Chen Gary Gang, Fiori Laura M, Mamer Orval A, Turecki Gustavo
McGill Group for Suicide Studies, Douglas Mental Health University Institute, McGill University, Montreal, QC, Canada.
Methods Mol Biol. 2011;720:427-36. doi: 10.1007/978-1-61779-034-8_27.
There is considerable evidence supporting a role of the polyamine system in the etiology and pathology of mental disorders. Changes in the expression and activity of polyamine anabolic/catabolic enzymes, as well as in the levels of individual polyamines, have been found in many psychiatric conditions, including schizophrenia, mood disorders, anxiety, and suicidal behavior. Recent microarray studies have found that spermidine/spermine-N¹-acetyltransferase (SAT1, SSAT), the key enzyme in charge of the polyamine catabolic pathway, is downregulated in brain tissue of individuals who were depressed and died by suicide. To provide further insight into the downstream effects of altered SAT1 expression, we developed a quantitative gas chromatography-mass spectrometry method for measurement of polyamine concentrations in postmortem human brain tissues. This protocol employs a conventional electron ionization method with total ion and selected ion monitoring. This method can accurately measure the levels of the polyamines putrescine, spermidine, and spermine from very small quantities (1-50 mg) of postmortem brain tissues, with quantitation limits down to 10 ng/g of wet tissue for putrescine and 100 ng/g for spermidine and spermine.
有大量证据支持多胺系统在精神障碍的病因学和病理学中发挥作用。在许多精神疾病中,包括精神分裂症、情绪障碍、焦虑症和自杀行为,都发现了多胺合成/分解代谢酶的表达和活性以及个体多胺水平的变化。最近的微阵列研究发现,负责多胺分解代谢途径的关键酶亚精胺/精胺-N¹-乙酰基转移酶(SAT1,SSAT)在抑郁且自杀死亡个体的脑组织中表达下调。为了进一步深入了解SAT1表达改变的下游效应,我们开发了一种定量气相色谱-质谱法来测量死后人类脑组织中的多胺浓度。该方案采用常规电子电离方法并进行全离子和选择离子监测。这种方法可以从极少量(1-50毫克)的死后脑组织中准确测量腐胺、亚精胺和精胺的多胺水平,腐胺的定量限低至10纳克/克湿组织,亚精胺和精胺为100纳克/克。