感谢S.Yokoyama协助收集鱼。我们也非常感谢瓦川村有关饲养方法的有用建议。本研究由日本科学促进会（KAKENHI）通过授予TS（24370006和16K07507）的资助

收集和运送活鱼注意:本协议描述了如何从具有深度≥15米的气囊收集具有气囊的鱼。快速输送到表面会导致气囊膨胀，压力变化会严重危害或杀死鱼类。谨慎是必要的，因为在第一步中对鱼类造成的损害将对其生存和后期行为产生不利影响。 <ol><li>在与SCUBA潜水之前，请收集以下材料:适合捕获水下目标物种的手网;双重聚乙烯袋，足够大的鱼;橡胶带（ø80 mm x 6 mm）;用于在表面充气收集袋的氧气瓶;作为抗微生物剂的Elbagin包含10％的硝基苯乙烯钠;聚苯乙烯泡沫箱，其中放置收集袋;移液管;并且如有必要（参见步骤1.4），长度为的绳子相当于最大潜水深度，以及至少2公斤的重量。 </li><li>潜水时用手网收集目标鱼类;同时仍然在水下，将捕获的鱼放在聚乙烯袋中，并用橡皮筋绑扎袋子的口。如果将鱼运送到水族馆需要≥1天的时间，还可以从其他袋子中的自然栖息地中保存水，以用于更新养鱼用水。 注意:为了安全起见，水下任务必须由至少两个人进行。 </li><li>以不超过1米/分钟的速度从收集点收集鱼的表面:在10米或更深的深度处，每2米停止上升1 - 5分钟;并从10米的深度到每1米的表面停止1 - 5分钟。当捕获的鱼不能保持其在袋中的浮力（ 即 ，如果它们在膨胀的气体膀胱浮起来的同时仍然试图向底部游泳时），或者保持相同的深度1-5分钟或de更深一点到2米。一旦鱼似乎恢复了浮力，恢复上升到地面。 注意:如果SCUBA罐中的空气有可能在上升时耗尽，则将绳索固定在聚乙烯袋上，并附加≥2kg的重量;在水下操作者安全上升之后，以与上述相同的速度将其拉到表面。 </li><li>在海滩或船上，表面处理后，每个聚乙烯袋的水中溶解10 ppm的Elbagin。 注意:虽然一些鱼可能在被带到地面之后可能无法保持其浮力，但大多数鱼可能会在一天之内恢复。 </li><li>如果收集袋中的鱼的密度很高，则可以在多袋之间划分鱼，以防止在运输过程中彼此相互碰撞而彼此相互损害。如果鱼是侵略性物种，请将它们分开包裹。 </li><li>使用氧气充气收集袋，并再次使用r密封袋的口ubberbands。当将氧气引入袋中时，将喷嘴降低到袋子的水中以增加溶解氧含量;完全膨胀的袋子应该是1/4满水。 </li><li>将收集袋放在聚苯乙烯泡沫箱中，以保持水温稳定，并在黑暗条件下降低鱼的压力。 </li><li>如果将鱼运送到水族馆需要≥1天的时间，每天一次可以在每个养鱼袋中换入1/4至1/3的水分，使用从栖息地保留的水加入10 ppm Elbagin ，并用氧气重新包装。每天用手网或移液管清除底部的死鱼和排泄物。 注意:如果涉及飞机运输，在运输之前等待至少1天将鱼适应到表面的空气压力，因为在短期内两步压力变化（从水下到地面，表面到上部空气）可能会产生负面影响鱼的生存。 </l<html>我&gt; </ol>将鱼适应水族馆<ol><li>将含有鱼的袋子在水族箱中漂浮30分钟以平衡水温。 </li><li>在10分钟的时间内，用水族箱水逐渐更换袋子水，以避免水分化学（ 例如 ，pH，盐度）突然变化引起的冲击。 注意:由于水分化学差异太大而引起震惊的鱼可能会显示身体颜色和/或行为的异常变化。适应期间仔细监测。 </li><li>将10 ppm Elbagin溶解在水族馆水中。 </li><li>此后，每天一次，持续3天，更新1/3的水族馆水，将10ppm的Elbagin添加到更换的水中。 </li><li>最后，通过一天一次交换一半的水族馆水，直到颜色消失，开始消除Elbagin。 </li></ol> 3.注射可见植入弹性体（VIE）标签以识别个体鱼T"&gt;注:在这项工作中，个别鱼使用VIE标签标识;对于示例，请参阅弗雷德里克·10，奥尔森和Vøllestad11，和勒布朗和诺克斯12此外，如果研究的物种是足够大的手握住，步骤3.2中使用的手术台将不是必需的。 <ol><li>确定每个人的注射位置和标签颜色。最安全的选择是在身体的厚背部或尾部部位选择一个注射点，并避免将其注射到可能刺穿内部器官的腹部。 注意: 图1中描述了编号系统:如果研究物种足够大以注入8个可能的位置，则该系统将允许使用单一颜色识别154个个体。 VIE标签的十种颜​​色可商购。根据鱼的身体颜色选择可区分的颜色。 </li><li> &lt;强&gt;对于小于可以握在手中的鱼，准备如下的外科手术台（也参见Kinkel 等人 13 ）（ 图2 ）。 <ol><li>切出5厘米长×5厘米宽的软海绵，比培养皿的高度至少低5-10毫米。 </li><li>在大约5 - 10 mm深度的海绵上切割一个凹槽，并将其宽度调整为鱼的近似身体宽度。将聚氯乙烯（PVC）板（0.3mm厚）切割成5cm×1.5cm宽，并将其弯曲成谷形折叠（或M形）。 </li><li>将开槽的PVC板放在开槽的海绵上，然后将海绵放入培养皿（ø160 mm，30 mm深）。使用回收罐的水填充培养皿，直到PVC板充分浸泡。 </li></ol></li><li>根据VIE标签手册准备VIE标签。混合弹性体材料，并将混合物加入到3-mL syri中配备29号针头，包含在套件中。 </li><li>在执行标签注射时准备使用两个水箱:一个用于麻醉，一个用于恢复;调整这些的温度和盐度浓度以匹配饲养水族馆的水。使用带有空气石头的空气泵将回水罐中的水温和循环。 </li><li>通过将2-甲基喹啉与等体积的99.5％乙醇混合来制备麻醉液;将其加入麻醉罐中以达到18ppm的浓度。 注意:麻醉液的最佳浓度将取决于个体的物种和体型。因此，提前检查研究物种的最佳浓度。 </li><li>将各个鱼转移到麻醉池，让它们变得麻醉:就是说，当鱼缸外面被点击时，等待鱼不会被触动或水的振动反应。当身体颜色在麻醉时变化在许多鱼中，也仔细监测身体颜色，并判断鱼是否从其变化中麻醉。因为鱼有可能跳出麻醉罐，使用透明的亚克力板覆盖，至少直到鱼开始缓慢移动。 注意:根据研究种类及其体型，微调麻醉和/或调整麻醉液浓度所需的时间。如果手术运动被麻醉所阻止，那么鱼会死亡的风险很高。 </li><li>如果身体变暖，鱼就会变弱，麻醉鱼时，手指会保持在比较冷的水中。 </li><li>从麻醉坦克提起鱼，并快速测量或记录必要的数据（总长度，性别等 ）。如果鱼在测量过程中似乎恢复，请立即将其转移回麻醉池。 </li><li>将鱼转移到手术台。将鱼腹侧放在t中他的PVC槽。如果体型非常小，则在注射时使用双目显微镜。如果研究种类足够大，可以握在手中，则在被保持时注入VIE标签。 </li><li>将针的斜面朝向外部并指向鱼头。皮下注射针，或多或少地平行于身体，并尽可能靠近皮肤下方。根据鱼体尺寸和最终看到标签的容易程度调整插入深度。 </li><li>在取针时注入VIE标签，并在针斜度到达针入口点之前停止注射（如果在相对较宽的区域注入标签，将很容易辨别）。 </li><li>完成标签注射后，立即将鱼转移到回收罐。如果恢复看起来太慢，用手轻轻地循环水。 </li><li>恢复后，将鱼返回饲养水族馆，并继续添加10 ppm Elbagin到水中3天。 注意:在VIE标签的低可见度条件下，UVA滤光灯将有助于识别标签。 </li></ol> 4.计数脱胶胶鸡蛋<ol><li>为了创建一个人造卵巢，鱼可以将粘合蛋沉积，将不透明的PVC管（ø5厘米，6厘米长）切成两半，垂直于直径。 注意:上述管道尺寸适用于产卵相对较小的虫虫T. marinae 。因此，调整适合研究种类的PVC管尺寸和形状。 </li><li>在防水片上打印5 mm x 5 mm格栅，并将其切割成适合PVC管道的内部区域。 </li><li>使用橡皮筋将防水片固定在PVC管的内部。 注意:如果女性难以在防水片上附上鸡蛋，并且鸡蛋从纸张上落下，则使纸张用砂纸进行纹理化。 </li><li>用一层沙子覆盖水族馆底部，1- 厚2厘米。将大约三分之一的PVC管倾斜地插入砂中，将板材固定侧向下。 </li><li> 鱼成功产卵后，从PVC管中取出蛋块，将其放入带有水族馆水的培养皿中。将一张新的纸张固定在PVC管内，并将其放回水族箱的底部。最后，通过拍摄鸡蛋块来计数鸡蛋。 <ol><li>或者，如果在产卵后进行亲子蛋保育的观察，小心不要将鸡蛋从水中取出。相反，使用培养皿与水族馆水一起舀起鸡蛋和床单，快速拍摄鸡蛋质量，然后仔细地将鸡蛋块重新吸收到同一个PVC管中，然后将其全部恢复到原来的位置水族箱底部。最后，使用图片计数鸡蛋。 </li></ol></li><li> 使用ImageJ计数鸡蛋（ 图3 </stro纳克&gt;）。这些自动和手动细胞计数方法在以下详细说明:基本细胞计数（芝加哥大学综合光显微核心设施http://digital.bsd.uchicago.edu/image_j.php）;粒子分析（http://imagej.net/Particle_Analysis）;单元格计数器（https://imagej.net/Cell_Counter）。 <ol><li>否则，如果鸡蛋密集沉积，则通过计算面积密度比估计蛋数:打印5×5 mm 2网格中的鸡蛋计算鸡蛋密度，用ImageJ测量鸡蛋覆盖的表面积，并估算每单位和表面积以上密度的鸡蛋总数。以下解释了ImageJ的上述表面积测量方法:Set scale ...（Research Services Branch，https://imagej.nih.gov/ij/docs/menus/analyze.html）;它解释了如何定义图像的空间尺度;使用ImageJ测量表面积（Keene州立大学，学术技术，https://dept.keene.edu/at/2011/04/15/by-matthew-ragan/）。 注意:如果鸡蛋是透明的，黑色防水片更适合自动计数方法，因为ImageJ可以单独识别鸡蛋的轮廓。 </li></ol></li></ol>

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I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behaviorally+induced+maternal+stress+in+a+fish+influences+progeny+quality+by+a+hormonal+mechanism.">Behaviorally induced maternal stress in a fish influences progeny quality by a hormonal mechanism.</a> Ecology. 79 (6), 1873-1883 (1998).</li><li>Knaepkens, G., Bruyndoncx, L., Coeck, J., Eens, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spawning+habitat+enhancement+in+the+European+bullhead+(Cottus+gobio),+an+endangered+freshwater+fish+in+degraded+lowland+rivers.">Spawning habitat enhancement in the European bullhead (Cottus gobio), an endangered freshwater fish in degraded lowland rivers.</a> Biodivers Conserv. 13 (13), 2443-2452 (2004).</li><li>Shiogaki, M., Dotsu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+life+history+of+the+gobiid+fish,+Zonogobius+boreus.">The life history of the gobiid fish, Zonogobius boreus.</a> Bulletin of the Faculty of Fisheries, Nagasaki University. 37, 1-8 (1974).</li><li>Meunier, B., Yavno, S., Ahmed, S., Corkum, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+documentation+of+spawning+and+nest+guarding+in+the+laboratory+by+the+invasive+fish,+the+round+goby+(Neogobius+melanostomus).">First documentation of spawning and nest guarding in the laboratory by the invasive fish, the round goby (Neogobius melanostomus).</a> J Great Lakes Res. 35 (4), 608-612 (2009).</li></ol>

作者没有什么可以披露的。

许多鱼类的生殖生态常常通过实验饲养来揭示。特别是，变性6，8，14，择偶15,16和种内竞争7,17一直使用水族馆保持鱼类详细调查的频繁的话题。此外，在水族箱观察到一些结果已经在后面的字段8，18</sup...

鱼类的形态学，生态学和行为学表​​现出明显的自适应进化1 。特别是与生殖相关的生态特征特别多样，其中大部分可以直接受个人健身的影响2 。为了深入了解导致不同鱼类独特特征演变的选择性压力，使用活鱼直接观察生殖和社会行为通常有助于证实理论假设。 然而，对鱼类的连续现场观察可能需要专门的难以维护的水下设备和设施。在这些情况下，野生捕捞的水族馆饲养的鱼的观察是有帮助3，4，5。此外，鱼类行为的有效观察，其他罕见或困难ULT在自然条件下，观察可以通过在水族箱6，7，8操纵实验成为可能。通过最大程度地减少人为压力和身体伤害，在良好条件下养鱼是准确生态调查的关键。 侏儒虾青y素总长 23-25毫米，分布在西太平洋，安静，庇护的海湾，深度9-26米9 。在这项工作中， T. marinae被用作一个模型，用于描述使用独立的水下呼吸器（SCUBA），鱼类运输和最终驯化鱼类进行直接观察的鱼类收集系列的基本技术的研究物种的生殖行为和生态学。

<table border="0" cellpadding="0" cellspacing="0" width="1037">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:192px;">Hand net</td>
			<td style="width:193px;">Nisso</td>
			<td style="width:108px;">AQ-17</td>
			<td style="width:544px;">Select for the target species size.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polyethylene bag</td>
			<td>San-U Fish Farm</td>
			<td>8194</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rubber band</td>
			<td>ESCO Co. LTD.</td>
			<td>78-0420-64</td>
			<td>&#248; 80 mm x 6 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen cylinder</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Oxgen cylinder for diving equipment suits.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Elbagin</td>
			<td>Japan Pet Design Co. Ltd.</td>
			<td>75950</td>
			<td>Pafurazine F (provided from same company) is equivalent drug to Elbagin.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polystyrene foam box</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pipette</td>
			<td>AS ONE</td>
			<td>1-8625-04</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rope</td>
			<td>Mizukami Kinzoku Co. LTD.</td>
			<td>95301601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weight</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Weight for diving equipment suits.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Water tank</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air pump</td>
			<td>KOTOBUKI</td>
			<td colspan="2">4972814 062115</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air stone</td>
			<td>KOTOBUKI</td>
			<td colspan="2">4972814 232204</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-Methylquinoline&#160;</td>
			<td>Wako</td>
			<td>170-00376</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol (99.5)</td>
			<td>Wako</td>
			<td>057-00456</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Visible implant elastomer tag kit</td>
			<td>Northwest Marine Technology</td>
			<td>N/A</td>
			<td>http://www.nmt.us/products/vie/vie.shtml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Soft sponge</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC board</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>0.3 mm thickness is easy to use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri dish</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>A large one, such as&#160; &#248; 160 mm and 30 mm depth, is convenient for the injection of the VIE tag.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transparent acrylic board</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UVA filtered light&#160;</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC pipe</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>&#248; 5 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Waterproof sheet</td>
			<td>SOMAR Corp.</td>
			<td>3EKW03</td>
			<td>The film for the plain copier.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sand</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereo microscope</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Camera</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

basic methods for the study of reproductive ecology of fish in aquaria

当持续的实地调查是不可能的时候，养殖观察对于揭示鱼行为和生态学方面是有价值的。在这里，描述了一系列基本技术，以便观察野生捕获的小型鱼的生殖行为，作为模型，保存在水族箱中。该方法着重于三个步骤:收集，运输和观察底物spavner的生殖生态学。活鱼收集和运输的基本方面是（1）防止对鱼的伤害，（2）仔细适应水族馆。收集活鱼时，预防伤害如划痕或突然变化的水压是必不可少的，因为任何物理伤害可能会对鱼的生存和后期行为产生负面影响。仔细适应水族箱可降低发病死亡率，减轻运输冲击。饲养期间的观察包括（1）识别个人双鱼和（2）监测产卵，对鱼或鸡蛋没有负面影响，从而对研究种的生殖生态进行详细的调查。皮下注射可见植入弹性体（VIE）标签是用于随后鉴定个体鱼的精确方法，并且可以与大范围的鱼一起使用，对其生存和行为影响最小。如果研究物种是沉积粘蛋的底物，那么由聚氯乙烯（PVC）管道构建的人造巢场，加上可拆卸的防水片将有助于计数和监测鸡蛋，减少调查人员对巢穴的影响和鱼的保卫行为。虽然这种基本方法需要在研究文章中很少提及的技术，但它们是进行需要养殖野生鱼类的实验的基础。

按照上述方法，分别于2014年4月，2015年和2016年在日本鹿儿岛县Am美大岛境外收集了41，15和96个海棠的个体（ 表1 ）。在每种情况下，25（61％），14（93％）和91（95％）个人住在直到在水族馆里存放鸡蛋。福田等人报道3 ，在2014年观察期结束前只有一只鱼死亡，鱼类产卵否则在捕获后7天开始出现，表明个体在良好条件下饲?...

Watch this Scientific Journal Video about Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria at JoVE.com

Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

描述了一系列基本方法来研究养殖在水族箱中的鱼类的生殖生态。这些是使用SCUBA收集鱼类，运送活鱼以及观察保持在水族箱中的野生捕捞鱼的繁殖行为的有用方案。

鱼类生殖生态学研究的基本方法

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a ...

basic-methods-for-the-study-of-reproductive-ecology-of-fish-in-aquaria

Research

JoVE Journal

Behavior

11.4K 次观看.  Tokyo University of Marine Science and Technology. 描述了一系列基本方法来研究养殖在水族箱中的鱼类的生殖生态。这些是使用SCUBA收集鱼类，运送活鱼以及观察保持在水族箱中的野生捕捞鱼的繁殖行为的有用方案。 

视频: Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

Assessment and Evaluation of the High Risk Neonate: The NICU Network Neurobehavioral Scale

There has been a long-standing interest in the assessment of the neurobehavioral integrity of the newborn infant. The NICU Network Neurobehavioral Scale (NNNS) was developed as an assessment for the at-risk infant. These are infants who are at increased risk for poor developmental outcome because of insults during prenatal development, such as substance exposure or prematurity or factors such as poverty, poor nutrition or lack of prenatal care that can have adverse effects on the intrauterine environment and affect the developing fetus. The NNNS assesses the full range of infant neurobehavioral performance including neurological integrity, behavioral functioning, and signs of stress/abstinence. The NNNS is a noninvasive neonatal assessment tool with demonstrated validity as a predictor, not only of medical outcomes such as cerebral palsy diagnosis, neurological abnormalities, and diseases with risks to the brain, but also of developmental outcomes such as mental and motor functioning, behavior problems, school readiness, and IQ. The NNNS can identify infants at high risk for abnormal developmental outcome and is an important clinical tool that enables medical researchers and health practitioners to identify these infants and develop intervention programs to optimize the development of these infants as early as possible. The video shows the NNNS procedures, shows examples of normal and abnormal performance and the various clinical populations in which the exam can be used.

The NICU Network Neurobehavioral Scale (NNNS) was developed as an assessment for the at-risk infant. The purpose of this article is to describe the NNNS, provide video examples of the NNNS procedures and discuss the ways in which the exam has been used.

评估和高危新生儿的评价:新生儿重症监护病房的网络神经行为量表

There has been a long-standing interest in the assessment of the neurobehavioral integrity of the newborn infant.  The NICU Network Neurobehavioral Scale ...

科研

行为学

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.

立体定向显微注射病毒载体表达Cre重组酶研究的靶基因的作用在可卡因条件性位置偏爱

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes ...

A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during abstinence. Moreover, various environmental challenges can exacerbate this effect, as well as increase ongoing drug intake.
The procedure we describe here highlights the impact of a common environmental challenge, food restriction, on drug craving that is expressed as an augmentation of drug seeking in abstinent rats.
Rats are implanted with chronic intravenous i.v. catheters, and then trained to press a lever for i.v. heroin over a period of 10-12 days. Following the heroin self-administration phase the rats are removed from the operant conditioning chambers and housed in the animal care facility for a period of at least 14 days. While one group is maintained under unrestricted access to food (sated group), a second group (FDR group) is exposed to a mild food restriction regimen that results in their body weights maintained at 90% of their nonrestricted body weight. On day 14 of food restriction the rats are transferred back to the drug-training environment, and a drug-seeking test is run under extinction conditions (i.e.&#160;lever presses do not result in heroin delivery).
The procedure presented here results in a highly robust augmentation of heroin seeking on test day in the food restricted rats. In addition, compared to the acute food deprivation manipulations we have used before, the current procedure is a more clinically relevant model for the impact of caloric restriction on drug seeking. Moreover, it might be closer to the human condition as the rats are not required to go through an extinction-training phase before the drug-seeking test, which is an integral component of the popular reinstatement procedure.

A procedure that allows a demonstration of robust augmentation of drug seeking in food-restricted rats is described. Following heroin self-administration training, rats go through an abstinence period, in a drug-free environment, during which they are mildly food restricted. Drug seeking is then tested in the drug-associated environment.

一个过程来研究延长食品的限制对海洛因寻求戒断大鼠的影响

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during ...

Methods to Explore the Influence of Top-down Visual Processes on Motor Behavior

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are deliberately planned, while others spontaneously occur below conscious awareness. The deliberate component of this dichotomy has been studied extensively in several contexts, while the spontaneous component remains largely under-explored. Moreover, how perceptual processes modulate these movement classes is still unclear. In particular, a currently debated issue is whether the visuomotor system is governed by the spatial percept produced by a visual illusion or whether it is not affected by the illusion and is governed instead by the veridical percept. Bistable percepts such as 3D depth inversion illusions (DIIs) provide an excellent context to study such interactions and balance, particularly when used in combination with reach-to-grasp movements. In this study, a methodology is developed that uses a DII to clarify the role of top-down processes on motor action, particularly exploring how reaches toward a target on a DII are affected in both deliberate and spontaneous movement domains.

It is unclear how top-down signals from the ventral visual stream affect movement. We developed a paradigm to test motor behavior towards a target on a 3D depth inversion illusion. Significant differences are reported in both deliberate, goal-directed movements and automatic actions under illusory and veridical viewing conditions.

方法探索自上而下视觉过程对电机性能的影响

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are ...

Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both laboratory and clinical settings. Currently, preclinical ERG and VEP measurements are performed with anesthesia to ensure stable electrode placements. However, the very presence of anesthesia has been shown to contaminate normal physiological responses. To overcome these anesthesia confounds, we develop a novel platform to assay ERG and VEP in conscious rats. Electrodes are surgically implanted sub-conjunctivally on the eye to assay the ERG and epidurally over the visual cortex to measure the VEP. A range of amplitude and sensitivity/timing parameters are assayed for both the ERG and VEP at increasing luminous energies. The ERG and VEP signals are shown to be stable and repeatable for at least 4 weeks post surgical implantation. This ability to record ERG and VEP signals without anesthesia confounds in the preclinical setting should provide superior translation to clinical data.

We show surgical implantation and Recording procedures to measure visual electrophysiological signals from the eye (electroretinogram) and brain (visual evoked potential) in conscious rats, which is more analogous to the human condition where recordings are conducted without anesthesia confounds.

植入和无线电图记录和视觉诱发电位清醒大鼠

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

叶切蚁病原体攻击行为防御研究的可视化指南

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the appearance and behavior of visual stimuli, compared to manipulating live animals. Here, we present the FishSim Animation Toolchain, a software framework developed to provide researchers with an easy-to-use method for implementing 3D computer animations in behavioral experiments with fish. The toolchain offers templates to create virtual 3D stimuli of five different fish species. Stimuli are customizable in both appearance and size, based on photographs taken of live fish. Multiple stimuli can be animated by recording swimming paths in a virtual environment using a video game controller. To increase standardization of the simulated behavior, the prerecorded swimming path may be replayed with different stimuli. Multiple animations can later be organized into playlists and presented on monitors during experiments with live fish.
In a case study with sailfin mollies (Poecilia latipinna), we provide a protocol on how to conduct a mate-choice copying experiment with FishSim. We utilized this method to create and animate virtual males and virtual model females, and then presented these to live focal females in a binary choice experiment. Our results demonstrate that computer animation may be used to simulate virtual fish in a mate-choice copying experiment to investigate the role of female gravid spots as an indication of quality for a model female in mate-choice copying.
Applying this method is not limited to mate-choice copying experiments but can be used in various experimental designs. Still, its usability depends on the visual capabilities of the study species and first needs validation. Overall, computer animations offer a high degree of control and standardization in experiments and bear the potential to &#39;reduce&#39; and &#39;replace&#39; live stimulus animals as well as to &#39;refine&#39; experimental procedures.

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Using the novel FishSim Animation Toolchain, we present a protocol for non-invasive visual manipulation of public information in the context of mate-choice copying in sailfin mollies. FishSim Animation Toolchain provides an easy-to-use framework for the design, animation and presentation of computer-animated fish stimuli for behavioral experiments with live test fish.

利用fishsim动画工具链调查鱼类行为--以鱼翅钼中的选物复制为例

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the ...

Using Facial Electromyography to Assess Facial Muscle Reactions to Experienced and Observed Affective Touch in Humans

"Affective" touch is believed to be processed in a manner distinct from discriminatory touch and to involve activation of C-tactile (CT) afferent fibers. Touch that optimally activates CT fibers is consistently rated as hedonically pleasant. Patient groups with impaired social-emotional functioning also show disordered affective touch ratings. However, relying on self-reported ratings of touch has many limitations, including recall bias and communication barriers. Here, we describe a methodological approach to study affective responses to touch via facial electromyography (EMG) that circumvents the reliance on self-report ratings. Facial EMG is an objective, quantitative, and non-invasive method to measure facial muscle activity indicative of affective responses. Responses can be assessed across healthy and patient populations without the need for verbal communication. Here, we provide two separate datasets demonstrating that CT-optimal and non-optimal touch elicit distinct facial muscle reactions. Moreover, facial EMG responses are consistent across stimulus modalities, e.g. tactile (experienced touch) and visual (observed touch). Finally, the temporal resolution of facial EMG can detect responses on timescales that supersede that of verbal reporting. Together, our data suggest that facial EMG is a suitable methodology for use in affective tactile research that can be used to supplement, or in some cases, supplant, existing measures.

We describe a protocol to assess facial muscle activity in response to experienced and observed tactile stimulation using facial electromyography.

面部肌电图对人体有经验和观察到的情感触觉的影响评估

"Affective" touch is believed to be processed in a manner distinct from discriminatory touch and to involve activation of C-tactile (CT) afferent fibers. ...

In Vivo Protocol of Controlled Subconcussive Head Impacts for the Validation of Field Study Data

Subconcussive hits pose a threat to neuronal health as they have shown to induce neuronal structural damage and functional impairment without causing outward symptomology and appear to be a key contributor to an irreversible neurodegenerative disease, chronic traumatic encephalopathy (CTE). In addition, athletes can incur more than 1,000 of these hits per season. The subconcussive soccer heading model (SSHM) is a relevant, reproducible, and leading method of isolating and examining the effects of these subconcussive head impacts. By controlling variables such as ball traveling speed, the frequency of impacts, interval, ball placement to the head, as well as by measuring head impact magnitude, the SSHM provides the scientific community with a superior avenue of investigating the acute subconcussive effects on neuronal health. In this paper, we demonstrate the utility of SSHM in studying a time-course expression of neurofilament-light polypeptide (NF-L) in plasma in a repeated measures fashion. NF-L is an axonal injury marker that has previously been shown to be elevated in boxers and football players following subconcussive head trauma. Thirty-four adult aged soccer players were recruited and randomly assigned to either a soccer heading (n = 18) or kicking (n = 16) group. The heading group executed 10 headers with soccer balls projected at a velocity of 25 mph over 10 min. The kicking group followed the same protocol with 10 kicks. Plasma samples were obtained before and at 0 h, 2 h, and 24 h after heading/kicking and assessed for NF-L expressions. The heading group showed a gradual increase in plasma NF-L expression and peaked at 24 h after the heading protocol, whereas the kicking group remained consistent across the time points. These results confirmed the NF-L data from clinical field studies, encouraging the use of SSHM to validate clinical subconcussion data.

The subconcussive soccer heading model is a safe and concise methodological approach to isolate and measure the effects of subconcussive head impacts.

控制下冲击头冲击的活体协议对现场研究数据的验证

Subconcussive hits pose a threat to neuronal health as they have shown to induce neuronal structural damage and functional impairment without causing ...

Assessing the Coherence of Parents' Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with parents. This paper focuses on such a method, namely Five-Minute Speech Sample-Coherence (FMSS-Coherence). In this method, a parent is asked to speak for five uninterrupted minutes about her/his child and their relationship. The resulted narrative is coded for coherence, namely the extent to which the parent provides&#160;in the narrative&#160;a clear, consistent, multidimensional and well-supported portrayal of the child. FMSS-Coherence is based on attachment research that shows that the coherence of parents&#8217; narratives is indicative of the quality of the parent-child relationship and child adjustment. It overcomes the limitations of attachment narrative measures which are typically labor intensive. FMSS-Coherence is less nuanced than extant attachment narrative measures. Yet, studies of families from different cultural backgrounds, across different child ages and in the context of typically developing children as well as children with special needs suggest that coherence can be reliably evaluated using the FMSS procedure. Furthermore, parents&#8217; FMSS-Coherence is associated with parenting quality and children&#8217;s socio-emotional adjustment. Thus, it holds promise for researchers and practitioners who seek a relatively time- and cost-effective method for assessing the coherence of parents&#8217; narratives regarding their child.

This paper introduces a method for assessing the coherence of parents' short narratives regarding their child and their relationship using the five-minute speech sample procedure.

使用五分钟言语样本程序评估家长对孩子的简短叙述的一致性

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with ...