本研究部分由国立卫生研究院 (R01 NS102209) 资助, 并由克莱尔. 福布斯信托基金捐赠。

我们的实验性议定书得到了马里兰大学动物保育和使用委员会的批准, 并遵循了国家卫生研究院的护理和使用实验动物指南。
注: 所有实验均采用成年雄性大鼠 (250–350 g)。不同的动物群被用于生化分析、行为实验和睡眠唤醒记录。这些动物被安置在一个温度控制的设施在马里兰精神病学研究中心。他们被保持在一个12/12 小时的光黑暗的周期, 与灯在授时时间 (ZT) 0 和熄灯在 ZT 12。在实验中, 这些动物收到了随意食物和水的机会。该设施得到了美国实验室动物护理认证协会的充分认可。
1. 腹腔犬尿氨管理对大鼠的作用
注: 在本议定书中, 犬尿氨是在 zt 0 (光相开始), 并收集在 zt 2 和 zt 4, 以确定一个时间路线的犬尿氨新陈代谢。用盐水注射的动物作为控制。例如, 如果一只老鼠重500克, 所需剂量为100毫克/千克, 则大鼠应接受5毫升注射10毫克/毫升溶液的犬尿氨。
<ol><li>犬尿氨硫酸 ("犬尿氨"), 并将其放入一个20毫升玻璃瓶。一定要把犬尿氨在冰上。</li>
	<li>
添加生理盐水以达到所需浓度, 并将 pH 值调整为 7.6, 使用 1 N 氢氧化钠 (氢氧化钠)。涡旋和油脂实验溶液直到犬尿氨完全溶解。 注意事项: 在处理氢氧化钠时, 遵循所有建议预防措施。<ol>
<li>例如, 要达到10毫克/毫升溶液, 重出200毫克的犬尿氨, 并把它放在玻璃瓶。加入15毫升盐水和漩涡和油脂实验 (20 赫为 5–10 s, 1/8 英寸直径 microtip) 它溶化。使用氢氧化钠, 调整溶液的 pH 值。最后, 用生理盐水调节体积到20毫升。</li>
	</ol>
</li>
	<li>根据动物的体重和所需的治疗剂量, 对每只实验鼠进行称量, 计算每种注射量。小心地将动物从笼子里取出, 注入溶液腹腔, 将动物送回笼子。</li>
</ol>2. 用高效液相色谱法测量犬尿氨
<ol><li>组织收集 注: 在本议定书中, 犬尿氨是在 zt 0 (光相开始), 并收集在 zt 2 和 zt 4, 以确定一个时间路线的犬尿氨新陈代谢。用盐水注射的动物作为控制。<ol>
<li>用二氧化碳弄死动物。呼吸症状停止1分钟后, 用斩首法进行二次安乐死。</li>
		<li>收集全树干血, 放置一个一次性漏斗到一个15毫升管含有 20 ul 的 K3-EDTA (0.5 米)。让血液从体内排出到管子里。反复翻转管子, 确保 EDTA 混合在一起。</li>
		<li>使用剪刀, 把头上的皮肤切开中线, 露出头骨。去掉头骨后部多余的颈部肌肉, 在脊髓的开口处, 从头骨向后切开。<ol>
<li>轻轻拉两侧的头骨打开, 以揭露大脑, 然后小心地删除的大脑与镊子, 以免损害它。用剃刀刀片取出嗅球, 并将大脑从中线的一半切开。使用钳, 解剖 hemibrain 到感兴趣的区域 (即海马和皮质)。</li>
		</ol>
</li>
		<li>把所有解剖过的脑部放在铝箔上干冰上。组织完全冻结后, 将其转移到20毫升的塑料瓶, 并将其存储在-80 摄氏度。</li>
		<li>将整个树干的血液离心在 300 x g 10 分钟, 取出上清 (等离子) 并将其转移到新的离心管中, 然后将其储存在-80 摄氏度。</li>
	</ol></li>
	<li>样品准备
	<ol>
<li>血浆: 从被处理的动物收集<ol>
<li>用等离子解冻冷冻管 (见步骤2.1 为组织收集) 在湿冰。</li>
			<li>在一个新的离心管, 稀释样品与超纯水 (1:2 为犬尿氨, 1:10 为 KYNA)。到 100 ul 稀释样品, 添加 25 ul 6% 高氯酸。 注意事项: 在处理高氯酸时遵循所有建议预防措施。</li>
			<li>涡流所有样品, 然后离心他们在 1.2万 x g为10分钟。一旦完成, 删除 100 ul 的上清 (从每管), 并把它们放入一个小0.25 毫升离心管犬尿氨或 KYNA 的决心。</li>
		</ol>
</li>
		<li>大脑: 从被处理的动物收集<ol>
<li>放置管与冰冻脑标本 (见步骤2.1 为组织收集) 在干冰。在精确的分析天平上单独称量每个脑样本。把它还给管子, 放回干冰上。</li>
			<li>在所有的样品被权衡后, 把管子移到湿冰上。稀释每个样品1:10 瓦特/v 与超纯水和融汇他们使用一个 sonicator (20 赫 5–10 s, 1/8 英寸直径 microtip)。例如, 对于100毫克的脑组织, 添加 900 ul 的超纯水。</li>
			<li>整除 100 ul 每稀释的样品到一个新的管子和酸化它与 25 ul 25% 高氯酸。涡旋, 然后离心 1.2万 x g 10 分钟。</li>
			<li>离心后, 去除 100 ul 的上清, 并把它放到小0.25 毫升离心管 KYNA 的决心。</li>
		</ol>
</li>
	</ol></li>
	<li>高效液相色谱仪的建立与运行
	<ol>
<li>准备50毫米醋酸钠移动相。在1升的超纯水中溶解6.81 克醋酸钠三水。用冰乙酸将 pH 值调整为 6.2, 然后将真空过滤成清洁玻璃器皿。将醋酸钠移动相转移到干净的1升玻璃瓶中。</li>
		<li>准备500毫米醋酸锌。在500毫升的超纯水中溶解54.88 克醋酸锌, 然后将其真空过滤。把它转移到一个干净的500毫升玻璃瓶。用超纯水和500毫升瓶填充1升瓶, 高效液相色谱级乙腈。</li>
		<li>将进气管从 HPLC 机分别放入流动相、超纯水和100% 乙腈。将醋酸锌溶液分别通过与移动相结合的蠕动泵, 并在衍生之前进行泵浦。</li>
		<li>使用该机器上的控制面板, 程序它运行5% 乙腈和95% 超纯水0.5 毫升/分钟. 允许它运行20–30分钟, 以达到稳定的压力和基线。</li>
		<li>建立稳定基线后, 将溶液组成改为5% 乙腈和95% 醋酸钠流动相。平衡30分钟。</li>
		<li>打开荧光探测器中的灯, 让它预热。</li>
		<li>同时, 开机后柱泵的流量可达到0.1 毫升/分. 允许它在大约20分钟后到达稳定压力约 500 psi。</li>
		<li>制定标准。<ol>
<li>对于 KYNA, 从1毫米的库存溶液开始, 稀释它以准备5标准浓度 (即10 nm、5 nm、2.5 nm、1 nm 和 500 pM)。 注: 这将产生一个标准曲线范围从 200 fmoles 到 10 fmoles 每 20 ul 注射液。</li>
			<li>对于犬尿氨, 从1毫米的库存溶液开始, 稀释它以制备5标准浓度 (即10 微米、5微米、2.5 微米、1微米和 500 nM)。 注: 这将产生一个标准曲线范围从 200 pmoles 到 10 pmoles 每 20 ul 注射液。</li>
		</ol>
</li>
		<li>建立高效液相色谱系统软件, 控制样本序列参数, 允许多样本 autoinjections。<ol>
<li>当在 "运行示例" 屏幕上, 单击 "文件" 并向下拖动以 "创建新的示例集"。当新窗口打开时, 选择 "空"。单独列出所有标准 (请参阅步骤 2.3.8) 或按应运行的顺序排列示例。</li>
			<li>在 "函数" 列中, 指定标准和示例。标准应在序列的开始和结束时进行测定。在每5个样品之间注入水。</li>
			<li>将每个样本的运行时间设置为15分钟. 从标准和等离子制剂中注入 20 ul, 或从大脑匀浆制剂中注入 30 ul 样品。</li>
			<li>设置激发和发射波长 (取决于被评估的化合物) 为犬尿氨到励磁: 365 毫微米, 发射: 480 毫微米和保留时间: ~ 6 分钟, 并且为 KYNA 到励磁: 344 毫微米, 发射: 398 毫微米, 并且保留时间: ~ 11 分钟。</li>
			<li>设置好所有参数后, 机器平衡, 运行该序列。</li>
			<li>要量化数据, 请导航到 "浏览项目" 屏幕。在 "样本集" 选项卡下, 双击要量化的样本集。从所有注射的列表中, 突出显示所有标准并右键单击。在新窗口中, 选择 "进程", 然后使用下拉菜单选择 "如何" ("如何") 旁边的 "校准和定量":。</li>
			<li>再次突出显示所有标准, 右键单击并选择 "审阅"。当图谱打开时, 使用顶部的按钮 "集成", 然后 "校准" 每个标准;这将自动创建标准曲线。</li>
			<li>返回到 "示例集" 选项卡, 然后双击示例集。其次, 突出所有标准。重复上面提到的过程, 而是从下拉菜单中选择 "定量"。再次突出显示所有示例, 然后右键单击并选择 "审阅"。当图谱打开时, 使用顶部的按钮 "集成", 然后 "定量" 每个示例。 注意: 这将根据先前创建的标准曲线输出每个样本的浓度。</li>
		</ol>
</li>
	</ol></li>
</ol>3. 被动回避范式
注意: 这些行为实验是根据我们的生化发现和急性犬尿氨挑战而设计的。为了最大限度地增加脑 KYNA, 犬尿氨 (100 毫克/千克) 是在 zt 0, 2 h 的训练课程之前, 在被动回避范式测试海马介导的学习, 这发生在 zt 2。该仪器包括2个同等大小的隔间 (21.3 厘米高, 20.3 厘米宽, 15.9 厘米深), 由断头台门隔开, 并包含在隔音盒内。测试仪器的两个隔间被称为 "光边" 和 "暗面"。光明面的墙壁是清晰的, 在试验期间, 灯会打开, 以进一步照亮这个车厢。黑暗的车厢的墙壁被完全覆盖, 以保持一个黑色的条件。
<ol><li>
1天: 训练试验
	<ol>
<li>在测试之前, 用70% 乙醇清洗仪器。</li>
		<li>把动物带到测试室去。</li>
		<li>轻轻地将实验动物放到仪器的光边, 然后关闭并闩锁设备门和隔音盒。</li>
		<li>使用关联的软件, 开始试用。从主屏幕中, 从下拉菜单中选择 "文件", 然后从 "打开会话"。在新窗口中, 确保选中了正确的过程和设备, 然后单击 "关闭"。接下来, 选择 "配置" 然后 "信号"。在新窗口中, 选择正确的设备, 然后单击 "问题"。 注: 该软件将启动灯打开在设备的轻的一面, 并打开断头台门之间的两个车厢。将启动计时器。</li>
		<li>当动物完全进入设备的黑暗面时, 断头台门就会关闭, 而一个不可避免的脚震动 (0.56 mA 为1秒) 将被交付。</li>
		<li>记录在动物进入黑暗的车厢之前已经过去了的时间。</li>
		<li>在动物受到惊吓后, 允许三十年代的恢复时间, 然后从仪器中取出动物。</li>
		<li>把动物放回笼子里。</li>
		<li>在与下一只动物开始试验之前, 用湿毛巾擦拭器具并处理任何粪便颗粒。</li>
		<li>所有的动物都经过训练后, 把它们送回动物设施。</li>
	</ol>
</li>
	<li>
2天: 测试试验
	<ol>
<li>训练试验结束后, 将动物带回测试室24小时。开始对第一个动物的测试试验, 把它放到仪器的光边, 关闭并闩上门和隔音盒。</li>
		<li>使用与前一天相同的软件命令启动测试试验;灯将在仪器的光边打开, 两个车厢之间的断头台门将会打开。将启动计时器。</li>
		<li>当动物进入黑暗的车厢, 断头台门将关闭。记录所经过的时间。 注: 如果动物留在测试仪器的光边上, 试验将在最大300s 后通过软件终止。</li>
		<li>将动物放回自己的笼子里, 清洁仪器 (如步骤3.1.9 中所述), 然后再开始与下一只动物的试验。</li>
	</ol>
</li>
</ol>4. 睡眠分析
<ol><li>无线脑电图/肌电图发射机的外科植入 注: 遥测发射机应在手术前进行消毒和准备。<ol>
<li>使用剃刀刀片, 轻轻地除去小长度的塑料涂层, 露出导线引线。将发射机放在一个50毫升的锥形管中, 充满杀菌液 [例如, Wavecide (2.65% 戊二醛)]。<ol>
<li>将发射机放在消毒液中至少12小时。然后, 在手术之前, 把溶液转移到一个废物容器里。用无菌盐水冲洗发射机。</li>
		</ol>
</li>
		<li>在手术当天, 在将动物置于麻醉室之前称其为体重。诱导3–5% 异氟醚 100% O2。</li>
		<li>成功的麻醉后, 取出动物并小心剃须, 使用电动剃须刀, 头部和颈部的顶部, 以及在身体背部的一小块头发, 只是稍微左和下方的胸腔。</li>
		<li>管理卡洛芬 (5 毫克/千克) 皮下术后镇痛。</li>
		<li>将恒温控制的温水加热垫设置在37摄氏度的动物下, 以保持动物的体温在手术期间。</li>
		<li>在立体定向的框架内, 将动物放在鼻锥上以保持麻醉状态, 并放置耳棒以稳定头部。用 betadine 擦洗和70% 乙醇的交替拭子消毒剃毛皮肤。将 opthalamic 软膏涂抹在眼睛上以润滑它们。</li>
		<li>为确保在第一切口前有足够的麻醉, 请使用脚趾捏试验。</li>
		<li>用一把无菌手术刀, 从眼睛后部切开到颈部后方。颅骨和颈背颈部肌肉应暴露。</li>
		<li>如果需要暴露头骨, 使用棉尖喷头去除任何膜。微型牛头犬钳可以用来保持皮肤远离头骨。定位并标记 bregma 的位置。钻2毛刺孔和插入金属螺丝2.0 毫米前/+ 1.5 毫米侧向和7.0 毫米后/-1.5 毫米侧面相对于 bregma。完成2次革命拧紧螺钉。用一块纱布遮住裸露的头骨。</li>
		<li>使用无菌手术剪刀, 使切口进入体腔, 就在肋骨下面。</li>
		<li>在手术单上, 一定要把要植入的发射机的序列号记录下来。</li>
		<li>将无菌发射机插入体腔内。导线引线应该仍然是在身体外面。</li>
		<li>使用可吸收缝线缝合肌肉壁, 确保所有的线索聚集到一个切口的一端。</li>
		<li>从头上取出纱布, 把切口下面的皮肤抬起来。从头部切口到侧面切口, 在皮肤下运行一双长的不育止血剂。保持对皮肤的压力, 以减少任何伤害。<ol>
<li>一旦止血剂在身体切口附近可见, 夹住任何可能用手术剪刀覆盖的膜, 并抓住四发射器引线与止血剂。慢慢地抽出止血剂, 使电线躺在皮肤下, 跑到头骨上。</li>
		</ol>
</li>
		<li>指定哪些2引线将连接到头骨。使用镊子, 用一只手抓住电线的一端, 并在塑料护套的末端下方用另一只。拉上电线, 直到单独的循环可以做出来。</li>
		<li>将暴露的导线紧裹在金属螺钉3–4x 的周围。一旦安全, 拧紧螺丝, 以防止电线解开和剪辑任何额外的电线。用第二个引线和螺钉重复此操作。</li>
		<li>拉两个脑电图线索从体腔, 使他们坐在紧贴头骨。</li>
		<li>使用牙科水泥覆盖螺钉和皮质引线, 在暴露的头骨上创建一个上限。确保没有牙科水泥粘在皮肤或肌肉, 没有锋利的边缘被创造。允许牙科水泥干燥。 注意: 瓶盖应该足够小, 可以在上面缝合皮肤。</li>
		<li>对于肌电图的引线, 抓住的线和塑料鞘的末端, 如步骤4.1.15 所述。伸展电线, 直到各个回路清晰可见。</li>
		<li>在2–3毫米的右侧, 在颈部肌肉下运行22克针, 然后才允许它出现。使用美容整形缝合, 放置一个单一的, 松散缝合周围的嵌入针。</li>
		<li>将肌电信号线插入针中, 拔出针, 使肌电信号在肌肉下线。拧紧缝合线, 以确保导线保持到位, 并夹住可能从肌肉中出现的多余导线。</li>
		<li>重复步骤4.1.20 和4.1.21 的第二个肌电信号引线, 约1毫米向下从第一个铅。</li>
		<li>拉两个肌电信号的线索从体腔, 使他们坐在舒适的肌肉。</li>
		<li>使用美容整形针闭合头部和颈部切口。把所有多余的电线塞在身体的皮肤下, 用伤口夹来闭合身体切口。</li>
		<li>在两个切口部位应用利多卡因软膏, 协助动物术后疼痛。</li>
		<li>把动物送回笼子, 让笼子坐在暖气垫上, 直到动物从麻醉中恢复。 注意: 动物应在开始录音前7–10天恢复。</li>
	</ol></li>
	<li>脑电图/肌电信号记录 注: 在本议定书中, 我们在 ZT 0 中管理生理盐水或犬尿氨, 然后记录动物的睡眠-唤醒模式为24小时。<ol>
<li>在指定的房间里完成所有的睡眠记录, 在这段时间内动物不会被打断。</li>
		<li>将动物的家庭笼子 (轴承植入的脑电图/肌电信号发射机) 放在接收器单元上。用磁铁打开手术植入的发射机。 注: 当发射器被激活时, 接收装置上应出现一盏灯。</li>
		<li>使用相关软件设置脑电图/肌电信号数据的记录。<ol>
<li>选择 "实验", 然后从下拉菜单中 "创建"。命名实验, 然后保存它。接下来, 选择 "硬件", 然后 "编辑 MX2 配置"。</li>
			<li>在新窗口中, 选择实验中使用的所有发射器, 并指示每个接收板的配对。准备就绪后, 按 "开始所有连续" 启动录制。</li>
		</ol>
</li>
		<li>在实验结束时, 选择 "停止所有连续" 在相关的软件和关闭发射机使用磁铁。</li>
		<li>弄死动物由 CO2窒息然后心脏穿刺。通过用手术剪刀在头部顶部重新打开切口, 并从牙科水泥和颈部肌肉中切开无引线, 小心地取出发射机。</li>
		<li>接着, 打开体腔, 找到发射器, 慢慢地将其从体内移除。</li>
	</ol></li>
	<li>脑电图/肌电信号分析
	<ol>
<li>使用关联的软件分析睡眠唤醒数据。打开程序并选择 "添加新位置"。在新窗口中, 选择要导入到评分软件中的数据。 注意: 这将在软件内创建一个新文件, 其中包含实验期间收集的所有原始波形。</li>
		<li>手动为所需的实验评分警戒状态。评分后, 分析的参数, 如总持续时间, 回合次数, 和平均回合期间的各种警戒状态 [快速眼动 (rem), 非 REM (NREM), 和唤醒]。 注意: 分析可以在整个记录或缩短到特定的时间点进行。在此, 对 zt 0 至 zt 2 之间的记录期进行了分析。</li>
	</ol></li>
</ol>

<ol>
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作者没有什么可透露的。

为了可靠地评估周围犬尿氨管理后的脑 KYNA, 必须结合和解释生化和功能性实验。在这里, 我们提出了一个详细的协议, 允许新用户建立有效的方法来测量血浆犬尿氨和脑 KYNA 的大鼠。血浆中犬尿氨的测定证实了准确的注射和代谢产物的测定 KYNA 证实了大脑中的从头合成。所描述的荧光液相色谱法有以下几个优点: 从柴田18到 derivatize KYNA, 在锌2 +的存在下, 包括在 nanom...

金伯利进程负责降解近95% 的必需氨基酸色氨酸1。在哺乳动物的大脑中, 犬尿氨被带入星形胶质细胞代谢成 neuroactive 小分子 KYNA 主要由酶犬尿氨转氨酶 (KAT) II2。KYNA 作为拮抗剂在 NMDA 和α7nACh 受体在大脑2,3,4, 也靶向信号受体包括芳烃受体 (AHR) 和 G 蛋白耦合受体 35 (GPR35)5 ,6。在实验动物中, 大脑 KYNA 的海拔在行为分析的2、7、8、9、10中被证明会损害他们的认知能力。.一个新的假说表明, KYNA 在调节认知功能方面起着不可或缺的作用, 它影响睡眠唤醒行为11, 从而进一步支持星形胶质细胞衍生分子在调节睡眠神经生物学方面的作用, 并认知12。
临床上, KYNA 的海拔在脑脊液和验尸后的脑组织中发现, 精神分裂症13,14,15,16, 一个衰弱的精神紊乱以认知障碍为特征的。精神分裂症患者也经常被睡眠紊乱所困扰, 这可能使病情恶化17。了解 KP 代谢和 KYNA 在调节睡眠与认知之间的关系, 特别是学习和记忆之间的作用, 可能会导致新的治疗精神分裂症和其他一些不良结局的治疗方法的发展。精神疾病。
一个可靠和一致的方法来测量金伯利的代谢物, 是重要的, 以确保从各个机构的研究可以融入到科学的理解, kp 生物学。目前, 我们描述了用高效液相色谱法测定大鼠脑血浆和 KYNA 中犬尿氨的方法。本议定书在锌2 +的存在下使用荧光检测, 最初是由柴田18和最近适应和优化到 derivatize 以500毫米醋酸锌作为后柱试剂, 从而使在脑内内源性、nanomolar 量 KYNA 的检测11。
为刺激本议定书所述的 KYNA 内源性生产, 直接 bioprecursor 犬尿氨在大鼠体内注入腹腔 (ip)。结合生化评估来确定 KYNA 生产的程度, 犬尿氨挑战对海马依赖性记忆 (被动回避范式) 和睡眠唤醒体系 (EEG 和肌电信号) 的影响也是调查11。这些技术的结合允许研究犬尿氨挑战在大鼠体内的生物化学和功能影响。

<table>
	<tbody>
		<tr>
			<td>Wistar rats</td>
			<td>Charles River Laboratories</td>
			<td></td>
			<td>adult male, 250-350 g</td>
		</tr>
		<tr>
			<td>L-kynurenine sulfate</td>
			<td>Sai Advantium</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ReproSil-Pur C18 column (4 x 150 mm)</td>
			<td>Dr. Maisch GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EZ Clips</td>
			<td>Stoelting Co.</td>
			<td>59022</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounting materials screws</td>
			<td>PlasticsOne</td>
			<td>00-96 X 1/16</td>
			<td></td>
		</tr>
		<tr>
			<td>Nonabsorbable Sutures</td>
			<td>MedRep Express</td>
			<td>699B</td>
			<td>CP Medical Monomid Black Nylon Sutures, 4-0, P-3, 18&#34;, BOX of 12</td>
		</tr>
		<tr>
			<td>Absorbable Sutures</td>
			<td>Ethicon</td>
			<td>J310H</td>
			<td>4-0 Coated Vicryl Violet 1X27&#39;&#39; SH-1</td>
		</tr>
		<tr>
			<td>Dental Cement</td>
			<td>Stoelting Co.</td>
			<td>51458</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill Bit</td>
			<td>Stoelting Co.</td>
			<td>514551</td>
			<td>0.45 mm</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Alliance HPLC system</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>E2695 separation module</td>
			<td>Waters</td>
			<td>176269503</td>
			<td></td>
		</tr>
		<tr>
			<td>2475 fluorescence detector</td>
			<td>Waters</td>
			<td>186247500</td>
			<td></td>
		</tr>
		<tr>
			<td>post-column reagent manager</td>
			<td>Waters</td>
			<td>725000556</td>
			<td></td>
		</tr>
		<tr>
			<td>Lenovo computer</td>
			<td>Waters</td>
			<td>668000249</td>
			<td></td>
		</tr>
		<tr>
			<td>Empower software</td>
			<td>Waters</td>
			<td>176706100</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Passive avoidance box for rat</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Extra tall MDF sound attenuating cubicle</td>
			<td>MedAssociates</td>
			<td>ENV-018MD</td>
			<td>Interior: 22&#34;W x 22&#34;H x 16&#34;D</td>
		</tr>
		<tr>
			<td>Center channel modulator shuttle box chamber</td>
			<td>MedAssociates</td>
			<td>ENV-010MC</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel grid floor for rat</td>
			<td>MedAssociates</td>
			<td>ENV-010MB-GF</td>
			<td></td>
		</tr>
		<tr>
			<td>Auto guillotine door</td>
			<td>MedAssociates</td>
			<td>ENV-010B-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Quick disconnect shuttle grid floor harness for rat</td>
			<td>MedAssociates</td>
			<td>ENV-010MB-QD</td>
			<td></td>
		</tr>
		<tr>
			<td>Stimulus light, 1&#34; white lens, mounted on modular panel</td>
			<td>MedAssociates</td>
			<td>ENV-221M</td>
			<td></td>
		</tr>
		<tr>
			<td>Sonalert module with volume control for rat chamber</td>
			<td>MedAssociates</td>
			<td>ENV-223AM</td>
			<td></td>
		</tr>
		<tr>
			<td>SmartCtrl 8 input/16 output package</td>
			<td>MedAssociates</td>
			<td>DIG-716P2</td>
			<td></td>
		</tr>
		<tr>
			<td>8 Channel IR control for shuttle boxes</td>
			<td>MedAssociates</td>
			<td>ENV-253C</td>
			<td></td>
		</tr>
		<tr>
			<td>Infrared source and dectector array strips</td>
			<td>MedAssociates</td>
			<td>ENV-256</td>
			<td></td>
		</tr>
		<tr>
			<td>Tabletop interface cabinet, 120 V 60 Hz</td>
			<td>MedAssociates</td>
			<td>SG-6080C</td>
			<td></td>
		</tr>
		<tr>
			<td>Dual range constant current aversive stimulation module</td>
			<td>MedAssociates</td>
			<td>ENV-410B</td>
			<td></td>
		</tr>
		<tr>
			<td>Solid state grid floor scrambler module</td>
			<td>MedAssociates</td>
			<td>ENV-412</td>
			<td></td>
		</tr>
		<tr>
			<td>Dual A/B shock control module</td>
			<td>MedAssociates</td>
			<td>ENV-415</td>
			<td></td>
		</tr>
		<tr>
			<td>2&#39; 3-Pin mini-molex extension</td>
			<td>MedAssociates</td>
			<td>SG-216A-2</td>
			<td></td>
		</tr>
		<tr>
			<td>10&#39; Shock output cable, DB-9 M/F</td>
			<td>MedAssociates</td>
			<td>SG-219G-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Shuttle shock control cable 15&#39;, 6</td>
			<td>MedAssociates</td>
			<td>SG-219SA</td>
			<td></td>
		</tr>
		<tr>
			<td>Small tabletop cabinet and power supply, 120 V 60 Hz</td>
			<td>MedAssociates</td>
			<td>SG-6080D</td>
			<td></td>
		</tr>
		<tr>
			<td>PCI interface package</td>
			<td>MedAssociates</td>
			<td>DIG-700P2-R2</td>
			<td></td>
		</tr>
		<tr>
			<td>Shuttle box avoidance utility package</td>
			<td>MedAssociates</td>
			<td>SOF-700RA-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
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			<td>Sleep-Wake Monitoring Equipment</td>
			<td></td>
			<td></td>
			<td></td>
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			<td>Ponehmah software</td>
			<td>Data Sciences International (DSI)</td>
			<td>PNP-P3P-610</td>
			<td></td>
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			<td>MX2 8 Source Acquisition interface</td>
			<td>Data Sciences International (DSI)</td>
			<td>PNM-P3P-MX204</td>
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			<td>Dell computer, Optiplex 7020, Windows 7, 64 bit</td>
			<td>Data Sciences International (DSI)</td>
			<td>271-0112-013</td>
			<td></td>
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			<td>Dell 19&#34; computer monitor</td>
			<td>Data Sciences International (DSI)</td>
			<td>271-0113-001</td>
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			<td>Receivers for plastic cages, 8x</td>
			<td>Data Sciences International (DSI)</td>
			<td>272-6001-001</td>
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			<td>Cisco RV130 VPN router</td>
			<td>Data Sciences International (DSI)</td>
			<td>RV130</td>
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			<td>Matrix 2.0</td>
			<td>Data Sciences International (DSI)</td>
			<td>271-0119-001</td>
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			<td>Network switch</td>
			<td>Data Sciences International (DSI)</td>
			<td>SG200-08P</td>
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			<td>Neuroscore software</td>
			<td>Data Sciences International (DSI)</td>
			<td>271-0171-CFG</td>
			<td></td>
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		<tr>
			<td>Two biopotential channels transmitter, model TL11M2-F40-EET</td>
			<td>Data Sciences International (DSI)</td>
			<td>270-0134-001</td>
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a high-performance liquid chromatography measurement of kynurenine and kynurenic acid: relating biochemistry to cognition and sleep in rats

色氨酸降解的犬尿氨通路 (KP) 与精神疾病有牵连。具体地说, 星形胶质细胞衍生代谢物 kynurenic 酸 (KYNA) 是n-甲基-d-天门冬氨酸 (NMDA) 和α7烟碱乙酰胆碱 (α7nACh) 受体的拮抗剂, 与健康和疾病的认知过程有牵连。随着精神分裂症患者大脑中 KYNA 水平的升高, 谷氨酸和胆碱能受体的故障被认为是与认知功能障碍相关的因果关系, 这是疾病精神病理学的核心领域。KYNA 可能在精神分裂症的个体中扮演 pathophysiologically 重要的角色。通过直接 bioprecursor 犬尿氨对动物进行治疗, 可以提高啮齿动物脑内内源性 KYNA, 大鼠的临床前研究表明, KYNA 的急性海拔可能会影响他们的学习和记忆过程。本协议详细描述了这一实验方法, 并结合 a) 对血液犬尿氨水平和脑 KYNA 形成 (使用高效液相色谱) 的生化分析, b) 行为测试, 以探讨与海马相关的上下文记忆 (被动回避范式) 和 c) 对睡眠唤醒行为的评估 [将脑电图 (EEG) 和肌电图 (肌电信号) 信号结合在一起的遥测记录] 在大鼠中。结合起来, 研究了高 KYNA、睡眠和认知之间的关系, 该协议详细描述了一种实验方法来了解大鼠体内犬尿氨海拔和 KYNA 形成的功能结果。通过这一协议的变化得到的结果将测试的假说, KP 和 KYNA 在调节睡眠和认知的健康和疾病状态的关键作用。

为了验证采用腹腔犬尿氨注射液作为提高脑 KYNA 的方法, 对组织进行了 HPLC 分析。标准曲线 (图 1) 是使用相关软件构造的, 并允许对组织样本进行量化。犬尿氨和 KYNA 的代表性图谱如图 2所示。犬尿氨在6分钟的保留时间内观察到, KYNA 的保留时间为11分钟。为了观察 KP 动力学, 在本协议中管理了100毫克/千克的犬尿氨, 并在注射后立...

Watch this Scientific Journal Video about A High-performance Liquid Chromatography Measurement of Kynurenine and Kynurenic Acid: Relating Biochemistry to Cognition and Sleep in Rats at JoVE.com

A High-performance Liquid Chromatography Measurement of Kynurenine and Kynurenic Acid: Relating Biochemistry to Cognition and Sleep in Rats

高效液相色谱法测定犬尿氨和 Kynurenic 酸: 对大鼠认知和睡眠的相关生物化学研究

犬尿氨通路 (KP) neuroactive 代谢物的改变与精神疾病有牵连。调查啮齿类动物体内改变的犬尿氨通路代谢的功能性结果可能有助于阐明新的治疗方法。目前的协议结合了生物化学和行为学的方法来研究急性犬尿氨挑战对大鼠的影响。

The kynurenine pathway (KP) of tryptophan degradation has been implicated in psychiatric disorders. Specifically, the astrocyte-derived metabolite ...

a-high-performance-liquid-chromatography-measurement-kynurenine

Research

JoVE Journal

Behavior

10.4K 次观看.  University of Maryland School of Medicine. 犬尿氨通路 (KP) neuroactive 代谢物的改变与精神疾病有牵连。调查啮齿类动物体内改变的犬尿氨通路代谢的功能性结果可能有助于阐明新的治疗方法。目前的协议结合了生物化学和行为学的方法来研究急性犬尿氨挑战对大鼠的影响。

视频: A High-performance Liquid Chromatography Measurement of Kynurenine and Kynurenic Acid: Relating Biochemistry to Cognition and Sleep in Rats

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

一致性的任务作为一种手段来获取目标行为的措施中的橡胶手错觉范式Crossmodal

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

科研

行为学

Eye Tracking, Cortisol, and a Sleep vs. Wake Consolidation Delay: Combining Methods to Uncover an Interactive Effect of Sleep and Cortisol on Memory

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these two factors may interact to influence consolidation. Here we present a protocol to examine the interactive influence of cortisol and sleep on memory consolidation, by combining three methods: eye tracking, salivary cortisol analysis, and behavioral memory testing across sleep and wake delays. To assess resting cortisol levels, participants gave a saliva sample before viewing negative and neutral objects within scenes. To measure overt attention, participants&#8217; eye gaze was tracked during encoding. To manipulate whether sleep occurred during the consolidation window, participants either encoded scenes in the evening, slept overnight, and took a recognition test the next morning, or encoded scenes in the morning and remained awake during a comparably long retention interval. Additional control groups were tested after a 20&#160;min&#160;delay in the morning or evening, to control for time-of-day effects. Together, results showed that there is a direct relation between resting cortisol at encoding and subsequent memory, only following a period of sleep. Through eye tracking, it was further determined that for negative stimuli, this beneficial effect of cortisol on subsequent memory may be due to cortisol strengthening the relation between where participants look during encoding and what they are later able to remember. Overall, results obtained by a combination of these methods uncovered an interactive effect of sleep and cortisol on memory consolidation.

We present a protocol used to discover an interactive effect between sleep and cortisol on memory consolidation, particularly for negative arousing images. Specifically, the experimental design utilizes eye tracking, salivary cortisol analysis, and behavioral memory testing &#8211; methods that can be used with both healthy and clinical participants.

眼睛跟踪，皮质醇和睡眠与唤醒延迟合并:合并方法来揭开睡眠和皮质醇的交互式影响记忆

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these ...

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

揭开节拍耳聋:检测节律紊乱与同步手指敲击和感知定时任务

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

一个过程来观察大鼠巴甫洛夫空调酒精寻求行为的背景下引发的重建

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

Modulating Cognition Using Transcranial Direct Current Stimulation of the Cerebellum

Numerous studies have emerged recently that demonstrate the possibility of modulating, and in some cases enhancing, cognitive processes by exciting brain regions involved in working memory and attention using transcranial electrical brain stimulation. Some researchers now believe the cerebellum supports cognition, possibly via a remote neuromodulatory effect on the prefrontal cortex. This paper describes a procedure for investigating a role for the cerebellum in cognition using transcranial direct current stimulation (tDCS), and a selection of information-processing tasks of varying task difficulty, which have previously been shown to involve working memory, attention and cerebellar functioning. One task is called the Paced Auditory Serial Addition Task (PASAT) and the other a novel variant of this task called the Paced Auditory Serial Subtraction Task (PASST). A verb generation task and its two controls (noun and verb reading) were also investigated. All five tasks were performed by three separate groups of participants, before and after the modulation of cortico-cerebellar connectivity using anodal, cathodal or sham tDCS over the right cerebellar cortex. The procedure demonstrates how performance (accuracy, verbal response latency and variability) could be selectively improved after cathodal stimulation, but only during tasks that the participants rated as difficult, and not easy. Performance was unchanged by anodal or sham stimulation. These findings demonstrate a role for the cerebellum in cognition, whereby activity in the left prefrontal cortex is likely dis-inhibited by cathodal tDCS over the right cerebellar cortex. Transcranial brain stimulation is growing in popularity in various labs and clinics. However, the after-effects of tDCS are inconsistent between individuals and not always polarity-specific, and may even be task- or load-specific, all of which requires further study. Future efforts might also be guided towards neuro-enhancement in cerebellar patients presenting with cognitive impairment once a better understanding of brain stimulation mechanisms has emerged.

Transcranial direct current stimulation (tDCS) over the cerebellum exerts a remote effect on the prefrontal cortex, which can modulate cognition and performance. This was demonstrated using two information-processing tasks of varying complexity, whereby only cathodal tDCS improved performance when the task was difficult, but not easy.

调节认知使用小脑经颅直流电刺激

Numerous studies have emerged recently that demonstrate the possibility of modulating, and in some cases enhancing, cognitive processes by exciting brain ...

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 ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little research to reveal the possible mechanisms behind these effects. To investigate the effect of acupuncture on epilepsy and sleep, several issues need to be addressed. The first is to identify the acupoints, which correspond between humans, rats, and mice. Furthermore, the depth of insertion of the acupuncture needle, the degree of needle twist in manual needle acupuncture, and the stimulation parameters for electroacupuncture (EA) need to be determined. To evaluate the effects of acupuncture on epilepsy and sleep, a feasible model of epilepsy in rodents is required. We administer pilocarpine into the left central nucleus of the amygdala (CeA) to simulate focal temporal lobe epilepsy (TLE) in rats. Intraperitoneal (IP) injection of pilocarpine induces generalized epilepsy and status epilepticus (SE) in rats. Five IP injections of pentylenetetrazol (PTZ) with a one-day interval between each injection successfully induces spontaneous generalized epilepsy in mice. Recordings of electrocorticograms (ECoGs), electromyograms (EMGs), brain temperature, and locomotor activity are used for sleep analysis in rats, while ECoGs, EMGs, and locomotor activity are employed for sleep analysis in mice. ECoG electrodes are implanted into the frontal, parietal, and contralateral occipital cortices, and a thermistor is implanted above the cerebral cortex by stereotactic surgery. EMG electrodes are implanted into the neck muscles, and an infrared detector determines locomotor activity. The criteria for categorizing vigilance stages, including wakefulness, rapid eye movement (REM) sleep, and non-REM (NREM) sleep are based on information from ECoGs, EMGs, brain temperature, and locomotor activity. Detailed classification criteria are stated in the text.

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

癫痫Electrocorticograms（ECoGs）的操作，并睡在针刺大鼠和小鼠

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

A New Method for Inducing a Depression-Like Behavior in Rats

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is no existing format for studying the mechanism of action, prevention, containment, and treatment of contagious depression. The purpose of this study, therefore, was to establish the first animal model of contagious depression.
Healthy rats can contract depressive behaviors if exposed to depressed rats. Depression is induced in rats by subjecting them to several manipulations of chronic unpredictable stress (CUS) over 5 weeks, as described in the protocol. A successful sucrose preference test confirmed the development of depression in the rats. The CUS-exposed rats were then caged with na&#239;ve rats from the contagion group (1 na&#239;ve rat/2 depressed rats in a cage) for an additional 5 weeks. 30 social groups were created from the combination of CUS-exposed rats and na&#239;ve rats.
This proposed depression-contagion protocol in animals consists mainly of cohabiting CUS-exposed and healthy rats for 5 weeks. To ensure that this method works, a series of tests are carried out - first, the sucrose preference test upon inducing depression to rats, then, the sucrose preference test, alongside the open field and forced-swim tests at the end of the cohabitation period. Throughout the experiment, rats are given tags and are always returned to their cages after each test.
A few limitations to this method are the weak differences recorded between the experimental and control groups in the sucrose preference test and the irreversible traumatic outcome of the forced swim test. These may be worth considering for suitability before any future application of the protocol. Nonetheless, following the experiment, na&#239;ve rats developed contagion depression after 5 weeks of sharing the same cage with the CUS-exposed rats.

This protocol describes a new model by which healthy rats could contract depression over a given time periodthrough contagion by exposure to chronic unpredictable stressed (CUS) rats.

一种诱导大鼠抑郁症状行为的新方法

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is ...

A Conditioned Place Preference Protocol for Measuring Incubation of Craving in Rats

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed incubation of drug craving. Here, we describe a morphine conditioned place preference (CPP) protocol for measuring the incubation of craving in rats. In this protocol, a CPP paradigm mainly employing somatosensory cues is used to establish a long-term reward memory of morphine. A three-chamber CPP box that differs in the texture of the chamber floor is constructed. First, the animals are tested for their baseline preference to the two side chambers for three consecutive days. Then, they are injected intraperitoneally with morphine/saline and put into their non-preferred/preferred chamber for 45 min. After 6 days of conditioning, their preference to the side chambers is tested for 15 min at different time points after the last conditioning session. With this paradigm, the reward memory of morphine could last for at least 18 days. To test whether the above-mentioned protocol can model increased craving, the number of entrances into the two side chambers are counted during the abstinence period. The results show that the entrances increased, suggesting that the CPP paradigm could mimic the incubation of craving. Future studies can employ this model to study neural mechanisms underlying long-term memory and incubation of craving.

Here, a morphine conditioned place preference (CPP) protocol is described to measure incubation of craving in rats.

一种用于大鼠渴望培养的条件场所偏好协议

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed ...

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and dysfunctions in mood and cognition are common and often shared symptoms in neurodegenerative diseases. Shown here is a behavioral test battery for motor, mood, and cognition, which can be repeatedly tested in a longitudinal study. This battery assesses the overall behavioral phenotype in mice by examining each domain of behavior with at least two independent well-accepted tests (i.e., open-field test and rotarod test for motor function, social interaction test, elevated plus maze test, and forced swim test for emotional function, and Morris water maze test and novel object recognition test for cognitive function). Therefore, this sensitive and comprehensive test battery is a powerful tool for the study of behavioral alternation in neurodegeneration.

A comprehensive behavioral test battery of motor skills, mood&#8212;including social interaction, depression, and anxiety&#8212;and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.