这项工作得到了国家卫生研究院/国家神经紊乱和中风补助金 1R01NS096225-01A1, 美国心脏协会授予 AHA-13SDG1395001413, AHA-17GRNT33660336, AHA-17POST33660174,路易斯安那州立大学资助研究委员会, 马尔科姆费斯特心血管研究奖学金, 和伊夫林 F. 麦克奈特脑研究所。

所有实验程序都是按照国家卫生研究院的指导方针进行的, 并得到机构动物保育和使用委员会 (路易斯安那州立会健康科学中心-什里夫波特) 的批准, 用于雄性大大鼠的使用 (300-350 克, 9-10 周大)。大鼠在 ACA 手术前一夜禁食。
1. 迷宫设备的设计与设置
注: 在执事和罗的2006模型31上的 T 迷宫设计基础。
<ol><li>利用之32设计迷宫的3D 结构。要创建 T 型迷宫的3D 结构, 请构造一个外部长度为 200 mm、宽度为 165 mm 和高度 148 mm 的起始臂, 以适应3D 打印机的打印尺寸。使用壁厚5.5 毫米, 地板厚度为8毫米。</li>
	<li>使用3D 打印机打印迷宫 (请参阅材料表)32。如果实验室中没有3D 打印机, 请使用其他材料, 如木材、中密度纤维板或塑料 (即聚氯乙烯), 可以从家居改善商店购买。<ol>
<li>由于高度限制在打印区域, 构造迷宫的墙壁在二个分开的3D 印刷品并且一起在迷宫汇编 (即, 第二个墙壁高度被增加到迷宫部分增加高度由140毫米, 为总墙壁高度280毫米)。每个独立的3D 打印基座包含一个 "T" 形锁定机构, 其中一个部分连接到下一个。</li>
		<li>在起点胳膊的连接点与目标胳膊, 创造一个165毫米宽部分加入开始胳膊的宽度与目标胳膊的那。用类似的设计方法作为启动臂来构造目标臂;但是, 根据执事和罗的设计, 将手臂的宽度减小到100毫米。</li>
		<li>有关 T 形迷宫的详细示意图/维度, 请参见图 1 。</li>
		<li>在开始臂和目标臂的接合处包括一个中心分区到设计中。从 T 迷宫和200毫米的后壁扩展这个分区到开始胳膊划分目标胳膊。此分区还扩展了迷宫的高度 (图 1)。</li>
	</ol>
</li>
</ol>2. 窒息心脏骤停 (ACA)
<ol><li>高压釜手术工具 (121 ° c 为15分钟) 在开始手术之前。用70% 的乙醇消毒手术表15分钟, 在手术现场刮毛动物毛发。将控告溶液应用于皮肤表面进行外科手术。</li>
	<li>麻醉
	<ol>
<li>麻醉大鼠与4% 异氟醚和30:70 混合物 o2和 N2o (300 毫升/分钟 o 2 和700毫升/分钟 n 2O) 通过面具。</li>
		<li>给大鼠气管插管进行机械通气 (插管后, 大鼠与呼吸机连接)。</li>
		<li>通过将异氟醚降低4% 到 2%, 用30:70 混合物 o2和 N2来维持麻醉, 使用捏响应法确定麻醉深度。</li>
		<li>将药膏涂抹在眼部, 防止麻醉时干燥。Regulae 的体温由啮齿动物加热垫和肛门探针作为温度参考。</li>
	</ol></li>
	<li>气管插管
	<ol>
<li>把老鼠放在感应室里。麻醉4% 异氟醚和30:70 混合 o2和 N2o 的大鼠。</li>
		<li>从感应室取出老鼠将麻醉动物置于仰卧位, 将鼠脸朝向麻醉面罩。</li>
		<li>用左手拇指和食指轻轻地向动物的左边或右边移动舌头。</li>
		<li>滑行一个14口径的挠性静脉导管 (49 毫米长) 超过17口径钝尖移针 (93 毫米长, 在针的尖端10度角)。将17口径钝尖移针插入气管。</li>
		<li>从气管轻轻拉出17口径的移针。将14针导管中枢连接到呼吸器。调整呼吸机冲程容积为 0.67 mL/100 g, 呼吸率60呼吸/分钟。</li>
		<li>在整个过程中, 将头部和体温保持在37° c, 由啮齿动物加热垫以肛门探针作为温度参考。</li>
	</ol></li>
	<li>股动脉和静脉插管
	<ol>
<li>在腹股沟附近 (两侧) 剃毛, 并将控告到皮肤表面进行手术。</li>
		<li>把老鼠放在仰卧位。用手术剪刀在腹股沟区做一个切口 (10 毫米)。</li>
		<li>用钝尖钳分离结缔组织, 直至腹股沟韧带外露。用止血来抓腹股沟韧带。股动脉和静脉位于腹股沟韧带下方。</li>
		<li>使用钝尖端钳分离结缔组织, 直到股动脉和静脉暴露。</li>
		<li>用细尖钳将股神经沿股动脉轻轻分离。仔细分离股动脉和静脉作为一个单位通过细尖钳。</li>
		<li>用细尖钳将股动脉与静脉分离。</li>
		<li>在静脉下放置2件5-0 丝缝合线 (一条朝向腿部, 另一个朝向身体)。</li>
		<li>在身体旁边绑一个松散的结。使用止血来保持和拉缝合尽可能接近身体的两侧。</li>
		<li>在腿旁边系一个松散的结。通过止血将缝线拉向腿部, 使静脉充满血液。</li>
		<li>用微解剖剪刀 (45 °角) 在静脉 (大约0.1 毫米) 上做一个小切口。用消毒纱布浸泡任何血液。</li>
		<li>附上一个钝尖端针头注射器 (充满盐水与20ü/毫升肝素) 到一个 PE-50 导管。用生理盐水填充 PE-50 导管, 20 U/毫升肝素。切割 PE-50 导管与解剖剪刀在45°的角度, 以创建一个点或尖锐的结束。用钝尖钳夹住 PE-50 导管的末端。轻轻地将 PE-50 导管插入股静脉。</li>
		<li>在导管完全插入后, 慢慢地管理0.1 毫升的肝素/生理盐水, 以确保没有泄漏。扎牢固缝合结 (single-knot) 以稳定 PE-50 导管。保持 PE-50 导管持续静脉注射 (IV) 各种药物的注射。</li>
		<li>使用1毫升注射器连接23口存根适配器管理维库溴化 (0.67 毫克/千克, 管理每10分钟) 通过股静脉, 以固定在整个过程中的老鼠。</li>
		<li>在动脉下放置2件5-0 丝缝线 (一条朝向腿部, 另一根朝向身体)。</li>
		<li>在腿旁边系一个松散的结。使用止血来保持和拉缝合尽可能对腿部。</li>
		<li>在身体旁边绑一个松散的结。通过止血保持和拉动缝合体, 使动脉充满血液。</li>
		<li>用微解剖剪刀 (45 °角) 在动脉 (大约0.1 毫米) 上做一个小切口。</li>
		<li>附上一个钝尖端针头注射器 (充满盐水与20ü/毫升肝素) 到一个 PE-50 导管。用生理盐水填充 PE-50 导管, 20 U/毫升肝素。切割 PE-50 导管与解剖剪刀在45度角, 创造一个点或尖锐的结束。用钝尖钳夹住 PE-50 导管的末端。用钝尖钳夹住 PE-50 导管的末端。轻轻地将 PE-50 导管插入股动脉。</li>
		<li>在导管完全插入后, 慢慢地抽回注射器, 确保导管功能正常。扎牢固缝合结 (single-knot) 以稳定 PE-50 导管。保持 PE-50 导管连续记录动脉压力和血气。</li>
	</ol></li>
	<li>窒息心脏骤停 (ACA) 程序
	<ol>
<li>调整行程音量, o2或 N2o 级别, 根据需要调节生理参数 (如 pO2、不正常的2、血压和 pH 值)。使用这些参数的正常生理范围: pO2: 100 柱, 肺混浊,2: 35-40 柱, 血压: 100 柱, pH: 7.4。</li>
		<li>使用1毫升注射器连接23口存根适配器管理维库溴化铵 (0.67 毫克/千克, 静脉注射) 经股静脉, 并等待2分钟. 在执行 ACA 之前, 请确保血压在100柱左右。</li>
		<li>通过从呼吸机上拔下气管插管 (14 针导管中枢), 诱发窒息 (6 分钟)。通过1毫升注射器进一步阻断气管插管, 以确保完全窒息。 注意: 6 分钟的窒息时间被定义为呼吸机断开和复苏开始之间的时期。完整的心跳骤停被定义为平均动脉压低于10柱。</li>
		<li>在最后一分钟的呼吸暂停, 调整呼吸频率的呼吸机80呼吸/分钟, 并增加 O2到2升/分钟与 0% N2O。此操作将会将其余的异氟醚或 N2O 在呼吸机中残留。</li>
		<li>最小后呼吸暂停, 从气管插管中取出1毫升注射器。将气管插管重新连接到呼吸机。</li>
		<li>使用1毫升注射器连接23口的存根适配器管理肾上腺素 (0.005 毫克/千克, 静脉注射) 通过股静脉和管理手动胸部按压的拇指, 指数, 和中指在动物的胸部在 x 和轻循环运动z-axis (200/分钟) 直到自发循环返回 (平均动脉压力≥50柱)33,34,35。</li>
		<li>使用另1毫升注射器与23口的存根适配器连接, 以管理碳酸氢钠 (1 当量/千克, 静脉注射) 经股静脉后, 立即返回自发循环 (50 柱或更高)33,34,35以缓解呼吸道酸中毒。</li>
		<li>在复苏后10分钟测量血气, 以确定酸碱状态 (ACA 后的 pH 值应在7.35 到7.40 左右)</li>
		<li>用止血钳夹住股动脉和静脉。缓慢, 轻轻地用钝尖钳清除动脉和静脉导管。结扎股动脉/静脉5-0 丝缝合, 防止出血。用3-0 丝缝线缝合手术部位的皮肤。使用间断缝合技术, 以尽量减少伤口重新打开的机会。</li>
		<li>等待, 直到鼠呼吸本身 (通常是30分钟到60分钟后复苏), 断开从呼吸机的老鼠, 并轻轻地取出气管插管。</li>
		<li>将老鼠放在婴儿孵化器 (27 ° c, 50% 湿度) 过夜。将软化的食物 (通过浸泡在水中) 和水放入婴儿孵化器过夜。</li>
		<li>把老鼠转移到单独的笼子里, 然后把老鼠带到动物设施里, 用普通的食物和水。T 迷宫测试开始3天后 ACA。</li>
	</ol></li>
</ol>3. T 型迷宫
<ol><li>
动物制剂
	<ol>
<li>手术前一天 (假或 ACA), 处理每只老鼠5分钟。在处理 (图 2) 时, 切勿将老鼠从笼中提升 (480 mm x 250 mm x 200 mm, 塑料透明保持架)。</li>
		<li>在处理老鼠之后, 用一只手轻轻地把老鼠从尾巴上捡起来, 另一只手支撑着它的腿。让他们从手跳到笼子 (100 毫米高度) 5 次。把每只老鼠分成单独的笼子, 这样它们就不会在食物和/或战斗中占主导地位。</li>
		<li>假或 ACA 手术后三天 (图 2), 在第一次运行开始之前, 将笼子里的老鼠转移到一个安静、黑暗的房间里。只打开一个低功率的台灯, 并把它放在测试室的角落, 以保持最低照明。让老鼠适应黑暗10分钟。</li>
		<li>下午进行所有实验, 以避免日变化对大鼠的影响。不要建议操作者接受假或 ACA 手术。</li>
	</ol>
</li>
	<li>
自发交替
	<ol>
<li>铺一层薄薄的被褥 (〜10毫米厚) 覆盖整个地板的迷宫。然后将鼠放在开始臂 ("T" 的底部), 这是每一个运行的出发点, 并允许每只老鼠3分钟探索左右目标手臂。</li>
		<li>一旦老鼠提交到一个特定的目标手臂 (所有四只爪子的老鼠已经进入目标臂), 阻止 "T" 连接的开始臂和对方的目标手臂 (图 1), 以防止老鼠进入对方的目标手臂。把老鼠留在迷宫里30秒, 然后把老鼠捡起来放回笼子里, 时间最短 (约30秒)。然后从 t 形迷宫中取出 "t" 型接合块 (125 mm x 230 mm x 65 mm, 由3D 打印机制作)。</li>
		<li>把老鼠放在开始的手臂上, 重复3.2.2。交替定义为: 当鼠进入相反的手臂, 与以前的运行36。老鼠每天执行4次跑步, 如下所示: 1st运行 2nd运行 10分钟休息 3rd运行 4th运行</li>
		<li>改变在10分钟的休息和动物之间的床上用品, 以消除气味偏见。在每个实验日结束时, 用75% 乙醇和蒸馏水清洗 T 迷宫。</li>
		<li>重复步骤3.2.1。-3.2.4。两天 (总共运行 12), 如图 2所示。</li>
	</ol>
</li>
	<li>
交替率和边偏好率计算
	<ol>
<li>计算% 的替换和% 的边首选项, 其中 L: 老鼠选择左臂 R: 老鼠选择右臂 正确的选择: 2nd运行与给定集中的 1st不同 (每个集合包含两个运行) 不正确的选择: 大鼠选择相同的手臂类似于以前的运行 <img alt="Equation 1" src="/files/ftp_upload/56694/56694eq1.jpg"> <img alt="Equation 2" src="/files/ftp_upload/56694/56694eq2.jpg"> 例如: 1天: 升升/升 2天: 升升/升 R 3天: R 升/升 l 替换: 2 (正确的选择)/6 (执行的总集合) * 100 = 33.33% 边偏爱:10 (L, 优选的边)/12 (总奔跑执行) * 100 = 83.33%</li>
	</ol>
</li>
	<li>
术后护理:
	<ol>
<li>给老鼠丁丙诺啡 (0.01 毫克/千克 IP) 每12小时, 在手术后2天。观察大鼠心脏骤停后1小时。</li>
		<li>将大鼠附着在呼吸机和加热垫上, 直到它恢复足够的意识来维持胸骨卧床。为了保持动物的体温, 在手术后, 把老鼠放在一个婴儿孵化器 (设置在27° c, 50% 湿度)。</li>
		<li>在手术后的前24小时内, 向动物提供软化的 chow (浸泡在水中)。如果老鼠不喝水, 管理抑菌盐水 (100 毫升/公斤/天, ip), 直到动物恢复和饮用水自由。</li>
		<li>给大鼠外用抗生素止痛 (肽和利多卡因软膏) 在所有的伤口。在它们完全恢复后, 将老鼠移回动物设施。</li>
	</ol>
</li>
	<li>
安乐死法
	<ol>
<li>在实验结束时使用5% 异氟醚和 100% N2O 安乐动物。</li>
	</ol>
</li>
</ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+deficits+associated+with+senescence:+a+neurophysiological+and+behavioral+study+in+the+rat.">Memory deficits associated with senescence: a neurophysiological and behavioral study in the rat.</a> J Comp Psychol. 93, 74-104 (1979).</li><li>Paul, C. M., Magda, G., Abel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+memory:+Theoretical+basis+and+comparative+review+on+experimental+methods+in+rodents.">Spatial memory: Theoretical basis and comparative review on experimental methods in rodents.</a> Behav Brain Res. 203, 151-164 (2009).</li><li>Vorhees, C. V., Williams, M. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Spontaneous alternation behavior. , (1989).</li><li>Deacon, R. M., Rawlins, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T-maze+alternation+in+the+rodent.">T-maze alternation in the rodent.</a> Nature protocols. 1, 7-12 (2006).</li><li>Wong, K. V., Hernandez, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+additive+manufacturing.">A review of additive manufacturing.</a> ISRN Mechanical Engineering. 2012, (2012).</li><li>Lin, H. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of biological statistics. 2, (2009).</li><li>Castellano, M. A., Diaz-Palarea, M. 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作者没有什么可透露的。

与执事和罗的协议31相比, 本研究做了一些修改。3D 打印机被用来建造 T 形迷宫。3的 d-印刷提供了可负担的和 cost-effective 的替代商业化 T 迷宫。为了减少实验过程中大鼠的焦虑, 在暗室中进行了最小光照的 T 迷宫。一旦鼠进入目标的武器之一, 我们轻轻地阻断对方的手臂。这避免了可能的压力, 从测试, 以及可能损害老鼠的尾巴, 而降低了断头台的门。任何不必要的压力都会抑制老...

根据美国心脏协会 (2017), 心脏骤停 (CA) 导致的死亡率每四分钟发生一次, 在美国每年影响超过40万人1。据记载, 由于血液灌注不足2,3,4, CA 可能导致神经元脑损伤。CA 诱发的脑损伤发生在海马的缺血敏感的 CA1 区5,6,7, 影响对学习和记忆至关重要的神经元8,9, 10、11、12。此外, 在海马 (即 CA1 神经元) 缺血条件下, 树突状脊柱密度的丧失在空间记忆损伤中起着重要作用13,14,15。由于这些病理变化的 CA, 行为障碍, 如: 焦虑, 抑郁症, 创伤后应激障碍和记忆丧失更普遍。尽管在医疗技术 (即有效的门诊服务) 方面取得了进步, 与提高的钙存活率相关, 但大多数的神经保护治疗 (低温除外) 在 ca 16 之后无法改善功能结果 ,17。CA 幸存者的生活质量通常很差, 并且背负着增加的医疗开支16。
通过行为测试对脑缺血的认知状态评估是重要的, 以确定药物疗效和最终发展成功的临床试验。在二十世纪四十年代, 爱德华托尔曼设计了第一个行为试验研究 hippocampus-based 空间记忆18。随后, 开发了不同的迷宫 (如莫里斯水迷宫、径向迷宫、T 或 Y 迷宫和巴恩斯迷宫) 来评估大鼠的 hippocampal-based 空间学习和记忆19,20,21,22 ,23。一个更广泛使用的行为测试是莫里斯水迷宫, 它检查空间学习和记忆在大鼠模型24。然而, 莫里斯水迷宫需要大鼠游泳和发挥充分的运动功能和控制。对于缺血实验, 如窒息心脏骤停 (ACA, 大鼠模型的钙) 模型, 需要插管股动脉/静脉, 以获得重要的血压, 血液气体和引进各种药物。由于股动脉/静脉插管可以抑制腿部移动, 使大鼠的游泳能力正常, 莫里斯水迷宫可能不是最适合测试认知损伤的 ACA。
巴恩斯迷宫是另一个被广泛使用的行为测试, 用来检查啮齿动物模型中的空间学习和记忆。巴恩斯迷宫不需要充分发挥马达功能和控制, 因此比莫里斯水迷宫的压力更小。在过去, 我们使用巴恩斯迷宫来进行实验, 以确定在控制或假与 ACA 诱导的大鼠之间是否发生功能性学习/记忆差异。由于巴恩斯迷宫广泛用于测量参考 (长期) 内存25,26, 而为巴恩斯迷宫所获得的数据并没有在轻度到中度 ACA 之后测试认知损伤的决心, 而aca 引起的认知缺陷与工作 (短期) 内存27、28、29、30有关, 这表明巴恩斯迷宫在我们的 aca 中评估内存功能是不可行的。模型.
因此, 我们开发了一个改进的 T 迷宫使用自发交替测试, 以评估工作 (短期) 记忆后 ACA。改进的 t 迷宫自发交替试验的主要优点是它的简单性和最小的压力, 与其他行为测试相比, 由于修改的 t 迷宫不需要事先的动物训练, 以及重计算根据莫里斯水迷宫和巴恩斯迷宫的要求, 分析或程序 (即大鼠的视频成像)。在这里我们表明, 改进的 T 迷宫自发交替试验是一个简单而高效的行为试验范例, 可以提供足够的分辨率, 以准确地检测和评价海马功能的疾病, 造成短期记忆丧失(即 ACA)。

<table>
	<tbody>
		<tr>
			<td>3D Printer</td>
			<td>MakerBot</td>
			<td>Replicator</td>
			<td>Fifth generation</td>
		</tr>
		<tr>
			<td>3D Printer Filament</td>
			<td>Hatchbox</td>
			<td></td>
			<td>PLA, 1.75 mm filament diameter</td>
		</tr>
		<tr>
			<td>200 Proof Pure Ethanol</td>
			<td>Koptec</td>
			<td>V1005SG</td>
			<td></td>
		</tr>
		<tr>
			<td>Sani-Chips</td>
			<td>PJ Murphy-Forest Products</td>
			<td></td>
			<td>Size: 8 to 20 mesh; 2.2 cubic foot/package; autoclavable bags</td>
		</tr>
		<tr>
			<td>Rat</td>
			<td>Charles River Laboratories</td>
			<td>Sprague-Dawley</td>
			<td></td>
		</tr>
		<tr>
			<td>Vecuronium bromide</td>
			<td>Sun Pharmaceutical</td>
			<td>47335-931-40</td>
			<td>10 mg</td>
		</tr>
		<tr>
			<td>Epinephrine</td>
			<td>Par Pharmaceutical</td>
			<td>42023-103-01</td>
			<td>Adrenalin Chloride Solution 1 mg/mL, 1:1000</td>
		</tr>
		<tr>
			<td>Buprenorphine Hydrochloride Injection</td>
			<td>Pfizer</td>
			<td>00409-2012-32</td>
			<td>0.3mg/mL</td>
		</tr>
		<tr>
			<td>SketchUp</td>
			<td>Trimble Inc.</td>
			<td></td>
			<td>3D modeling software</td>
		</tr>
		<tr>
			<td>VentElite Small Animal Ventilator</td>
			<td>Harvard Apparatus</td>
			<td>55-7040</td>
			<td>Animals raging in size from mouse to guinea pig (10g to 1kg)</td>
		</tr>
		<tr>
			<td>PowerLab 8/35</td>
			<td>Adinstruments</td>
			<td>PL3508</td>
			<td>8 analog input channels &#8211; 4 of which can be used in differential mode.</td>
		</tr>
		<tr>
			<td>Bio Amps</td>
			<td>Adinstruments</td>
			<td>FE132</td>
			<td>The Bio Amp is a galvanically isolated, high-performance differential bio amplifier optimized for the measurement of a wide variety of biological signals such as ECG, EMG and EEG recordings.</td>
		</tr>
		<tr>
			<td>Quad Bridge Amp</td>
			<td>Adinstruments</td>
			<td>FE224</td>
			<td>A four-channel, non-isolated bridge amplifier designed to allow the PowerLab to connect to most DC bridge transducers.</td>
		</tr>
		<tr>
			<td>LabChart 8</td>
			<td>Adinstruments</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ABL80 FLEX CO-OX blood gas analyzer</td>
			<td>Radiometer</td>
			<td></td>
			<td>pH / p CO2 / p O2</td>
		</tr>
		<tr>
			<td>SURFLO Teflon I.V. Catheter</td>
			<td>Terumo</td>
			<td>sc-361556</td>
			<td>Only use the flexible thin wall catheter (49-mm long)</td>
		</tr>
		<tr>
			<td>Pipet/Infusion Needle</td>
			<td>Hamilton</td>
			<td>7748-03</td>
			<td>17-gauge; 93-mm long; 10-degree angle</td>
		</tr>
		<tr>
			<td>Classic T3 Vaporizer</td>
			<td>SurgiVet</td>
			<td>VCT302</td>
			<td>Classic T3 Isoflurane Funnel Fill</td>
		</tr>
		<tr>
			<td>ENVIRO-PURE Charcoal Canister</td>
			<td>SurgiVet</td>
			<td>32373B10</td>
			<td>Designed to absorb waste anesthetic gas</td>
		</tr>
		<tr>
			<td>O2 single flowmeter</td>
			<td>SurgiVet</td>
			<td>32375B1</td>
			<td>0-1000 mL</td>
		</tr>
		<tr>
			<td>N2O Flowmeter</td>
			<td>VetEquip</td>
			<td>401721</td>
			<td>0-4LPM</td>
		</tr>
		<tr>
			<td>Clay Adams Intramedic Luer-Stub Adapter (Sterile)</td>
			<td>Becton Dickinson</td>
			<td>427565</td>
			<td>23 gauge</td>
		</tr>
		<tr>
			<td>Micro Forceps</td>
			<td>Black and Black surgical</td>
			<td>B3FRC-18 RM-8</td>
			<td>7 1/4&#34; (18 cm), 8mm RH, counterweight w/ guide pin 2mm, platform 6 x .3 mm, curved.</td>
		</tr>
		<tr>
			<td>Halstead Mosquito Forceps</td>
			<td>Roboz</td>
			<td>RS-7111</td>
			<td>Curved; 5&#34; Length, 1.3 mm tip diameter, 2.1 mm jaw width</td>
		</tr>
		<tr>
			<td>Mixter Forceps</td>
			<td>Roboz</td>
			<td>RS-7291</td>
			<td>5.25&#34; Curved Extra Delicate, 1.1 mm tips</td>
		</tr>
		<tr>
			<td>Castroviejo Micro Dissecting Spring Scissors</td>
			<td>Roboz</td>
			<td>RS-5650</td>
			<td>Straight, Sharp Points; 9 mm Cutting Edge; 0.15 mm Tip Width; 3 1/2&#34; Overall Length</td>
		</tr>
		<tr>
			<td>Mayo-Stille Scissors</td>
			<td>Roboz</td>
			<td>RS-6891</td>
			<td>5.5&#34; Round Curved</td>
		</tr>
		<tr>
			<td>Dumont #5 Forceps</td>
			<td>Roboz</td>
			<td>RS-5058</td>
			<td>45 Deg Dumoxel Tip Size .10 x .06 mm</td>
		</tr>
		<tr>
			<td>Olsen-Hegar Combination Scissor And Needle Holder</td>
			<td>Roboz</td>
			<td>RS-7884</td>
			<td>Cross Serration Tip; 5.5&#34; Length</td>
		</tr>
		<tr>
			<td>Moloney Forceps</td>
			<td>Roboz</td>
			<td>RS-8254</td>
			<td>Serrated; Slight Curve; 4.5&#34; Length</td>
		</tr>
	</tbody>
</table>

utilizing the modified t-maze to assess functional memory outcomes after cardiac arrest

背景: 评估轻度到中度认知损害在全球脑缺血 (即心脏骤停) 模型可能是困难的, 因为在手术后运动不良。例如, 接受外科手术的大鼠和莫里斯水迷宫可能无法游泳, 从而使实验无效。
新方法: 建立了改进的行为自发交替 T 迷宫试验。修改后的 T 迷宫协议的主要优点是它的相对简单的设计, 是强大到足以评估功能学习/记忆后缺血。此外, 数据分析简单明了。我们使用 T 迷宫来确定在轻度到中度 (6 分钟) 窒息心跳骤停 (ACA) 时, 老鼠的学习/记忆力缺陷。大鼠有一个自然的勘探倾向, 并将探索在 T 迷宫的替代武器, 而海马损伤大鼠倾向于采取侧偏爱, 导致减少自发交替率, 揭示海马相关功能性学习/记忆在存在或缺席 ACA。
结果: ACA 组有较高的侧偏爱比和较低的交替对照。
与现有方法比较: 莫里斯水和巴恩斯迷宫在评估学习/记忆功能方面更为突出。然而, 莫里斯水迷宫比其他迷宫更有压力。巴恩斯迷宫被广泛用于测量参考 (长期) 记忆, 而 ACA 诱发的认知赤字则与工作 (短期) 记忆更密切相关。
结论: 我们已经制定了一个简单的, 但有效的策略来描绘工作 (短期) 记忆通过 T 迷宫在我们的全球脑缺血模型 (ACA)。

ACA (全球脑缺血) 主要导致工作 (短期) 记忆缺陷28,29。为了评估在 ACA 之后的学习和记忆功能, 我们使用了改进的自发交替测试来评估工作 (短期) 内存30。自发交替试验结果表明, ACA 组 (26.19 ± 4.96%) 连续三天的交替率明显低于对照组 (62.96 ± 6.07%)(* p0.05)35由于提交给 ACA 的大鼠与对照组相比 (82.14 ?...

Watch this Scientific Journal Video about Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest at JoVE.com

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

应用改良 T 迷宫评估心脏骤停后的功能记忆结果

本协议描述了使用一个改进的 T 迷宫评估功能学习/记忆在窒息心脏停搏诱发脑缺血。

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor ...

utilizing-modified-t-maze-to-assess-functional-memory-outcomes-after

Research

JoVE Journal

Behavior

12.0K 次观看.  Louisiana State University Health Science Center. 本协议描述了使用一个改进的 T 迷宫评估功能学习/记忆在窒息心脏停搏诱发脑缺血。

视频: Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

科研

行为学

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris水迷宫实验:优化小鼠品系和测试环境

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

评估儿童工作记忆:儿童综合评估电池 - 工作记忆（CABC-WM）

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...

Using Wavelet Entropy to Demonstrate how Mindfulness Practice Increases Coordination between Irregular Cerebral and Cardiac Activities

In both the East and West, traditional teachings say that the mind and heart are somehow closely correlated, especially during spiritual practice. One difficulty in proving this objectively is that the natures of brain and heart activities are quite different. In this paper, we propose a methodology that uses wavelet entropy to measure the chaotic levels of both electroencephalogram (EEG) and electrocardiogram (ECG) data and show how this may be used to explore the potential coordination between the mind and heart under different experimental conditions. Furthermore, Statistical Parametric Mapping (SPM) was used to identify the brain regions in which the EEG wavelet entropy was the most affected by the experimental conditions. As an illustration, the EEG and ECG were recorded under two different conditions (normal rest and mindful breathing) at the beginning of an 8-week standard Mindfulness-based Stress Reduction (MBSR) training course (pretest) and after the course (posttest). Using the proposed method, the results consistently showed that the wavelet entropy of the brain EEG decreased during the MBSR mindful breathing state as compared to that during the closed-eye resting state. Similarly, a lower wavelet entropy of heartrate was found during MBSR mindful breathing. However, no difference in wavelet entropy during MBSR mindful breathing was found between the pretest and posttest. No correlation was observed between the entropy of brain waves and the entropy of heartrate during normal rest in all participants, whereas a significant correlation was observed during MBSR mindful breathing. Additionally, the most well-correlated brain regions were located in the central areas of the brain. This study provides a methodology for the establishment of evidence that mindfulness practice (i.e., mindful breathing) may increase the coordination between mind and heart activities.

This manuscript describes how to use the wavelet entropy index to analyze high-density electroencephalography (EEG) and electrocardiography (ECG) data. We show that the irregularity of cerebral and cardiac activities became more coordinated during mindfulness-based stress reduction practice.

使用小波熵来证明正念练习如何增加不规则脑与心脏活动之间的协调

In both the East and West, traditional teachings say that the mind and heart are somehow closely correlated, especially during spiritual practice. One ...

A Within-subjects Experimental Protocol to Assess the Effects of Social Input on Infant EEG

Despite the importance of social interactions for infant brain development, little research has assessed functional neural activation while infants socially interact. Electroencephalography (EEG) power is an advantageous technique to assess infant functional neural activation. However, many studies record infant EEG only during one baseline condition. This protocol describes a paradigm that is designed to comprehensively assess infant EEG activity in both social and nonsocial contexts as well as tease apart how different types of social inputs differentially relate to infant EEG. The within-subjects paradigm includes four controlled conditions. In the nonsocial condition, infants view objects on computer screens. The joint attention condition involves an experimenter directing the infant's attention to pictures. The joint attention condition includes three types of social input: language, face-to-face interaction, and the presence of joint attention. Differences in infant EEG between the nonsocial and joint attention conditions could be due to any of these three types of input. Therefore, two additional conditions (one with language input while the experimenter is hidden behind a screen and one with face-to-face interaction) were included to assess the driving contextual factors in patterns of infant neural activation. Representative results demonstrate that infant EEG power varied by condition, both overall and differentially by brain region, supporting the functional nature of infant EEG power. This technique is advantageous in that it includes conditions that are clearly social or nonsocial and allows for examination of how specific types of social input relate to EEG power. This paradigm can be used to assess how individual differences in age, affect, socioeconomic status, and parent-infant interaction quality relate to the development of the social brain. Based on the demonstrated functional nature of infant EEG power, future studies should consider the role of EEG recording context and design conditions that are clearly social or nonsocial.

This novel protocol is designed to assess the neural bases of social interaction in infants. The paradigm is designed to tease apart how various social inputs such as language, joint attention, and face-to-face interaction relate to infant neural activation. Infant EEG power is recorded during both social and nonsocial conditions.

一个被试内实验协议，以评估对婴幼儿脑社会投入的影响

Despite the importance of social interactions for infant brain development, little research has assessed functional neural activation while infants ...

A Community-based Stress Management Program: Using Wearable Devices to Assess Whole Body Physiological Responses in Non-laboratory Settings

A pragmatic breath-based intervention to benefit human performance and stress management is timely and valuable to individuals seeking holistic approaches for emotional regulation and optimizing compensatory reserve mechanisms. This protocol is designed to not only teach mind-body awareness but also to provide feedback utilizing physiological data and survey results. The primary findings of this study showed that heart coherence and alpha variables were significantly correlated after four weeks of the breath-based meditation stress protocol. Meditation and rhythmic breathing produced significant increases in alpha brain activity. These brain physiological responses conformed to the Pleth Variability Index (PVI) changes, suggesting the ability of the human body to enter into a meditative state and effectively manage stress. When assessed after four weeks of daily practicing the techniques employed in the stress management protocol, based on the Five Facet Mindfulness Questionnaire, subjects improved in applying mindfulness skills. The overall mindfulness score, Pleth variability index (PVI), and perfusion index (PI) increased after the 4-week intervention period. Results from electroencephalography (brain waves) were consistent with a meditative state during the post-study follow-up session. This provides evidence that wearable devices are feasible for data collection during a breath-based stress management intervention. This protocol can be easily and efficiently implemented into any study design in which physiological data are desired in a non-laboratory-based setting.

Stress is an unavoidable and persistent component of life and holistic approaches for its management are being considered. A standardized methodology was created to demonstrate the feasibility of a breath-based stress management protocol that can be used with commercially available portable technology.

基于社区的压力管理计划: 在非实验室环境中使用可穿戴设备评估全身生理反应

A pragmatic breath-based intervention to benefit human performance and stress management is timely and valuable to individuals seeking holistic approaches ...

Reversible Cooling-induced Deactivations to Study Cortical Contributions to Obstacle Memory in the Walking Cat

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For example, in the cat, visual information about an impending obstacle can modulate stepping for avoidance. Locomotor adaptation can also occur independent of vision, as sudden tactile inputs to the leg by an expected obstacle can modify the stepping of all four legs for avoidance. Such complex locomotor coordination involves supraspinal structures, such as the parietal cortex. This protocol describes the use of reversible, cooling-induced cortical deactivation to assess parietal cortex contributions to memory-guided obstacle locomotion in the cat. Small cooling loops, known as cryoloops, are specially shaped to deactivate discrete regions of interest to assess their contributions to an overt behavior. Such methods have been used to elucidate the role of parietal area 5 in memory-guided obstacle avoidance in the cat.

Complex locomotion in naturalistic environments requiring careful coordination of the limbs involves regions of the parietal cortex. The following protocol describes the use of reversible cooling-induced deactivation to demonstrate the role of parietal area 5 in memory-guided obstacle avoidance in the walking cat.

可逆冷却诱导 Deactivations 对行走猫的障碍记忆的皮层贡献研究

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For ...

The Modified Temptation Resistance Task: A Paradigm to Elicit Children's Strategic Lie-telling

The protocol for the temptation resistance paradigm was designed to elicit 2- to 8-year-old children's strategic lie-telling behaviors. The reward of transgression was intended to be too tempting to resist, so that children's spontaneous lie-telling behavior in the presence of irreversible evidence due to the transgression could be observed. Conducted in a developmental psychology laboratory behind a one-way mirror and video recorded with a hidden camcorder, the protocol starts with a guessing game where the child is given three trials to guess the identities of hidden objects under an upside down cup. The experimenter gives hints in the first two trials to make sure the child "guess" them correctly. At the beginning of the final trial, the experimenter leaves the room briefly and asks the child not to peek under the cup. If the child could not resist the temptation and peeks, small particles previously filled in the cup would scatter on a grooved surface. Upon the experimenter's return, the child is asked if he/she has peeked. If he/she denies transgression contrary to the presence of the physical evidence, he/she is asked to explain why the particles are on the table. Three responses could be observed in the procedure, including the initial transgression, truth-telling or lie-telling behavior if the child peeked, and the strategic lie-telling behavior in the presence of the physical evidence.

The protocol for the temptation resistance paradigm was designed to elicit 2- to 8-year-old children&#39;s strategic lie-telling behaviors. The reward of transgression was intended to be too tempting to resist, so that children&#39;s spontaneous lie-telling behavior in the presence of irreversible evidence due to the transgression could be observed.

修正诱惑力的任务: 一种诱导儿童战略说谎的范式

The protocol for the temptation resistance paradigm was designed to elicit 2- to 8-year-old children's strategic lie-telling behaviors. The reward of ...

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper understanding of the neurobiological causes underlying the decision-making problems. A cost-benefit based T-maze task is used for measuring decision making in which rodents choose between a high reward arm (HRA) and a low reward arm (LRA). There are two paradigms of the T-maze decision-making task, one in which the cost is a time delay and the other in which it is physical effort. Both paradigms require a tedious and labor-intensive management of experimental animals, multiple doors, pellet reward, and arm choice recordings. In the current work, we invented an apparatus based on traditional T-maze with full automation for pellet delivery, door management and choice recordings. This automated setup can be used for the evaluation of both delay- and effort-based decision making in rodents. With the protocol described here, our lab investigated the decision-making phenotypes of multiple genetically modified mice. In the representative data, we showed that the mice with ablated medial habenular showed aversions of both delay and effort and tended to choose the immediate and effortless reward. This protocol helps to decrease the variability caused by experimenter intervention and to enhance experiment efficiency. In addition, chronic silicon probe or microelectrode recording, fiber-optic imaging and/or manipulation of neural activity can be easily applied during the decision-making task using the setup described here.

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

基于自动 T 迷宫的自由移动啮齿动物延迟和努力决策分析装置及协议

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

Biomechanical Analysis Methods to Assess Professional Badminton Players' Lunge Performance

Under the condition of simulating a badminton court in the laboratory, this study used the injury mechanism model to analyze the maximal right lunge movements of eight professional badminton players and eight amateur players. The purpose of this protocol is to study the differences in kinematics and joint moment of the right knee and ankle. A motion capture system and force plate were used to capture data of the joint movements of the lower extremity and the vertical ground reaction force (vGRF). Sixteen young men who did not have any sports injuries in the past 6 months took part in the study. The subjects performed a maximal right lunge from the start position with their right foot, stepping on and fully contacting with the force plate, hit the shuttlecock with an underhand stroke to the designated position in the backcourt, and then returned to the start/end position. All subjects wore the same badminton shoes to avoid a difference in impact from different badminton shoes. The amateur players showed a greater range of ankle movement and reverse joint moment on the frontal plane, and a larger internal joint rotation moment on the horizontal plane. The professional badminton players exhibited greater knee moment on the sagittal and frontal planes. Therefore, these factors should be considered in the development of the training program to reduce the risk of sports injuries in knee and ankle joints. This study simulates the real badminton court and calibrates the range of activities of each movement of the subjects so that the subjects complete the experimental action in a natural state with high quality. A limitation of this study is that it does not combine joint load and muscle activity. Another limitation is that the sample size is small and should be expanded in future studies. This research method can be applied to the lower limb biomechanical research of other footwork in the badminton project.

Here, we present a protocol to evaluate the differences in injury mechanisms between professional and amateur players when performing a badminton maximal right lunge movement by analyzing lower limb kinematics.

生物力学分析方法,以评估职业羽毛球运动员的隆起表现

Under the condition of simulating a badminton court in the laboratory, this study used the injury mechanism model to analyze the maximal right lunge ...