作者希望感谢本科生丹尼尔韩为他的帮助运行 Matlab 代码和本科生亚历克斯 Boettcher 为他的协助运行行为实验。这个项目是由 NSF IOS 1256799 支持 R.L.M., 并由国家卫生研究院的药物滥用的国家研究所在获奖数字 T32DA007234。

此处所述的所有程序均由明尼苏达大学机构动物保育和使用委员会 (IACUC) 批准, 并符合《实验室动物护理和使用指南》 13.
 1. 动物和插管手术 <ol> <li> 在大约55天的时间内从一个普通动物供应商获得叙利亚仓鼠. 
	注意: 虽然动物的年龄会因各种实验范式的限制而变化, 但对于性行为来说, 获得性成熟的动物是非常重要的. </li>
	<li> 在受控温度 (22 和 #176; C) 和照明 (14 h 灯后10小时暗灯亮出13:00 小时) 环境中, 与食物和水可用 ad 随意 除了在实验测试期间. 
	注: 由于仓鼠季节性繁殖, 这14:10 光/暗循环模仿它们的自然繁殖条件. </li>
	<li> 对实验室进行1周的驯化后, 在全身麻醉下进行无菌双侧 ovariectomies 和颅内立体定向 cannulations ( 如 戊巴比妥), 并管理适当的经组织批准的术后镇痛和抗生素治疗如前所述 14 , 15 . 
	注意: 对于性行为的研究, ovariectomies 是必要的, 以消除主要的内源性激素, 使动物可以统一诱导的性接受与外源激素的实验时间。
	<ol> <li> 对于多巴胺能和谷氨酸探针植入, stereotaxically 种植指南插管 (0.7 毫米直径; 图 3 )进入感兴趣的领域 ( 例如 , 本实验室以右核伏隔 (NAc) 为目标), 如在其他地方的 15 中所描述的那样。
		注: 虽然所提供的数据来自每只动物的单个谷氨酸或多巴胺探针的单传感器记录, 但该系统允许在单个动物中同时记录多达4传感器;因此, 1-4 插管可以植入根据所需的实验设计和结果。
		<ol> <li> 使用立体定向图谱 ( 例如 , 由桑色素和 #38 的金黄仓鼠大脑 ) 确定感兴趣区域的种植座坐标, 请记住, 传感器将套管下面的1毫米扩展到脑组织。
		</ol> </li> <li> 在将导管 stereotaxically 降低到所需的背腹位置后, 将其贴到头骨上, 使用由牙科丙烯酸构建的帽, 用不锈钢骨螺钉和塑料头固定 ( 图3;有关更多详细信息, 请参见 15 . </li>
		<li> 对于碳纤维测试, 插入一个参考电极 (350 和 #956; m 直径, 7.5 毫米总长度) 到对侧半球. 
		注: 不需要与套管的具体位置或距离;参考电极可以放置在对侧半球方便的任何地方. </li>
	</ol> </li> </ol> 2。生物传感器和碳纤维检测 <ol> <li> 手术后1周的恢复, 管理激素启动方案, 诱导性接受。
	<ol> <li> 注入10和 #181; 在0.1 毫升棉籽油皮下 (南卡罗来纳州), 约48ĥ和 24 h 雌二醇 g, 在性行为测试之前, 对男性的性接受度为 11 。
		<li> 注入500和 #181; 0.1 毫升棉籽油中的黄体酮 g, 在引入雄性诱导性接受前 4 h. 11 . </li>
	</ol> </li> <li> 在其每日循环的黑暗阶段开始时测试所有动物, 因为啮齿动物的社会行为会在白光和红光测试条件下受到改变 16 17 , 18 . </li> <li> 用于酶生物传感器的谷氨酸测试, 校准传感器 体外 ( 图 4 ), 然后使用之前所述 15 , 19 . <ol> <li> 使校准解决方案在20毫升玻璃离心管的测试中更加新鲜。对于分析溶液, 将7.4 毫克 l-谷氨酸分解为10毫升的超纯 H 2 O 通过轻轻倒置管。使干扰溶液溶解176.1 毫克 AA 在10毫升的超纯 H 2 O. </li>
		<li> 通过在基本实验室环架底座上放置磁力搅拌器来设置校准。在磁力搅拌器的顶部放置一个20毫升的夹套烧杯, 并使用中型2爪钳夹住。
		<ol> <li> 将磁性搅拌棒和20毫升的100毫米磷酸盐缓冲盐水 (PBS) 添加到夹套烧杯中。将夹套烧杯连接到循环水浴, 将缓冲液加热到37和 #176; C. 
			注意: 酶生物传感器的校准必须在体温下进行, 因为酶活性受温度影响. </li>
		</ol> </li> <li> 将传感器校准托架放在夹套烧杯的顶部, 并在顶部设置4通道校准前置放大器, 使用直角钳固定到位。将前置放大器连接到数据调理和采集装置, 以记录校准数据. </li>
		<li> 测试每个酶生物传感器对谷氨酸 (或其他商业可用生物传感器探头的其他分析) 的灵敏度, 例如, 在实验录制之前, 将探针放置在夹套烧杯, 淹没传感腔和部分氯化银 (氯化) 参考线在缓冲液中。
		<ol> <li> 在将每个传感器 (最多 4) 连接到4通道校准前置放大器的端口上之前, 在计算机接口上使用免费下载的采集软件启动新的记录。允许传感器达到稳定的基线. </li>
		当传感器到达稳定基线时, </ol> </li> <li> 开始添加分析物注入。使用校准架上的孔将吸管尖端引入缓冲液。使用 "录制软件" 中的 "快速密钥注释" 工具在进行注入时进行批注。
		<ol> <li> 使10和 #181; M 将移40和 #956 的谷氨酸添加到缓冲区解决方案中。等待传感器在加法之间稳定。添加3注射的分析物溶液, 然后加入干扰 AA 溶液 (50 和 #956; 250 和 #956 的注入量; M 浓度的变化). 
			注: 校准允许转换安培信号的变化 (在步骤2.12 中的酶实验记录中看到), 以改变谷氨酸浓度, 如果需要。有关逐步指令, 请参见参考 15 。该实验室使用的多巴胺能探针在制造过程中由商业供应商进行校准。谷氨酸探针必须在使用时进行校准, 因为它们的灵敏度随着时间的推移而降低, 因为酶的成分会降低 19 。由于碳纤维电极不经过降解, 该公司在装运前进行的校准就足够了。这里是增加的可能性损坏稀薄的碳纤维 (350 和 #956; m 直径) 通过另外的处理. </li>
		</ol> </li> </ol> </li> <li> 从动物和 #39 的家笼中取出带床上用品的测试室. 
	注意: 这增加了女性和 #39; 对测试室的熟悉程度, 以及暴露雄性对雌性在雌二醇治疗中散发的性刺激嗅觉交配的提示. </li>
	<li> 在插入探针之前, 用异氟醚或在诱导室中的另一种挥发性麻醉剂轻轻麻醉动物. </li>
	<li> 从导套管中取出咬合 obdurator, 然后在导向轴上插入碳纤维电极或酶探针. </li>
	<li> 探头插入后, 将动物放在测试室中. 
	注: 本实验室使用10加仑玻璃水族箱 (24.5 x 48.9 x 29.4 厘米), 但也可采用相等面积的圆形腔室. </li>
	<li> 开始在商业上可用的软件程序中记录视频和 time-locked 安培信号, 在其他地方详细描述 15 . </li>
	<li> 将传感器连接到记录系统: 电位通过电屏蔽电缆和电动旋转。通过拧紧固定在头上的针脚, 稳定传感器连接。必要时使用实验室胶片加强连接. 
	注: 需要一个定制的连接器来连接碳纤维电极和参考电极的电位, 而酶谷氨酸传感器可以直接连接到电位. </li>
	<li> 在实验测试前允许传感器在大脑中平衡. 
	注意: 平衡时间的不同取决于记录传感器 ( 例如 , 酶传感器需要 2-4 h 平衡, 而碳纤电极只需要大约30分钟)。如果两种类型的传感器被植入, 平衡的更长的酶传感器时间. </li>
	<li> 在传感器平衡后, 将刺激男性引入测试室. </li>
	<li> 在第一次安装后, 由刺激男性插入 (称为 intromission), 继续录制额外的 10-30 分钟, 具体取决于实验目标. </li>
	<li> 在进行行为测试后, 断开女性和 #39 的传感器. </li>
	<li> 将两个仓鼠从测试室中取出。使用70% 乙醇去除床上用品和清洁房间. 
	注: 由于碳纤维记录所需的短暂平衡, 步骤 2.3-2.14 可以重复, 以测试多个动物在同一天。相比之下, 由于酶传感器需要大约 4 h 的平衡, 每天只测试一只动物, 以限制可能的学科变异由于动物和 #39 的差异; 昼夜时间在测试. </li>
</ol> 3。牺牲和灌注 <ol> <li> 在测试期结束后, 用安乐试剂 ( (如 , 本实验室使用腹腔注射0.2 毫升苯妥英和戊巴比妥溶液) 深入麻醉女性试验对象。.</li>
	<li> Transcardially 灌注的动物与25毫米 PBS, pH 7.6, 以消除所有循环血液, 其次是20分钟的固定使用4% 甲醛 pbs. </li>
	<li> 在一夜间50毫升锥形管中, 将4% 甲醛溶液中的30毫升的大脑和后缀移除。
	<ol> <li> 将大脑存储在10% 蔗糖-PBS 溶液中, 直到准备好串行部分. 
		注意: 大脑可以储存2周, 每周进行蔗糖溶液的改变, 以避免污染物的潜在生长. </li>
	</ol> </li> <li> 串行部分40和 #181 中的大脑; m 切片通过植入区域, 如前面所描述的冻结切片或低温 19 。
	<li> 在商业上可用的粘附涂层幻灯片上连续安装切片 (或参见 20 进行)。允许干燥, 至少在一夜. </li>
	<li> 用甲酚紫色染料、清除和片着色幻灯片, 如前所述 20 . </li>
	<li> 使用明亮的现场显微镜对幻灯片进行图像验证, 以确认传感器的解剖位置. </li>
</ol> 4。行为编码 <ol> <li> 使用免费的商用可下载软件查看和注释视频 (参见 材料的表 ) 慢动作, 以精确的代码行为 time-locked 安培信号. 
	注意: 要在分析中使用 MATLAB 代码 (见下面的 ) , 请对每个女性和 #39 的 开始 和 结束 分别进行批注; s 和 #39 的行为。
	<ol> <li> 在框架中对 startLordosis 进行批注时, 雌性启动了 dorsoflexion 的背部并将其尾部向上缺陷。当女性终止这种姿势时, 当雌性调整到测试室的另一个位置时, 通常会发生这种情况. </li>
		<li> 批注 startAI 当雌性处于前姿势时, 雄性会将他的鼻子移向雌性和 #39; 生殖器区域, 在那里可能会发生嗅吸、舔舐或鼻子她的会阴区域。注释 endAI 当雄性从靠近雌性和 #39 的生殖器区域移开他的鼻子时. </li>
		<li> 批注 startMount 当男性接近并将其前爪置于女性的位置时, 无论安装尝试的方向如何 (如 、侧面、臀部). </li>
		<li> 批注 startIntromission 当成功的插接时, 会使安装的男性生殖器进入女性和 #39 的阴道. 
		注: intromission 是行为可区分的, 在 intromission 之前发生, 男性明显拉他的上肢对他的身体, 推力他的骨盆向前, 并卷曲他的尾巴向上, 表明渗透 (见强类 = "xfig" > 图 4 C )。
		<ol> <li> 如果适用, 则批注 射精 。
			注意: 在交配回合结束时, 与成功的 intromission 结合, 雄性会射精。虽然你无法想象实际的排放, 雄性仓鼠有一个典型的踩踏运动与他们的后腿在 intromission 21 , 从而注释的射精发生的脚运动. </li>
		</ol> </li> <li> 批注 endIntromission 和 endMount 同时在框架中, 当男性已经删除了他的前两个爪子, 因此, 没有与女性的接触。由于经常无法想象他的撤退, 同时编码这两种行为, 以使一致性和可靠性跨交配回合. 
		注意: 交配回合要么遵循一个成功的射精, 要么是当女性打破了前的姿势后, 至少有一个 mount . </li>
	</ol> </li> </ol> 5。示例数据分析 注意: 使用固定电位碳纤和酶记录多巴胺 a谷氨酸, 分别, 允许的能力, 以测量的补品和阶段性模式的电化学释放。该实验室使用一个免费的商业采集系统和软件来记录和转换从模拟到数字的记录信号 (偏差 0.600 V, 采样率 = 1 Hz). 
<ol> <li> 示例数据分析协议的说明 <ol> <li> 尽管可以使用其他程序, 但用 MATLAB 对记录的电化学信号中的主音和相位 (瞬态) 变化进行量化。
		<ol> <li> 对于所提供的数据, 使用50点移动平均滤波器计算主音信号。通过从原始数据中减去主音信号, 计算出信号的瞬态唯一归一化表示。在识别相位峰值时, 有各种算法选项 ( 例如 , 将峰值标记为超过平均值 1 SD 的信号的变化). </li>
			<li> 确定这些峰值的位置, 即 , 局部极大值, 从正常化信号使用 MATLAB 函数 peakfinder , 将最小峰值振幅设置为信号的根平均平方。从归一化信号中检测出峰值的位置, 提取各峰值的振幅。检测峰值的能力允许识别任何可能与之相符的 time-locked 行为, 或在被调查的神经中驾驶相对峰值. </li>
		</ol> </li> <li> 按照行为注释对处理的数据进行分段 (请参见步骤 3); 学生和 #39; s t 测试用于评估强直性神经递质水平、瞬变的发生和振幅在交配回合期间, pre-mating 回合前与前的瞬变。为了测量电化学释放的补剂模式的变化, 在交配回合 ( 即 , pre-mating 回合前) 之前, 在 前 位置直接发生的数据点与交配回合中的数据点进行了比较。使用 t 测试的会话. </li>
	</ol> </li> <li> 为所描述的数据分析协议准备原始数据 注: 如上所述, 虽然其他程序或分析可能是同样可行的选项, 但为了使用上面描述的 MATLAB 代码, 必须准备原始数据以下方式: <ol> <li> 导出注释和电压文件。要做到这一点, 在 #39; 文件和 #39; 选项卡上, 选择 #39; 导出和 #39; 函数并选择 #39; 批注和 #39; 选项卡以保存批注。在 "#39; 文件和 #39; 选项卡上, 选择 #39; 导出和 #39; 函数并选择 #34; TSV 和 #39; 选项卡将电压测量保存为 TSV 扩展文件. </li>
		<li> 使用电子表格程序打开每个文件并保存为和 #39; xls/xlsx 和 #39; 使文件与 MATLAB 兼容. 
		注: 对于和 #39; 注释和 #39; 文件, 还有其他注释必须在电子表格程序中手动添加 post-export, 以口述 pre-mating 前的 开始 和 end (批注为 PreMBlordorsis ) 和交配回合, 使这些时间点可以直接比较, 在步骤5.1.2 中提到. </li>
		<li> 注释 pre-mating 回合前 ( startPreMBlordosis ) 的开始, 同时前行为开始 ( startLordosis ) 也包括 装入 行为, 并且在新的交配回合开始时, 如果女性保持在前的姿势. </li>
		<li> 在第一个 装入 开始时, 对 pre-mating 回合前 ( endPreMBlordosis ) 的末尾进行批注. </li>
		<li> 在 endPreMBlordosis 的同时对交配回合 ( startMB ) 的开始进行注解, 因为这意味着交配回合的开始. </li>
		<li> 在 装入 包括 射精 (请参见步骤 4.1.4.1) 的末尾 (endMB) 或在前姿势 ( endLordosis ) 的终止后, 对交配回合的结束进行批注. </li>
	</ol>
	</li>
</ol>

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作者没有什么可透露的。

虽然相对简单, 但在使用此技术时可能会出现一些问题。首先, 探针的立体定位必须是精确的: 不像微透析, 样本更广泛的细胞外环境半径周围的探针, 这种技术只允许测量的神经递质进入直接接触用探针其次, 在碳纤维记录的情况下, 由于纤维的宽度小, 断裂可能发生, 并且探针必须插入与故意关心。在谷氨酸生物传感器的情况下, 如果探针不能在3周的保证时限内使用, 就会发生酶降解。幸运的是, 所有这些问题都是可寻址的, 并且可以通过适当的护理和注意细节来消除。
潜在的问题警告出现在固定潜在的碳纤维录音的特殊性。因为多巴胺的应用电位与其他胺, 如去甲肾上腺素, 如果录音是在区域释放两个神经递质, 比这种缺乏化学特异性可能限制结果的解释。在 NAc 中, 这并不构成一个主要问题, 多巴胺是主要的儿茶酚胺发射器24。由于 NAc 中约有98% 的细胞是对多巴胺神经24反应的中刺神经元, 因此信号偏向于检测多巴胺, 并有助于使用这种更关键的方法。
这种酶生物传感器和固定电位碳纤维记录系统的另一个好处是, 可以进行双探针记录, 使活性和 nonelectroactive 神经递质, 如多巴胺和谷氨酸可以测量同一动物同时。虽然这里提供的数据来自于单独的动物的录音, 但这些技术可以同时使用, 这样就可以在多个脑区或同一个区域中评估神经的收敛性。在同一动物的录音组合, 或在这里显示的动物的录音比较, 都是在网络连接和信号背后阐明机制的有力工具。
总之, 本文展示了酶传感和固定电位记录技术, 利用单个实验装置来测量多活性和 nonelectroactive 神经递质的功能。尽管这里所提供的数据来自于多个动物的单个神经递质记录, 但这个系统可以同时在一个动物中记录多达4传感器, 同时15。样本的结果共同提供了洞察的快速神经的雌性仓鼠的具体模式的交配刺激从男性。此外, 这些实验表明, time-locked 反应的多巴胺能和谷氨酸瞬变在新发现的女性的核心, 从男性 intromission, 这表明这个版本可能负责编码不同的奖励性质的性行为。我们的目标是继续利用雌性叙利亚仓鼠的交配模式, 以发展更全面的图片的关系,在体内多巴胺和谷氨酸释放和基础电路的单个组成部分的交配刺激从男性。我们认为, 对这些神经递质的特征进行定性的能力不仅从基础科学的角度来看是有益的23, 而且在阐明可能的治疗方法的病理形式的奖励行为吸毒成瘾25。

在清醒行为的动物中测量神经递质释放的能力使研究人员能够将特定行为与神经的时空模式联系起来--这是研究自然的机制和电路的有力工具。和 real-time 的行为。从历史上看, 微透析用于测量脑细胞外环境中的电反应和反应物质1。这项技术使用一个类似的离子组成的水溶液的连续流动的胞外流体, 通过一个微透析探针组成一个小轴与尖端制成的透中空纤维膜2。在插入探针后, 神经递质或其他分析的利益可以通过被动扩散在透膜之前被收集, 然后用高效液相色谱法 (HPLC) 进行分析,分析化学技术通常用于分离、识别和量化异构混合物中的组分3。
虽然微透析是一种敏感的技术, 可以用来测量几乎任何感兴趣的分析物, 时间分辨率很低, 最大采样率在几分钟到几十分钟的顺序1,2。快速扫描循环伏安法 (FSCV) 的发明, 一种依赖于活性种的氧化还原电位的技术, 可以在细胞外液中的感兴趣的分析物的瞬时浓度附近进行澄清。简要 (参见罗宾逊et al.4为了进行广泛的审查, 在快速的时间刻度4中, 使用一个电极来提高和降低三角波的电压。当电压在正确的范围内, 复利的化合物被反复氧化和减少。这种氧化和还原导致电子的运动产生一个小的交流电流。扫描率发生在秒的规模与氧化和减少的化合物发生在微秒。通过从产生的电流中减去探针所产生的背景电流, 就可以产生电压与每个化合物所特有的电流图。由于电压振荡的时间尺度是已知的, 这些数据可以用来计算作为一个时间函数的电流的情节。因此, 只要在每次氧化和还原反应中转移的电子数已知为4, 就可以确定化合物的相对浓度。
这种化学特异性和高的时间分辨率使 FSCV 成为一种强有力的技术来检测变化的化学浓度在体内。然而, 尽管有这些多方面的优势, 这项技术需要广泛的技术专长和昂贵的设备和设置。此外, nonelectroactive 神经递质 (例如, 谷氨酸) 不能用这种技术来衡量。幸运的是, 在电化学领域的技术进步5, 以及这些发明的商业化, 引入了一个相对简单的方法来测量清醒行为动物的 non-electroactive 神经递质不影响时间精度--一种称为酶生物传感器技术的技术。这种技术使用酶的 nonelectroactive 神经递质转化为两个底物, 其中之一是活性过氧化氢被检测为安培氧化电流产生的应用电位5.商用生物传感器探头 (参见图 1) 通过竞争性地减少内源 interferents 的贡献, 有选择地测量分析的兴趣。在谷氨酸的情况下, 共同的干扰抗坏血酸 (aa) 的贡献是竞争性地减少到被测量的电流由 co-localizing aa 氧化酶在传感器的活跃酶表面上, 转换 aa 为 non-electroactivedihydroascorbate 和水。此外, 在酶层下存在的负电荷磺聚合物层排除了内源性阴离子化合物。
这种生物传感器实验装置可以测量活性神经递质, 如在 FSCV, 但它采用了固定电位记录6。与在 FSCV 中应用的振荡电压相反, 在固定电位记录中, 电压保持在对感兴趣的分析物的氧化还原电位上。尽管它的化学选择性比 FSCV 低, 因为多个神经递质可能具有相同的氧化还原潜能, 但在绝大多数情况下, 大脑区域的神经递质是扭曲的, 这种方法的交钥匙性质胜过了缺乏化学特异.
能够测量活性和 nonelectroactive 神经递质释放在近 real-time, 并链接到特定的行为事件提供了一个机会, 研究收敛神经递质释放。这份手稿详细介绍了这一系统的使用, 以询问多巴胺和谷氨酸神经响应自然奖励在清醒行为仓鼠。本文的目的是详细介绍在雌性仓鼠性行为期间, 测量这种神经递质释放的过程, 目的是展示其检查其他行为和实验范式的可行性。
仓鼠是用于电化学记录的理想模型 历史上, 老鼠和老鼠模型在性行为的研究中被使用。这些啮齿类动物参与一个动态的交配序列, 涉及许多女性邀约行为, 包括跳跃, 穿梭, 和耳朵摆动引诱男性追逐, 并最终安装女性7。由男性装载 (有或没有阴道渗透) 仅持续几秒钟, 在期间女性参与她的性行为姿态 (被称为前) 也仅几秒钟在恢复活跃邀约行为之前。这种行为模式, 由高水平的活动穿插, 短暂的不动的时期, 是一个问题, 以测量神经行为的动物。首先, 在与神经活动无关的安培记录中可以有运动工件。第二, 运动与特定神经递质在某些脑区的释放有关。例如, 多巴胺释放已被耦合到背部和腹侧纹状体的运动活动8,9, 这一发现构成了微透析测量多巴胺后精神的基础管理10。因为在 mos 中女性典型的邀约行为啮齿类动物涉及高水平的运动活动, 并由10分钟的性行为测试的大部分, 这使得很难把神经的变化归咎于性行为的显式成分, 集体最后只分钟.
为了分析女性性行为的神经特征, 该实验室寻找了一个在性行为中有最小运动活动的物种。交配序列在叙利亚仓鼠 (Mesocricetus 鲫) 是理想的神经录音由于缺乏邀约行为通常见于老鼠和老鼠11。因此, 雌性仓鼠将进入并保持前的姿势, 在10分钟的测试会话12中超过9分钟。由于女性缺乏外来的运动动作 ,在体内可以获得与雄性互成分相关的电化学记录。
仓鼠交配回合 在将雄性刺激动物引入试验室后, 雄性将在安装前先进行雌生殖器调查 (AI) (图 2a)。为了雄鸟的坐骑, 雌鸟必须接受一种被称为前的性体位, 在这一姿势中, 她会拱起背部, 使其尾巴偏转, 这样, 雄性就能获得阴道的生殖器。雄鸟会装上雌性, 用两只爪子 (图 2B) 将她的后躯紧握起来, 并开始尝试获取 intromission (图 2C)。雄鸟在最终达到射精前, 会将雌性 (不插入) 和 intromit 多次安装在一起。这一系列的坐骑和 intromissions 导致射精被称为 "交配回合"。男性将有几个交配回合在一个单一的会议。

<table><tbody><tr><td>Nembutal</td><td>Oak Pharmaceuticals Inc.</td><td>76478-501-50</td><td>Pentobarbital sodium injection, USP. This lab uses 8.5 mg/100 g body weight, injected intraperotineally.&amp;nbsp;</td></tr><tr><td>Loxicom analgesic&amp;nbsp;</td><td>Norbrook Laboratories&amp;nbsp;</td><td>6451603670</td><td>NSAID antinflammatory and analgesic used for post-operative pain control. Generic: meloxicam.&amp;nbsp;</td></tr><tr><td>Enroflox antibiotic&amp;nbsp;</td><td>Norbrook Laboratories&amp;nbsp;</td><td>5552915411</td><td>Fluoroquinolone antibiotic for post-operative infection prevention. Generic: Enrofloxacin.</td></tr><tr><td>Beuthanasia-D&amp;nbsp;</td><td>Merck Animal Health</td><td>00061047305</td><td>Pentobarbital Sodium, Phenytoin Sodium euthanasia agent.</td></tr><tr><td>Bone screws</td><td>Pinnacle Technologies, Inc.</td><td>8111-16</td><td>1/8&quot; bone screw (Pkg. of 16) used to affix skull cap to skull.&amp;nbsp;</td></tr><tr><td>Dental acrylic (Bosworth Duz-All)</td><td>Bosworth&amp;nbsp;</td><td>166261C&amp;nbsp;</td><td>Self curing dental acrylic is used in construction of a skull cap to affix cannula and head mount to skull.</td></tr><tr><td>Hardware biosensor setup&amp;nbsp;</td><td>Pinnacle Technologies, Inc.</td><td>8400-K2</td><td>Pinnacle offers complete hardware kits for new users of our tethered biosensor system for rats. Kits include a commutator, preamplifier, and data conditioning and acquisition system</td></tr><tr><td>Base video computer package</td><td>Pinnacle Technologies, Inc.</td><td>9000-K1</td><td>The base computer package includes a preconfigured computer with ample hard disk storage, a high-definition monitor, a keyboard and mouse, an uninterruptible power supply, and all necessary cables.&amp;nbsp;</td></tr><tr><td>Video EQ700 EverFocus camera&amp;nbsp; package</td><td>Pinnacle Technologies, Inc.</td><td>&amp;nbsp;9000-K10&amp;nbsp;</td><td>EQ700 night vision capable box camera with independent IR source was obtained as part of Pinnacle video computer package. Dome camera (9000-K9) and HD camera (9000-K11) options are also available.&amp;nbsp;</td></tr><tr><td>Sirenia Acquisition software</td><td>Pinnacle Technologies, Inc.</td><td>Free--available to download from pinnaclet.com</td><td>Sirenia&amp;nbsp;Acquisition provides a single platform for recording data from any Pinnacle hardware system. The software features synchronization of all data streams, user-configurable settings, data consolidation, and multiple export options. In addition, the software includes basic review and analysis modules for biosensor recordings. Sirenia&amp;nbsp;delivers free ll-in-one software that is ideal for data acquisition and review.</td></tr><tr><td>Tethered rat&amp;nbsp;in vitro&amp;nbsp;calibration kit</td><td>Pinnacle Technologies, Inc.</td><td>7000-K2-T-BAS&amp;nbsp;</td><td>In order to relate the current changes measured by a biosensor to actual changes in analyte concentration, it is necessary to calibrate the biosensor prior to implantation into the animal. The process also confirms the integrity and selectivity of the sensors.&amp;nbsp;Calibration kit includes 20 mL jacketed beaker (#7058), 1/2&quot; by 1/8&quot; magnetic stir bar (#7059), right angle clamp (#7056), 2 prong single-adjustment clamp (#7055), 4-channel calibration preamplifer (#7053), and calibration holder (#7051).&amp;nbsp;</td></tr><tr><td>Stir plate&amp;nbsp;</td><td>Corning</td><td>6795-410D</td><td>Corning digital Stirrer, 5&quot; x 7&quot;, 120 VAC used to spin magnetic stirrer in jacketed beaker during &lt;em&gt;in vitro&lt;/em&gt; calibration of glutamate biosensors.</td></tr><tr><td>Water bath capable of closed loop circulation</td><td>PolyScience</td><td>8006A11B</td><td>PolyScience 8006A11B 6L Standard Digital Heated Circulating Bath, 120VAC water bath was used with plastic tubing to heat jacketed beaker to physiological temperature.&amp;nbsp;</td></tr><tr><td>Carbon fiber sensor with BASi rat cannulae</td><td>Pinnacle Technology, Inc.</td><td>7002-CFS</td><td>Carbon fiber electrode used for recording dopamine neurotransmission.</td></tr><tr><td>Ag/AgCl reference electrode</td><td>Pinnacle Technology, Inc.</td><td>7065</td><td>Necessary for carbon fiber recordings.</td></tr><tr><td>Glutamate biosensors&amp;nbsp;</td><td>Pinnacle Technology, Inc.</td><td>7001&amp;nbsp;</td><td>Enzymatic biosensor probe used for recording glutamatergic neurotransmission.</td></tr><tr><td>BASi guide cannulae</td><td>Pinnacle Technologies, Inc.</td><td>7030&amp;nbsp;</td><td>Guide cannulae implanted into brain region of interest to guide probe.</td></tr><tr><td>BASi cannula plastic headpiece for rats&amp;nbsp;</td><td>Pinnacle Technologies, Inc.</td><td>7011&amp;nbsp;</td><td>Headmount stabilizes probe and attaches to potentiostat.</td></tr></tbody></table>

measuring in vivo changes in extracellular neurotransmitters during naturally rewarding behaviors in female syrian hamsters

在快速的时间尺度上测量神经递质释放的能力允许神经的模式与特定的行为或操纵相联系;阐明底层机制和电路的有力工具。虽然微透析技术已被用于几十年来测量几乎任何对大脑感兴趣的分析物, 但这种技术在时间分辨率上是有限的。另外, 快速扫描循环伏安法既精确又灵敏;然而, 由于这一技术上困难的方法依赖于 electroactivity 的分析物的兴趣, 有可能检测 nonelectroactive 物质 (例如, 神经递质谷氨酸) 被消除。本文详细介绍了结合固定电位安培和酶传感的交钥匙系统, 用于测量活性和 nonelectroactive 神经递质的时间精度。这两种强大的技术配对, 使测量的补品和相位神经相对轻松, 并允许记录多个神经递质同时。这篇手稿的目的是演示在雌性仓鼠中使用自然奖赏行为 (即, 性行为) 来测量多巴胺和谷氨酸神经在体内的过程, 最终目标是显示本试验的技术可行性, 用于检测其他行为和实验范例。

利用上述的电化学和行为编码方法, 该实验室已经开始在体内性行为的记录中, 对多巴胺和谷氨酸中的补品和阶段性波动进行表征。由于这个方法的世俗地确切的方式, 我们能更加准确地刻画神经在性行为期间;以及在交配回合期间, 将特定的释放模式变化归因于相应的补药变化, 以及在女性接受个体交配行为期间的相应的瞬时波动 (例如, intromission, 射精)从安装男性。
具体来说, 我们使用这种方法的重点是测量多巴胺和谷氨酸在不同动物的 NAc 中的释放模式。NAc 是一个关键的地区参与奖励处理22 , 并已被描述为一个积分区域的奖励方面的性行为行为23。
我们观察到, 在性行为期间, 雌性在交配期间, 表现出在 NAc 核心的多巴胺能水平增加 (图 5)。虽然这些补品的变化显示了所有交配回合, 上述方法可以测量在每一个交配回合之间的补药变化, 从而增加了对神经递质释放的总变化的时间识别整个整个会话。此外, 这一描述的方法可以精确地确定相位变化, 并暂时地映射可能与这些阶段性神经递质波动相关的行为。具体来说, 这些初步结果表明, 在交配回合期间, NAc 的多巴胺能瞬变出现在阴道插入 (intromission) 的男性 (图 6)。此外, 这种与 intromission 的联系是特定的, 对其他交配行为, 如 AI, 可忽略的多巴胺能反应。
类似的模式已经观察到其他动物的谷氨酸释放, 与快速瞬变, 对应于个别 intromissions 在交配回合 (图 7)。这种模式是区域特异的, 没有明显的信号发生在 NAc 壳或内侧尾状的其他动物。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56135/56135fig1.jpg"> 
图 1: 酶生物传感器技术. 标准制造的商业可用酶生物传感器探针, 通过酶介导的处理来检测神经递质 (左图)。在谷氨酸传感器的情况下, 谷氨酸氧化酶是受聘于酶层 (右图) 转换 nonelectroactive 神经递质的活性过氧化氢 (H2O2), 被氧化检测到铂铱 (铂-Ir) 电极。抗坏血酸 (aa), 一个共同的干扰, 是被氧化的相同的电位, 通过添加 AA 氧化酶在酶层, 转换干扰 nonelectroactive 水。其他负活性 interferents 存在于大脑中被排除通过被动选择性膜。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56135/56135fig2.jpg"> 
图 2: 仓鼠交配回合的组成部分. 在仓鼠, 交配回合包括一系列的交配行为, 在此期间, 女性保持在不动的前姿势, 而男性 anogenitally 调查(a), 装载(B), 并达到 intromission (C).注意 intromission 是行为区别于安装, 男性拉他的上肢对他的身体, 推力他的骨盆向前, 并卷曲他的尾巴向上, 表明渗透。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56135/56135fig3.jpg"> 
图 3: 头骨盖结构. 指南插管, 骨螺钉, 和牙科丙烯酸是用来建造一个头骨帽, 以允许插入碳纤维或酶探针 (参见15逐步详细)。这个数字描绘了一个单一的套管植入在右 NAc 的酶传感器 (没有植入的参考电极) 和塑料头安装传感器的稳定性。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56135/56135fig4.jpg"> 
图 4: 谷氨酸生物传感器校准. 每个生物传感器对谷氨酸的敏感性在实验记录之前被测试了在体外10 µm 添加谷氨酸 (绿色) 在37° c (由于酵素活动受温度的影响)。每个传感器被证实是响应到抗坏血酸 (AA; 红色) 或多巴胺 (DA; 蓝色)。这种校准允许在实验记录中看到的安培信号的变化转化为谷氨酸浓度的变化。 <a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56135/56135fig5.jpg"> 
图 5: 在交配回合中, 多巴胺的补品增加。 premating 回合前信号与交配回合前信号的程式化例子.为了测量补剂多巴胺的释放, 在交配回合 (mb) 之前前的安培反应与多巴胺水平相比, 在一个 mb。灰色方框表示在 MB 前前。blue 框表示在 MB 期间前。B.从一种动物的代表性定量比较 preMB 前和 MB 前之间的平均电压。误差线表示平均值 (SEM) 的标准误差, 星号表示显著差异 (p 值和 #60; 0.05)。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56135/56135fig6.jpg"> 
图 6: 具有代表性的多巴胺跟踪. 多巴胺的阶段性释放是 time-locked 的 intromissions 从雄性的背部 NAc 核心。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56135/56135fig7.jpg"> 
图 7: 具有代表性的谷氨酸跟踪. 谷氨酸瞬变对应于个别 intromissions 从男性在背部 NAc 核心在延长性经验。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56135/56135fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters at JoVE.com

Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

本文详细介绍了使用碳纤电极和酶生物传感器技术的固定电位安培记录, 以测量在自然奖励行为中的高时间分辨率的多巴胺和谷氨酸的释放。雌性仓鼠

雌性叙利亚仓鼠自然奖赏行为中细胞外神经递质的测量体内变化

The ability to measure neurotransmitter release on a rapid time scale allows patterns of neurotransmission to be linked to specific behaviors or ...

measuring-vivo-changes-extracellular-neurotransmitters-during

Nanyang Technological University

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Introduction to Psychology

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Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

Biopsychology

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10.1K 次观看.  University of Minnesota. 本文详细介绍了使用碳纤电极和酶生物传感器技术的固定电位安培记录, 以测量在自然奖励行为中的高时间分辨率的多巴胺和谷氨酸的释放。雌性仓鼠

视频: Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

Operant Sensation Seeking in the Mouse

Operant methods are powerful behavioral tools for the study of motivated behavior. These 'self-administration' methods have been used extensively in drug addiction research due to their high construct validity. Operant studies provide researchers a tool for preclinical investigation of several aspects of the addiction process. For example, mechanisms of acute reinforcement (both drug and non-drug) can be tested using pharmacological or genetic tools to determine the ability of a molecular target to influence self-administration behavior1-6. Additionally, drug or food seeking behaviors can be studied in the absence of the primary reinforcer, and the ability of pharmacological compounds to disrupt this process is a preclinical model for discovery of molecular targets and compounds that may be useful for the treatment of addiction3,7-9. One problem with performing intravenous drug self-administration studies in the mouse is the technical difficulty of maintaining catheter patency. Attrition rates in these experiments are high and can reach 40% or higher10-15. Another general problem with drug self-administration is discerning which pharmacologically-induced effects of the reinforcer produce specific behaviors. For example, measurement of the reinforcing and neurological effects of psychostimulants can be confounded by their psychomotor effects. Operant methods using food reinforcement can avoid these pitfalls, although their utility in studying drug addiction is limited by the fact that some manipulations that alter drug self-administration have a minimal impact on food self-administration. For example, mesolimbic dopamine lesion or knockout of the D1 dopamine receptor reduce cocaine self-administration without having a significant impact on food self-administration 12,16. 
Sensory stimuli have been described for their ability to support operant responding as primary reinforcers (i.e. not conditioned reinforcers)17-22. Auditory and visual stimuli are self-administered by several species18,21,23, although surprisingly little is known about the neural mechanisms underlying this reinforcement. The operant sensation seeking (OSS) model is a robust model for obtaining sensory self-administration in the mouse, allowing the study of neural mechanisms important in sensory reinforcement24. An additional advantage of OSS is the ability to screen mutant mice for differences in operant behavior that may be relevant to addiction. We have reported that dopamine D1 receptor knockout mice, previously shown to be deficient in psychostimulant self-administration, also fail to acquire OSS24. This is a unique finding in that these mice are capable of learning an operant task when food is used as a reinforcer. While operant studies using food reinforcement can be useful in the study of general motivated behavior and the mechanisms underlying food reinforcement, as mentioned above, these studies are limited in their application to studying molecular mechanisms of drug addiction. Thus, there may be similar neural substrates mediating sensory and psychostimulant reinforcement that are distinct from food reinforcement, which would make OSS a particularly attractive model for the study of drug addiction processes. The degree of overlap between other molecular targets of OSS and drug reinforcers is unclear, but is a topic that we are currently pursuing. While some aspects of addiction such as resistance to extinction may be observed with OSS, we have found that escalation 25 is not observed in this model24. Interestingly, escalation of intake and some other aspects of addiction are observed with self-administration of sucrose26. Thus, when non-drug operant procedures are desired to study addiction-related processes, food or sensory reinforcers can be chosen to best fit the particular question being asked. 
In conclusion, both food self-administration and OSS in the mouse have the advantage of not requiring an intravenous catheter, which allows a higher throughput means to study the effects of pharmacological or genetic manipulation of neural targets involved in motivation. While operant testing using food as a reinforcer is particularly useful in the study of the regulation of food intake, OSS is particularly apt for studying reinforcement mechanisms of sensory stimuli and may have broad applicability to novelty seeking and addiction.

In this protocol we describe a method of operant learning using sensory stimuli as a reinforcer in the mouse. It requires no prior training or food restriction, and it allows the study of motivated behavior without the use of a pharmacological or natural reinforcer such as food.

在鼠标的操作性感觉寻求

Operant methods are powerful behavioral tools for the study of motivated behavior.  These 'self-administration' methods have been used extensively in drug ...

科研

神经科学

Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and amphetamine), the role it plays in psychiatric illnesses (e.g. schizophrenia and Attention Deficit Hyperactivity Disorder), and its involvement in degenerative disorders like Parkinson's and Huntington's disease. Under normal physiological conditions, dopamine is known to regulate locomotor activity, cognition, learning, emotional affect, and neuroendocrine hormone secretion. One of the largest densities of dopamine neurons is within the striatum, which can be divided in two distinct neuroanatomical regions known as the nucleus accumbens and the caudate-putamen. The objective is to illustrate a general protocol for slice fast-scan cyclic voltammetry (FSCV) within the mouse striatum. FSCV is a well-defined electrochemical technique providing the opportunity to measure dopamine release and uptake in real time in discrete brain regions. Carbon fiber microelectrodes (diameter of ~ 7 &mu;m) are used in FSCV to detect dopamine oxidation. The analytical advantage of using FSCV to detect dopamine is its enhanced temporal resolution of 100 milliseconds and spatial resolution of less than ten microns, providing complementary information to in vivo microdialysis.

Using fast-scan cyclic voltammetry to measure electrically evoked presynaptic dopamine dynamics in striatal brain slices.

突触前多巴胺纹状体脑片的快速扫描循环伏安动态

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

一个多认知功能测试，并记录在啮齿动物的神经元活动的完全自动化和高度灵活的系统

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

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 ...

行为学

A Conflict Model of Reward-seeking Behavior in Male Rats

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual stimulus) that represents a high-value reward, and the aversive stimuli (pins), are concurrently presented. The male rats have to climb or jump over the obstacle full of pins to approach and investigate the sexual partner. If the animal persists in their approaching behavior regardless of the aversive stimuli, it is considered as a maladaptive or risky reward-seeking behavior. The conflict task permits the evaluation of deficit in inhibitory control resulting from exposure to abused drug, such as morphine, or a stressful event. 
The main advantage of this model is that it provides a simple and quick way to discover the deficit in inhibitory control after exposure to opiate drugs or other stressful events. In addition to opiates, this behavioral model would also be useful for quickly discovering the inhibitory control deficits induced by other addictive drugs. However, the limitation is that the male rats' performance may be subject to exercising effects with repeated testing under this conflict task. In the future, one can hope that the individuals with the compulsive phenotype of reward-seeking behavior after exposure to opiates will be identified based on modifying this conflict model.

This conflict model is used to measure the impairment of inhibitory control after exposure to addictive drugs, or other factors that may influence inhibitory control. A sexual stimulus and an aversive obstacle are concurrently presented, thus male rats have to conquer the obstacle to approach the sexual reward.

雄性大鼠寻救行为的冲突模型

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual ...

Chronic Stress Shifts Effort-Related Choice Behavior in a Y-Maze Barrier Task in Mice

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that measures motivation to expend effort and obtain reward. In mice, chronic stress exposure significantly impacts motivation to work for a higher value reward when a lesser value reward is freely available compared to unstressed mice. Here we describe the chronic corticosterone administration paradigm, which produces a shift in effortful responding in the Y-maze barrier task. In the Y-maze task, one arm contains 4 food pellets, while the other arm contains only 2 pellets. After mice learn to select the high reward arm, barriers with progressively increasing height are then introduced into the high reward arm over multiple test sessions. Unfortunately, most chronic stress paradigms (including corticosterone and social defeat) were developed in male mice and are less effective in female mice. Therefore, we also discuss chronic non-discriminatory social defeat stress (CNSDS), a stress paradigm we developed that is effective in both male and female mice. Repeating results with multiple distinct chronic stressors in male and female mice combined with increased usage of translationally relevant behavior tasks will help to advance the understanding of how chronic stress can precipitate mood disorders.

The Y-maze barrier task is a behavior test that examines motivation to expend effort for reward. Here, we discuss testing multiple well-validated chronic stressors including chronic corticosterone and social defeat stress with this behavior, as well as the novel chronic non-discriminatory social defeat stress (CNSDS), which is effective in females.

在小鼠的 Y-Maze 障碍任务中，慢性应力转移与工作量相关的选择行为

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that ...

Environmental Modulations of the Number of Midbrain Dopamine Neurons in Adult Mice

Long-lasting changes in the brain or &#8216;brain plasticity&#8217; underlie adaptive behavior and brain repair following disease or injury. Furthermore, interactions with our environment can induce brain plasticity. Increasingly, research is trying to identify which environments stimulate brain plasticity beneficial for treating brain and behavioral disorders. Two environmental manipulations are described which increase or decrease the number of tyrosine hydroxylase immunopositive (TH+, the rate-limiting enzyme in dopamine (DA) synthesis) neurons in the adult mouse midbrain. The first comprises pairing male and female mice together continuously for 1 week, which increases midbrain TH+ neurons by approximately 12% in males, but decreases midbrain TH+ neurons by approximately 12% in females. The second comprises housing mice continuously for 2 weeks in &#8216;enriched environments&#8217; (EE) containing running wheels, toys, ropes, nesting material, etc., which increases midbrain TH+ neurons by approximately 14% in males. Additionally, a protocol is described for concurrently infusing drugs directly into the midbrain during these environmental manipulations to help identify mechanisms underlying environmentally-induced brain plasticity. For example, EE-induction of more midbrain TH+ neurons is abolished by concurrent blockade of synaptic input onto midbrain neurons. Together, these data indicate that information about the environment is relayed via synaptic input to midbrain neurons to switch on or off expression of &#8216;DA&#8217; genes. Thus, appropriate environmental stimulation, or drug targeting of the underlying mechanisms, might be helpful for treating brain and behavioral disorders associated with imbalances in midbrain DA (e.g. Parkinson&#8217;s disease, attention deficit and hyperactivity disorder, schizophrenia, and drug addiction).

This protocol describes two different environmental manipulations and a concurrent brain infusion protocol to study environmentally-induced brain changes underlying adaptive behavior and brain repair in adult mice.

中脑多巴胺神经元的数目在成年小鼠环境的调变

Long-lasting changes in the brain or &#8216;brain plasticity&#8217; underlie adaptive behavior and brain repair following disease or injury. Furthermore, ...

<meta charset="utf8"></head><body>使用可口的食物获取小鼠模型研究暴饮暴食症

Intermittent Binge-Intake Model in Mice

Obesity affects one in eight individuals globally. Overeating, especially high-calorie foods, plays a significant role in obesity. Binge eating disorder (BED) is a complex condition caused by a combination of biological, psychological, and social factors. It involves excessive consumption of high-calorie foods in a short period and is often linked to anxiety and cravings.
Here, we present a protocol employing continuous and intermittent access to a new highly palatable food (HPF) -- M&amp;M&#39;s -- to develop a mouse model for investigating binge eating disorder (BED). The HPF, chosen for its high fat and sugar content, represents an optimal food source to study due to its strong palatability, which makes it particularly suited for examining compulsive eating behaviors. Using C57BL/6 mice, we provided continuous or intermittent access to M&amp;M&#39;s while allowing unrestricted access to standard chow and water. By the 8th day, the mice in the intermittent access group exhibited pronounced binge-eating behaviors, which persisted through the 26th day. These mice also consumed significantly more calories -- predominantly from the confectionary -- than those in the continuous access and control groups. The contrast between continuous and intermittent access underscores the critical role of feeding schedules in promoting the overconsumption of palatable foods.
Furthermore, behavioral assessments revealed intermittent access to the HPF-induced anxiety-like behaviors, highlighting the psychological impact of access patterns on both eating behavior and emotional states. By incorporating these two distinct feeding paradigms, this study offers valuable insights into how the availability of highly palatable foods can exacerbate binge-eating tendencies. This model provides a more realistic approach to studying binge-eating behaviors and their metabolic consequences. It highlights the importance of standardized models in biomedical research. Our findings offer insights into the physiological and neural mechanisms underlying BED, which could pave the way for developing effective therapeutic interventions for BED and obesity.

<meta charset="utf8"></head><body>作者聚焦:用于研究暴食症的基于 M&M 的可访问小鼠模型 - 对饮食行为、焦虑和神经机制的见解

A binge-eating model was established in mice using intermittent access to M&amp;M's (a highly palatable food made of fat and sugar). Mice with intermittent access to this high-fat, high-sugar food showed stress-like behaviors, increased caloric intake, and a preference for that food over standard chow compared to mice with continuous or no access to the food.

小鼠间歇性暴饮暴食模型

Obesity affects one in eight individuals globally. Overeating, especially high-calorie foods, plays a significant role in obesity. Binge eating disorder ...

雌性叙利亚仓鼠自然奖赏行为中细胞外神经递质的测量体内变化

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在小鼠的 Y-Maze 障碍任务中，慢性应力转移与工作量相关的选择行为

中脑多巴胺神经元的数目在成年小鼠环境的调变

小鼠间歇性暴饮暴食模型

小鼠间歇性暴饮暴食模型

小鼠间歇性暴饮暴食模型