Organotypic Hippocampal Slice Cultures
Organotypic Hippocampal Slice Cultures
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DOI：
10.3791/2462
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发表时间：
2011-02-01
影响因子：
1.2
通讯作者：
Barria, Andres
中科院分区：
文献类型：
--
作者：
Opitz-Araya, Ximena;Barria, Andres
海马体是边缘系统的一个组成部分，在长期记忆和空间导航中起着重要作用(1)。海马体神经元在短暂的强烈激活后可以改变其连接的强度。这种现象被称为长期增强（LTP），可以持续数小时或数天，并已成为学习和记忆的最佳候选机制(2)。此外，海马良好的解剖结构和连通性(3)使其成为研究突触传递和突触可塑性的经典模型系统(4)。随着我们对海马体突触生理学的理解不断加深，以及分子参与者的识别，操纵突触蛋白的需求变得势在必行。器官型海马培养提供了简单的基因操作和精确的药物干预的可能性，但保持突触组织对于在更自然的背景下理解突触功能至关重要，而不是常规培养解离神经元方法。在这里，我们提出了一种制备和培养海马切片的方法，这种方法可以很容易地适应于其他大脑区域。这种方法允许使用不同的方法（如病毒感染（5,6）或生物学(7)）轻松访问切片进行遗传操作。此外，切片可以很容易地恢复生化分析(8)，或转移到显微镜成像(9)或电生理实验（10）。
The hippocampus, a component of the limbic system, plays important roles in long-term memory and spatial navigation(1). Hippocampal neurons can modify the strength of their connections after brief periods of strong activation. This phenomenon, known as long-term potentiation (LTP) can last for hours or days and has become the best candidate mechanism for learning and memory(2). In addition, the well defined anatomy and connectivity of the hippocampus(3) has made it a classical model system to study synaptic transmission and synaptic plasticity(4).As our understanding of the physiology of hippocampal synapses grew and molecular players became identified, a need to manipulate synaptic proteins became imperative. Organotypic hippocampal cultures offer the possibility for easy gene manipulation and precise pharmacological intervention but maintain synaptic organization that is critical to understanding synapse function in a more naturalistic context than routine culture dissociated neurons methods.Here we present a method to prepare and culture hippocampal slices that can be easily adapted to other brain regions. This method allows easy access to the slices for genetic manipulation using different approaches like viral infection(5,6) or biolistics(7). In addition, slices can be easily recovered for biochemical assays(8), or transferred to microscopes for imaging(9) or electrophysiological experiments(10).