Name: murine drinking models in the development of pharmacotherapies for alcoholism: drinking in the dark and two-bottle choice
Uploaded: 2019-01-07T14:00:00
Description: Watch this Scientific Journal Video about Murine Drinking Models in the Development of Pharmacotherapies for Alcoholism: Drinking in the Dark and Two-bottle Choice at JoVE.com

这项工作得到了 ctsi nhcrr/ncat驻 ul1tr000130 (d. l. d.)、aa022448 (d. l. d.) 和 usc 药学学校研究赠款的部分支持。

这里描述的所有程序都已得到南加州大学健康科学校区动物护理和使用机构委员会的批准。
1. 实验设置和组装
<ol><li>在研究开始前获取以下所有用品和化学品: 小鼠、cages/金属笼上衣、床上用品、食物、水、乙醇、钢琴、拉链、收缩包装、实用刀、拉链、胶带、bunsen 燃烧器、秤、头灯。</li>
	<li>从商业来源或内部群体获得 c57bl/6j 小鼠, 请记住, 小鼠可以被分组, 直到测试时间...

<ol>
	<li>Litten, R. Z., Falk, D. E., Ryan, M. L., Fertig, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discovery,+Development,+and+Adoption+of+Medications+to+Treat+Alcohol+Use+Disorder:+Goals+for+the+Phases+of+Medications+Development.">Discovery, Development, and Adoption of Medications to Treat Alcohol Use Disorder: Goals for the Phases of Medications Development.</a> Alcoholism: Clinical and Experimental Research. 40 (7), 1368-1379 (2016).</li><li>Grant, B. F., Dawson, D. A., Stinson, F. S., Chou, S. P., Dufour, M. C., Pickering, R. 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+12-month+prevalence+and+trends+in+DSM-IV+alcohol+abuse+and+dependence:+United+States,+1991-1992+and+2001-2002.">The 12-month prevalence and trends in DSM-IV alcohol abuse and dependence: United States, 1991-1992 and 2001-2002.</a> Drug and Alcohol Dependence. 74 (3), 223-234 (2004).</li><li>Sacks, J. J., Gonzales, K. R., Bouchery, E. E., Tomedi, L. E., Brewer, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=National+and+State+Costs+of+Excessive+Alcohol+Consumption.">National and State Costs of Excessive Alcohol Consumption.</a> American Journal of Preventive Medicine. 49 (5), 73-79 (2015).</li><li>Litten, R. Z., 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=Medications+development+to+treat+alcohol+dependence:+a+vision+for+the+next+decade.">Medications development to treat alcohol dependence: a vision for the next decade.</a> Addiction Biology. 17 (3), 513-527 (2012).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Johnson+Medication+Treatment+of+Different+Types+of+Alcoholism.">Johnson Medication Treatment of Different Types of Alcoholism.</a> American Journal of Psychiatry. 167 (6), 630-639 (2010).</li><li>Litten, R. Z., Wilford, B. B., Falk, D. E., Ryan, M. L., Fertig, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+medications+for+the+treatment+of+alcohol+use+disorder:+An+evaluation+of+clinical+efficacy+and+safety.">Potential medications for the treatment of alcohol use disorder: An evaluation of clinical efficacy and safety.</a> Substance Abuse. 37 (2), 286-298 (2016).</li><li>Litten, R. Z., Ryan, M. L., Falk, D. E., Reilly, M., Fertig, J. B., Koob, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+of+Alcohol+Use+Disorder:+Understanding+Mechanisms+to+Advance+Personalized+Treatment.+Alcoholism:+Clinical+and+Experimental.">Heterogeneity of Alcohol Use Disorder: Understanding Mechanisms to Advance Personalized Treatment. Alcoholism: Clinical and Experimental.</a> Research. 39 (4), 579-584 (2015).</li><li>Schuckit, M. A., 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+Genome-Wide+Search+for+Genes+That+Relate+to+a+Low+Level+of+Response+to+Alcohol.">A Genome-Wide Search for Genes That Relate to a Low Level of Response to Alcohol.</a> Alcoholism: Clinical and Experimental Research. 25 (3), 323-329 (2001).</li><li>Batki, S. L., Pennington, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+Personalized+Medicine+in+the+Pharmacotherapy+of+Alcohol+Use+Disorder:+Targeting+Patient+Genes+and+Patient+Goals.">Toward Personalized Medicine in the Pharmacotherapy of Alcohol Use Disorder: Targeting Patient Genes and Patient Goals.</a> American Journal of Psychiatry. 171 (4), 391-394 (2014).</li><li>Koob, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theoretical+frameworks+and+mechanistic+aspects+of+alcohol+addiction:+alcohol+addiction+as+a+reward+deficit+disorder.">Theoretical frameworks and mechanistic aspects of alcohol addiction: alcohol addiction as a reward deficit disorder.</a> Current topics in behavioral neurosciences. 13, 3-30 (2013).</li><li>Yoneyama, N., Crabbe, J. C., Ford, M. M., Murillo, A., Finn, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voluntary+ethanol+consumption+in+22+inbred+mouse+strains.">Voluntary ethanol consumption in 22 inbred mouse strains.</a> Alcohol. 42 (3), 149-160 (2008).</li><li>Rhodes, J. S., Best, K., Belknap, J. K., Finn, D. A., Crabbe, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+simple+model+of+ethanol+drinking+to+intoxication+in+C57BL/6J+mice.">Evaluation of a simple model of ethanol drinking to intoxication in C57BL/6J mice.</a> Physiology &amp; Behavior. 84 (1), 53-63 (2005).</li><li>Thiele, T. E., Navarro, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term="Drinking+in+the+dark"+(DID)+procedures:+A+model+of+binge-like+ethanol+drinking+in+non-dependent+mice.">"Drinking in the dark" (DID) procedures: A model of binge-like ethanol drinking in non-dependent mice.</a> Alcohol. 48 (3), 235-241 (2014).</li><li>Crabbe, J. C., Spence, S. E., Brown, L. L., Metten, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alcohol+preference+drinking+in+a+mouse+line+selectively+bred+for+high+drinking+in+the+dark.">Alcohol preference drinking in a mouse line selectively bred for high drinking in the dark.</a> Alcohol. 45 (5), 427-440 (2011).</li><li>Sprow, G. M., Thiele, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurobiology+of+binge-like+ethanol+drinking:+Evidence+from+rodent+models.">The neurobiology of binge-like ethanol drinking: Evidence from rodent models.</a> Physiology &amp; Behavior. 106 (3), 325-331 (2012).</li><li>Neasta, J., Hamida, B., Yowell, Q., Carnicella, S., Ron, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+for+mammalian+target+of+rapamycin+complex+1+signaling+in+neuroadaptations+underlying+alcohol-related+disorders.">Role for mammalian target of rapamycin complex 1 signaling in neuroadaptations underlying alcohol-related disorders.</a> Proceedings of the National Academy of Sciences. 107 (46), 20093-20098 (2010).</li><li>Huynh, N., 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=Preclinical+development+of+moxidectin+as+a+novel+therapeutic+for+alcohol+use+disorder.">Preclinical development of moxidectin as a novel therapeutic for alcohol use disorder.</a> Neuropharmacology. 113, 60-70 (2017).</li><li>Egli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+experimental+paradigms+and+animal+models+be+used+to+discover+clinically+effective+medications+for+alcoholism.">Can experimental paradigms and animal models be used to discover clinically effective medications for alcoholism.</a> Addiction Biology. 10 (4), 309-319 (2005).</li><li>Huynh, N., Arabian, N., Lieu, D., Asatryan, L., Davies, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilizing+an+Orally+Dissolving+Strip+for+Pharmacological+and+Toxicological+Studies:+A+Simple+and+Humane+Alternative+to+Oral+Gavage+for+Animals.">Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals.</a> Journal of Visualized Experiments. (109), (2016).</li><li>Yardley, M. 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=Ivermectin+reduces+alcohol+intake+and+preference+in+mice.">Ivermectin reduces alcohol intake and preference in mice.</a> Neuropharmacology. 63 (2), 190-201 (2012).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> Journal of Pharmacology and Pharmacotherapeutics. 1 (2), 87-93 (2010).</li></ol>

全世界的估计表明, 多达7600万人符合酒精使用障碍 (aud) 诊断的标准。不幸的是, 目前可用的药物治疗基本无效, 需要进一步发展, 以抵消这一临床人群的需求20。为此, 以下协议旨在通过举例说明两种最基本的啮齿类动物饮酒模式来促进这一努力: 双瓶选择 (tbc) 和黑暗中饮酒 (did)。这两种模型都测量乙醇的非操作性自管理, 即小鼠口服乙醇。在 tbc 范式中, 乙醇 (10% vv) 和水都是连续?...

在过去的25年多时间里, 在开发治疗酒精使用障碍 (aud)1的药物方面做出了巨大的努力。尽管取得了许多进展, 但澳元仍然是一个重大的公共卫生问题, 影响到 1 800多万美国人, 每年花费 2, 200多亿美元。目前只有三种 fda 批准的药物, 二硫酮、纳曲酮和阿卡莫酸, 所有这些药物在临床试验中都产生了不一致的结果, 即使与临床环境中的心理社会干预相结合, 也取得了有限的成功4 个,5,6,7。
目前 aud 治疗失败的一个主要原因与 aud8的异质性有关。虽然环境和遗传因素都有助于澳元的发展, 但遗传力估计占发病风险的 50-60%.与抑郁症的治疗类似, 人们普遍认为, 患有澳元的患者将需要各种量身定制的药物, 以满足每个患者的需求 1 0.
显然, 迫切需要更有效的治疗方法, 如果简化本已艰巨和耗时的药物发现过程3, 将有助于这种治疗。为此, 下面的协议证明了两种广泛用于检测 aud 11 的神经生物学基础的啮齿类动物饮酒模型的临床前适用性.更具体地说, 本文介绍的方法可以评估候选化合物在减少酒精消费的有效性, 在 "适度" 和 "暴饮暴食" 的情况下, 利用两瓶选择 (tbc) 和在黑暗中饮酒 (did) 的范式,分别。这两种范式都考察了非操作乙醇自我管理, 即小鼠口头和随意摄入乙醇, 从而说明了高脸和构建有效性作为人类酒精中毒的模型11。
在 tbc 饮酒, 也被称为免费饮酒、偏好饮酒或社交饮酒, 两瓶溶液在家里的笼子里不断提供。一个瓶子含有水, 另一个含有稀释的乙醇溶液, 因此乙醇的浓度可以改变 (例如, 5-30% v/v)11,12。老鼠可以不断地接触到这两个瓶子, 因此, 可以选择从每个瓶子里喝多少酒。
该模型评估每只小鼠的乙醇消费量 (g/kg), 以及乙醇偏好比 (乙醇消费量; 总液体消费量)。它通常被用来比较不同品种小鼠的饮酒水平, 或者在特定的基因操纵 (如基因敲除或敲除) 之后, 导致血液乙醇浓度 (bec) 类似于在人类身上发现的水平。适量、14岁。
在 did 过程中, 在黑暗循环开始3小时后, 用 20% (v/v) 的乙醇溶液交换家庭笼瓶, 以进行有限的饮用。饮酒是连续4天的周期, 在4-day 持续 2小时, 第4天持续4小时。第4天作为第4天测试前的酒精适应期。因此, 小鼠将可靠地消耗足够的乙醇, 以达到 bec & gt;100 mg/dl, 因此, 表现出中毒的行为影响, 在人类中发现的暴饮暴食 13,14,15.除饮酒场所之外, 任何时候都可以获得水。
有限的获取饮料有几种变化。例如, 在间歇性访问模型中, 老鼠仅在周一、周三和周五收到两瓶 (一瓶含有20% 的水, 另一瓶含有20% 乙醇), 平日和周末分别有24小时和48小时的戒断期, 为 16.经过几个星期的间歇性访问, 小鼠将逐渐和自愿上升的饮酒水平, 最终达到相关的 dod 模型中观察到的 beck。然而, did 似乎是评估暴饮暴食行为最常用的模型。其他间歇性饮酒的模式也存在, 但它们依赖于限制获得食物或蒸汽室导致自愿自我管理增加的情况, 这使得它们对自愿饮酒的代表性降低了16。

<table border="0" cellpadding="0" cellspacing="0" style="width:1617px;" width="1616">
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	<tbody>
		<tr height="25">
			<td height="25" style="height:25px;width:483px;">1 L&#160; graduated cylinder</td>
			<td style="width:315px;">VWR</td>
			<td style="width:232px;">https://us.vwr.com/store/product/20935285/marisco-single-scale-cylinder-graduates-john-m-maris-co</td>
			<td style="width:588px;">To prepare ethanol solution.</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">1 L glass bottle</td>
			<td>Pyrex (Fisher Scientific)</td>
			<td>https://www.fishersci.com/shop/products/pyrex-reusable-media-storage-bottles-12/p-42752</td>
			<td>To prepare ethanol solution.</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">100 mL graduated cylinder</td>
			<td>Fisher Scientific</td>
			<td>https://www.fishersci.com/shop/products/kimble-chase-kimax-class-a-to-contain-graduated-cylinders-8/p-4369311</td>
			<td>To prepare ethanol solution.</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Analox</td>
			<td></td>
			<td></td>
			<td>One potential method of analyzing DID blood samples is by using the analox machine</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">ball-bearing sipper tubes</td>
			<td>Ancare Corp.</td>
			<td>http://www.ancare.com/products/watering-equipment/open-drinking-tubes/straight-tubes-ball-point</td>
			<td>Length: 2.5 inches, Diameter: 5/16 inches, Model: TD100</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">C57BL/6J Mice</td>
			<td>Jackson lab</td>
			<td>https://www.jax.org/strain/000664</td>
			<td>May also come from internal breeding colony</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">disposable serological pipets</td>
			<td>VWR International (VWR)</td>
			<td>https://us.vwr.com/store/product/4760455/vwr-disposable-serological-pipets-polystyrene-sterile-plugged</td>
			<td>10 mL, 18 mL, or 25 mL&#160;</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">ethanol, pure, 190 proof (95%), USP, KOPTEC</td>
			<td>Decon Labs (VWR)</td>
			<td>https://us.vwr.com/store/product/4542412/ethanol-pure-190-proof-95-usp-koptec</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">heat gun&#160;</td>
			<td>Master Appliances Corp.</td>
			<td colspan="2">http://www.masterappliance.com/master-heat-guns-kits/</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">heat shrink tubing</td>
			<td>---</td>
			<td>---</td>
			<td>Diameter: 3/8 inches</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">industrial knife/blade</td>
			<td>---</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">metal cage plate</td>
			<td>---</td>
			<td>---</td>
			<td>Should be available through the university/institutional vivarium</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">mouse RO water</td>
			<td>---</td>
			<td>---</td>
			<td>Should be available through the university/institutional vivarium</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">portable electronic scale</td>
			<td>Ohaus (VWR)</td>
			<td>https://us.vwr.com/store/product/4789377/portable-electronic-cs-series-scales-ohaus</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">red light headlamp</td>
			<td>nyteBright (Amazon)</td>
			<td>https://www.amazon.com/LED-Headlamp-Flashlight-Red-Light/dp/B00R0LMMF8/ref=sr_1_1?ie=UTF8&#38;qid=1499591137&#38;sr=8-1-spons&#38;keywords=red+lamp+headlamp&#38;psc=1</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">silicone stoppers</td>
			<td>Fisher</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">thermometer</td>
			<td>Fisher Scientific</td>
			<td>https://www.fishersci.com/shop/products/fisher-scientific-hygro-thermometer-clock-large-display-2/p-4077232</td>
			<td>---</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">weigh boat</td>
			<td>VWR International (VWR)</td>
			<td>https://us.vwr.com/store/product/16773534/vwr-pour-boat-weighing-dishes</td>
			<td>The lid from a pipete tip box is an appropriate alternative</td>
		</tr>
	</tbody>
</table>

murine drinking models in the development of pharmacotherapies for alcoholism: drinking in the dark and two-bottle choice

酒精使用障碍 (aud) 是一个主要问题, 估计全世界有超过7600万人符合诊断标准。目前的治疗方法仅限于三种 fda 批准的药物, 即使与心理社会干预相结合, 这些药物也基本无效, 高复发率就证明了这一点。因此, 寻找更多的新治疗方法是一个重要的公共卫生目标。为此, 下面的协议使用两个简单的啮齿类动物饮酒模型来评估铅抗酒精化合物的临床前疗效: 两瓶选择 (tbc) 和在黑暗中饮酒 (did)。前者允许小鼠适度自愿饮酒, 而后者则诱导小鼠在短时间内自愿饮酒, 模仿暴饮暴食。这两种模式的简单和高吞吐量特性允许快速筛选药理制剂或识别表现出某些自愿饮酒行为的小鼠菌株。

在以下具有代表性的调查中, 采用两瓶选择 (tbc) 范式对社交饮酒进行了建模。简单地说, 老鼠可以接触到两瓶溶液, 其中一瓶含有水, 另一瓶含有 10% (vv) 乙醇溶液。研究对象随后被分割并均匀地分配给药物治疗组, 莫西丁 (mox) 与盐水控制, 因此每个组的平均乙醇摄入量水平将以最小的变化。
最初基线10e 摄入量在24小时期间稳定?...

Watch this Scientific Journal Video about Murine Drinking Models in the Development of Pharmacotherapies for Alcoholism: Drinking in the Dark and Two-bottle Choice at JoVE.com

Murine Drinking Models in the Development of Pharmacotherapies for Alcoholism: Drinking in the Dark and Two-bottle Choice

小鼠饮酒模型在酒精中毒药物治疗发展中的应用: 黑暗中的饮酒和两瓶选择

酒精使用障碍 (aud) 是一个主要的国家健康问题, 需要开发更有效的治疗方法来抵消这些病人群体的需求。为此, 下面的协议使用两个简单的啮齿类动物饮酒模型来评估铅抗酒精化合物的临床前疗效。

Alcohol Use Disorder (AUD) is a major problem with more than an estimated 76 million people worldwide meeting the diagnostic criteria. Current treatments ...

这一范式的总体目标是使用酒精使用障碍的啮齿动物模型评估潜在的抗酒精药物的有效性。酒精使用障碍是一个重大的公共卫生危机。目前的治疗是无效的，需要持续的临床前研究。我们的协议使我们能够评估潜在治疗的安全性和效用，并了解疾病本身的基本神经生物学。在 Bunsen 燃烧器上加热实用刀开始该步骤。使用这把刀，切掉大约一英寸的塑料18毫升血清移液器的顶部和底部。接下来在热枪下加热移液器。将滚珠轴承吸管插入移液器的底端。然后使用市售的收缩包装枪，用收缩包装将吸管密封到位。用硅胶塞盖住另一个开口。对于动物习惯，在抵达时，每只老鼠在标准聚碳酸酯聚糖酮笼内，配有床上用品、食物和水和金属网笼顶。从一周单间住房适应的第四天开始，测量和记录小鼠的日常身体和食物重量。使用倒置瓶旁边的蚀刻测量进水量，记录半月板的最高点。一旦水摄入量稳定到过去三天均值的正负 10%变化，开始使用 TBC 或 DID 获得乙醇。要进行TBS，在1000毫升中准备10%乙醇溶液，在894.7毫升水中加入105.3毫升的190证明谷物乙醇，然后彻底摇动。鉴于乙醇迅速蒸发，每周更换两次溶液。在TBC的第一天，在每个笼子里，清空两个水瓶中的一个，用新鲜准备的乙醇溶液填充到边缘。鉴于乙醇和水很难在视觉上区分，请清楚地标记瓶子及其相应的内容。根据需要向水瓶添加更多溶液。然后将瓶子放回笼子中，确保盖子牢固关闭，没有任何气泡或喷口泄漏。只需轻拍瓶子即可清除任何气泡，使空气能够逃离瓶子。每隔一天交替瓶子的位置，以纠正在饮酒活动中与条件位置偏好相关的影响。记录乙醇摄入量水平，分析10%乙醇摄入量和偏好比。一旦乙醇摄入量变得稳定，在日常测量过程中对每只小鼠进行单一控制内内注射，以习惯小鼠注射。一旦一个低变异性的乙醇基线重新建立，用乙醇摄入量值将小鼠分成对照组和实验组，以便所有组都有大致相似的平均乙醇摄入量值。开始每天的药物药位，无论是急性或多天持续时间。要进行 DID，在预定的乙醇获取的每一天，记录水量、食物摄入量和体重的测量结果，并进行药物给药。在根据药物的药代动力学选择的光周期中，在预选期间执行此操作，使化合物在饮酒期间接近大脑的最大浓度。接下来，在1000毫升中，将210.5毫升的190证明谷物乙醇加入789.5毫升的水中，准备20%乙醇溶液。然后彻底摇动它。在饮用期开始前填充乙醇瓶，以便一旦 DID 开始，可以简单地将水瓶替换为酒精瓶。在整个 DID 会话期间，使用红灯前照灯不打扰动物。在 DID 饮用会议开始时，记录每只小鼠的积量，该会议计划从光循环前三小时进入黑暗循环。然后用一瓶20%乙醇溶液更换每个水瓶，并记录起始乙醇体积。两小时后，在第一天到第三天到四小时后，在第四天饮酒会议结束时阅读并记录最后的乙醇量。然后分析20%乙醇摄入量。随后，用水瓶代替乙醇瓶并记录水量。这一数字显示了对急性剂量莫西德金在每公斤5毫克对乙醇摄入量和偏好的影响的评估。单剂量的莫西德汀显著减少酒精摄入量超过45%以上相比，与莫西汀注射和偏好超过30%10%乙醇摄入量和偏好都保持明显低于盐水后，立即在MOX治疗的一天。在这里， 狂欢饮酒是使用 DID 程序建模的。老鼠在一天到第四天被施用盐水，基线20%乙醇在第四天建立。在第二个每周周期的第一天到第三天，所有小鼠每天接受另一次盐水注射。然后，第三天的乙醇摄入量值被分成两组，随后以每公斤5毫克的速度注射一剂莫西德金，或在第四天接受盐水注射。下一周，所有小鼠在一天到第四天每天接受盐水注射，第四天再次测量乙醇摄入量。对MOX的急性给药进行了分析，发现与药物注射相比，酒精摄入量显著减少54%以上。请记住，今后必须使用本协议进行实验，以评估潜在药物的机制、剂量和毒性，并评估其与当前治疗方案相关的有效性。

Research

JoVE Journal

Medicine

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

小鼠胰岛的分离和包膜下肾移植的方法

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

科研

医学

A Strategy to Identify de Novo Mutations in Common Disorders such as Autism and Schizophrenia

一项战略，以确定在常见的疾病，如自闭症和精神分裂症的新突变

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, ...

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

利用X射线计算机断层扫描在小鼠肥胖模型的FAT卷的分割和测量

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

闭塞性细支气管炎在原位肺移植小鼠模型的发展

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

基于音频的虚拟游戏环境的发展，以帮助盲人的导航技巧

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Technical Aspects of the Mouse Aortocaval Fistula

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Collection and Extraction of Saliva DNA for Next Generation Sequencing

收集和唾液DNA提取用于新一代测序

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality ...

Mouse Kidney Transplantation: Models of Allograft Rejection

鼠肾移植：移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

伤口的牙髓评价一个直接盖髓模型的开发和治疗小鼠中修复性牙本质形成

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...