Name: recording behavioral responses to reflection in crayfish
Uploaded: 2010-05-14T17:00:00.000+00:00
Duration: 11 min 30 s
Description: Watch this Scientific Journal Video about Recording Behavioral Responses to Reflection in Crayfish at JoVE.com

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。这是目前尚不清楚是否小龙虾，专门由一个小龙虾图像或运动所吸引。然而，这是可以探讨的视觉输入的作用，并确定使用镜子，并通过提供一个透明的分区的图像的视觉线索。 以前的结果表明，主导和从属的小龙虾在本质上是相同的方式回应反光表面只有30分钟的配对会议后，出现行为上的差异与3 - 14天的配对，因为失去小龙虾的行为的变化（5月和名士，2007年）。因此，在视觉线索的行为之一的观察取决于遇到的结果和社会的持续时间。输赢斗殴遇到被称为影响未来遇到的成功，赢得再次获胜的可能性增加，亏损增加随后失去（许狼，1999年）的概率。行为可塑性的内在机制，表现出失去反应，可能与神经系统的变化，伴随着加强从属地位（5月名士，2006年，2007年）的视觉线索的小龙虾。 这里所描述的实验，可以进行修改，以评估感官线索，引出方法的行为的复杂性。这可以通过改变背后一个透明的分区，并结合化学和触觉因素等视觉线索提供视觉线索的各个方面。空调水可以添加到镜面/磨砂水族馆添加化学成分。此外，孔可添加到透明阻隔在年底室坦克从生活来源的小龙虾，以便化学信号的传输。

<table><tbody><tr><td>Single chamber test tank</td><td>Brock University Machine Shop</td><td></td><td></td></tr><tr><td>End-chamber test tank</td><td>Brock University Machine Shop</td><td></td><td></td></tr><tr><td>Isolation container</td><td>Rubbermaid</td><td></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></td><td></td><td>48 cm x 25 cm x 13 cm</td></tr><tr><td>Social housing chamber</td><td></td><td></td><td>58 cm x 30 cm x 35 cm</td></tr><tr><td>small plastic flower pot</td><td></td><td></td><td>to serve as shelter</td></tr><tr><td>Coolpix 4500 camera</td><td>Nikon Instruments</td><td></td><td></td></tr><tr><td>T120 video cassette recorder</td><td>RCA</td><td></td><td></td></tr><tr><td>Computer</td><td></td><td></td><td>Windows Movie Maker or other video acquiring software</td></tr><tr><td>webcam</td><td>Logitech</td><td></td><td></td></tr><tr><td>modelling clay</td><td>Sculptee</td><td></td><td>to make control cube</td></tr><tr><td>red latex paint</td><td>FolkArt</td><td></td><td>to make control cube</td></tr><tr><td>waterbase varnish</td><td>FolkArt</td><td></td><td>to make control cube</td></tr></tbody></table>

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

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

10.7K 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

Physiological Recordings of High and Low Output NMJs on the Crayfish Leg Extensor Muscle

We explain in detail how to expose and conduct electrophysiological recordings of synaptic responses for high (phasic) and low (tonic) output motor neurons innervating the extensor muscle in the walking leg of a crayfish. Distinct differences are present in the physiology and morphology of the phasic and tonic nerve terminals. The tonic axon contains many more mitochondria, enabling it to take a vital stain more intensely than the phasic axon. The tonic terminals have varicosities, and the phasic terminal is filiform. The tonic terminals are low in synaptic efficacy but show dramatic facilitated responses. In contrast, the phasic terminals are high in quantal efficacy but show synaptic depression with high frequency stimulation. The quantal output is measured with a focal macropatch electrode placed directly over the visualized nerve terminals. Both phasic and tonic terminals innervate the same muscle fibers, which suggests that inherent differences in the neurons, rather than differential retrograde feedback from the muscle, account for the morphological and physiological differentiation.

This article demonstrates how to conduct electrophysiological recordings of synaptic responses on the extensor muscle in the walking leg of a crayfish and how the nerve terminals are visualized to show the gross morphological differences of high- and low-output nerve terminals.

小龙虾腿伸肌高和低输出NMJs生理录音

We explain in detail how to expose and conduct electrophysiological recordings of synaptic responses for high (phasic) and low (tonic) output motor ...

科研

神经科学

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 ...

Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception 

The primary purpose of this experiment is to demonstrate primary sensory neurons conveying information of joint movements and positions as proprioceptive information for an animal. An additional objective of this experiment is to learn anatomy of the preparation by staining, dissection and viewing of neurons and sensory structures under a dissecting microscope. This is performed by using basic neurophysiological equipment to record the electrical activity from a joint receptor organ and staining techniques. The muscle receptor organ (MRO) system in the crayfish is analogous to the intrafusal muscle spindle in mammals, which aids in serving as a comparative model that is more readily accessible for electrophysiological recordings. In addition, these are identifiable sensory neurons among preparations. The preparation is viable in a minimal saline for hours which is amenable for student laboratory exercises. The MRO is also susceptible to neuromodulation which encourages intriguing questions in the sites of modulatory action and integration of dynamic signals of movements and static position along with a gain that can be changed in the system.

Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception

The primary purpose of this experiment is to understand how primary sensory neurons convey information of joint movements and positions as proprioceptive information for an animal. An additional objective of this report is present the anatomy of the preparation by dissection and viewing of neurons under a dissecting microscope.

小龙虾腹部肌肉受体机关：在本体的学生实验室练习

The primary purpose of this experiment is to demonstrate primary sensory neurons conveying information of joint movements and positions as proprioceptive ...

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities 

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the visual scene. However, the OKR requires that an animal detect moving stripes and it is possible that fish that fail to exhibit an OKR may not be completely blind. One zebrafish mutant, the no optokinetic response c (nrc) has no OKR under any light conditions tested and was reported to be completely blind. Previously, we have shown that OFF-ganglion cell activity can be recorded in these mutants. To determine whether mutant fish with no OKR such as the nrc mutant can detect simple light increments and decrements we developed the visual motor behavioral assay (VMR). In this assay, single zebrafish larvae are placed in each well of a 96-well plate allowing the simultaneous monitoring of larvae using an automated video-tracking system. The locomotor responses of each larva to 30 minutes light ON and 30 minutes light OFF were recorded and quantified. WT fish have a brief spike of motor activity upon lights ON, known as the startle response, followed by return to lower-than baseline activity, called a freeze. WT fish also sharply increase their locomotor activity immediately following lights OFF and only gradually (over several minutes) return to baseline locomotor activity. The nrc mutants respond similarly to light OFF as WT fish, but exhibit a slight reduction in their average activity as compared to WT fish. Motor activity in response to light ON in nrc mutants is delayed and sluggish. There is a slow rise time of the nrc mutant response to light ON as compared to WT light ON response. The results indicate that nrc fish are not completely blind. Because teleosts can detect light through non-retinal tissues, we confirmed that the immediate behavioral responses to light-intensity changes require intact eyes by using the chokh (chk) mutants, which completely lack eyes from the earliest stages of development. In our VMR assay, the chk mutants exhibit no startle response to either light ON or OFF, showing that the lateral eyes mediate this behavior. The VMR assay described here complements the well-established OKR assay, which does not test the ability of zebrafish larvae to respond to changes in light intensities. Additionally, the automation of the VMR assay lends itself to high-throughput screening for defects in light-intensity driven visual responses.

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities

We developed the Visual-Motor Response to quantitate the motor output of larval zebrafish in response to light increments and decrements. We also examined zebrafish vision mutants, including the no optokinetic response (nrc) mutants, which were thought to be completely blind when tested by another vision assay, the optokinetic reflex.

斑马鱼的响应行为的检测，以测量光强的变化

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the ...

生物学

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Continuous Noninvasive Measuring of Crayfish Cardiac and Behavioral Activities

A crayfish is a pivotal aquatic organism that serves both as a practical biological model for behavioral and physiological studies of invertebrates and as a useful biological indicator of water quality. Even though crayfish cannot directly specify the substances that cause water quality deterioration, they can immediately (within a few seconds) warn humans of water quality deterioration via acute changes in their cardiac and behavioral activities.
In this study, we present a noninvasive method that is simple enough to be implemented under various conditions due to a combination of simplicity and reliability in one model.
This approach, in which the biological organisms are implemented into environmental evaluation processes, provides a reliable and timely alarm for warning of and preventing acute water deterioration in an ambient environment. Therefore, this noninvasive system based on crayfish physiological and ethological parameter recordings was investigated for the detection of changes in an aquatic environment. This system is now applied at a local brewery for controlling&#160;quality of the water used for beverage production, but it can be used at any water treatment and supply facility for continuous, real-time water quality evaluation and for regular laboratory investigations of crayfish cardiac physiology and behavior.

This article presents a noninvasive biomonitoring system for the continuous recording and analyses of crayfish cardiac and locomotor activities. This system consists of a near-infrared optical sensor, a video-tracking module, and software for evaluating crayfish heartbeats that reflects its physiological condition and characterizes crayfish behavior during heartbeat fluctuations.

小龙虾心脏的连续无创测量与行为活动

A crayfish is a pivotal aquatic organism that serves both as a practical biological model for behavioral and physiological studies of invertebrates and as ...

行为学

Recordings of Neural Circuit Activation in Freely Behaving Animals

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. Traditional non-invasive methods require at least partially restrained research subjects, and they only allow identification of large numbers of simultaneously activated neurons. On the other hand, small ensembles of neurons or individual neurons can only be measured using single-cell recordings obtained from largely reduced preparations. Since the expression of natural behavior is limited in restrained and dissected animals, the underlying neural mechanisms that control such behavior are difficult to identify. 
Here, I present a non-invasive physiological technique that allows measuring neural circuit activation in freely behaving animals. Using a pair of wire electrodes inside a water-filled chamber, the bath electrodes record neural and muscular field potentials generated by juvenile crayfish during natural or experimentally evoked escape responses. The primary escape responses of crayfish are mediated by three different types of tail-flips which move the animals away from the point of stimulation. Each type of tail-flip is controlled by its own neural circuit; the two fastest and most powerful escape responses require activation of different sets of large &#8220;command&#8221; neurons. In combination with behavioral observations, the bath electrode recordings allow unambiguous identification of these neurons and the associated neural circuits. Thus activity of neural circuitry underlying naturally occurring behavior can be measured in unrestrained animals and in different behavioral contexts.

Non-invasive measurements of neural activity patterns in freely behaving animals are obtained by combining neurophysiological recordings with high speed videography.

自由表现动物的神经回路活化的录音

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. ...

Measures of Heart and Ventilatory Rates in Freely Moving Crayfish

The fear, flight or fight response serves as the fundamental physiological basis for examining an organism's awareness of its environment under an impending predator attack.  Although it is not known whether invertebrates posses an autonomic nervous system identical to that of vertebrates, evidence shows invertebrates have a  sympathetic-like  response to regulate the internal environment and ready the organism to act behaviorally to a given stimuli.  Furthermore, this physiological response can be feasibly measured and it acts as a biological index for the animal's internal state.  Measurements of the physiological response can be directly related to internal and external stressors through changes in the central nervous system controlled coordination of the cardio-vascular and respiratory systems.   More specifically, monitoring heart and ventilation rates provide quantifiable measures of the stress response not always behaviorally observed. Crayfish are good model organisms for heart and ventilatory rate measurements due to the feasibility of recording, as well as the rich history known of the morphology of the crayfish, dating back to Huxley in 1888, and the well-studied typical behaviors.

Invertebrates show an autonomic sympathetic-like response similar to that described for vertebrates. The coordination of the cardio-vascular and ventilatory systems allows for measurement of a biological index in which to assess an organism internal state.

在自由移动的小龙虾的心脏和换气率的措施

The fear, flight or fight response serves as the fundamental physiological basis for examining an organism's awareness of its environment under an ...

Historical View and Physiology Demonstration at the NMJ of the Crayfish Opener Muscle

Here we present some of the key important discoveries made with the opener neuromuscular (NMJ) preparation of crustaceans and illustrate that there is still much to learn from this model preparation. In understanding the history one can appreciate why even today this NMJ still offers a rich playground to address questions regarding pre- and post-synaptic function and plasticity. The viability and ease of access to the terminal for intracellular as well as extracellular electrophysiology and imaging are significant advantages. The mechanisms behind the modulation of vesicular kinetics and fusion within the high- and low-output terminals are begging for investigation. The preparation also offers a testable model system for computational assessments and manipulations to examine key variables in theoretical models of synaptic function, for example calcium dynamics during short-term facilitation. The synaptic complexity of active zone and statistical nature of quantal release is also an open area for future investigation both experimentally and computationally.

The opener muscle of the crayfish leg is presented for its historical importance and experimental versatility in muscle phenotype, synaptic physiology and plasticity.

小龙虾开瓶器肌肉NMJ的历史观和生理学示范

Here we present some of the key important discoveries made with the opener neuromuscular (NMJ) preparation of crustaceans and illustrate that there is ...

Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises

The primary purpose of these procedures is to demonstrate for teaching and research purposes how to record the activity of living primary sensory neurons responsible for proprioception as they are detecting joint position and movement, and muscle tension. Electrical activity from crustacean proprioceptors and tension receptors is recorded by basic neurophysiological instrumentation, and a transducer is used to simultaneously measure force that is generated by stimulating a motor nerve. In addition, we demonstrate how to stain the neurons for a quick assessment of their anatomical arrangement or for permanent fixation. Staining reveals anatomical organization that is representative of chordotonal organs in most crustaceans. Comparing the tension nerve responses to the proprioceptive responses is an effective teaching tool in determining how these sensory neurons are defined functionally and how the anatomy is correlated to the function. Three staining techniques are presented allowing researchers and instructors to choose a method that is ideal for their laboratory.

Physiological and anatomical techniques are demonstrated to address function and structure for joint proprioceptors and muscle tension receptors in crustacean walking limbs.

本体感觉和张力感受器在蟹四肢:学生实验室练习

The primary purpose of these procedures is to demonstrate for teaching and research purposes how to record the activity of living primary sensory neurons ...