Johnson S A, Morgan D G, Finch C E
J Neurosci Res. 1986;16(1):267-80. doi: 10.1002/jnr.490160123.
The postmortem stability of brain RNA was measured in rat and human samples for up to 48 hr. Whole rat brains, cooled at a rate approximating that of human brains awaiting autopsy, were collected at intervals from 0 to 48 hr after death and frozen. These samples were compared with freshly obtained, unfrozen rat brains. Two potentially independent characteristics of these RNA populations were measured: the recovery or yield of RNA/gram tissue (quantity) and the integrity or extent of degradation (quality). Total RNA yields were similar after all postmortem delays. Hybridization of [32P]-labeled cDNA probes to nitrocellulose filter blots of electrophoretically separated rat brain RNA failed to reveal degradation of the specific rat brain mRNAs during the postmortem period. Similarly, in vitro translation of these same rat total RNA samples produced high molecular weight translation products with no differences between long and short postmortem times. Human cerebral cortex RNA was prepared by the same methods as those used for rat brain from a neurosurgical sample and four other donors with postmortem intervals from 7 to 36 hr. Typically, the yield of total RNA from human brain was 40-50% of the yield from rat brain. When analyzed by RNA gel blot hybridization studies, as for rat brain RNA, human cortical RNA appeared slightly degraded. However, the degree of apparent RNA degradation was not related to the postmortem interval. In vitro translation products of human cortical RNA revealed high molecular weight peptides at all postmortem intervals, but slightly less [35S]incorporation into these bands was found at the longer postmortem intervals relative to the shorter times. Together, these results demonstrate an extensive stability of brain RNA that invites aggressive use of molecular genetic techniques for the study of human neurodegenerative diseases.
在大鼠和人类样本中测量了脑RNA长达48小时的死后稳定性。将全大鼠脑以接近等待尸检的人类脑的速率冷却,在死后0至48小时每隔一段时间收集并冷冻。将这些样本与新鲜获取的未冷冻大鼠脑进行比较。测量了这些RNA群体的两个潜在独立特征:RNA/克组织的回收率或产量(数量)以及降解的完整性或程度(质量)。所有死后延迟后总RNA产量相似。用[32P]标记的cDNA探针与电泳分离的大鼠脑RNA的硝酸纤维素滤膜杂交,未发现死后期间特定大鼠脑mRNA的降解。同样,这些相同大鼠总RNA样本的体外翻译产生高分子量翻译产物,死后时间长短之间没有差异。人类大脑皮质RNA通过与用于大鼠脑相同的方法从神经外科样本和另外四名死后间隔为7至36小时的供体中制备。通常,人类脑总RNA的产量是大鼠脑产量的40 - 50%。当通过RNA凝胶印迹杂交研究分析时,与大鼠脑RNA一样,人类皮质RNA似乎略有降解。然而,明显的RNA降解程度与死后间隔无关。人类皮质RNA的体外翻译产物在所有死后间隔都显示出高分子量肽,但相对于较短时间,在较长死后间隔时发现这些条带中[35S]掺入量略少。总之,这些结果表明脑RNA具有广泛的稳定性,这促使人们积极使用分子遗传学技术来研究人类神经退行性疾病。