Delaney S M, Geiger J D
Department of Pharmacology and Therapeutics, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
J Neurosci Methods. 1996 Feb;64(2):151-6. doi: 10.1016/0165-0270(95)00119-0.
A high-energy focused microwave system for killing experimental animals was used to rapidly inactivate enzymes and prevent postmortem breakdown of adenine nucleotides and adenosine, thereby enabling accurate measurements of AMP, ADP, ATP and adenosine in rat brain. For comparison, purine levels were measured in brains of rats killed by decapitation, decapitation into liquid nitrogen, or in situ freezing of the brain with liquid nitrogen. Of the three microwave irradiation power levels used, 10, 6.0 or 3.5 kW, rats killed by 10 kW had the highest ATP levels (28.8 nmol/mg protein) and cellular energy charge value (0.8), and the lowest levels of AMP (2.2 nmol/mg protein) and adenosine (19.7 pmol/mg protein). Of the 6 brain regions studied, adenosine levels (pmol/mg protein) ranged from 10 in cerebral cortex to 170 in cerebellum of rats killed using 10 kW microwave irradiation and, for comparison, ranged from 840 in cerebral cortex to 2498 in striatum of rats killed by decapitation. Focused microwave killing permits precise and accurate measurements of purines in discrete regions of rat brain.
一种用于处死实验动物的高能聚焦微波系统被用于快速使酶失活，并防止腺嘌呤核苷酸和腺苷在死后分解，从而能够准确测量大鼠脑中的AMP、ADP、ATP和腺苷。作为比较，对通过断头、断头后置入液氮或用液氮原位冷冻大脑处死的大鼠脑内嘌呤水平进行了测量。在所使用的三个微波辐照功率水平（10、6.0或3.5千瓦）中，被10千瓦微波辐照处死的大鼠ATP水平最高（28.8纳摩尔/毫克蛋白质），细胞能荷值为0.8，而AMP（2.2纳摩尔/毫克蛋白质）和腺苷（19.7皮摩尔/毫克蛋白质）水平最低。在所研究的6个脑区中，使用10千瓦微波辐照处死的大鼠，腺苷水平（皮摩尔/毫克蛋白质）在大脑皮层为10，在小脑中为170，作为比较，断头处死的大鼠大脑皮层腺苷水平为840，纹状体中为2498。聚焦微波处死法能够精确、准确地测量大鼠脑离散区域中的嘌呤。
