作者要感谢以下：约瑟夫Bronzino博士，博士哈米斯阿布 - 哈萨布拉，RJ奥斯汀LAFRANCE先生，和杰西卡Koranda女士。

这个协议是适当的为3至18个月的年龄和近似体重30-50克）的小鼠。老鼠可以从杰克逊实验室（缅因州巴港）获得。所有手术及实验方案已获本圣三一学院动物护理和使用委员会是根据美国国立卫生研究院指南实验动物的护理和使用。 1。动物的制备及外科手术<ol><li>制备含有氯胺酮（25毫克/毫升），甲苯噻嗪（2.5毫克/毫升）和乙酰丙嗪（0.5毫克/毫升）麻醉鸡尾酒。注射1毫升/公斤体重0.2毫升/公斤注射补充，每45分钟之后维持麻醉的使用踏板撤回反射稳定的深度腹部右下象限。 </li><li>通过使用电动剃须刀剃须皮草在其头盖骨准备麻醉的小鼠进行手术。剃光面积应包括眼睛的中间，耳朵中间。确保不刮胡子的whiske因为它们是鼠标桶的感官系统的一部分的RS。 </li><li>用棉签异丙醇其次是聚维酮碘适用于剃区域的头部。接着，也用棉签，适用于眼睛少量矿物油以防止眼球从干燥。 </li><li>使用婴儿鼠耳袖口，而不是常规耳棒装载动物的头部立体框架设备上。要做到这一点，首先安装在左耳上袖口左耳。然后轻轻地支持动物的头，用一只手，推进耳袖口用另一只手轻轻地右耳到右袖口。下一步，放置一个加热垫动物的身下，并将其设置到86°F，以保持体温。 </li><li>然后，尝试移动头部一侧到另一侧检查双侧刚性。动物被正确地安装在立体定位框架上时，头部是刚性的，不能移动一边到另一边，但可以很容易地转动上下运动。 </li><li>轻轻休息鼠的鼻/齿列上的口鼻部装配在确保门牙轻轻地但稳固地搁在鼻子/齿杆的齿孔径内。 </li><li>用无菌解剖刀，使中线切口从眼睛的中间开始向耳朵的中间。确保手术刀是在一个45°角举行，并施加足够的压力通过底层筋膜，但不切透颅骨因为这会导致过量出血。 </li><li>用棉签分离筋膜和暴露颅骨。地标颅骨前囟和lambda应清晰可见（ 图1A）。确保该动物的头骨平面位置通过调整鼻/齿栏，使得前囟和lambda的地标都在同一个背腹（DV）位置（+0.1毫米）。 </li><li>取少量无菌生理盐水到暴露的部位，用棉签轻轻擦拭头骨。然后等待2-3分钟的头骨完全风干然后再继续。 </li><li>安装在左手臂立体定位针测量前囟和lambda的DV位置。这两个标志性建筑的DV定位应在0.1 mm对方。如果不是，则立体框架的鼻部栏可以上下调节，以实现这一目标。 </li><li>定位于前囟针来记录它的前 - 后（AP），横向（LAT）和背腹（DV）的测量。可以肯定的针头刚触及颅骨和不穿透它。 </li><li>使用鼠标脑图谱，如“老鼠大脑中的立体坐标”1，确定目标结构的坐标：海马内侧穿通通路（MPP）的束角和齿状回（DG），相对于拉姆达和前囟，分别为（也见表1）。然后，使用一个细点钢笔或铅笔上的骷髅标记这些点。 </li><li>此外，标上骷髅头的对侧两点。这些点WHICH将作为接地（GND）和基准（REF）应该是从中线约3毫米和纵向定位彼此相对（ 图1A，也见表1）。 </li><li>从左侧立体定位臂取下针头标记并将它与一个电动牙钻搭载了立体定位安装更换。定位钻头在每个标记点，使大约0.5毫米的直径小毛刺的孔。钻孔时，使用重复向上和向下运动，并经常检查钻头是否已经渗透颅骨暴露脑表面。 </li><li>轻轻地但坚定地螺丝钉电极（＃0-80不锈钢机螺钉1/8，开槽圆头）在每个对侧孔（GND和REF），使它们刚刚接触皮质表面无穿透它（ 图1A ）。这两个螺旋电极作为地面和参考。 </li><li>在电摩刺激和记录电极德持有人的左和右立体定位臂，分别。 </li><li>连接的刺激电极端子的电刺激器具有电流隔离和记录电极端子电生理学差分放大器（增益= 1,000，带通滤波器= 1赫兹-3千赫），然后到一个数字示波器诱发反应的目视检查。此外，连接GND和REF电极端子连接到差分放大器。 </li><li>轻轻地，慢慢地降低双方的刺激和记录电极以0.5 mm递增，同时在视觉上监测到600-800微安的刺激震撼的诱发反应。继续降低每个电极，直到他们达到各自目标的DV位置和刻板信号示波器（ 图1B）是观察。 </li><li>随后，用牙科丙烯酸水泥作盖固定到位的电极端子;任何牙科水泥，触及皮肤，应抹去马上。一旦牙科水泥完全固化的立体框架转移动物到一个干净的啮齿动物的笼子。用加热灯或垫以保持核心温度。 </li><li>监测动物每隔一小时，直到它恢复意识。一个注射氟尼辛可以施用作为镇痛药。 </li></ol> 2。 LTP的诱导<ol><li>让动物5-7天，从手术中恢复。将动物组成的法拉第笼（122厘米x 43厘米x 43厘米）配备隔音材料和5通道换向器旋转连接电极导致刺激/记录设置的录音环境。 </li><li>让动物的电极连接到刺激和记录工具（参见步骤1.17），在之前的录音环境适应1-2小时。 </li><li>设置刺激器控制来输出400微安，并记录平均10的振幅诱发反应，确保第在刺激之间至少10秒的经过。使用图1C所示的方法进行量化的诱发反应。重复此过程，600，800，1,000，1,200，1,400和微安。 </li><li>通过绘制诱发反应与刺激强度的平均振幅构造的输入/输出曲线。从该曲线确定它对应于测得的最大振幅的50％的刺激强度。使用50％强度的试验的其余部分。 </li><li>然后，使用50％的刺激强度，通过记录5的平均振幅诱发反应每分钟15分钟得到基线。再次确保刺激之间至少有10秒的流逝。 </li><li>随后交付强直刺激组成的10连发10个脉冲传递在400赫兹，5 Hz到内侧穿通通路的爆率。密切监测动物扣押的迹象，包括湿狗甩头。如果动物有癫痫发作的experim耳鼻喉科应终止并从该动物的所有数据应该被排除在研究成果。 </li><li>然后，继续录得5的平均振幅诱发反应每分钟30分钟posttetanization。在此之后，这个幅度比较通过计算从基线的百分比变化（ 图2）在步骤2.5中得到的基准振幅。 </li><li>下列实验结束时，动物被吸入异氟醚的方法是与美国兽医医学协会安乐死的指导方针相一致的方式实施安乐死。 </li></ol>

<ol>
	<li>Paxinos, G., Franklin, K. B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The mouse brain in stereotaxic coordinates. , (2004).</li><li>Bliss, T. V. P., Lomo, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-lasting+potentiation+of+synaptic+transmission+in+the+dentate+area+of+the+anesthetized+rabbit+following+stimulation+of+the+perforant+path.">Long-lasting potentiation of synaptic transmission in the dentate area of the anesthetized rabbit following stimulation of the perforant path.</a> J. Physiol. 232, 331-356 (1973).</li><li>Blaise, J. H., Bronzino, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+stimulus+frequency+and+age+on+bidirectional+synaptic+plasticity+in+the+dentate+gyrus+of+freely+moving+rats.">Effects of stimulus frequency and age on bidirectional synaptic plasticity in the dentate gyrus of freely moving rats.</a> Exp. Neurol. 182, 497-506 (2003).</li><li>Blaise, J. H., Koranda, J. L., Chow, U., Haines, K. E., Dorward, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+isolation+stress+alters+bidirectional+long-term+synaptic+plasticity+in+amygdalo-hippocampal+synapses+in+freely+behaving+adult+rats.">Neonatal isolation stress alters bidirectional long-term synaptic plasticity in amygdalo-hippocampal synapses in freely behaving adult rats.</a> Brain Res. 1193, 25-33 (2008).</li><li>Koranda, J. L., Masino, S. A., Blaise, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bidirectional+synaptic+plasticity+in+the+dentate+gyrus+of+the+awake+freely+behaving+mouse.">Bidirectional synaptic plasticity in the dentate gyrus of the awake freely behaving mouse.</a> J. Neurosci. Methods. 167, 160-166 (2008).</li><li>Cooke, S. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autophosphorylation+of+alphaCaMKII+is+not+a+general+requirement+for+NMDA+receptor-dependent+LTP+in+the+adult+mouse.">Autophosphorylation of alphaCaMKII is not a general requirement for NMDA receptor-dependent LTP in the adult mouse.</a> J. Physiol. 574, 805-818 (2006).</li><li>Davis, S., Bliss, T. V., Dutrieux, G., Laroche, S., Errington, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+and+duration+of+long-term+potentiation+in+the+hippocampus+of+the+freely+moving+mouse.">Induction and duration of long-term potentiation in the hippocampus of the freely moving mouse.</a> J. Neurosci. Methods. 75, 75-80 (1997).</li><li>Jones, M. W., Peckham, H. M., Errington, M. L., Bliss, T. V., Routtenberg, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+plasticity+in+the+hippocampus+of+awake+C57BL/6+and+DBA/2+mice:+interstrain+differences+and+parallels+with+behavior.">Synaptic plasticity in the hippocampus of awake C57BL/6 and DBA/2 mice: interstrain differences and parallels with behavior.</a> Hippocampus. 11, 391-396 (2001).</li><li>Zhang, T. A., Tang, J., Pidoplichko, V. I., Dani, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Addictive+nicotine+alters+local+circuit+inhibition+during+the+induction+of+in+vivo+hippocampal+synaptic+potentiation.">Addictive nicotine alters local circuit inhibition during the induction of in vivo hippocampal synaptic potentiation.</a> J. Neurosci. 30, 6443-6453 (2010).</li><li>Tang, J., Dani, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+enables+in+vivo+synaptic+plasticity+associated+with+the+addictive+drug+nicotine.">Dopamine enables in vivo synaptic plasticity associated with the addictive drug nicotine.</a> Neuron. 63, 673-682 (2009).</li></ol>

作者什么都没有透露。

在这个协议中，我们已经证明了在自由活动的小鼠研究LTP在DG可靠，简便的方法。而LTP的清醒大鼠许多研究已经进行3,4，很少有清醒小鼠主要是由于在小鼠颅有限，房地产和相对平均重量电极的探头的重量所带来的技术复杂性进行小鼠5。的几项研究已证明LTP的DG在自由活动利用任一微驱动电极系统或结型场效应晶体管集成在这必然增加了电极的有效载荷负担的动物6-10的探?...

技术的发展来操纵基因产生了几乎每一个神经变性疾病和神经系统疾病的转基因和基因敲除小鼠模型。因此有必要在以前在较大的啮齿动物到小鼠动物模型中使用的电生理研究技术翻译。一个这样的神经电生理研究技术是利用长时程增强（LTP）来测试参与各种神经病理疾病的神经元网络中的突触连接的有效性。本协议描述技术LTP的自由活动的小鼠可靠的电生理研究。此协议的比其他的优点在于，它是简单和容易实现的，它也是相当少昂贵，因为它不需要也不使用昂贵的计算机控制的微驱动系统也不场效应晶体管的探头的，并且，据我们所知，是慢性电生理记录吨第一视频协议Ø研究LTP在自由活动的小鼠。为此，我们在本文中描述的简单方法用于研究长时程增强中自由活动的小鼠。这些方法可以很容易地转换成神经病理性疾病的转基因和基因敲除小鼠模型。

<table border="0" cellpadding="0" cellspacing="0" width="514">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Ketamine (100 mg/ml)</td>
			<td style="width:104px;">Henry Schein</td>
			<td style="width:137px;">10177</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Xylazine (20 mg/ml)</td>
			<td style="width:104px;">Henry Schein</td>
			<td style="width:137px;">33197</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Acepromazine (10 mg/ml)</td>
			<td style="width:104px;">Henry Schein</td>
			<td style="width:137px;">2177</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Dental acrylic powder</td>
			<td style="width:104px;">Lang Dental Manufacturing Co.</td>
			<td style="width:137px;">1330CLR</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Dental acrylic liquid</td>
			<td style="width:104px;">Lang Dental Manufacturing Co.</td>
			<td style="width:137px;">1306CLR</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Tungsten wire (0.127 mm)</td>
			<td style="width:104px;">World Precision Instruments</td>
			<td style="width:137px;">TGW0515</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Stainless Steel Hypodermic Tubing (0.286 mm)</td>
			<td style="width:104px;">World Precision Instruments</td>
			<td style="width:137px;">832400</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Flunixin (50 mg/ml)</td>
			<td style="width:104px;">Henry Schein</td>
			<td style="width:137px;">14165</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Epoxilyte</td>
			<td style="width:104px;">Superior Essex</td>
			<td style="width:137px;">EP 6001-M</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Stainless steel wire insert (0.2 mm)</td>
			<td style="width:104px;">World Precision Instruments</td>
			<td>792900</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:195px;">Stereotaxic frame apparatus</td>
			<td style="width:95px;">Kopf Instruments</td>
			<td style="width:137px;">Model 902</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:195px;">Ear cuffs (ear cups)</td>
			<td style="width:95px;">Kopf Instruments</td>
			<td style="width:137px;">Model 921&#160;</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:195px;">Electrophysiological stimulator</td>
			<td style="width:95px;">Astro-Med, Inc.</td>
			<td style="width:137px;">S88</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:195px;">Digital oscilloscope</td>
			<td style="width:95px;">B &#38; K Precision Corp.</td>
			<td style="width:137px;">2542</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:195px;">Current isolation unit</td>
			<td style="width:95px;">Astro-Med, Inc.</td>
			<td style="width:137px;">PSIU-6</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:195px;">Differential amplifier</td>
			<td style="width:95px;">World Precision Instruments, Inc.</td>
			<td style="width:137px;">DAM-50</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Commutator</td>
			<td style="width:95px;">Plastics One</td>
			<td style="width:137px;">SLC6</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">Dental drill</td>
			<td style="width:95px;">Stoelting</td>
			<td style="width:137px;">58650</td>
			<td style="width:79px;"></td>
		</tr>
	</tbody>
</table>

long-term potentiation of perforant pathway-dentate gyrus synapse in freely behaving mice

的突触效能的长时程增强的研究，其特性使其作为一个潜在的细胞机制基本学习和信息存储吸引力的活动依赖的突触现象，早已被用来阐明各种神经回路中的海马，杏仁核的生理和其他边缘和皮质结构。考虑到这一点，神经系统疾病的转基因小鼠模型代表有用的平台上进行长时程增强（LTP）的研究开发基因在正常和异常的突触通信中参与学习，情感和信息的神经元网络中的作用更深入的了解处理。本文介绍的方法进行可靠地诱导LTP在自由活动的小鼠。这些方法可以在转基因的研究中使用和敲除自由活动的神经退行性疾病的小鼠模型。

表1示出了如在此协议中使用的坐标为DG和MPP 图1A示出的标记为在头骨的目标结构;。还示出的是接地电极和参考电极的位置， 图1B示出了代表性的诱发反应的痕迹都预和posttetanization在相同的动物。注意，posttetanization诱发反应比pretetanization响应其表示LTP诱导2。 图1C示出了用于量化的振幅响应的方法大。实际上， 图2示出了响应...

Watch this Scientific Journal Video about Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice at JoVE.com

Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

神经系统疾病的转基因和基因敲除小鼠模型可用于研究基因在正常和异常的神经生理学中的作用是有用的。本文介绍的方法可用于研究长时程增强，它可依据学习和记忆，在转基因和基因敲除自由活动的神经病理学的小鼠模型细胞机制。

穿通通路 - 齿状回突触在自由活动的小鼠长时程增强

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential ...

long-term-potentiation-perforant-pathway-dentate-gyrus-synapse-freely

Research

JoVE Journal

Behavior

14.0K 次观看.  Trinity College. 神经系统疾病的转基因和基因敲除小鼠模型可用于研究基因在正常和异常的神经生理学中的作用是有用的。本文介绍的方法可用于研究长时程增强，它可依据学习和记忆，在转基因和基因敲除自由活动的神经病理学的小鼠模型细胞机制。 

视频: Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice and other rodents. Improvements in video tracking software (VTS) have allowed it to be coupled to LMA testing, dramatically improving specificity and sensitivity when compared to the line crossings method with manual scoring. In addition, VTS enables high-throughput experimentation. While similar to automated video tracking used for the open field test (OFT), LMA testing is unique in that it allows mice to remain in their home cage and does not utilize the anxiogenic stimulus of bright lighting during the active phase of the light-dark cycle. Traditionally, LMA has been used for short periods of time (mins), while longer movement studies (hrs-days) have often used implanted transmitters and biotelemetry. With the option of real-time tracking, long-, like short-term LMA testing, can now be conducted using videography. Long-term LMA testing requires a specialized, but easily constructed, cage so that food and water (which is usually positioned on the cage top) does not obstruct videography. Importantly, videography and VTS allows for the quantification of parameters, such as path of mouse movement, that are difficult or unfeasible to measure with line crossing and/or biotelemetry. In sum, LMA testing coupled to VTS affords a more complete description of mouse movement and the ability to examine locomotion over an extended period of time.

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice. Coupling of video tracking software (VTS) and LMA allows for the improvement of specificity and sensitivity, especially when compared with the manual, line crossing method of LMA analysis. Additionally VTS allows long-term tracking of mouse LMA.

鼠标短期和长期活动性分析视频跟踪软件

Locomotor  activity (LMA) is a simple and easily performed measurement of behavior in mice  and other rodents. Improvements in video tracking software ...

科研

行为学

Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice

State-of-the-art electrophysiological recordings from the brains of freely behaving animals allow researchers to simultaneously examine local field potentials (LFPs) from populations of neurons and action potentials from individual cells, as the animal engages in experimentally relevant tasks. Chronically implanted microdrives allow for brain recordings to last over periods of several weeks. Miniaturized drives and lightweight components allow for these long-term recordings to occur in small mammals, such as mice. By using tetrodes, which consist of tightly braided bundles of four electrodes in which each wire has a diameter of 12.5 &mu;m, it is possible to isolate physiologically active neurons in superficial brain regions such as the cerebral cortex, dorsal hippocampus, and subiculum, as well as deeper regions such as the striatum and the amygdala. Moreover, this technique insures stable, high-fidelity neural recordings as the animal is challenged with a variety of behavioral tasks. This manuscript describes several techniques that have been optimized to record from the mouse brain. First, we show how to fabricate tetrodes, load them into driveable tubes, and gold-plate their tips in order to reduce their impedance from M&Omega; to K&Omega; range. Second, we show how to construct a custom microdrive assembly for carrying and moving the tetrodes vertically, with the use of inexpensive materials. Third, we show the steps for assembling a commercially available microdrive (Neuralynx VersaDrive) that is designed to carry independently movable tetrodes. Finally, we present representative results of local field potentials and single-unit signals obtained in the dorsal subiculum of mice. These techniques can be easily modified to accommodate different types of electrode arrays and recording schemes in the mouse brain.

The design and assembly of microdrives for in vivo electrophysiological recordings of brain signals from the mouse is described. By attaching microelectrode bundles to sturdy driveable carriers, these techniques allow for long-term and stable neural recordings. The lightweight design allows for unrestricted behavioral performance by the animal following drive implantation.

清醒的行为小鼠慢性神经录音的微型硬盘阵列建设

State-of-the-art electrophysiological recordings from the brains of freely behaving animals allow researchers to simultaneously examine local field ...

Trace Fear Conditioning in Mice

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings between a neutral stimulus and an unconditioned stimulus. There is a 20 sec trace period that separates each conditioning trial. On the following day freezing is measured during presentation of the conditioned stimulus (CS) and trace period. On the third day there is an 8 min test to measure contextual memory. The representative results are from mice that were presented with the aversive unconditioned stimulus (shock) compared to mice that received the tone presentations without the unconditioned stimulus. Trace fear conditioning has been successfully used to detect subtle learning and memory deficits and enhancements in mice that are not found with other fear conditioning methods. This type of fear conditioning is believed to be dependent upon connections between the medial prefrontal cortex and the hippocampus. One current controversy is whether this method is believed to be amygdala-independent. Therefore, other fear conditioning testing is needed to examine amygdala-dependent learning and memory effects, such as through the delay fear conditioning.

In the following experiment we describe a protocol for trace fear conditioning in mice. This type of associative memory includes a trace period that separates the neutral stimulus and the unconditioned stimulus.

跟踪恐惧条件小鼠

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

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

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

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

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

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

A Novel Experimental and Analytical Approach to the Multimodal Neural Decoding of Intent During Social Interaction in Freely-behaving Human Infants

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental paradigms and analysis tools have been limited to constrained laboratory tasks and contexts due to technical limitations imposed by the available set of measuring and analysis techniques and the age of the subjects. These limitations severely limit the study of developmental neural dynamics and associated neural networks engaged in cognition, perception and action in infants performing &#8220;in action and in context&#8221;. This protocol presents a novel approach to study infants and young children as they freely organize their own behavior, and its consequences in a complex, partly unpredictable and highly dynamic environment. The proposed methodology integrates synchronized high-density active scalp electroencephalography (EEG), inertial measurement units (IMUs), video recording and behavioral analysis to capture brain activity and movement non-invasively in freely-behaving infants. This setup allows for the study of neural network dynamics in the developing brain, in action and context, as these networks are recruited during goal-oriented, exploration and social interaction tasks.

This protocol presents a novel methodology for the neural decoding of intent from freely-behaving infants during unscripted social interaction with an actor. Neural activity is acquired using non-invasive high-density active scalp electroencephalography (EEG). Kinematic data is collected with inertial measurement units and supplemented with synchronized video recording.

在自由，表现人类婴儿一种新的实验和分析方法意向的多模态神经解码期间社会互动

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental ...

A Pressure Injection System for Investigating the Neuropharmacology of Information Processing in Awake Behaving Macaque Monkey Cortex

The top-down modulation of feed-forward cortical information processing is functionally important for many cognitive processes, including the modulation of sensory information processing by attention. However, little is known about which neurotransmitter systems are involved in such modulations. A practical way to address this question is to combine single-cell recording with local and temporary neuropharmacological manipulation in a suitable animal model. Here we demonstrate a technique combining acute single-cell recordings with the injection of neuropharmacological agents in the direct vicinity of the recording electrode. The video shows the preparation of the pressure injection/recording system, including preparation of the substance to be injected. We show a rhesus monkey performing a visual attention task and the procedure of single-unit recording with block-wise pharmacological manipulations.

Here, we show the pressure injection of neuropharmacological substances during single-cell recording in an awake, behaving macaque monkey. This procedure allows pharmacological manipulation in the direct vicinity of a cortical recording site.

对调查信息处理的神经药理学清醒行为猕猴皮质压力喷射系统

The top-down modulation of feed-forward cortical information processing is functionally important for many cognitive processes, including the modulation ...

Multi-layer Cortical Ca2+ Imaging in Freely Moving Mice with Prism Probes and Miniaturized Fluorescence Microscopy

In vivo circuit and cellular level functional imaging is a critical tool for understanding the brain in action. High resolution imaging of mouse cortical neurons with two-photon microscopy has provided unique insights into cortical structure, function and plasticity. However, these studies are limited to head fixed animals, greatly reducing the behavioral complexity available for study. In this paper, we describe a procedure for performing chronic fluorescence microscopy with cellular-resolution across multiple cortical layers in freely behaving mice. We used an integrated miniaturized fluorescence microscope paired with an implanted prism probe to simultaneously visualize and record the calcium dynamics of hundreds of neurons across multiple layers of the somatosensory cortex as the mouse engaged in a novel object exploration task, over several days. This technique can be adapted to other brain regions in different animal species for other behavioral paradigms.

Multi-layer Cortical Ca2+ Imaging in Freely Moving Mice with Prism Probes and Miniaturized Fluorescence Microscopy

Here, we present a procedure for performing large-scale Ca2+ imaging with cellular-resolution across multiple cortical layers in freely moving mice. Hundreds of active cells can be observed simultaneously using a miniature, head-mounted microscope coupled with an implanted prism probe.

多层皮质Ca<sup&gt; 2+</sup&gt;使用棱镜探针和小型荧光显微镜成像自由移动的小鼠

In vivo circuit and cellular level functional imaging is a critical tool for understanding the brain in action. High resolution imaging of mouse cortical ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

用于体内的无线双向接口记录和刺激自由行为大鼠的神经活动

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer&#039;s Disease, Using IntelliCage

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

基于 IntelliCage 的阿尔茨海默病多基因模型认知功能的自动、长期行为检测

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and human studies to investigate the cortical processing of nociceptive information. These laser-evoked brain potentials (LEPs) consist of several transient responses that are time-locked to the onset of laser stimuli. However, the functional properties of the LEP responses are still largely unknown, due to the lack of a sampling technique that can simultaneously record neural activities at the surface of the cortex (i.e., electrocorticogram [ECoG] and scalp electroencephalogram [scalp EEG]) and inside the brain (i.e., local field potential [LFP]). To address this issue, we present here an animal protocol using freely moving rats. This protocol is composed of three main procedures: (1) animal preparation and surgical procedures, (2) a simultaneous recording of ECoG and LFP in response to nociceptive laser stimuli, and (3) data analysis and feature extraction. Specifically, with the help of a 3D-printed protective shell, both ECoG and LFP electrodes implanted on the rat&#39;s skull were securely held together. During data collection, laser pulses were delivered on the rat&#39;s forepaws through gaps in the bottom of the chamber when the animal was in spontaneous stillness. Ongoing white noise was played to avoid the activation of the auditory system by the laser-generated ultrasounds. As a consequence, only nociceptive responses were selectively recorded. Using the standard analytical procedures (e.g., band-pass filtering, epoch extraction, and baseline correction) to extract stimulus-related brain responses, we obtained results showing that LEPs with a high signal-to-noise ratio were simultaneously recorded from ECoG and LFP electrodes. This methodology makes the simultaneous recording of ECoG and LFP activities possible, which provides a bridge of electrocortical signals at the mesoscopic and macroscopic levels, thereby facilitating the investigation of nociceptive information processing in the brain.

We developed a technique that simultaneously records both electrocorticography and local field potentials in response to nociceptive laser stimuli from freely moving rats. This technique helps establish a direct relationship of electrocortical signals at the mesoscopic and macroscopic levels, which facilitates the investigation of nociceptive information processing in the brain.

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...