Melendez-Ferro Miguel, Perez-Costas Emma, Roberts Rosalinda C
Department of Psychiatry and Behavioral Neurobiology, University of Alabama at Birmingham, Sparks Center 865, 1720 7th Avenue South, Birmingham, AL 35294, United States.
J Neurosci Methods. 2009 Jan 30;176(2):72-7. doi: 10.1016/j.jneumeth.2008.08.021. Epub 2008 Aug 26.
The study of dendritic spine shape and number has become a standard in the analysis of synaptic transmission anomalies since a considerable number of neuropsychiatric and neurological diseases have their foundation in alterations in these structures. One of the best ways to study possible alterations of dendritic spines is the use of Golgi impregnation. Although usually the Golgi method implies the use of fresh or fixed tissue, here we report the use of Golgi-Cox for the staining of human and animal brain tissue kept frozen for long periods of time. We successfully applied the Golgi-Cox method to human brain tissue stored for up to 15 years in a freezer. The technique produced reliable and reproducible impregnation of dendrites and dendritic spines in different cortical areas. We also applied the same technique to rat brain frozen for up to 1 year, obtaining the same satisfactory results. The fact that Golgi-Cox can be successfully applied to this type of tissue adds a new value for hundreds of frozen human or animal brains kept in the freezers of the laboratories, that otherwise would not be useful for anything else. Researchers other than neuroanatomists, i.e. in fields such as biochemistry and molecular biology can also benefit from a simple and reliable technique that can be applied to tissue left from their primary experiments.
由于相当多的神经精神疾病和神经疾病都基于这些结构的改变,树突棘的形状和数量研究已成为分析突触传递异常的标准方法。研究树突棘可能改变的最佳方法之一是使用高尔基染色法。虽然通常高尔基方法意味着使用新鲜或固定组织,但在此我们报告使用高尔基-考克斯法对长时间冷冻保存的人和动物脑组织进行染色。我们成功地将高尔基-考克斯法应用于在冰箱中保存长达15年的人脑组织。该技术在不同皮质区域产生了可靠且可重复的树突和树突棘染色。我们还将相同技术应用于冷冻长达1年的大鼠脑,获得了同样令人满意的结果。高尔基-考克斯法能够成功应用于这类组织这一事实,为保存在实验室冰箱中的数百个人类或动物冷冻大脑增添了新价值,否则这些大脑将毫无用处。除神经解剖学家之外的研究人员,即在生物化学和分子生物学等领域的研究人员,也可以从这种可应用于其初步实验剩余组织的简单可靠技术中受益。