Groves Thomas R, Wang Jing, Boerma Marjan, Allen Antiño R
Division of Radiation Health, University of Arkansas for Medical Sciences; Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences; Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences.
Division of Radiation Health, University of Arkansas for Medical Sciences; Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences.
J Vis Exp. 2017 Jun 22(124):55696. doi: 10.3791/55696.
Dendritic spines are the protuberances from the neuronal dendritic shafts that contain excitatory synapses. The morphological and branching variations of the neuronal dendrites within the hippocampus are implicated in cognition and memory formation. There are several approaches to Golgi staining, all of which have been useful for determining the morphological characteristics of dendritic arbors and produce a clear background. The present Golgi-Cox method, (a slight variation of the protocol that is provided with a commercial Golgi staining kit), was designed to assess how a relatively low dose of the chemotherapeutic drug 5-flurouracil (5-Fu) would affect dendritic morphology, the number of spines, and the complexity of arborization within the hippocampus. The 5-Fu significantly modulated the dendritic complexity and decreased the spine density throughout the hippocampus in a region-specific manner. The data presented show that the Golgi staining method effectively stained the mature neurons in the CA1, the CA3, and the dentate gyrus (DG) of the hippocampus. This protocol reports the details for each step so that other researchers can reliably stain tissue throughout the brain with high quality results and minimal troubleshooting.
树突棘是神经元树突轴上的突起，含有兴奋性突触。海马体内神经元树突的形态和分支变化与认知和记忆形成有关。有几种高尔基染色方法，所有这些方法都有助于确定树突分支的形态特征，并产生清晰的背景。目前的高尔基-考克斯方法（是随商业高尔基染色试剂盒提供的方案的一个轻微变体）旨在评估相对低剂量的化疗药物5-氟尿嘧啶（5-Fu）如何影响树突形态、棘突数量以及海马体内分支的复杂性。5-Fu以区域特异性方式显著调节树突复杂性，并降低整个海马体的棘突密度。所呈现的数据表明，高尔基染色方法有效地对海马体CA1、CA3和齿状回（DG）中的成熟神经元进行了染色。本方案报告了每个步骤的详细信息，以便其他研究人员能够以高质量的结果和最少的故障排除可靠地对整个大脑的组织进行染色。
