Quinan Virginia, Dugar Anushree, Bauer Deborah
Neuroscience Program, Wellesley College, Wellesley, MA 02481.
J Undergrad Neurosci Educ. 2019 Jun 30;17(2):A111-A118. eCollection 2019 Spring.
Long-term potentiation (LTP) is thought to be a critical mechanism underlying learning and memory. Although LTP is now widely performed in neuroscience research laboratories and the theory behind it is taught in many undergraduate courses, it is rare for undergraduate students to have the opportunity to perform LTP experiments themselves. Here, we describe a series of two laboratory sessions in which upper level students learn how to perform LTP experiments in acute hippocampal slices from wild type mice. In Laboratory 1, students practice the techniques necessary to set up the experiments. These techniques include making solutions, pulling glass recording electrodes, performing brain removal, preparing hippocampal slices, and positioning electrodes in area CA1. For Laboratory 2, hippocampal slices are prepared in advance by the instructors. Students record LTP by stimulating the Schaffer collateral axons and recording postsynaptic field potential responses in the apical dendritic region of area CA1. Once the students determine appropriate stimulus strength, they collect baseline responses, deliver a tetanic stimulus, and then collect responses 10 and 30 minutes following tetanic stimulation. Students analyze the data in LabChart 7 (ADInstruments - North America, Colorado Springs, CO, 2011) and perform appropriate statistical tests to determine whether potentiation has occurred. These laboratory exercises provide a unique opportunity for students to gain an appreciation for the techniques that are fundamental to studies of neural electrophysiology and plasticity as evidenced through a learning assessment tool.
长时程增强(LTP)被认为是学习和记忆的关键机制。尽管LTP现在在神经科学研究实验室中广泛开展,其背后的理论也在许多本科课程中讲授,但本科生很少有机会亲自进行LTP实验。在此,我们描述了一系列的两个实验课程,高年级学生在其中学习如何在野生型小鼠的急性海马切片中进行LTP实验。在实验课程1中,学生练习开展实验所需的技术。这些技术包括配制溶液、拉制玻璃记录电极、进行脑切除、制备海马切片以及将电极置于CA1区。在实验课程2中,海马切片由教师提前制备。学生通过刺激海马伞轴突并记录CA1区顶端树突区域的突触后场电位反应来记录LTP。一旦学生确定了合适的刺激强度,他们收集基线反应,给予强直刺激,然后在强直刺激后10分钟和30分钟收集反应。学生在LabChart 7(ADInstruments - North America, Colorado Springs, CO, 2011)中分析数据,并进行适当的统计测试以确定是否发生了增强。这些实验练习为学生提供了一个独特的机会,让他们通过一种学习评估工具来了解神经电生理学和可塑性研究的基础技术。