Support provided by grant AA019462 to DAH and AA019884 to DDS.

在这里和在所附的视频中描述的所有程序已经批准的健康科学中心的机构动物护理和使用委员会和新墨西哥大学的主校区。 1.产前乙醇暴露<ol><li>获得所有需要的材料和化工产品：糖精钠盐水合物，190证明乙醇（95％的酒精体积比），玻璃珠饮水管，天然橡胶白色＃4塞一个孔，1个“弯管球点，纸统治者从WWW印刷.vendian.org / mncharity / DIR3 / paper_rulers /。 </li><li>获得成熟的成年大鼠饲养员从供应商或替代来源。使用长埃文斯饲养者在该协议提出的方法和代表性的数据。雌性体重125-150克，并到达使得它们大约9-10周的繁殖时老时6-7周龄（步骤1.5）。确保所有的准水坝都是首次母亲。确保男性是12周龄的到来和15周龄在养殖协议的开始。 </li><li>房子单独塑料笼中的动物，在22°C在反向12小时亮/暗时间表（灯从21:00到09:00小时）。为获得食物和自来水随意在整个研究中，包括所有的饮用水会议期间。进到步骤1.4之前允许至少1周驯化至设施。 </li><li>获得基线体重对每个雌性鼠。 </li><li>评价在雌性大鼠孕前饮用水平2周。按照步骤1.5.1-1.5.6秩序。 <ol><li>天1和2填写包含在自来水中0.066％糖精溶液（无酒精）饮用水管。为了量化消费贴上一纸尺，毫米精度的饮水管之前，加油吧。使用这种方法，因为它与重管量化乙醇消费相关的测量误差降低。填充管标尺上20mm的大关。开始四小时drinki纳克会话（步骤1.5.1-1.5.6和1.7）在黑暗阶段（10:00小时）时的活性和饮酒水平是最高的发病后1小时。 </li><li>在每次饮用会议结束量化被消耗的糖精溶液的体积。为了便于测量，确定每毫米的解决方案在标尺上的提前量和转换毫米到卷。对于这里推荐的管子每个MM对应0.366毫升。 </li><li>天3和4填充有2.5％的乙醇（体积/体积），并在自来水中0.066％糖精溶液的管。在每届会议结束量化的乙醇溶液（每公斤体重的重量），这是消费量。 </li><li>第5天，然后填充管，用5％乙醇（V / V），并在自来水中0.066％糖精溶液。在每届会议结束量化的乙醇溶液（每公斤体重的重量），这是消费量。 </li><li>在孕前饮酒阶段完成权衡大鼠和计算平均乙醇消费ption并为整个组的标准偏差。除去大鼠的量意味着消耗大于1个标准差以上或以下基团从研究意味着减少怀孕期间在自愿饮用的可变性。 </li><li>分配剩余的大鼠要么使得孕前饮酒水平匹配尽可能为两组糖精控制或PAE的条件。 </li></ol></li><li>在1-14天对每个雌性大鼠与一个成熟的男性饲养员。雌性大鼠在繁殖期不消耗乙醇。确认妊娠阴道塞的存在下，权衡雌性大鼠，并分别容纳她。这被定义为妊娠天1.（可选）离开雌性大鼠与男性饲养员长达5天，之后从研究去除女性。 注：由于怀孕期间乙醇消费始于妊娠期第1天的检查10:00小时的阴道塞的存在应该执行在此之前的时间。 </li><li>提供乙醇或糖精溶液为每天4小时（1000到1400小时），进行妊娠的持续时间。 <ol><li>自妊娠第1天提供的雌性大鼠与0％或5％的乙醇和基于该组指配在自来水0.066％糖精溶液。确保0％乙醇溶液提供到糖精对照大鼠的体积相匹配的由乙醇大鼠消耗5％乙醇溶液的平均体积。在每届会议结束量化的乙醇溶液（每公斤体重重）或糖精被消耗的数量。 </li><li>称取大鼠水坝每周评估孕妇体重增加。 </li><li>提供食物和自来水在任何时候，包括饮用水会议。 </li><li>停止乙醇暴露过程时，后代是天生的。录制现场幼崽和短重的数量在出生。指定诞生之日起为0日龄。 </li><li>剔除垃圾约2-3日龄10只。尝试吨Ø维持男性与女性的动物在每个垃圾相等比例。如果这是不可能的，剔除至10幼仔与男性和女性的不等号。 </li><li>记录的小狗出生的权重后2-3天。 </li><li>在断奶约21-24日龄和房屋同性配对的动物从相同的产前治疗条件的动物。不要使用超过1-2只从每个实验每窝以限制潜在的垃圾影响。 </li></ol></li></ol> 2.社会行为<ol><li>获取和准备所有必要的材料和设备。 <ol><li>获得社会互动的装置。用的腔室与材料构成一个敞开的顶部，易于清洗和消毒。该装置的前部，应覆盖有刚性透明塑料（〜2毫米的厚度）进行拍摄。行的内壁和地板用透明塑料（〜2毫米厚），以帮助清洁和气味控制。代替反射镜沿着背面内部わLL以帮助分析。 注意：对于该代表性数据这里报告的自定义腔室（95厘米长×47厘米宽×43厘米高），具有开放的顶部和刚性透明塑料（2毫米厚）覆盖所述装置的内部侧面和地面使用。镜子被置于沿后壁。的具体尺寸和材料都没有对社会行为的测量关键的，然而，我们建议的尺寸足够大，以确保真正的社会互动可以从其他的行为区分开来。也就是说，该装置应足够大，使动物并不总是接近彼此。 </li><li>获取摄像机能低或无光照条件下拍摄，因为所有的拍摄时，很少或没有环境光在可见光谱范围内进行的。确保相机具有高分辨率的红外光谱范围内，但是，能够记录所有的相机在其本土黑暗条件下夜间模式或用另外的红外照明就足够了。 </li><li>该装置的周围的位置的红外照明器，以改善该视频记录装置的照明。 </li><li>获得实验室级木片（白杨芯片）。 </li><li>得到一种刷子，簸箕，二氧化氯和异丙醇（70％）用于清洁会话和气味控制之间的装置。 </li></ol></li><li>设备驯化和社会行为<ol><li>在实验的这一阶段，使成年大鼠是年龄至少90天测量。 </li><li>此前每一届删除任何树林芯片，擦拭干净设备用异丙醇控制会话之间的气味，并提供新鲜的木屑。确保该木片完全覆盖该装置的底部。用适当的试剂清洗和消毒的装置中，诸如二氧化氯，是必要的。 </li><li>连续三天，把动物和笼子的队友进入腔室30分钟，以适应动物到装置。在适应会议室的所有灯都关掉。 </li><li>在第三次会议驯化的房子年底分别在新笼子新鲜的床上用品，食物和水24小时的动物来激励社会交往。 </li><li>在动物后，次日24小时创纪录的社会交往分离。 <ol><li>从该装置中删除木屑，清洁和消毒用二氧化氯，用异丙醇擦拭，以控制气味，并取代在会议之前的木片。 </li><li>定位一个或多个摄像机来记录的相互作用。位置的至少一个照相机，在装置的前部，以便在该装置的后壁的反射镜可以提供用于分析的其他视角。 </li><li>检索动物一次并保持动物在摄像机前面，使得皮毛图案的独特功能可以注意到。这些识别特征毛发可用于在分析过程中，而不是人为地标记的动物区分的影响。 注：由于啮齿动物社会交往的许多方面涉及嗅觉信号和闻到的合作伙伴，无论应避免可能引进国外气味。 Long-Evans大鼠通常具有可被利用来区分任何给定的一对动物的皮毛图案的一些特征。对于其它菌株（ 例如 ，Sprague-Dawley大鼠）的替代方法，如标记用无味染料尾部也可以使用。认识到，许多感兴趣的行为是朝向具体的目标是非常重要的（ 例如 ，肛门生殖器嗅探定向尾部的根部附近，冲着颈部，或朝向侧面或腹部侵略性攻击的颈背嬉戏攻击）。标志着动物接近尾部远离感兴趣这些目标的前端被推荐。 </li><li>至少12分钟的社会互动的录像。</li><li>监测动物的整个会话战斗。如果可能的话，观看经由显示器或窗口中的动物，以便实验者是不是在会话期间的房间。 注：战斗一直与成年大鼠的研究很少观察到的，但是，动物应该在整个会话监测。会议应，如果有过多的打斗或者不再有伤害或伤害的迹象的动物。 </li></ol></li></ol></li><li>行为编码和分析。 <ol><li>找出感兴趣的下列行为按照以前的工作与PAE 1,18。量化的持续时间，频率和延迟下列行为之一的第一次出现;摔跤（包括钉扎），拳击，爬行（渡）过/合作伙伴下，肛门生殖器嗅，合作伙伴的身体（身体嗅探）等嗅探，相互理（伙伴的修饰），饲养和嗅探/挖木片。每种行为的例子是说明D在这篇文章中的视频组件。 </li><li>量化来自视频感兴趣的社会行为。获得的频率，总的持续时间和延迟，以第一个实例为每个感兴趣的行为。 注：获得这些措施可以手动实现，然而，使用数字化视频的计算机化分析这些措施量化值得推荐。一个MATLAB（www.mathworks.com）脚本播放视频和行为的定量是作为补充本文。 </li><li>编码被结束对所有的大鼠后生成的持续时间，频率和延迟数据与统计软件包进行分析。 </li></ol></li></ol>

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-dependent+effects+of+prenatal+ethanol+exposure+on+synaptic+plasticity+and+learning+in+mature+offspring.">Dose-dependent effects of prenatal ethanol exposure on synaptic plasticity and learning in mature offspring.</a> Alcohol. Clin. Exp. Res. 26 (11), 1752-1758 (2002).</li><li>Rasmussen, D. D., 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=Chronic+daily+ethanol+and+withdrawal:+1.+Long-term+changes+in+the+hypothalamo-pituitary-adrenal+axis.">Chronic daily ethanol and withdrawal: 1. Long-term changes in the hypothalamo-pituitary-adrenal axis.</a> Alcohol. Clin. Exp. Res. 24 (12), 1836-1849 (2000).</li><li>Savage, D. D., 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=Effects+of+a+Novel+Cognition-Enhancing+Agent+on+Fetal+Ethanol-Induced+Learning+Deficits.">Effects of a Novel Cognition-Enhancing Agent on Fetal Ethanol-Induced Learning Deficits.</a> Alcohol. Clin. Exp. Res. 34 (10), 1793-1802 (2010).</li><li>Goodlett, C. R., Johnson, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+binge+ethanol+exposure+using+intubation:+Timing+and+dose+effects+on+place+learning.">Neonatal binge ethanol exposure using intubation: Timing and dose effects on place learning.</a> Neurotoxicol. Teratol. 19 (6), 435-446 (1997).</li><li>Staples, M. C., Rosenberg, M. J., Allen, N. A., Porch, M. W., Savage, D. 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> Impact of Combined Prenatal Ethanol and Prenatal Stress Exposure on Anxiety and Hippocampal-Sensitive Learning in Adult. 37 (12), 2039-2047 (2013).</li><li>Champagne, F. A., Francis, D. D., Mar, A., Meaney, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+maternal+care+in+the+rat+as+a+mediating+influence+for+the+effects+of+environment+on+development.">Variations in maternal care in the rat as a mediating influence for the effects of environment on development.</a> Physiol. Behav. 79 (3), 359-371 (2003).</li><li>Hamilton, D. 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=Patterns+of+social-experience-related+c-fos+and+Arc+expression+in+the+frontal+cortices+of+rats+exposed+to+saccharin+or+moderate+levels+of+ethanol+during+prenatal+brain+development.">Patterns of social-experience-related c-fos and Arc expression in the frontal cortices of rats exposed to saccharin or moderate levels of ethanol during prenatal brain development.</a> Behav. Brain Res. 214 (1), 66-74 (2010).</li><li>Barnett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A study in behaviour: Principles of ethology and behavioural physiology displayed mainly in the rat. , (1963).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Play-fighting+differs+from+serious+fighting+in+both+target+of+attack+and+tactics+of+fighting+in+the+laboratory+rat+Rattus-Norvegicus.">Play-fighting differs from serious fighting in both target of attack and tactics of fighting in the laboratory rat Rattus-Norvegicus.</a> Aggressive Behav. 13 (4), 227-242 (1987).</li><li>Meaney, M. J., Stewart, 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+descriptive+study+of+social-development+in+the+rat+(Rattus-Norvegicus).">A descriptive study of social-development in the rat (Rattus-Norvegicus).</a> Anim. Behav. 29 (1), 34-45 (1981).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prejuvenile+onset+of+play+fighting+in+laboratory+rats+(Rattus+norvegicus).">The prejuvenile onset of play fighting in laboratory rats (Rattus norvegicus).</a> Dev. Psychobiol. 31 (3), 193-205 (1997).</li><li>Himmler, B. T., Pellis, V. C., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peering+into+the+dynamics+of+social+interactions:+measuring+play+fighting+in+rats.">Peering into the dynamics of social interactions: measuring play fighting in rats.</a> J. Vis. Exp. (71), e4288-e4288 (2013).</li><li>Panksepp, J., Siviy, S., Normansell, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+psychobiology+of+play+:+Theoretical+and+methodological+perspectives.">The psychobiology of play : Theoretical and methodological perspectives.</a> Neurosci. Biobehav. Rev. 8 (4), 465-492 (1984).</li><li>Siviy, S. M., Panksepp, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+modulation+of+juvenile+play+in+rats.">Sensory modulation of juvenile play in rats.</a> Dev. Psychobiol. 20 (1), 39-55 (1987).</li><li>Pellis, S. 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=The+effects+of+orbital+frontal+cortex+damage+on+the+modulation+of+defensive+responses+by+rats+in+playful+and+nonplayful+social+contexts.">The effects of orbital frontal cortex damage on the modulation of defensive responses by rats in playful and nonplayful social contexts.</a> Behav. Neurosci. 120 (1), 72-84 (2006).</li><li>Blanchard, R. J., Blanchard, D. C., Takahashi, T., Kelley, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attack+and+defensive+behavior+in+albino-rat.">Attack and defensive behavior in albino-rat.</a> Anim. Behav. 25 (3), 622-634 (1977).</li></ol>

The authors have no conflicts of interest to disclose.

此处所描述的产前酒精暴露范例涉及在妊娠期间的乙醇自愿食用（5％V / V）的大鼠水坝。有许多用于在文献中，这相对于乙醇给药时间，剂量，时间和路线，以及所研究的物种不同表示脑发育过程中暴露的非人类的动物到乙醇的协议。虽然这里不提供彻底治疗各种暴露协议的优点，自愿饮用方法PAE的几个优点在本协议中描述的高亮显示。以前我们使用的流质饮食协议，常用于研究这方面的做法，?...

儿童估计有1-5％被诊断患有胎儿酒精谱系障碍（FASDs）2，其中包括胎儿酒精综合症（FAS），部分FAS（疫区），与酒精相关的神经发育障碍（ARNDs）3。在社会行为和认知赤字之间的儿童FASDs 4-7观察到的最常见的不良后果。消极后果不限于重产前酒精暴露（PAE），中度PAE不导致FAS的显眼形态，行为和认知缺陷特性可引起比较微妙，但仍然持续存在，缺陷在人类与FASDs 8-10和大脑发育11在非人类的动物暴露于乙醇。理解适度PAE的行为和相应的神经生物学后果的重要性是由这表明绝大多数的胎儿酒精中毒综合症病例属于少秒内目前的估计强调艾威范围频谱12。 几个独立的实验室报告了啮齿类动物的社会行为改变大脑发育过程中乙醇暴露相关，包括减少调查和互动1,13-15，改变打法14,16,17，增加响应积极的互动17,18，改建为社会刺激19-21，和赤字在社会上获得的食物的喜好和社会的认可存储器22。社会行为缺陷已观察暴露于重（血液乙醇浓度（BEC中）〜300毫克/分升）22,23或乙醇更温和的水平（BEC中〜80毫克/分升）1，并在广泛的范围内的其他显著参数因素，包括曝光定时，曝光的持续时间，并在行为测定时的年龄。 以前的研究已经证明，改变社会INT的具体方面eraction在成年大鼠歧视暴露在中等水平的酒精从控制动物暴露于糖精1,18。特别是，适度PAE一直被与强劲增长的摔跤，这表明增加的攻击行为，并在成年后社会调查（ 例如 ，嗅探合作伙伴）水平较低有关。因为在社会行为改变是PAE的可靠的后果，以下的PAE社会行为的量化可以保持为前进朝向PAE相关的改变中的社会行为的神经基础的更完整的理解和介入方法的发展的重要性。本文和相关视频的目标是提供指令的温和PAE协议和方法，用于在成年子女社会行为具有的量化可靠地分辨产前醇暴露从非暴露鼠后代。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Saccharin sodium salt hydrate</td>
			<td>Sigma</td>
			<td>S1002</td>
			<td></td>
		</tr>
		<tr>
			<td>190 proof ethanol</td>
			<td>Sigma</td>
			<td>493538</td>
			<td></td>
		</tr>
		<tr>
			<td>Beaded glass drinking tubes</td>
			<td>Fisher</td>
			<td>14-955K</td>
			<td></td>
		</tr>
		<tr>
			<td>Natural rubber white #4 stopper one hole</td>
			<td>Plasticoid</td>
			<td>LSG4M181</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#34; bend tubes-ball point</td>
			<td>Ancare</td>
			<td>TD-199-3&#34;</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper rulers</td>
			<td>N/A</td>
			<td>N/A</td>
			<td><a href="http://www.vendian.org/mncharity/dir3/paper_rulers">www.vendian.org/mncharity/dir3/paper_rulers</a></td>
		</tr>
		<tr>
			<td>Apparatus for social interaction</td>
			<td>Custom built</td>
			<td>N/A</td>
			<td>95 cm X 47 cm X 43 cm</td>
		</tr>
		<tr>
			<td>Video cameras</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Capable of recording low/no light conditions</td>
		</tr>
		<tr>
			<td>Infrared illuminators</td>
			<td>Vitek</td>
			<td>VT-IR1-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Teklad laboratory grade sani-chips</td>
			<td>Harlan</td>
			<td>7090A</td>
			<td></td>
		</tr>
		<tr>
			<td>Brush and dustpan</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>Sigma</td>
			<td>W292907</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorine Dioxide (1.5 mg Tablets)</td>
			<td>Quiplabs</td>
			<td>N/A</td>
			<td>Prepare per manufacturer&#39;s recommendation</td>
		</tr>
	</tbody>
</table>

moderate prenatal alcohol exposure and quantification of social behavior in adult rats

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

以上的许多育种轮的过程中雌性大鼠的乙醇条件一致喝平均约2.1克/公斤的每4小时饮用会话乙醇。鼠水坝引入饮用管后的第一个15至30分钟期间消耗大约一半的四个小时的总的，产生了峰值母体血清乙醇浓度为约60毫克/分升，在45分钟的时间点测量。在剩余3.5小时的饮用期间，他们继续消耗5％乙醇0.4克/公斤体重/小时的低，但相对稳定的速率。自愿乙醇消费的这种水平和模式，对孕妇体重增加，后?...

Watch this Scientific Journal Video about Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats at JoVE.com

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

适度的产前酒精暴露在成年大鼠社会行为的量化

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

moderate-prenatal-alcohol-exposure-quantification-social-behavior

Research

JoVE Journal

Behavior

9.6K 次观看.  University of New Mexico. The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood. 

视频: Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

一个过程来观察大鼠巴甫洛夫空调酒精寻求行为的背景下引发的重建

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

科研

行为学

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

啮齿动物脑显微注射学习动机行为的分子基板

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

应力的社会问题：社会支持和社会认同的特里尔社会的实验操作压力测试

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods require saccharin/sucrose fading, water deprivation and/or extended training to initiate operant responding in rats. This paper describes a novel and efficient method to quickly initiate operant responding for ethanol that is convenient for experimenters and does not require water deprivation or saccharin/sucrose fading, thus eliminating the potential confound of using sweeteners in ethanol operant self-administration studies. With this method, Wistar rats typically acquire and maintain self-administration of a 20% ethanol solution in less than two weeks of training. Furthermore, blood ethanol concentrations and rewards are positively correlated for a 30 min self-administration session. Moreover, naltrexone, an FDA-approved medication for alcohol dependence that has been shown to suppress ethanol self-administration in rodents, dose-dependently decreases alcohol intake and motivation to consume alcohol for rats self-administering 20% ethanol, thus validating the use of this new method to study the reinforcing properties of alcohol in rats.

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

一种用于评价大鼠乙醇的补强性能没有禁水，糖精衰落或扩展访问训练方法

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

使用仿引起范式检查召回内存在婴幼儿时期

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

鼠类精神运动警觉测试（rPVT）：一个在大鼠和小鼠的神经行为评估性能的方法

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

用小鼠训练和社会观察教老鼠主动改变笼子

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

Integrating Computerized Linguistic and Social Network Analyses to Capture Addiction Recovery Capital in an Online Community

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery capital development in online communities of recovery from alcohol and drug (AOD) addiction. Recovery capital was conceptualised as both engagement in the online recovery community and identification with the community. To measure recovery capital development, naturally occurring data were extracted from the social media page of a specific recovery program, with the page being set up as a resource for a face-to-face recovery program. To map engagement with the online community, social network analysis (SNA) capturing online social interaction was performed. Social interaction was measured through the linkages between the online contributors/members of the online community as represented by program clients, staff, and supporters from the broader community. To capture markers of social identification with the online community, computerised linguistic analysis of the textual data (content from posts and comments) was conducted. Recovery capital captured in this way was analysed against retention data (a proxy outcome indicator), as days spent in the (face-to-face) recovery program. The online data extracted was linked to participant data in regards to program retention to test prediction of a key recovery outcome. This approach allowed the examination of the role of online support communities and assessment of the association between recovery capital (developed via the online community of recovery) and recovery outcomes.

The article describes a novel approach for analyzing dynamic online social interactions (in an online context) exemplified by a study of an online community of recovery from alcohol and drug addiction.

整合计算机语言和社交网络分析,在在线社区中捕捉成瘾恢复资本

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery ...

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include dominant-submissive relationships are common in both animal and human societies. These relationships can be affected by interactions with others and with their environment, making them difficult to analyze in a controlled study. Rather than a simple dominance hierarchy, this formation has a complicated presentation that allows rats to avoid aggression. Status can be stagnant or mutable, and results in complex societal stratifications. Here we describe a complex diving-for-food task to investigate rodent social hierarchy and behavioral interactions. This animal model may allow us to assess the relationship between a wide range of mental illnesses and social organization, as well as to study the effectiveness of therapy on social dysfunction.

This protocol describes a method of examining social hierarchy in a rat model. Rats perform a complex diving-for-food task in which they form a distinct hierarchy according to their willingness to dive underwater and swim to obtain a food pellet. This method is used to understand decision making and social relationships among highly social animals in small groups.