由美国国立卫生研究院授予AG027297，从肯塔基脊髓和颅脑损伤的研究信托奖，并从Kleberg基金会的礼物支持的工作。

<ol>
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生产这种视频文章是徕卡Microsystems公司赞助。

在本议定书中概述的步骤将有助于确保至少为迅速有效地开展脑解剖年龄在年轻成年大鼠，。我们还提供足够的细节，对于初学者LTP和LTD成立自己的切片研究。如果老化和突触功能和可塑性AD变化的进一步探索，是您的目标之一，有至少有两个其他的方法问题，上面提到的，值得进一步考虑。首先，一些实验室已经表明的Ca 2 +：Mg2 +的比例在录音学联可以在2,10,24,25海马诱导突触可塑性的一个显着的效果。在哺乳动物脑脊液中的 Ca 2 +：Mg2 +的比例大约是（如见 26） 。然而，学联的Ca 2 + Mg2 +的比例接近2片突触功能和可塑性研究。在早期的研究中，这种做法很可能是调整，优化诱导的LTP，随后成为日常所有的可塑性研究。然而，这种做法可以在老化和AD的研究问题，因为在神经元的Ca 2 +调节的特点差异。具体来说，CA 2 +内流和/或Ca 2 +诱导的Ca 2 +释放是升高的老年大鼠和/或在AD模型小鼠神经元激活3,27-31。有限公司感应是在学联的Ca 2 +水平的微妙的变化特别敏感。我们的协议，采用2mm的钙2 +和2 mM的镁，在公司通常结果年龄，但不年轻的成年动物2，同时研究利用一个钙2 +：镁+比例接近两个，已观察到的成年人强大的公司在该没有年龄差异2,10结合减少有限公司或在32岁的大鼠。这些意见强调需要仔细考虑学联钙2 +和Mg2 +水平进行比较时，在中青年成年动物的Ca 2 +依赖可塑性。 第二个方法问题， 涉及 33突触后膜的去极化和突触强度的差异可能老化/基因型的LTP的强烈依赖。在一个典型的LTP的实验，基线和LTP的刺激强度是一般调整，以生产半最大（或三季度最大）EPSP的幅度。潜在的问题是，通常表明，老年大鼠和APP/PS1小鼠突触强度降低相对年轻和/或野生型对口，这意味着，基线EPSP值也将被更小，老年大鼠和APP/PS1小鼠。较小的EPSPS可能转化为在LTP的刺激少的去极化，导致减少的概率为诱导LTP的33。由于这种潜在的混淆，这是很难以确定是否这些动物表现出吞吐量赤字，可塑性赤字或两者。也就是说，年龄和/或APP/PS1小鼠LTP的诱导机制可能是功能完好（无可塑性赤字），但在这些条件下不够刺激（吞吐量赤字）。这个区别是至关重要的，吞吐量和可塑性机制机制的反应可能非常不同的一个特定的药物治疗。我们尽量减少吞吐量降低的LTP诱导的影响，通过规范的所有片LTP的刺激前EPSP的幅度相同的水平（如为1 mV）。其他策略可能会有效，以及（如电压或电流钳的使用，以平衡各群体之间在膜电位的LTP的刺激），并调查在这些动物模型的LTP时，应考虑。

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>氯化钠</td><td>费舍尔</td><td> BP358 - 1 </td></tr><tr><td>氯化钾</td><td>费舍尔</td><td> BP366 - 500 </td></tr><tr><td> KH 2 PO 4（一元） </td><td>西格玛</td><td> P5379 - 100G </td></tr><tr><td> 硫酸镁 </td><td>西格玛</td><td> M2643 - 500G </td></tr><tr><td> 氯化钙 （二水） </td><td>西格玛</td><td> C3306 - 250G </td></tr><tr><td>碳酸氢钠3 </td><td>费舍尔</td><td> S233 - 500 </td></tr><tr><td> C 6 H 12 O 6（葡萄糖） </td><td>费舍尔</td><td> BP350 - 1 </td></tr></tbody></table>试剂要求见表1。 <table border="1"><tbody><tr><td> 设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>锥形瓶</td><td>费舍尔</td><td> FB - 500 - 2000 FB - 500 - 1000 </td><td></td></tr><tr><td>水族馆鼓泡</td><td></td><td></td><td>用于充氧媒体。可在大多数宠物商店</td></tr><tr><td> 50毫升的玻璃烧杯</td><td>费舍尔</td><td> 02，约440克</td><td>对于脑学联storarge </td></tr><tr><td>封口膜</td><td>费舍尔</td><td> 13-374-10 </td><td></td></tr><tr><td>小动物断头台</td><td>世界精密仪器（WPI） </td><td> DCAP - M </td><td></td></tr><tr><td>平纸巾</td><td></td><td></td><td></td></tr><tr><td> ＃11羽手术刀片</td><td>费舍尔</td><td> 08 - 916 - 5B </td><td></td></tr><tr><td> Beebee骨剪刀</td><td>精细科学的工具（FST）的</td><td> 16044-10 </td><td></td></tr><tr><td> Lempert Rongeurs </td><td> Roboz </td><td> RS - 8321 </td><td>使用老鼠</td></tr><tr><td>弗里德曼 - 皮尔森Rongeurs </td><td>福斯特</td><td> 16020-14 </td><td>使用小鼠或大鼠</td></tr><tr><td>海马工具</td><td>福斯特</td><td> 10099-15 </td><td></td></tr><tr><td>勺子</td><td></td><td></td><td>一个塑料小勺将尽</td></tr><tr><td>抹刀</td><td>费舍尔</td><td> 21 - 401 - 25A </td><td>抹刀</td></tr><tr><td> 0虹膜剪刀</td><td>福斯特</td><td> 14058-09 </td><td></td></tr><tr><td>塑料转移吸管</td><td>费舍尔</td><td> 13-711-43 </td><td></td></tr><tr><td> 110毫米的Whatman滤纸</td><td>费舍尔</td><td> 09 - 805E </td><td>滤纸猫。 1001-110 </td></tr><tr><td>玻璃培养皿</td><td>费舍尔</td><td></td><td></td></tr><tr><td>徕卡VT1000P手册振动切片机</td><td> Vibratome </td><td></td><td></td></tr><tr><td> 0.1毫米FA - 10羽毛小号刀片</td><td>特德佩拉</td><td> 121-9 </td><td> 0.1毫米FA - 10羽毛小号刀片</td></tr><tr><td>硼硅玻璃巴斯德吸取（橡胶球） </td><td>费舍尔</td><td> 13 - 678 - 20A </td><td>对于转让切片提示折断更大的开放和热抛光</td></tr><tr><td> 35毫米Polysterine的培养皿</td><td>康宁</td><td> 430588 </td><td>用于收集解剖后切片</td></tr></tbody></table> 表2工具和材料解剖<table border="1"><tbody><tr><td> 设备/材料 </td><td> 公司 </td><td> 评论（可选） </td></tr><tr><td>控股室</td><td>客户定制</td><td></td></tr><tr><td> P - 97水平移液器拖轮</td><td>萨特仪器有限公司</td><td></td></tr><tr><td>振动隔离表</td><td>技术制造公司（TMC） </td><td></td></tr><tr><td>法拉第笼</td><td>自定义内置</td><td></td></tr><tr><td>耐热吸气瓶底部手枪（产品编号1220 - 1L） </td><td>康宁</td><td></td></tr><tr><td>重力contolled四集稳压器（产品编号2C8891） </td><td>巴克斯特</td><td></td></tr><tr><td>中央真空系统</td><td></td><td>可在最现代化的实验室</td></tr><tr><td> 95％的O 2 / 5％的CO 2气体混合</td><td>苏格兰人T -总值有限公司</td><td></td></tr><tr><td> TygonTM实验室管 为O 2 / CO 2交付</td><td> Fisher Scientific则</td><td>无毒，非氧化性，有各种尺寸的。 </td></tr><tr><td> Eclipse的E600FN显微镜</td><td>尼康</td><td> 10倍和40倍的目标近红外线过滤器，和GFP，DS - red2的过滤器， </td></tr><tr><td>酷捕捉ES数码相机</td><td> Photometrics </td><td>酷捕捉ES数码相机</td></tr><tr><td>的X - Cite荧光照明灯</td><td> EXFO </td><td>的X - Cite荧光照明灯</td></tr><tr><td>显微镜平台</td><td>锡斯基尤</td><td>自定义组装</td></tr><tr><td> RC - 22潜水录音室（产品编号64-0228） </td><td>华纳仪器（威斯康星州） </td><td>需要P - 1型平台和舞台适配器（产品从华纳＃64-0277） </td></tr><tr><td> TC2BIP 2/3Ch温度控制器 </td><td>细胞Microcontrols </td><td> TC2BIP 2/3Ch温度控制器 </td></tr><tr><td> 4轴手动微型机械臂</td><td>锡斯基尤</td><td></td></tr><tr><td>铂铱丝（0.002） （＃PTT0203项目） </td><td> WPI </td><td></td></tr><tr><td> A365刺激隔离器</td><td> WPI </td><td> A365刺激隔离器</td></tr><tr><td> Multiclamp 700B放大器</td><td> Axon仪器</td><td></td></tr><tr><td> Digidata 1322A的A / D转换转换器</td><td> Axon仪器</td><td></td></tr><tr><td> PClamp软件</td><td> Axon仪器</td><td></td></tr><tr><td>个人电脑（奔腾4） </td><td>戴尔</td><td></td></tr></tbody></table> 表3。电设备和材料

preparation of acute hippocampal slices from rats and transgenic mice for the study of synaptic alterations during aging and amyloid pathology

啮齿类动物的海马脑片制备也许是最广泛使用的工具为调查哺乳动物突触功能和可塑性。海马可快速，轻松地从大鼠和小鼠中提取和切片仍然可行含氧人工脑脊液小时。此外，基本electrophysisologic的技术很容易应用到海马突触功能的调查，并提供了一​​些认知障碍的最好的生物标志物。海马脑片是在学习和记忆有关的突触可塑性机制的研究，特别是流行的。诱导长时程增强和突触效能的抑郁症（LTP和LTD）在海马（或缺乏）的变化是经常被用来描述神经表型的认知受损的动物和/或评估行动的机制益智化合物。本文概述了我们准备从老鼠和转基因小鼠海马突触改变与大脑衰老和阿尔茨海默氏病（AD）1-3相关的研究中使用的程序。使用的老年大鼠和AD模型小鼠可呈现出一套独特的挑战，习惯于在他们的研究中使用年轻的大鼠和/或小鼠的研究人员。老年大鼠有较厚的比年轻大鼠和小鼠的头骨和更严厉的结缔组织，可延缓脑提取和/或夹层，并因此否定或夸大真正的突触功能和可塑性的年龄差异。老龄化和淀粉样蛋白的病理还可能加剧在解剖过程持续的海马损伤，再复杂的生理评估得出的任何推论。在这里，我们讨论了在解剖过程中采取的步骤，以尽量减少这些问题。在“健康”和“不健康”的片从大鼠和小鼠获得的突触反应的例子，以及有代表性的突触可塑性实验。在这些动物模型（如录音解决方案组件，刺激参数）的突触功能的其他方法因素可能产生的影响进行了讨论。虽然本文的重点是对老年大鼠和转基因小鼠，新手使用切片生理，要找到足够的细节，在这里开始自己的研究，使用各种啮齿类动物模型。

Watch this Scientific Journal Video about Preparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology at JoVE.com

Preparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology

本文概述了程序，准备从老鼠和转基因小鼠海马突触改变与大脑衰老和与年龄有关的神经退行性疾病如阿尔茨海默氏症，相关的研究。

突触改变老化过程中淀粉样蛋白病理研究从大鼠和转基因小鼠海马急性片的制备

The rodent hippocampal slice preparation is perhaps the most broadly used tool for investigating mammalian synaptic function and plasticity. The ...

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93.6K 次观看.  University of Kentucky College of Public Health. 本文概述了程序，准备从老鼠和转基因小鼠海马突触改变与大脑衰老和与年龄有关的神经退行性疾病如阿尔茨海默氏症，相关的研究。