NSERC研究的支持。

1。社会化<ol><li>扑灭现有的社会行列，每个小龙虾是采用7 - 14天，在一个单独的塑料容器（长30厘米× 17.5厘米（宽）x 13厘米深），过滤，汽水和一个小塑料花盆作为填充栖身之所。如果使用透明的容器，一个分频器（如纸巾）被放置在容器之间，以确保小龙虾是视觉隔离。 </li><li>孤立地停留在一个额外的3 - 14天的“隔离”组中的小龙虾。水是改变和小龙虾是美联储每周3次。 </li><li>要建立社会地位，小龙虾被放置在同性对初始隔离。小龙虾的重量，并测量体长（讲台尾节）;对大小在10％以内的重量和长度。白色丙烯酸涂料适用于个人识别每个小龙虾的甲壳。 （通常是一个点是适用于一个成员和其他线路。） </li><li>经过30分钟的休息期，每个小龙虾对允许在一个干净的，白色的塑料容器（48厘米长x宽25厘米× 13厘米深）充满了过滤，汽水（室温）中途，交互。无论遇到的是记录到相机连接到录像机的VHS录像带，或它是一个摄像头，记录到一台计算机。这种互动使统治地位行列中来发展。 </li><li>继最初的互动，每个小龙虾对住在一起在一个较大的塑料容器（长58厘米× 30厘米（宽）x 35厘米深），另外3 - 14天。这个容器接收来自循环水过滤系统。一个单一的PVC管（约10厘米长）提供住房。 </li></ol> 2。测试水族馆和控制的建设方法一<ol><li>要研究一个反射的环境中，玻璃或有机玻璃水族箱（52厘米长x宽25厘米× 30厘米的高），使用（图1）的反应。 </li><li>一面镜子（长25厘米× 20厘米高）被放置在水族箱的一端，并用胶水或安装的蜡担保。两镜（长26厘米× 20厘米高），放置在两侧，毗邻端镜，并用胶水或蜡担保。 </li><li>其余的墙壁上挂满粉嫩窗膜。这部影片中删除的反思，但不干预的照明条件下。可能需要一个光打磨的电影，以确保电影是完全不反光的。 </li></ol>方法二<ol start="4"><li>要检查两个终端室提供了一个活的小龙虾（“源小龙虾”），有机玻璃水族箱的视觉刺激的反应（图2）。水族馆包含一个大的中央车厢（48厘米长x宽26厘米× 26厘米的高）和两个较小的大小相等的车厢（长12厘米× 26厘米，宽26厘米高的的）。最终车厢是由透明的有机玻璃，这是密封，以防止水和化学信号传递之间的车厢脱离中心车厢。 </li><li>整个水族馆的外围涂在无反射的磨砂塑料，留下透明的两端各有两个障碍。 </li><li>作为对照，造型粘土构建了一个多维数据集（例如Sculptee）是用来提供惰性物体的视觉输入。立方体（3.5厘米× 3.0厘米× 3.3厘米）是源小龙虾的近似量，并描绘了一幅类似的颜色红色。 </li></ol> 3。视觉训练方法一<ol><li>小龙虾是录像镜面/磨砂罐3 - 14天以下的斗争，以确定主导地位的状态。 </li><li>小龙虾都放在各一个，成一个特百惠曝气，过滤后的水的容器和一个塑料花盆住房。小龙虾被带到考场，并给予测试前30分钟，以适应环境。 </li><li>测试水族馆，镜子和半不反光的塑料内衬，曝气，过滤后的水充满深度约20厘米的半衬里。 </li><li>数码相机是通过音频/视频电缆连接到VHS录像机，并放置足够高，以查看整个水族馆水族馆以上。视频记录到VHS录像带。另外，可用于连接到电脑摄像头，并且可以录制视频与电影制作计划，并存储在DVD中。 </li><li>到使用花盆的水族馆中心，是轻轻地放在一对小龙虾和它的活动是20分钟的记录。任何人在录音室中，以减少多余的视觉和听觉输入干扰。正尽一切努力，以确保录音环境安静。 </li><li>以下测试，小龙虾被删除，清空坦克，洗净，用清水加气。 </li><li>实验条件下，此消彼长，水族馆转身的地方对面的房间的镜子。这有助于弥补未知因素的影响，可能会导致小龙虾喜欢水族馆的一面。 </li><li>在相同的方式作为第一对的第二个成员，然后进行测试。 </li></ol>方法二<ol start="9"><li>方法我所描述的，小龙虾带来的测试室，并使用相同的录音设备录像。 </li><li>钱伯斯（中间试验室和两个终端室）曝气，过滤后的水充满高度约20厘米。 </li><li>一个小龙虾，大约同样大小的小龙虾被测试，是摆在一端室​​，放置在其他控制立方体。 </li><li>使用的花盆住房，被测试的小龙虾轻轻放入水族箱的中心和20分钟的录像。 </li><li>以下测试，水族馆被掏空，洗净，用清水加气。 </li><li>测试前的一对第二个成员，源小龙虾和控制多维数据集移动到另一端商会，制衡的条件。 </li></ol> 4。视觉刺激的反应分析方法一<ol><li>因此，实验者小龙虾的社会地位，在镜面/磨砂水族馆的行为是盲目的，是分析前，分析战斗行为的审判。 </li><li>水族馆的每一侧的小龙虾上花费的时间量被记录下来。小龙虾被认为是从一个侧面传递到下一个时的螯足髋通过中线的坦克（图3）。 </li><li>转弯，转弯，隧道，冻结和反向行走（请参阅表1描述）的行为的频率也记录下来。 </li><li>行为上展出的坦克反射面比较使用配对t -检验每个社会群体的非反射方表现出的行为。 </li></ol>方法二<ol start="5"><li>至于方法，我，小龙虾行为的录像带分析前，分析斗争试验。 </li><li>水族馆两侧的小龙虾上花费的时间量的记录方法一</li><li>过弯的频率，最终面临，转弯，穿越，饲养记录和反向行走（见表1）。 </li><li>源小龙虾端上执行的行为进行比较使用配对t -检验每个社会群体的控制立方体方执行的行为。 </li></ol> 5。打架分析<ol><li>查看，并取得了配对，在最初的30分钟的战斗行为的录音。每30分钟的录音会议，包括几个交锋中，小龙虾，使用它们的爪子和/或天线进行接触。在这些接触中，chelae通常用于推，引人注目或抓住对手，和触角，有时鞭打对手。一个小龙虾，可能会尝试翻转它的对手，并保持他在地板上。每次遇到结束时，反向行走或翻转的尾巴来实现的一个小龙虾务虚“逃生游泳。” </li><li>斗殴遇到被记录下来，和小龙虾的身份（由胸部上的标线涂料决定），每次遇到撤退也被记录下来。失去了每30分钟会议的大多数遇到的小龙虾被视为失败者，和其他小龙虾被视为获胜者。 </li></ol> 6。代表性的成果在30分钟的战斗期间，所有对表现出好斗的行为。这些行为包括窃听天线，冲刺，推，抓，chelae引人注目，并翻转对手，据报道在其他地方（如5月名士2006年，2007年）。最后一个小龙虾在每对一贯务虚和避免进一步的接触。撤退的小龙虾被视为失败者，是作为下属的排名，获奖的小龙虾为主导排名。 排名也可以通过观察小龙虾的行为在保温水箱，其中包含只有一个塑料管住房配对房屋期间整个监控。占主导地位的小龙虾总是占有住房，不服从小龙虾。下属的小龙虾往往与chelae各界举行并拢，在前面，和占主导地位的的小龙虾通常各界提出chelae的进一步分开（Guiasu邓纳姆，1998年）。当添加新的食物保温水箱，占主导地位的小龙虾，通常首先需要的食物。在我们的研究与体育虾 （如5月名士2006年，2007年），通常有没有排名逆转后三天连续配对。 在极少数情况下，对一名成员死亡配对期前完成。这是更可能发生，如果配对期长（如14天）和小龙虾，如果过于密切的大小和重量相匹配。因此，死亡是可以避免的允许重量大约在10％的差异。如果一对的一名成员死亡，其他成员的数据是不包括在这项研究。 主导和从属的小龙虾一般反射环境的不同反应。始终占主导地位的小龙虾花更多的时间上的反射面，但下属和孤立的小龙虾不（5名士，2006年，2007年）。主导的小龙虾，也表现出过弯和转向上比非反射面的反射面更频繁，但隔离和下属通常不。主导和从属的小龙虾之间的这种差异，然而，并非立即建立，但发展在3-14天的配对期间（2007年5月名士）。 初步证据表明，主导和从属的小龙虾的反应不同，一个活的小龙虾，经过3天的配对（源小龙虾 ）（May等人，2008年）所提供的视觉线索。水族馆方主导的小龙虾花更多的时间与源小龙虾和那边的角落，但下属的小龙虾没有表现出双方之间的这种差异。 <table border="1"><tbody><tr><td> 行为 </td><td> 说明 </td></tr><tr><td> 转弯 </td><td>面临的水族箱的角落，其余两个爪子提示，触摸不同的墙壁（图4）至少5小号</td></tr><tr><td> 年底面临 </td><td>在水箱的一端面临至少5秒触摸墙底（图5）至少有一爪的尖端墙</td></tr><tr><td> 转折点 </td><td>改变行走的路径方向从顺时针逆时针或反之亦然 （图6） </td></tr><tr><td> 路口 </td><td>离开水族馆周边和步行至少一个车身长度，任何其他的墙壁（图7） </td></tr><tr><td> 反向行走 </td><td>倒着走路至少一个车身长度</td></tr><tr><td> 冻结 </td><td>突然停止所有可见的运动，包括附属物和天线，至少有5秒</td></tr><tr><td> 饲养起来 </td><td>人大常委会第四次和第五次腿走路，同时抬起靠墙胸部腹面，至少有一个在墙壁上行走的腿</td></tr></tbody></table> 表1： 由 P. 展出 。行为虾在中央室的测试箱。 <img src="/files/ftp_upload/1956/1956fig1.jpg" alt="图1" /> 图1。原理方法一半的水族箱中使用的水族馆是林立的镜子，而另一半则是不反光的塑料内衬。 <img src="/files/ftp_upload/1956/1956fig2.jpg" alt="图2" /> 图2方法二使用水族箱示意图。这种坦克是由测试小龙虾被放在一个中心区，和两个终端室除以一个完整的有机玻璃屏障。一个活的小龙虾是摆在一端室​​，并在作为对照的另一端室放置一个立方体。 <img src="/files/ftp_upload/1956/1956fig3.jpg" alt="图3" /> 图3。测试水族馆作为一个垂直的虚线描绘中线示意图。小龙虾，从一个侧面传递到其他的螯足髋通过中线。 <img src="/files/ftp_upload/1956/1956fig4.jpg" alt="图4" /> 图4。转弯时表示小龙虾的位置图。请注意，两个chelae提示触摸不同的墙壁。 <img src="/files/ftp_upload/1956/1956fig5.jpg" alt="图5" /> 图5图显示小龙虾的位置，同时也面临墙底。请注意，两个chelae接触相同的墙壁。 <img src="/files/ftp_upload/1956/1956fig6.jpg" alt="图6" /> 图6。示意图表明转折点。扭转运动的方向构成转动。打开可能会发生在一个角落里（如方向1方向2方向4方向5）或侧面（未显示）。进入转弯和留在同一方向（例如方向1方向3）不转一转。 <img src="/files/ftp_upload/1956/1956fig7.jpg" alt="图7" /> FigurE 7。示意图显示过路行为。隧道发生时的小龙虾在至少一个车身长度，从最近的墙壁（箭头）的距离，叶一墙和旅行到另一个墙上。 <img src="/files/ftp_upload/1956/1956fig8.jpg" alt="图8" /> 图8。照片描绘养育起来的行为。小龙虾，看台上的最后两个腿走路和升降机身体靠墙，至少有一个步行腿接触。

<ol>
	<li>Bruski, C. A., Dunham, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+importance+of+vision+in+agonistic+communication+of+the+crayfish+Orconectes+rusticus.+I:+an+analysis+of+bout+dynamics.">The importance of vision in agonistic communication of the crayfish Orconectes rusticus. I: an analysis of bout dynamics.</a> Behav. 103, 83-107 (1987).</li><li>Delgado-Morales, G., Hernandez-Falcon, J., Ramon, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Agnostic+behavior+in+crayfish:the+importance+of+sensory+inputs.">Agnostic behavior in crayfish:the importance of sensory inputs.</a> Crustaceana. 77 (1), 1-24 (2004).</li><li>Guiasu, R. C., Dunham, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inter-form+agonistic+contests+in+male+crayfishes,+Cambarus+robustus+(Decapoda,+Cambaridae).">Inter-form agonistic contests in male crayfishes, Cambarus robustus (Decapoda, Cambaridae).</a> Invert. Biol.. 117 (2), 144-154 (1998).</li><li>May, H. Y., Mercier, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+crayfish+to+a+reflective+environment+depend+on+dominance+status.">Responses of crayfish to a reflective environment depend on dominance status.</a> Can. J. Zool. 84 (8), 1104-1111 (2006).</li><li>May, H. Y., Mercier, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Joffre+Duration+of+socialization+influences+responses+to+a+mirror:+responses+of+dominant+and+subordinate+crayfish+diverge+with+time+of+pairing.">Joffre Duration of socialization influences responses to a mirror: responses of dominant and subordinate crayfish diverge with time of pairing.</a> J. Exp. Biol. 210, 4428-4436 (2007).</li><li>May, H. Y., Luebbert, J., Mercier, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+Crayfish+to+a+Visual+Stimulus+Provided+by+a+Conspecific.">Responses of Crayfish to a Visual Stimulus Provided by a Conspecific.</a> , (2008).</li></ol>

占主导地位的小龙虾花更多的时间在一个反射的环境观察表明他们认识到作为一个同种反射的图像，和足够的视觉线索引出方法的行为。使用活的小龙虾，而不是一个反映图​​像的类似的研究结果支持这样的解释。从属和孤立的小龙虾没有表现出这种反射环境的偏好，也不做下属的小龙虾似乎更愿意在一个立方体的一个活的小龙虾。因此，视觉线索的反应取决于主导地位排名。这是目前尚不清?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Single chamber test tank</td>
<td>&nbsp;</td>
<td>Brock University Machine Shop</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>End-chamber test tank</td>
<td>&nbsp;</td>
<td>Brock University Machine Shop</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isolation container</td>
<td>&nbsp;</td>
<td>Rubbermaid</td>
<td>&nbsp;</td>
<td>30 cm x 17.5 cm x 13 cm holes in lid to allow air and light to pass through when lid is on</td>
</tr>
<tr>
<td>Fighting chamber</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>48 cm x 25 cm x 13 cm</td>
</tr>
<tr>
<td>Social housing chamber</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>58 cm x 30 cm x 35 cm</td>
</tr>
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<td>small plastic flower pot</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>to serve as shelter</td>
</tr>
<tr>
<td>Coolpix 4500 camera</td>
<td>&nbsp;</td>
<td>Nikon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>T120 video cassette recorder</td>
<td>&nbsp;</td>
<td>RCA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Computer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Windows Movie Maker or other video acquiring software</td>
</tr>
<tr>
<td>webcam</td>
<td>&nbsp;</td>
<td>Logitech</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>modelling clay</td>
<td>&nbsp;</td>
<td>Sculptee</td>
<td>&nbsp;</td>
<td>to make control cube</td>
</tr>
<tr>
<td>red latex paint</td>
<td>&nbsp;</td>
<td>FolkArt</td>
<td>&nbsp;</td>
<td>to make control cube</td>
</tr>
<tr>
<td>waterbase varnish</td>
<td>&nbsp;</td>
<td>FolkArt</td>
<td>&nbsp;</td>
<td>to make control cube</td>
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recording behavioral responses to reflection in crayfish

社会行为取决于从视觉，机械和嗅觉系统的感觉输入。一个重要的问题涉及每个指导行为感官方式的相对作用。已审查通过隔离的机械和化学感受刺激的视觉刺激的视觉输入的作用。在一些研究中的视觉输入（Bruski与邓纳姆，1987 年 。德尔加多莫拉莱斯等，2004）已删除眼罩或低光的强度，并已阐明剩余的感官模式的影响。另一种方法是研究在没有任何适当的机械和化学感受线索的视觉输入的影响。这种做法的目的是确定的视觉输入的独家作用。 我们使用两种方法，提供的视觉刺激，不提供化学和机械线索，以小龙虾。在第一种方法，小龙虾是录像的墙壁有一半是在镜子覆盖提供反射环境的水族馆，和另一半则是在非反射（哑光）塑料盖。这使小龙虾的反射和非反射环境之间的选择。反射环境提供视觉线索，在小龙虾的反映图像的形式，因为它整个坦克的一半移动，这些视觉线索是从非反射一半的坦克失踪。另一种方法是到录像带的小龙虾在每年年底由更小室分隔水族馆的行为，在一个小室在对面的小室提供视觉线索和惰性对象，提供一个非移动的视觉输入的小龙虾，非小龙虾的来源。 我们公布的结果表明，小龙虾的反射环境的反应取决于社会和主导地位排名。社会化的小龙虾花更多的时间在反射环境，并表现出比在非反射的环境中的某些行为，更经常有孤立的小龙虾没有表现出这种差异。小龙虾，在同性对安置开发的主导或从属的社会地位。反射的反应不同的主导和从属的小龙虾（2006年5月名士，名士，2007年）。反射面占主导地位的小龙虾花更多的时间，更频繁地进入反射的角落，在反射的角落里花费更多的时间比非反射面。下属的小龙虾走反向往往比非反射面的反射面。初步数据表明，在毗邻的一个小室的小龙虾（May 等人，2008年）所提供的视觉线索类似的效果。

Watch this Scientific Journal Video about Recording Behavioral Responses to Reflection in Crayfish at JoVE.com

Recording Behavioral Responses to Reflection in Crayfish

我们已经开发出两种方法，研究触觉和化学信号的情况下对行为视觉线索的影响。一种方法涉及在水族馆反光外墙摄录小龙虾的反应;其他检查提供一个透明的分区后面的一个活的小龙虾的视觉输入的影响。

小龙虾的反思记录行为反应

Social behavior depends on sensory input from the visual, mechanical and olfactory systems.  One important issue concerns the relative roles of each ...

recording-behavioral-responses-to-reflection-in-crayfish

Research

JoVE Journal

Neuroscience

10.6K 次观看.  Brock University. 我们已经开发出两种方法，研究触觉和化学信号的情况下对行为视觉线索的影响。一种方法涉及在水族馆反光外墙摄录小龙虾的反应;其他检查提供一个透明的分区后面的一个活的小龙虾的视觉输入的影响。

视频: Recording Behavioral Responses to Reflection in Crayfish

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises

The purpose of this report is to help develop an understanding of the effects caused by ion gradients across a biological membrane. Two aspects that influence a cell's membrane potential and which we address in these experiments are: (1) Ion concentration of K+ on the outside of the membrane, and (2) the permeability of the membrane to specific ions. The crayfish abdominal extensor muscles are in groupings with some being tonic (slow) and others phasic (fast) in their biochemical and physiological phenotypes, as well as in their structure; the motor neurons that innervate these muscles are correspondingly different in functional characteristics. We use these muscles as well as the superficial, tonic abdominal flexor muscle to demonstrate properties in synaptic transmission. In addition, we introduce a sensory-CNS-motor neuron-muscle circuit to demonstrate the effect of cuticular sensory stimulation as well as the influence of neuromodulators on certain aspects of the circuit. With the techniques obtained in this exercise, one can begin to answer many questions remaining in other experimental preparations as well as in physiological applications related to medicine and health. We have demonstrated the usefulness of model invertebrate preparations to address fundamental questions pertinent to all animals.

The experiments demonstrate an easy approach for students to gain experience in examining muscle structure, synaptic responses, the effects of ion gradients and permeability on membrane potentials. Also, a sensory-CNS-motor-muscle circuit is presented to show a means to test effects of compounds on a neuronal circuit.

膜电位，突触反应，神经回路，神经调节和肌肉组织学用小龙虾：学生实验室练习

The purpose of this report is to help develop an understanding of the effects caused by ion gradients across a biological membrane. Two aspects that ...

科研

神经科学

Correlating Behavioral Responses to fMRI Signals from Human Prefrontal Cortex: Examining Cognitive Processes Using Task Analysis

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar tasks. Participants' behavior during task performance in an fMRI scanner can then be correlated to the brain activity using the blood-oxygen-level-dependent signal. We measure behavior to be able to sort correct trials, where the subject performed the task correctly and then be able to examine the brain signals related to correct performance. Conversely, if subjects do not perform the task correctly, and these trials are included in the same analysis with the correct trials we would introduce trials that were not only for correct performance. Thus, in many cases these errors can be used themselves to then correlate brain activity to them. We describe two complementary tasks that are used in our lab to examine the brain during suppression of an automatic responses: the stroop1 and anti-saccade tasks. The emotional stroop paradigm instructs participants to either report the superimposed emotional 'word' across the affective faces or the facial 'expressions' of the face stimuli1,2. When the word and the facial expression refer to different emotions, a conflict between what must be said and what is automatically read occurs. The participant has to resolve the conflict between two simultaneously competing processes of word reading and facial expression. Our urge to read out a word leads to strong 'stimulus-response (SR)' associations; hence inhibiting these strong SR's is difficult and participants are prone to making errors. Overcoming this conflict and directing attention away from the face or the word requires the subject to inhibit bottom up processes which typically directs attention to the more salient stimulus. Similarly, in the anti-saccade task3,4,5,6, where an instruction cue is used to direct only attention to a peripheral stimulus location but then the eye movement is made to the mirror opposite position. Yet again we measure behavior by recording the eye movements of participants which allows for the sorting of the behavioral responses into correct and error trials7 which then can be correlated to brain activity. Neuroimaging now allows researchers to measure different behaviors of correct and error trials that are indicative of different cognitive processes and pinpoint the different neural networks involved.

The goal of our research is to correlate behavior to brain activity. Accurate behavioral measures and imaging techniques allow us to elucidate brain-behavior relationships.

关联从人类前额叶皮层的fMRI信号的行为反应：检查的认知过程，使用任务分析

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

运行轮活动在啮齿类动物的昼夜节律的记录和分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in regulating the intracellular calcium concentration. Changing environmental conditions, such as the light-dark cycle, can modify neuronal and neural network activity and the expression of a family of circadian clock genes within the suprachiasmatic nucleus (SCN), the location of the master circadian clock in the mammalian brain. Excitatory synaptic transmission leads to an increase in the postsynaptic Ca2+ concentration that is believed to activate the signaling pathways that shifts the rhythmic expression of circadian clock genes. Hypothalamic slices containing the SCN were patch clamped using microelectrodes filled with an internal solution containing the calcium indicator bis-fura-2. After a seal was formed between the microelectrode and the SCN neuronal membrane, the membrane was ruptured using gentle suction and the calcium probe diffused into the neuron filling both the soma and dendrites. Quantitative ratiometric measurements of the intracellular calcium concentration were recorded simultaneously with membrane potential or current. Using these methods it is possible to study the role of changes of the intracellular calcium concentration produced by synaptic activity and action potential firing of individual neurons. In this presentation we demonstrate the methods to simultaneously record electrophysiological activity along with intracellular calcium from individual SCN neurons maintained in brain slices.

Procedures are described to perform simultaneous recordings of membrane potential or current and changes of intracellular calcium concentration. Suprachiasmatic nucleus neurons are filled with the calcium indicator bis-fura-2 using a patch clamp electrode in the whole cell patch clamp configuration.

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house single or multiple neurons inside, are considered to make the major contribution in responding to the chemical stimuli. By directly recording action potential in the olfactory sensillum after exposure to stimuli, single sensillum recording (SSR) technique provides a powerful approach for investigating the neural responses of insects to chemical stimuli. For the bed bug, which is a notorious human parasite, multiple types of olfactory sensillum have been characterized. In this study, we demonstrated neural responses of bed bug olfactory sensilla to two chemical stimuli and the dose-dependent responses to one of them using the SSR method. This approach enables researchers to conduct early screening for individual chemical stimuli on the bed bug olfactory sensilla, which would provide valuable information for the development of new bed bug attractants or repellents and benefits the bed bug control efforts.

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

使用单一感器录音来检测臭虫的嗅觉神经元的反应，以信息化学物质

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

一种新的行为分析调查个人的味觉，可自由移动的熊蜂（<em&gt;欧洲熊蜂</em&gt;）

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

An Emerging Target Paradigm to Evoke Fast Visuomotor Responses on Human Upper Limb Muscles

To reach towards a seen object, visual information has to be transformed into motor commands. Visual information such as the object&#8217;s color, shape, and size are processed and integrated within numerous brain areas, then ultimately relayed to the motor periphery. In some instances, a reaction is needed as fast as possible. These fast visuomotor transformations, and their underlying neurological substrates, are poorly understood in humans as they have lacked a reliable biomarker. Stimulus-locked responses (SLRs) are short latency (&#60;100 ms) bursts of electromyographic (EMG) activity representing the first wave of muscle recruitment influenced by visual stimulus presentation. SLRs provide a quantifiable output of rapid visuomotor transformations, but SLRs have not been consistently observed in all subjects in past studies. Here we describe a new, behavioral paradigm featuring the sudden emergence of a moving target below an obstacle that consistently evokes robust SLRs. Human participants generated visually guided reaches toward or away from the emerging target using a robotic manipulandum while surface electrodes recorded EMG activity from the pectoralis major muscle. In comparison to previous studies that investigated SLRs using static stimuli, the SLRs evoked with this emerging target paradigm were larger, evolved earlier, and were present in all participants. Reach reaction times (RTs) were also expedited in the emerging target paradigm. This paradigm affords numerous opportunities for modification that could permit systematic study of the impact of various sensory, cognitive, and motor manipulations on fast visuomotor responses. Overall, our results demonstrate that an emerging target paradigm is capable of consistently and robustly evoking activity within a fast visuomotor system.

Presented here is a behavioral paradigm that elicits robust fast visuomotor responses on human upper limb muscles during visually guided reaches.

唤起人类上肢肌肉快速视觉运动反应的新兴目标范式

To reach towards a seen object, visual information has to be transformed into motor commands. Visual information such as the object&#8217;s color, shape, ...