气味3次连续，每次2分钟。使用显示顺序，可在表1中找到。 <ol><li>吸移管将50μl加臭剂的气味的被测试到棉棒的前端和重新盖上的气味源。提起笼子的顶部电线插入棉签的无味，木制的一端插入水瓶开盘，推着克通过从金属丝顶端的下侧的长度。避免接触棉签任何东西，包括手指，使异味污染是可以避免的。 </li><li>继续通过水的瓶口画棉签直到大约1英寸的棉花年末下垂入笼。加盖棉签使用实验室磁带笼边的木结束。 注意:小心不要让盖子回落到笼子，造成一个响亮的噪声会干扰测试鼠标。 </li><li>立即开始2分钟时间。用一个定时器时间2分钟演讲。记录花了2分钟，试验期间嗅气味的累计时间。使用单独的秒表计数动物的嗅探行为。主动嗅探定义为当动物的鼻子是面向棉签用它的鼻子〜2 cm或接近尖。 </li><li>在完成每次试验后，抬高电线盖并通过联合国向下拉动取出棉签盖子derside（这避免了气味的污染防止棉尖端接触到导线）。密封在500毫升的废液瓶的使用棉签。 </li><li>准备并插入下棉签，并开始计时。让每一个试验间隔约之间1分钟。继续直到每个气味已被提出的三次连续，对于每次2分钟。 </li></ol></li><li>一旦鼠标已经完成了所有的试验，清空含有用棉签在一个垃圾桶试验室可以在外面的废液瓶。返回鼠标它的笼子。 </li></ol> 6.分析<ol><li>用于使用双向ANOVA，然后进行Newman-Keuls检验或类似的事后检验，以确定显著习惯（更短的时间嗅探连续相同的气味），并且dishabitutation（更多时间嗅探一个新闻），每个组每气味条件分析数据。 </li></ol>

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<li>Pan, Y. W., Kuo, C. T., Storm, D. R., Xia, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inducible+and+targeted+deletion+of+the+ERK5+MAP+kinase+in+adult+neurogenic+regions+impairs+adult+neurogenesis+in+the+olfactory+bulb+and+several+forms+of+olfactory+behavior%5BTitle%5D)+AND+%22PLoS+ONE%22%5BJournal%5D)">Inducible and targeted deletion of the ERK5 MAP kinase in adult neurogenic regions impairs adult neurogenesis in the olfactory bulb and several forms of olfactory behavior.</a> PLoS ONE. 7, e49622(2012).</li>
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The authors have nothing to disclose.

本文提供的结果是最优的小鼠。小鼠表现出一种新的气味了强烈的反响，然后迅速，观察者。之一，在该方法中的关键步骤是气味的制备。调查人员需要采取非常谨慎，以彼此隔离的气味，以防止污染。另一个重要方面是棉签的呈现。研究者需要解决的棉签的最佳位置，以便鼠标可以调查尖，但不撕裂的尖端。测试的一个重要组成部分是气味的选择。我们提出了三个非社会的气味，但它不会是很?...

小鼠是依赖于嗅觉用于导航新环境，寻找食物，用于识别其它个人，和性行为1-3。重要的是，研究人员建立实验动物是否不管理涉及食品，社会交往，或意在引起从鼠标的响应任何气味行为测试前的嗅觉功能感。嗅觉缺失或无法不同的气味区分，可能会导致在各种行为范例误报或底片，所以其它类型的行为测试之前，执行一个鼠标的检测和辨别气味的能力应该建立。 嗅觉习惯/ dishabituation试验是在20世纪80年代4首先描述。测试已适应于由Drs小鼠使用。穆押嗯和杰奎琳·克劳利5。这是一种简单而廉价的测试，允许调查者ESTAblish一个鼠标可以检测和气味区分。除了测试嗅觉，该测试允许研究者观察小鼠的一般行为，应在测试方案的早期完成。关于鼠标的运动定性观察，焦虑的迹象，活动水平，并回应社会对气味的食物的气味可能预示着在测试可能进行的新领域。 在该试验中，浸入各种气味棉签呈现给小鼠三次在一排。与每个重复介绍的气味，将鼠标，观察者向棉棒，花费更少的时间与每个随后呈现调查它。当一个新的气味呈现，dishabituation发生，并且一个典型的鼠标会花费更多的时间来调查拭子，这表明它可在当前和以前的气味5区分。此测试被施用给一只小鼠在一时间，并包括45分钟ACClimation期间随后测试45分钟。 虽然这个测试很容易进行可能被用于调查有关鼠标的嗅觉系统的复杂问题。嗅觉的其他流行的测试，如掩埋食品测试，简单地建立嗅觉缺失的存在或不存在。然而，嗅觉辨别和习惯测试允许研究者确定鼠标不仅具有检测气味，但不同的气味之间可以区分的能力。习惯和dishabituation的格局已被用于建立新的突变体6,7气味区分。在一个令人惊讶的研究，Fadool及其同事利用气味的复杂混合物，表明小鼠基因靶向删除的Kv1.3通道都"的嗅觉超级"，可以区分非常相似的气味比正常老鼠更8。 当检查了新的基因敲除小鼠模型是有用的establis^ h的正常行为存在基本的感官任务。当早期的测试方案完成后，气味辨别和习惯测试给出了一个调查员有机会观察到任何异常行为。这些观察结果可以防止在随后的测试假阳性或阴性结果归因于混淆的突变体的特征。研究人员继续调查社会行为，需要验证基本的嗅觉功能变得越来越重要。除了检查嗅觉在突变系，这种测试可用于解决特定的问题，例如一个药物治疗是否特异性增加动物的响应社会刺激气味，或者如果它们的反应增大到所有刺激气味。

<table><tbody><tr><td>Mouse cage</td><td>Allentown</td><td>Standard mouse cage</td><td></td></tr><tr><td>Wire lid</td><td>Allentown</td><td>BCU Mouse WBL 2500</td><td></td></tr><tr><td>Bedding</td><td>Harlan</td><td>7090 Sani-Chips</td><td></td></tr><tr><td>Cotton swabs</td><td>VWR</td><td>89031-270</td><td>6&amp;rdquo; wooden handle</td></tr><tr><td>Banana extract</td><td>McCormick</td><td></td><td></td></tr><tr><td>Almond extract</td><td>McCormick</td><td></td><td></td></tr><tr><td>Laboratory tape</td><td>VWR</td><td>89098-062</td><td></td></tr><tr><td>Stop watch</td><td>VWR</td><td>62374-000</td><td></td></tr><tr><td>Nitrile gloves</td><td>VWR</td><td>82026</td><td></td></tr><tr><td>Timing device</td><td>VWR</td><td>61161-350</td><td></td></tr><tr><td>15 ml conical tubes</td><td>VWR</td><td>89003-294</td><td></td></tr><tr><td>2 L beakers&amp;nbsp;</td><td>Pyrex</td><td>1003</td><td></td></tr><tr><td>Parafilm</td><td>Parafilm</td><td>PM-992</td><td>4&amp;rdquo; x 250&amp;rsquo;</td></tr><tr><td>1 L bottle with cap</td><td>VWR</td><td>89000-240</td><td></td></tr></tbody></table>

testing for odor discrimination and habituation in mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

以下描述的方案，结果是使用10个成人（90-120日龄）的雄性C57BL / 6J小鼠和八个雄性FVB混合背景品系（50-70日龄）记录。测试如下进行多天的单一研究者。图形上的每个点代表花费调查气味的平均时间。 当第一次引入的棉签浸渍在水，小鼠花费了大量的时间调查这个新物体（ 图1:前三个数据点）。随着每个连续的演讲，更少的时间花在调查棉签，表示习惯。当一个?...

Watch this Scientific Journal Video about Testing for Odor Discrimination and Habituation in Mice at JoVE.com

Testing for Odor Discrimination and Habituation in Mice

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

在小鼠试验的气味辨别和习惯化

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

testing-for-odor-discrimination-and-habituation-in-mice

Research

JoVE Journal

Behavior

17.7K 次观看.  Baylor University. This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor. 

视频: Testing for Odor Discrimination and Habituation in Mice

The Attentional Set Shifting Task: A Measure of Cognitive Flexibility in Mice

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many neuropsychiatric and neurodevelopmental disorders, including depression, post-traumatic stress disorder, schizophrenia and autism spectrum disorder. Deficits in cognitive function also represent the most difficult symptom domain&#160;to successfully treat, as serotonin reuptake inhibitors and tricyclic antidepressants have only modest effects. Functional neuroimaging studies and postmortem analysis of human brain tissue implicate the PFC as being a primary region of dysregulation in patients with these disorders. However, preclinical behavioral assays used to assess these deficits in mouse models which can be readily manipulated genetically and could provide the basis for studies of new treatment avenues have been underutilized. Here we describe the adaptation of a behavioral assay, the attentional set shifting task (AST), to be performed in mice to assess prefrontal cortex mediated cognitive deficits. The neural circuits underlying behavior during the AST are highly conserved across humans, nonhuman primates and rodents, providing excellent face, construct and predictive validity.

The goal of this protocol is to perform a behavioral assay such as the attentional set shifting task (AST) to assess prefrontal cortex-mediated cognitive flexibility in mice.

在注意定势转移任务:测量小鼠认知的灵活性

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many ...

科研

行为学

Simple and Computer-assisted Olfactory Testing for Mice

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected with aging and is a strong predictor of neurodegenerative diseases. Thus, olfaction testing is used as a non-invasive diagnostic method to detect neurological deficits early on. In order to understand the mechanisms underlying olfactory network susceptibility, olfactory research in rodents has gained momentum in the past decade.
Here, we present a very simple, time efficient and reproducible olfactory testing method of innate odor perception and sensitivity in mice without the need of any prior food or water restriction. The tests are performed in a familiar environment to the mice, require only the scents and a 2 min session of odorant exposure. The analysis is performed, post-hoc, using computer-assisted commands on ImageJ and can be, therefore, carried out from start to end by one researcher.
This protocol does not require any special hardware or setup and is indicated for any laboratory interested in testing olfactory perception and sensitivity.

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

简单和计算机辅助测试嗅觉的小鼠

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

小鼠学习记忆研究的新对象识别实验

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

Olfactory Behavioral Testing in the Adult Mouse

The rodent olfactory system is of increasing interest to scientists, studied, in part, in systems biology because of its stereotyped, yet accessible circuitry. In addition, this area's unique ability to generate new neurons throughout an organism's lifetime makes it an attractive system for developmental and regenerative biologists alike. Such interest necessitates a means for a quick, yet reliable assessment of olfactory function. Many tests of olfactory ability are complex, variable or not specifically designed for mice. Also, some tests are sensitive to memory deficits as well as defects in olfactory abilities, confounding obtained results.
Here, we describe a simple battery of tests designed to identify defects in olfactory sensitivity and preference. First, an initial general health assessment allows for the identification of animals suitable for further testing. Second, mice are exposed to various dilutions of scents to ascertain whether there is a threshold difference. Third, mice are presented with various scents, both attractive and aversive, that allow for the assessment of olfactory preference. These simple studies should make the initial characterization of olfactory behavior accessible for labs of varied resources and expertise.

Fundamental, yet unique properties of the rodent olfactory system have led to its increasing study among biologists. A relatively simple assessment of its function is then also needed. Here we describe sensitive tests for the characterization of mouse olfactory sensitivity and preference.

在成年小鼠的嗅觉行为测试

The rodent olfactory system is of increasing interest to scientists, studied, in part, in systems biology because of its stereotyped, yet accessible ...

生物学

评估吸入环境污染物导致小鼠嗅觉损害

Studying the Effects of Inhaled Environmental Pollutants on Olfactory Function in Mice

Olfactory impairment is a significant public health problem and independently predicts the risk of neurodegenerative diseases. Inhaled environmental pollutants exposure may impair olfaction; thereby, there is an urgent need for methods to evaluate the effects of inhaled environmental pollutants exposure on olfaction. Mice are ideal models for olfactory experiments because of their highly developed olfactory system and behavioral characteristics. To assess the effects of inhaled environmental pollutants exposure on olfactory function in mice, a detailed buried food test and social odor discrimination experiment is provided, including the experiment preparation, the selection and construction of experimental facilities, the testing process, and indexes of time. Meanwhile, timekeeping equipment, operational details, and the experimental environment are discussed to ensure the success of the assay. Zinc sulfate is used as the treatment to demonstrate the feasibility of the experimental approach. The protocol provides a simple and clear operational process for assessing the effects of inhaled environmental pollutants on olfactory function in mice.

作者聚焦：评估空气污染物的嗅觉影响 - 埋藏的食物和社会气味测试

This paper provides a detailed description of the buried food test and social odor discrimination experiment to assess the effects of inhaled environmental pollutants exposure on olfactory function in mice.

研究吸入环境污染物对小鼠嗅觉功能的影响

Olfactory impairment is a significant public health problem and independently predicts the risk of neurodegenerative diseases. Inhaled environmental ...

环境科学

Investigating Migraine-Like Behavior Using Light Aversion in Mice

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as photophobia. Whilst it is impossible to confirm that a mouse is experiencing migraine, light aversion can be used as a behavioral surrogate for the migraine symptom of photophobia. To test for light aversion, we utilize the light/dark assay to measure the time mice freely choose to spend in either a light or dark environment. The assay has been refined by introducing two critical modifications: pre-exposures to the chamber prior to running the test procedure and adjustable chamber lighting, permitting the use of a range of light intensities from 55 lux to 27,000 lux. Because the choice to spend more time in the dark is also indicative of anxiety, we also utilize a light-independent anxiety test, the open field assay, to distinguish anxiety from light-aversive behavior. Here, we describe a modified test paradigm for the light/dark and open field assays. The application of these assays is described for intraperitoneal injection of calcitonin gene-related peptide (CGRP) in two mouse strains and for optogenetic brain stimulation studies.

Rodents are not able to report migraine symptoms. Here, we describe a manageable test paradigm (light/dark and open field assays) to measure light aversion, one of the most common and bothersome symptoms in patients with migraines.

使用轻度厌恶研究小鼠偏头痛样行为

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as ...

Assessment of Mouse Judgment Bias through an Olfactory Digging Task

Judgment biases (JB) are differences in the way that individuals in positive and negative affective/emotional states interpret ambiguous information. This phenomenon has long been observed in humans, with individuals in positive states responding to ambiguity 'optimistically' and those in negative states instead showing 'pessimism'. Researchers aiming to assess animal affect have taken advantage of these differential responses, developing tasks to assess judgment bias as an indicator of affective state. These tasks are becoming increasingly popular across diverse species and fields of research. However, for laboratory mice, the most widely used vertebrates in research and a species heavily relied upon to model affective disorders, only one JB task has been successfully validated as sensitive to changes in affective state. Here, we provide a detailed description of this novel murine JB task, and evidence of its sensitivity to mouse affect. Though refinements are still necessary, assessment of mouse JB opens the door for answering both practical questions regarding mouse welfare, and fundamental questions about the impact of affective state in translational research.

This article provides a detailed description of a novel mouse judgment bias protocol. Evidence of this olfactory digging task's sensitivity to affective state is also demonstrated and its utility across diverse research fields is discussed.

通过嗅觉挖掘任务评估小鼠判断偏差

Judgment biases (JB) are differences in the way that individuals in positive and negative affective/emotional states interpret ambiguous information. This ...

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.

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 ...

在小鼠试验的气味辨别和习惯化

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