Davenport April T, Grant Kathleen A, Szeliga Kendall T, Friedman David P, Daunais James B
Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, NC, USA.
Cell Tissue Bank. 2014 Mar;15(1):99-110. doi: 10.1007/s10561-013-9380-2. Epub 2013 May 25.
Appropriate animal models are critical to conduct translational studies of human disorders without variables that can confound clinical studies. Such analytic methods as patch-clamp electrophysiological and voltammetric recordings of neurons in brain slices require living brain tissue. In order to obtain viable tissue from nonhuman primate brains, tissue collection methods must be designed to preserve cardiovascular and respiratory functions for as long as possible. This paper describes a method of necropsy that has been used in three species of monkeys that satisfies this requirement. At necropsy, animals were maintained under a deep surgical plane of anesthesia while a craniotomy was conducted to expose the brain. Following the craniotomy, animals were perfused with ice-cold, oxygenated artificial cerebrospinal fluid to displace blood and to reduce the temperature of the entire brain. The brain was removed within minutes of death and specific brain regions were immediately dissected for subsequent in vitro electrophysiology or voltammetry experiments. This necropsy method also provided for the collection of tissue blocks containing all brain regions that were immediately frozen and stored for subsequent genomic, proteomic, autoradiographic and histological studies. An added benefit from the design of this necropsy method is that all major peripheral tissues were also collected and are now being utilized in a wide range of genomic, biochemical and histological assays. This necropsy method has resulted in the establishment and growth of a nonhuman primate alcohol tissue bank designed to distribute central nervous system and peripheral tissues to the larger scientific community.
合适的动物模型对于开展人类疾病的转化研究至关重要,因为这些模型不存在可能混淆临床研究的变量。诸如脑片神经元的膜片钳电生理记录和伏安法记录等分析方法需要活的脑组织。为了从非人灵长类动物大脑获取存活的组织,必须设计组织采集方法以尽可能长时间地维持心血管和呼吸功能。本文描述了一种已在三种猴子中使用的尸检方法,该方法满足这一要求。在尸检时,动物在深度手术麻醉平面下维持状态,同时进行开颅手术以暴露大脑。开颅手术后,给动物灌注冰冷的、充氧的人工脑脊液以置换血液并降低整个大脑的温度。在动物死亡后几分钟内取出大脑,并立即解剖特定脑区用于后续的体外电生理或伏安法实验。这种尸检方法还能收集包含所有脑区的组织块,这些组织块会立即冷冻并储存起来用于后续的基因组学、蛋白质组学、放射自显影和组织学研究。这种尸检方法设计的一个额外好处是,所有主要外周组织也被收集起来,目前正用于广泛的基因组学、生物化学和组织学检测。这种尸检方法促成了一个非人灵长类动物酒精组织库的建立和发展,该组织库旨在向广大科学界分发中枢神经系统和外周组织。