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P97 Pipette puller

Flaming/Brown Micropipette Puller

Company: Sutter Instrument
Catalog#: P97
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Protocols to Study Declarative Memory Formation in Mice and Humans: Optogenetics and Translational Behavioral Approaches
Author:
Date:
2018-06-20
[Abstract]  Declarative memory formation depends on the hippocampus and declines in aging. Two functions of the hippocampus, temporal binding and relational organization (Rawlins and Tsaltas, 1983; Eichenbaum et al., 1992; Cohen et al., 1997), are known to decline in aging (Leal and Yassa, 2015). However, in the literature distinct procedures have been used to study these two functions. Here, we describe the experimental procedures used to investigate how these two processes are related in the formation of declarative memory and how they are compromised in aging (Sellami et al., 2017). First, we studied temporal binding using a one-trial learning procedure: trace fear conditioning. It is classical Pavlovian conditioning requiring temporal binding since a brief temporal gap ... [摘要]  陈述性记忆的形成取决于海马体和老化程度的下降。已知海马,时间结合和关系组织的两种功能(Rawlins和Tsaltas,1983; Eichenbaum等人,1992; Cohen等人,1997),已知老龄化下降(Leal和Yassa,2015)。但是,在文献中已经使用不同的程序来研究这两个功能。在这里,我们描述了用于研究这两个过程如何与陈述性记忆的形成以及它们如何在衰老中受到损害有关的实验程序(Sellami et。,2017)。首先,我们使用单一试验性学习程序研究了时间绑定:追踪恐惧条件反射。这是经典的巴甫洛夫条件需要时间约束,因为短暂的时间间隔将条件刺激(CS)和无条件刺激(US)呈现分开。我们将追踪恐惧条件程序与光遗传学方法相结合,并且我们表明时间依赖性依赖于背侧(d)CA1活性在时间间隙上的依赖性。然后,我们研究了时间绑定和关系组织之间在声明性记忆形成中的相互作用,这种相互作用使用了小鼠中的两阶段径向迷宫任务及其在人类中的虚拟类比。行为过程包括一个初始学习阶段,在这个阶段中,受试者获得每个手臂持续的奖励/无奖赏价值,然后是测试阶段,其中武器中的奖励突发事件保持不变,但武器重新组合以评估灵活性,陈述性记忆。我们证明了dCA1依赖的时间结合对于记忆的关系组织的发展是必需的,这允许灵活的陈述记忆表达。此外,在衰老过程中,陈述性记忆的退化是由于时间约束能力的减少而阻碍了关系组织。 ...

Optogenetic Mapping of Synaptic Connections in Mouse Brain Slices to Define the Functional Connectome of Identified Neuronal Populations
Author:
Date:
2017-01-05
[Abstract]  Functional connectivity in a neural circuit is determined by the strength, incidence, and neurotransmitter nature of its connections (Chuhma, 2015). Using optogenetics the functional synaptic connections between an identified population of neurons and defined postsynaptic target neurons may be measured systematically in order to determine the functional connectome of that identified population. Here we describe the experimental protocol used to investigate the excitatory functional connectome of ventral midbrain dopamine neurons, mediated by glutamate cotransmission (Mingote et al., 2015). Dopamine neurons are made light sensitive by injecting an adeno-associated virus (AAV) encoding channelrhodopsin (ChR2) into the ventral midbrain of DATIREScre mice. The efficacy and ... [摘要]  神经回路中的功能连通性由其连接的强度,发病率和神经递质特性决定(Chuhma,2015)。使用光遗传学,可以系统地测量识别的神经元群体和定义的突触后靶神经元之间的功能性突触连接,以便确定该识别群体的功能性连接群。这里我们描述了用于研究由谷氨酸共转播介导的腹侧中脑多巴胺神经元的兴奋性功能性连接体的实验方案(Mingote等,2015)。通过将编码通道视紫红质(ChR2)的腺相关病毒(AAV)注射到DATIREScre小鼠的腹侧中脑中,使多巴胺神经元变得光敏。多巴胺合成酶酪氨酸羟化酶的免疫荧光证实ChR2表达在多巴胺神经元中的功效和特异性。然后,切片膜片钳记录由接受多巴胺神经元投影的区域中的神经元产生,并且确定兴奋性连接的发生率和强度。所有接受多巴胺神经元投射的区域的连接发生率和强度的总结构成功能性连接体。
【背景】为了建立特定神经回路的功能,有必要确定解剖连接,解剖连接的映射及其功能连接,连接的强度,发病率和神经递质性质的映射。使用单因素限制的病毒性突触后追踪技术,可以描述包括多巴胺系统在内的神经回路的复杂解剖连接(Callaway and Luo,2015; ...

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