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. 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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. 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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. 